TPHCWG Volume 4

Total Petroleum Hydrocarbon Criteria Working Group Series
Volume 4
Development of Fraction Specific Reference Doses (RfDs) and Reference Concentrations (RfCs) for Total Petroleum Hydrocarbons (TPH)
Total Petroleum Hydrocarbon Criteria Working Group Series
Volume 4
PREPARED FOR: Chevron, British Petroleum, and the Total Petroleum Hydrocarbon Criteria Working Group PREPARED BY: Exxon Biomedical Sciences, Inc.: D.A. Edwards, Ph.D. M.D. Andriot, Ph.D. M.A. Amoruso, Ph.D. A.C. Tummey C.J. Bevan, Ph.D. A. Tveit, Ph.D. L.A. Hayes, M.S., MLS EA Engineering, Science, and Technology, Inc.: S.H. Youngren, Ph.D. Remediation Technologies, Inc.: D.V. Nakles, Ph.D.
Amherst Scientific Publishers 150 Fearing Street Amherst, Massachusetts 01002 1997 by Amherst Scientific Publishers. All rights reserved. ISBN 1-884-940-13-7 The material contained in this document was obtained from independent and highly respected sources. Every attempt has been made to ensure accurate, reliable information, however, the publisher cannot be held responsible for the information or how the information is applied. Opinions expressed in this book are those of the Total Petroleum Hydrocarbon Criteria Working Group and do not reflect those of the publisher. This document was prepared by the Total Petroleum Hydrocarbon Criteria Working Group. Neither the Working Group nor members of the Working Group : a. Makes any warranty or representation, expressed or implied, with respect to the accuracy, completeness, or usefulness of the information contained in this report, or that the use of any apparatus, method, or process disclosed in this report may not infringe privately owned rights; or b. Assumes any liability with respect to the use of, or for damages resulting from the use of, any information, apparatus, method, or process disclosed in this report. Authorization to photocopy items for internal or personal use or the internal or personal use of specific clients is granted by Amherst Scientific Publishers, provided that $.50 per photocopied page is paid directly to Copyright Clearance Center, 222 Rosewood Drive, Danvers, MA 01923. Prior to photocopying items for educational classroom use, please contact the Copyright Clearance Center, Customer Service, 222 Rosewood Drive, Danvers, MA 01923. (508-750-8400) A portion of the proceeds from the sale of this book will be donated to the Plant-a-Tree Program, a reforestation program managed by the U.S. Forest Service. Printed in the United States of America
CONTENTS
PREFACE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vii
ACKNOWLEDGMENTS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ix I. INTRODUCTION/BACKGROUND . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 A. Hazard Assessment for TPH . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 1. Methodology for Human Health . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 2. Ecological Considerations and Assessment of New Toxicity Data . . . . 4 B. Evaluation of Toxicity Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 1. USEPA Methodology for RfD/RfC Development . . . . . . . . . . . . . . . . . 5 2. Prioritization of Toxicity Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 II. SUMMARY/CONCLUSIONS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
III. EVALUATION OF AROMATIC FRACTIONS . . . . . . . . . . . . . . . . . . . . . . . . . . . 10 A. C5 - C6 and C>7 - C8 Aromatic Fraction . . . . . . . . . . . . . . . . . . . . . . . . . . 10 1. Oral . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10 2. Inhalation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12 B. C8 - C10, C>10 - C12, and C>12 - C16 Aromatic Fraction . . . . . . . . . . . . . . . . 12 1. Oral . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .12 2. Inhalation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 C. C>16 - C21 and C>21 - C35 Aromatic Fraction . . . . . . . . . . . . . . . . . . . . . . . 14 1. Oral . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 2. Inhalation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 D. Overall Recommendations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15 IV. EVALUATION OF ALIPHATIC FRACTIONS . . . . . . . . . . . . . . . . . . . . . . . . . . . 15 A. C5 - C6 and C>7 - C8 Aliphatic Fraction . . . . . . . . . . . . . . . . . . . . . . . . . . 15 1. n-Heptane . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15 2. Commercial Hexane . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17 3. Other C5 - C8 Alkane/Cycloalkane Compounds . . . . . . . . . . . . . . . . . 17 4. Proposed Composition-Weighted RfD for TPH Fraction Containing C5 - C8 or C6 - C8 Aliphatics . . . . . . . . . . . . . . . . . . . . . . 19
B. C8 - C10, C>10 - C12, and C>12 - C16 Aliphatic Fraction . . . . . . . . . . . . . . . . 21 1. Summary of Inhalation Studies on Dearomatized Petroleum Streams and JP-8 . . . . . . . . . . . . . . . . . . 22 2. Summary of Oral Gavage Studies on Petroleum Streams and JP-8 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24 3. Summary and Conclusions for Oral RfDs and Inhalation RfCs . . . . . . 28 C. C>16 - C21 and C>21 - C35 Aliphatic Fraction . . . . . . . . . . . . . . . . . . . . . . . 29 1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29 2. Data Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29 3. Rationale for RfD Values . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31 REFERENCES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35 APPENDICES A. Literature Review (Individual Compounds) . . . . . . . . . . . . . . . . . . . . . . . 43 Attachment I - Results of Total Petroleum Hydrocarbons (TPH) Literature Search . . . . . . . . . . . . . . . . . . . . . . . . 47 Attachment II - EBSI Modified Deliverable . . . . . . . . . . . . . . . . . . . . . . 53 B. Toxicity Summaries for Both Aromatic and Aliphatic Constituents in the C4 to C22 Carbon Range . . . . . . . . . . . . . . . . . . . . . . 81 C. Review of American Petroleum Institute (APIs) Toxicity Data on Selected Refinery Streams . . . . . . . . . . . . . . . . . . . . 101 Attachment I - Composition Data on Selected Refinery Streams . . . . . 123
vi
PREFACE
This document is fourth in a series from the Total Petroleum Hydrocarbon Criteria Working Group (TPHCWG). The Working Group was formed in 1993 based on the observation that widely different clean-up requirements were being used by states at sites that were contaminated with hydrocarbon materials such as fuels, lubricating oils, and crude oils. These requirements were usually in the form of concentrations of total petroleum hydrocarbon, otherwise known as TPH, and ranged from 10 to over 10,000 milligrams of hydrocarbons per kilogram of soil. Members of the group jointly recognized that the numerical standard was not based on a scientific assessment of human health risk and they established the following goal for their effort: To develop scientifically defensible information for establishing soil cleanup levels that are protective of human health at hydrocarbon contaminated sites. The Working Group is guided by a steering committee consisting of representatives from industry, government, and academia. Some of the active participants among the more than 400 involved, include the Gas Research Institute, the Petroleum Environmental Research Forum, several major petroleum companies including Chevron, Exxon, and Shell, the American Petroleum Institute, the Association of American Railroads, several state governments (i.e., Washington, Texas, Colorado, Hawaii, Louisiana, New Mexico), the U.S. Environmental Protection Agency, the Department of Defense, and many consulting firms such as EA Engineering, Science and Technology. An overlying theme to this document is the importance of exposure potential when defining human health risk. The fate and transport of a chemical or mixture defines the exposure route and, in conjunction with receptor properties, concentrations at receptors. If fate and transport is not considered, unrealistic human health risks could be calculated, resulting in misinformed decisions about site clean-up, regulatory guidance, etc. This document summarizes the methods used to delineate TPH into equivalent carbon number fractions based on fate and transport considerations. The input into the fraction method included composition data on many common fuels and petroleum products. This information is provided in detail in Volume 2 of the Working Group reports. Once the fractions were defined, fraction-specific values of relevant physical-chemical properties were calculated based on correlations to boiling point. Companion volumes include Volume 1 which provides an overview of the complexities of petroleum hydrocarbon characterization and risk assessment and a discussion on the analytical methods available. In addition to descriptions about general analytical methods we have also provided a summary of a proposed GC-based analytical method developed by the Working Group that reports hydrocarbon results in equivalent carbon number groups or fractions.
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To complete the risk-based approach, the Working Group has also selected toxicity criteria (e.g., Reference Doses) for each of the defined fate and transport fractions. The evaluation of the toxicology research database and rationale behind the toxicity criteria selected is described in detail in Volume 4, Development of Fraction-Specific Toxicity Criteria for Total Petroleum Hydrocarbons (TPH) (in preparation). The analytical method, fate and transport considerations and toxicity criteria are the technical elements which fit into a risk-based framework for determining human health based criteria at petroleum hydrocarbon contaminated sites. The group selected the American Society for Testing and Materials (ASTM) Risk Based Corrective Action - RBCA framework as an example of how these elements can be used to calculate risk-based screening levels driven by noncancer human health risk for petroleum contaminated sites. We hope you find this document to be useful in your efforts to evaluate and determine acceptable riskbased criteria at petroleum sites.
viii
ACKNOWLEDGMENTS
The publication of this volume of the Total Petroleum Hydrocarbon Criteria Working Group Methodology would not have been possible without the hard work and dedication of individuals and organizations across the public and private sectors. Specifically, we would like to acknowledge the following members of the TPHCWG steering committee. Beth Albertson*, Friedman and Bruya Roger Andes and Christopher P.L. Barkan, Association of American Railroads Bruce Bauman, Roger Claff, Judith Shaw and Lorraine Twedock, API Barbara Brooks, Hawaii Department of Health Deborah Edwards and Joan Tell, Exxon Biomedical Sciences, Inc. John Fitzgerald, Massachusetts Department of Environmental Protection Kathy Garland*, New Mexico EMNRD Joseph Greenblott and Bruce Peirano, US EPA John Gustafson, Bruce Krewinghaus, Ross MacDonald, Ileana Rhodes, Shell Development Company Keith Hoddinott, US Army Jim Holler, Agency for Toxic Substances and Disease Registry Paul Kostecki and Tom Potter, University of Massachusetts, Amherst William Kucharski*, GECCO Inc. Mark Laughman, Louisiana Department of Environmental Quality Katherine Kurtz, Navy Environmental Health Center David Linz*, LinzTech David Nakles, RETEC Doug Orem and Susan Youngren, EA Environmental Science and Engineering Wade Weisman*, US Air Force Armstrong Lab. Robert Wilkenfeld*, Chevron * members of Executive Committee The TPHCWG would like to thank the following reviewers for their helpful suggestions and comments: Jane Sutherlin, LADEQ; James Evans, GRI; Peter Miasek, Imperial Oil; Bill Lowery, NJDEP; Adolfo Silva, Petro Canada; Daniel Smith, USAF; and Fred Reitman, Texaco. Additionally, we would like to commend Donna Voorhees (Menzie-Cura) and Tamlyn Oliver (AEHS) for their adept assistance in editing and publication of this document.
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I.
INTRODUCTION/BACKGROUND
The purpose of Volume 4 is to provide the basis for the development of fraction-specific reference doses/concentrations (RfDs/RfCs) for total petroleum hydrocarbon (TPH). The development of fraction-specific RfDs/RfCs provides for the hazard assessment of the TPH in the TPHCWGs risk assessment methodology (Figure 1). The selection of the most appropriate method for evaluating the risk of TPH is described below. The methodology selected was termed the indicator/surrogate approach, which is consistent with U.S. Environmental Protection Agency (USEPA) methodology and is illustrated in Figure 2 (USEPA, 1986). The indicators referred to are the single compounds within petroleum which are known to be carcinogens and which are evaluated/regulated individually (either federally or at state level). The surrogates for which RfDs/RfCs are developed are noncarcinogenic mixtures (fractions) which represent the mass of petroleum remaining after evaluation of the carcinogenic indicators. Indicators are evaluated first because their presence (even in relatively low concentrations) will drive a cleanup, due to their greater relative potency. The hazard assessment for TPH fractions would be utilized where indicator compounds are not present or are below regulatory criteria.
A. HAZARD ASSESSMENT FOR TPH 1. Methodology for Human Health
In order to develop a human health-based method for deriving cleanup levels for petroleum hydrocarbons in soil and groundwater, the authors surveyed the scientific literature to identify methods used to assess the potential health effects of petroleum hydrocarbons. Over 30 scientific papers and reports were reviewed and the citations can be found in the General section of the reference list. Based on this review, the approaches fit into two general categories: (1) use of toxicity data for the whole mixture or parent product (e.g., diesel fuel, gasoline, jet fuel, etc.) and (2) use of an indicator surrogate approach to assess the risk and toxicity posed by the mixture. The strengths, weaknesses, and the applicability of these scientific approaches were evaluated. Ideally, a hazard assessment should be conducted utilizing data on the mixture to which the receptor of concern is exposed. Utilizing data on the actual mixture present accurately accounts for the interactive effects of all components in the mixture. (Michelson and Boyce, 1993; Krewski and Thomas, 1992; WSDE, 1994; Warshawsky et al., 1993). Currently, these data are not available. The data which are available are (1) data on some whole mixtures or parent products (e.g., gasoline, jet fuel and mineral oil), (2) data on some individual compounds (indicators; e.g., benzene, benzo[a]pyrene) and (3) data on some fraction-specific mixtures. Toxicity data on whole mixtures or parent products are only available for gasoline, jet fuel, and mineral oil. Thus for other parent products (such as bunker fuel, diesel, lube oils, etc.) a whole mixture approach is not appropriate. In addition, once in the environment the parent product separates into fractions based on differences in fate and transport (Volume 3, TPHCWG methodology). The mixture to which a receptor is exposed will vary with space, time, and by media. Thus, a whole mixture approach would not be appropriate for a weathered release.
Figure 1. Risk Assessment for Petroleum Contaminated Media
Finally, there are no toxicity data on weathered fraction-specific mixtures or mixtures of parent products (such as mixtures of diesel and gasoline). Therefore, whole mixture toxicity data are only appropriate for the hazard assessment of a fresh release of a single, known product. Toxicity data on individual compounds were evaluated (see Appendix A). Of the 250 individual compounds identified in petroleum (TPHCWG Volume 2), only 95 had toxicity data. Of the 95 compounds with toxicity data, only 25 have sufficient data to develop toxicity criteria. Most of these compounds have USEPA-derived RfDs/RfCs or slope factors. Since there are thousands of individual compounds within petroleum, the toxicity data on individual compounds are only appropriate to evaluate the toxicity of those specific compounds, not the toxicity of mixtures such as TPH. The interactive effects of all the compounds present in TPH cannot be determined by data on 25 individual compounds. The toxicity data which are available on fraction-specific mixtures cover the aromatic fraction (> C5 - C8) and the aliphatic fractions of TPH. Again, the fractions of TPH referred to are those developed by the TPHCWG (Volume 3). Mixture data on the > C9 - C16 and > C16 - C35 aromatic fractions consist of data on C8 - C11 range only. In addition, data on petroleum components > C35 are nonexistent. However, compounds above C20 are not volatile or soluble in groundwater and will remain at the release site. In addition, compounds > C35 are not likely to be bioavailable by the oral or dermal routes (Brainard and Beck, 1992).
Figure 2. Toxicity Assessment Methods

(Most technically defensible approach that can be utilized with current data.)
Based on the lack of whole product toxicity data and the interest in assessing the impact of fate and transport on the risk of petroleum mixtures, the indicator/surrogate approach was chosen as the best available method for the hazard assessment of TPH. Another consideration in the selection of the indicator/surrogate approach was the use of this approach by two regulatory agencies: Massachusetts Department of Environmental Protection (MADEP) and British Columbia Environment (BCE) (MADEP, 1994; BCE, 1995). The TPHCWG method differs from the methods of these agencies in the way in which TPH is fractionated and the use of toxicity data on mixtures to derive the fraction-specific RfDs/RfCs. Again, the use of mixture data is preferable since it accounts for the interaction of compounds within the fraction. In summary, based on the uncertainties discussed above, a combination of data on individual compounds (indicators) and fraction-specific mixtures (surrogates) was chosen as the TPHCWG hazard assessment methodology. This methodology evaluates carcinogenic indicators to which the receptor is exposed individually. This is consistent with USEPA methodology for carcinogens. If these indicators are not present or are present below levels of concern, the remaining mass of petroleum is evaluated using fraction-specific surrogates. The fraction-specific composition of the mixture to which the receptor is exposed is determined, and surrogate RfDs/RfCs are utilized to determine risk or develop cleanup goals. The use of fraction-specific surrogates accounts for the effect of fate and transport on the whole mixture or parent product in that changes in the relative mass of each fraction at the receptor will be accounted for in the risk assessment. The assumptions (uncertainties) that remain in this method are as follows: (1) the method assumes that the toxicity of the fraction as tested does not significantly change with weathering in the environment, (2) that the composition of the fraction will not vary significantly from the surrogate tested, and (3) that the interaction of various fractions can be assumed to be additive. In Volume 5, a comparison of the cleanup goals derived for fresh gasoline and fresh jet fuel using the whole product and the TPHCWG method will be presented. For medicinal grade mineral oil, the whole product data match the TPHCWG method because medicinal grade mineral oil data was used to develop the > C16 C35 RfD. MADEP conducted a similar exercise for their fractionation method with gasoline (MADEP, 1994). The two cleanup goals were within an order of magnitude, and the level of uncertainty was deemed acceptable by MADEP.
2. Ecological Considerations and Assessment of New Toxicity Data
It should be noted that only the human health hazard has been evaluated in this project. Ecological receptors may or may not be protected by utilizing the RfDs/RfCs proposed in this document. The oral and inhalation RfDs developed in this report are for the sum of all constituent compounds that make up each fraction of TPH. The RfDs are based on the toxicity data for the surrogates (single compounds or, preferably, mixtures) which best represent the composition of each fraction. The RfDs have been developed based on all available toxicity data for both individual compounds and mixtures. This data was the best currently available; no new data were generated in this
project. However, it is recognized that new data are being generated which should be evaluated when available. In fact, new data will continue to be generated on petroleum compounds and mixtures and the fraction-specific RfDs should be reevaluated periodically. The RfDs/RfCs in combination with exposure concentrations are then utilized to develop hazard quotients for each fraction. The hazard quotients are summed according to USEPA methodology to develop a Hazard Index for the mixture actually present on site (USEPA, 1986). A description of the use of the RfDs/RfCs and fate and transport parameters (Volume 3) to evaluate the risk of TPH can be found in Volume 5. Both oral and inhalation (for volatile fractions) criteria have been developed applying USEPA methods which are described below. USEPA methods were strictly followed unless otherwise noted.
B. EVALUATION OF TOXICITY INFORMATION 1. USEPA Methodology for RfD/RfC Development
The USEPA issued The Risk Assessment Guidelines of 1986 in which methods for developing reference dose (RfD) values were given. Since then, Proposed Guidelines for Neurotoxicity; Guidelines for Developmental Toxicity Risk Assessment; and Guidelines for Reproductive Toxicity Risk Assessment have been published, which also discuss the development of RfDs. In addition, the IRIS database provides guidance for developing RfD values in Background Document 1 (Reference Dose: Use in Health Risk Assessment), last revised March 15, 1993. Information on the development and use of RfDs is also provided in Risk Assessment Guidance for Superfund Volume 1 Human Health Evaluation. A review of the available information shows that the USEPAs methods for determining RfDs remain unchanged. The USEPA issued Methods for Derivation of Inhalation Reference Concentrations and Application of Inhalation Dosimetry in which the methodology for developing inhalation reference concentrations (RfCs) is presented.
a. Definition of the Reference Dose/Concentration (RfD/RfC)
The RfD is an estimate (with uncertainty spanning perhaps an order of magnitude) of daily exposure to the human population, including sensitive subgroups, that is likely to be without appreciable risk of deleterious effects during a lifetime (USEPA, 1989). The RfC is an estimate (with uncertainty spanning perhaps an order of magnitude) of continuous inhalation exposure to the human population, including sensitive subgroups, that is likely to be without appreciable risk of deleterious effects during a lifetime (USEPA, 1994). Both the RfD and RfC are used to evaluate potential noncarcinogenic effects of exposure to a given compound. It is not used to evaluate carcinogenic endpoints. The RfD/RfC is operationally derived from a no-observed-adverse-effect-level (NOAEL) by application of uncertainty factors (UFs) that reflect various types of data sets used to estimate a reference value and by application of a modifying factor (MF) which reflects the completeness of the database.
b. Appropriate Data for Development of an RfD/RfC
The first step in developing an RfD/RfC is to choose a critical study and determine the NOAEL. The NOAEL is the highest dose at which no adverse effects are observed. If a NOAEL is not available, a lowest-observed-adverse-effect-level (LOAEL) can be used; however, this adds an additional uncertainty factor into the equation. The most appropriate source of the NOAEL for the oral RfD (or LOAEL) is from a chronic oral study. If there are no chronic oral data, subchronic oral data can also be used, but this adds another level of uncertainty into the derivation of the RfD. In some cases, chronic and subchronic inhalation studies can be used to develop oral RfDs when no oral data are available. Similarly, the most appropriate studies for the development of inhalation RfCs are chronic inhalation studies. Oral studies are not used in the development of inhalation RfCs. Most other toxicity data (i.e., dermal, acute, or genotoxic) are not recommended for use in the development of RfDs/RfCs.
c. Derivation of an RfD/RfC in This Document i. Oral RfD
The oral RfD is calculated using the following equation: RfD = NOAEL (or LOAEL)/(UF x MF) where the oral RfD is expressed in mg/kg/day; the NOAEL (or LOAEL) represents a critical effect; the uncertainty factor (UF) can range from 1 to 10,000.
ii. Inhalation RfC
According to USEPA methodology for Category 3 gases (exhibit their toxic effects outside of the respiratory tract), the inhalation RfC is calculated using the following equations: NOAELADJ = E x D (h/24 h) x W (days/7 days) where the NOAELADJ is expressed in mg/m3; E is the exposure level; D is the number of hours exposed; and W is the number of days of exposure. This equation is used to convert to a continuous exposure. NOAELHEC = NOAELADJ x (Hb/g)A/(Hb/g)H where the NOAELHEC is expressed in mg/m3; NOAELADJ is calculated from the equation above; (Hb/g)A/(Hb/g)H is the ratio of blood:gas (air) partition coefficient of the chemical for the laboratory animal species to the human value (if these values are unknown, the default ratio is 1). RfC = NOAELHEC (or LOAEL)/(UF x MF) where the inhalation RfC is expressed in mg/m3; the uncertainty factor (UF) can range from 1 to 10,000.
d. Uncertainty and Modifying Factors
Following are explanations of the different uncertainty factors used in the derivation of RfDs/RfCs: Use a 10-fold factor when extrapolating from valid experimental results in studies using prolonged exposure to average healthy humans. This factor is intended to account for the variation in sensitivity among the members of the human population and is referenced by USEPA as 10H. Use an additional 10-fold factor when extrapolating from valid results of long term animal studies when results of human studies are either not available or inadequate. This factor accounts for the uncertainty involved in extrapolating from animal data to humans and is referenced by USEPA as 10A. Use an additional 10-fold factor when extrapolating from less than chronic results on experimental animals when there are no useful long-term human data. This factor is intended to account for the uncertainty involved in extrapolating from less than chronic NOAELs to chronic NOAELs and is referenced by USEPA as 10S. Use an additional 10-fold factor when deriving an RfD from a LOAEL instead of a NOAEL. This factor is intended to account for the uncertainty involved in extrapolating from LOAELs to NOAELs and is referenced by USEPA as 10L. The modifying factor is an additional uncertainty factor that has been occasionally applied based on the strength of the database and professional judgment. The MF ranges from 0 to 10. The development of benchmark doses (vs. NOAEL-based) was considered; however, the methodology for deriving a benchmark dose was believed to be less established/more controversial at this time (USEPA, 1995). Thus, the NOAELbased methodology was utilized based on current practice/level of acceptance.
2. Prioritization of Toxicity Studies
The toxicity data evaluated were any subchronic, chronic, reproductive/ developmental, immunotoxicity or neurotoxicity data available on the 250 individual compounds identified in unweathered petroleum (multiple products and crudes) by the analytical section of the TPHCWG (Rhodes and Albertson, 1996). In addition, all available data on fraction-specific mixtures were identified and evaluated. In the case of multiple studies, preference was given to the mixture data for the following reasons. Based on studies using mixtures of benzene and toluene, polynuclear aromatic hydrocarbons (PAHs), human data relevant to the Gulf War, etc., it is obvious that the toxic potency of individual compounds can be influenced by the presence of other materials. Since petroleum is a mixture, it is most appropriate to evaluate it as such. In addition, only 250 of the thousands of compounds within petroleum have been identified. Of the 250 identified, only approximately 40 have enough
toxicity data available to develop RfDs/RfCs and only 95 have any toxicity data whatsoever (see Appendices A and B). It would be extremely costly to complete toxicity studies on all individual components in petroleum even if it were possible to analytically identify them. It is the belief of the TPHCWG that what is appropriate to study from a risk perspective is the toxicity of fraction-specific mixtures (as defined on the basis of environmental transport), since these are the mixtures that receptors will be exposed to in the environment. In developing oral RfDs or inhalation RfCs, route to route extrapolation was minimized; i.e., oral studies were used for oral criteria and inhalation studies for inhalation criteria wherever possible, based on US EPA preference (above). Finally, where more than one appropriate study was available, a weight-of-evidence approach was applied to develop the reference dose. The weight-of-evidence approach used for each fraction is described in detail in the respective sections of this report.
II. SUMMARY/CONCLUSIONS
The fraction-specific RfDs/RfCs recommended by the TPHCWG are listed in Table 1. The development of the fractions, which are based on environmental fate and transport characteristics of the identified constituents, is described in detail in the TPHCWG Volume 3 (Gustafson et al., 1997). The technical basis for the individual criteria may be found in Sections III and IV. Note that the criteria for the aromatic fractions are at least an order of magnitude lower than the criteria for the aliphatic fractions. This is based on uncertainty as well as toxicity. The availability of mixtures data is much greater for the aliphatic fractions than for the aromatic fractions; however, the toxicity data on aromatics indicate greater potency. Thus the aromatic fractions (for carbon numbers C9-C16 and C17-C34) are the fractions for which more uncertainty regarding potency exists. Prioritization of further testing should also account for the relative composition of fractions present. However, based on quality/quantity of toxicity data alone, uncertainty in this methodology could be reduced to the greatest extent by testing of mixtures representative of the C9-C16 and C17-C34 aromatic fractions. Testing of fraction-specific mixtures is being considered by British Columbia Environment(for ecological receptors) and by the U. S. Air Force (for human health). The Gas Research Institute and Petroleum Environmental Research Forum (PERF) are conducting testing of hydrocarbon contaminated soils, as well as developing fraction- specific analytical data which may soon be available to calibrate the TPHCWG methodology. There are also new data being developed on cyclohexane, mineral oil, and n-nonane which may directly impact the fraction-specific RfDs which are most appropriate for TPH. Finally, it is appropriate to mention that the TPHCWG did not originate the idea of utilizing fractions to evaluate the risk of TPH. The Massachusetts Department of Environmental Protection appears to have developed the first fractional approach (MADEP, 1994). In 1995, British Columbia Environment modified the MADEP approach to include fate and transport of fractions and to be specific for ecological receptors of concern in the province (BCE, 1995). The TPHCWG would like to thank these agencies for their leadership in this area.
Table 1. Preliminary TPHCWG Toxicology Fraction-Specific Oral RfDs (mg/kg/day) and Inhalation RfCs (mg/m3)
Carbon Range a Critical Effect Hepatoxicity, Nephrotoxicity 5.0 18.4 0.4
Aromatic Oral RfD (mg/kg/day) Neurotoxicity
Aromatic Inhalation RfC (mg/m3)
Aliphatic Oral RfD (mg/kg/day)
Aliphatic Inhalation RfC (mg/m3)
Critical Effect
Aliphatic C5 - C6 C>6 - C8 Aromatic C>7 - C8 0.2 Decreased body weight 0.1 1.0
0.20
C>8 - C10 C>10 - C12 C>12 - C16 NA Nephrotoxicity 2.0 NA
0.04
Hepatic and hematological changes Hepatic (foreign body reaction) granuloma
C>16- C21 C>21 - C35
0.03 b
Carbon range - actually equivalent carbon range (Gustafson et al., 1997). This is the pyrene (C16) value. NA = Not Available.
III. EVALUATION OF AROMATIC FRACTIONS

In this section, the oral RfDs and inhalation RfCs were evaluated along with other available data for chemical constituents in each carbon fraction range. Sources of this information included IRIS (1996), HEAST (1995), and the EBSI literature search (see Appendix A). The general approach determined by the TPHCWG which was used in this section involved the development of RfDs and RfCs for an entire fraction and not for a single compound. These recommended fraction- specific RfDs/RfCs are based on all available data which includes information on both individual compounds and mixtures within the given carbon range. The fractionspecific RfD/RfC is relevant at real-world sites if any compounds which make up the particular fraction are present. For each aromatic fraction, the rationale for the fraction-specific RfDs and inhalation RfCs is presented below. Summaries of the data used to determine these oral RfDs and inhalation RfCs can be found in Appendix B. One important point to remember is that as new data become available for aromatic compounds, the surrogate RfDs/RfCs should be reevaluated.
A. C>7 - C8 AROMATIC FRACTION
1. Oral
According to the analytical information presented in Volume 3 (Gustafson et al., 1997), seven compounds were identified in petroleum products in this carbon range. Oral RfDs (on IRIS) are available for six of these compounds: ethylbenzene, styrene, toluene, m-xylene, o-xylene, and p-xylene. From the EBSI literature review (see Appendix A), there were no other studies on the other compounds in this fraction that could be used in the development of RfDs. Table 2 lists the single compound RfDs for this fraction. These values range from 0.1 to 2.0 mg/kg/day. Summaries of the data used in the development of these RfDs can be found in Appendix B. Because there are six RfDs available for the seven compounds identified in this range, these values can be considered to be representative of the entire fraction. After reviewing the available toxicity and compositional information for this fraction, it was determined that the oral RfD of 0.2 mg/kg/day is appropriate. Although ethylbenzene has a lower RfD, it is the same order of magnitude as the fraction RfD and the RfD for other constituents in the fraction (i.e., toluene). The relative portions of ethylbenzene to other constituents in petroleum is such that the toluene concentration is approximately 10 times that of ethylbenzene in most unweathered products. Also, to use a value of 2 mg/kg/day (xylenes) would not be consistent with the other chemical-specific RfDs for the fraction. Thus, an RfD of 0.2 mg/kg/day was deemed protective. This surrogate value represents the fraction-specific RfD for aromatics in the C5 - C8 carbon range.
10
Table 2. RfDs/RfCs Associated with Compounds in the Equivalent Carbon Ranges

Available Oral RfDs (mg/kg/day) Available Inhalation RfCs (mg/m3) Fraction Oral RfD (mg/kg/day) Fraction Inhalation RfC (mg/m3)
Equivalent Carbon Ranges
Compounds w/Toxicity Data
C>7 - C8 0.2
Toluene (C7) Ethylbenzene (C8) Styrene (C8) Xylenes (o-, m- ,and p-) (C8) 0.4
0.2 0.1 0.2 2.0
0.4 1.0 1.0 NA
C>8 - C10 C>10 - C12 C>12 - C16 0.04
Isopropylbenzene (C9) Naphthalene (C10) Acenaphthene (C12) Biphenyl (C12) Fluorene (C13) Anthracene (C14) Fluoranthene (C16) Pyrene (C16) 0.04 0.04 0.06 0.05 0.04 0.3 0.04 0.03 NA 0.03 0.2 NA 0.09 0.0013 NA NA NA NA NA NA
0.2
C9 Aromatics Naphthalenes/ Methylnaphthalenes
C>16 - C21 C>21 - C35 NA NA NA
0.03
NA
NA - None Available
11
2. Inhalation
There are three compounds in this fraction which currently have inhalation RfCs on IRIS: toluene (0.4 mg/m3), ethylbenzene (1 mg/m3), and styrene (1 mg/m3) (Table 2). There were no additional inhalation data available on the other compounds in this range that could be used to develop RfCs. Again, these RfCs are felt to be representative of the entire fraction. The recommended inhalation RfC is 0.4 mg/m3. This value should be protective for the entire range of compounds within this fraction.
B. C>8 - C10, C>10 - C12, AND C>12 - C16 AROMATIC FRACTION
1. Oral
Within this carbon range, 77 individual compounds have been identified by the fate and transport section (Tell et al., 1997). Of these identified compounds, oral RfDs have already been developed for 8 of these compounds: isopropylbenzene, acenaphthene, biphenyl, fluorene, anthracene, fluoranthene, naphthalene, and pyrene. After reviewing the information from the literature search, there were no additional studies on individual compounds that could be used to develop additional RfDs. Table 2 lists the aromatic compounds in the C9 - C16 range and the associated oral RfDs. The RfDs range from 0.03 to 0.3 mg/ kg/day. Summaries of the data to develop these RfDs can be found in Appendix B. There are also oral data available on a mixture within this carbon range: naphthalene/ methylnaphthalenes. Data from this mixture were used to develop an RfD which was included in determining the fraction-specific RfD. This value can also be found in Table 2. Following is a brief summary on the development of the RfD for this mixture. Naphthalene/methylnaphthalenes 1. Rats were dosed orally with 0, 300, 600, or 1000 mg/kg for 13 weeks (unpublished data). Mean body weights and food consumption were significantly decreased in male rats at 1000 mg/kg. Histopathologic changes were centrilobular hepatocellular hypertrophy in both sexes at all dose levels; hyperplasia and hypertrophy of the thyroid in both sexes at all dose levels; and hyperplasia of the urinary bladder in male rats at all dose levels and in female rats at 300 mg/kg. NOEL <300 mg/kg LOAEL = 300 mg/kg/day Using an uncertainty factor of 10,000 (10 most sensitive, 10 animal to human, 10 subchronic to chronic, 10 LOAEL to NOAEL) (see Section B.1.c.i in section F): RfD = 0.03 mg/kg/day
12
2. Rats were dosed orally with 0, 75, 150, or 450 mg/kg during gestational days 6-15 (unpublished data). At 450 mg/kg, maternal body weight gain and food consumption were significantly decreased during the first three days of treatment. No adverse developmental effects. maternal NOEL = 150 mg/kg developmental NOEL >450 mg/kg These data on developmental toxicity were not used in the calculation of an RfD for this fraction. The RfD for this mixture is consistent with the RfDs for the other individual compounds in this equivalent carbon range. After reviewing the information on the available individual compounds and mixtures, it was determined that the oral RfD of 0.04 mg/kg/day would be an appropriate fraction-specific RfD for the three fractions in the C9 - C16 carbon range. Most of the available RfDs for individual compounds in this fraction are approximately 0.04 mg/kg/day (of the eight available RfDs, four of the compounds have RfDs of 0.04 mg/kg/day). The only exception is fluorene with a value of 0.3 mg/kg/day. The RfDs on individual compounds represent about 10% of the components that have been identified within the fractions. Data are also available on mixtures. An RfD for the naphthalenes/methylnaphthalenes was calculated as 0.03 mg/kg/day. This value supports the 0.04 mg/kg/day value. Because TPH is a mixture, emphasis needs to be placed on these available mixtures data.
2. Inhalation
Inhalation data are extremely limited in this carbon range. Inhalation RfCs have previously been developed for two individual compounds in this carbon range: isopropylbenzene (C9) (0.09 mg/m3) and naphthalene (C10) (0.0013 mg/m3). These data are not at all representative of the entire fraction. There were several inhalation studies on C9 aromatic mixtures that could be used to develop RfCs. Following is a description of these studies and the development of the RfCs. C9 Aromatics 1. Rats were exposed to 0, 100, 500, or 1500 ppm 6 hours/day, 5 days/week for 13 weeks (Douglas et al., 1993). There was reduced weight gain at 1500 ppm; no neurotoxicity. NOEL = 1500 ppm (7362 mg/m3) Converting to continuous exposure and using an uncertainty factor of 1000 (10 most sensitive, 10 animal to human, 10 subchronic to chronic) (See section B.1.c.ii in section I): RfC = 1.3 mg/m3
13
2. Mice were exposed by inhalation to 0, 100, 500, or 1500 ppm 6 hours/day during gestational days 6-15 (McKee et al., 1990). maternal NOEL = 100 ppm developmental NOEL = 100 ppm These data on developmental toxicity were not used in the calculation of an RfD for this fraction. 3. Rats were exposed by inhalation to 0, 450, 900, or 1800 mg/m3 6 hours/day, 5 days/week for 12 months (Clark et al., 1989). There were increased liver and kidney weights in male rats at 1800 mg/m3. No treatment-related histopathologic effects were observed. NOEL = 900 mg/m3 Converting to continuous exposure and using an uncertainty factor of 1000 (10 most sensitive, 10 animal to human, 10 subchronic to chronic) (see Section B.1.c.ii of section I): RfC = 0.2 mg/m3 The data on the C9 mixtures are more appropriate to use than the information on individual compounds because they are more representative of TPH, which is also a mixture. The mixtures data represent more compounds than the single compound information. The more conservative value, 0.2 mg/m3, was determined to be representative of this entire fraction.
C. C>16 - C21 AND C>21 - C35 AROMATIC FRACTION
1. Oral
There are no previously developed RfDs for chemicals in this equivalent carbon range. The literature search was reviewed and there were no available data to develop an RfD. The majority of the data on compounds in this range consisted of dermal application studies. Dermal studies are not appropriate for use in the development of oral RfDs. After reviewing available information and determining that there are no available RfDs for this group of compounds, the RfD for pyrene will be used as the surrogate for the fraction RfD. Pyrene is considered a conservative surrogate because it has a lower carbon number than any of the compounds in this fraction. This value (0.03 mg/kg/day) represents the fraction-specific RfD for the C17+ carbon range. See Appendix B for these data.
2. Inhalation
There are no appropriate data available for the development of RfCs in this carbon range. Also, the development of an inhalation RfC from this fraction was deter-
14
mined to be inappropriate because the compounds in this carbon range are not volatile and inhalation will not be a relevant exposure pathway.
D. OVERALL RECOMMENDATIONS (SEE TABLE 2)
1. Use the RfD of 0.2 mg/kg/day for C5 to C8 aromatic fraction. Use the RfC of 0.4 mg/m3 for C5 to C8 aromatic fraction. 2. Use the RfD of 0.04 mg/kg/day for C9 to C16 aromatic fraction. Use the RfC of 0.2 mg/m3 for C9 to C16 aromatic fraction. 3. Use the RfD of 0.03 mg/kg/day for C17 to C35 aromatic fraction. No RfC is recommended because the fraction is not volatile.
IV. EVALUATION OF ALIPHATIC FRACTIONS

In this section, the oral RfDs and inhalation RfCs for the aliphatic fractions are developed. As mentioned previously, the values for the aliphatic fractions are at least an order of magnitude greater than those for the aromatic fractions. This is a result of both a difference in uncertainty and potency. The data used to develop aliphatic RfDs were derived from multiple studies on representative mixtures. Therefore, although few USEPA-derived RfDs exist for individual compounds within these fractions, the RfDs for the aliphatic fractions are believed to have a much higher level of certainty than those for the C9 - C16 and C16 - C35 aromatic fractions.
A. C5 - C6 AND C>76- C8 ALIPHATIC FRACTION
n-Hexane is the only aliphatic compound in this fraction for which the USEPA has developed an inhalation Reference Concentration (RfC). Because of its unique toxicity, the use of n-hexane as the basis for the fraction-specific RfD considerably overestimates the health risks of hydrocarbons in this fraction. This section will present two toxicity data sets that are more representative of the potency of the C5-C8 aliphatic fraction. The first data set is on n-heptane, which has been extensively studied because it is structurally similar to n-hexane, and it can be metabolized to a gamma diketone metabolite, which is neurotoxic. The second data set includes toxicity studies on a solvent mixture containing hexane isomers. It is proposed that the health-based criteria for the C5-C8 alkane fraction be based on a percentage basis of n-hexane in relation to the rest of the hydrocarbons in this fraction.
1. n-Heptane
Animal studies have failed to demonstrate peripheral neuropathy (the critical effect of n-hexane exposure) from n-heptane exposure. Frontali et al (1981) showed that n-hexane produced neurotoxic effects, including axonal degeneration after 30 weeks of exposure, whereas n-heptane, as well as n-pentane and other hexane isomers did not. Takeuchi et al. (1980, 1981) also found no signs of abnormal neurobehavioral effects in rats exposed to 3000 ppm n-heptane or n-pentane
15
12 hours/day, 7 days/week for 16 weeks. There was no evidence of peripheral neuropathy and motor activity was normal. Rats exposed to 400 or 3000 ppm nheptane 6 hours/day, 5 days/week for 26 weeks showed no signs of neurotoxicity (API, 1980). However, a possible metabolite of n-heptane, the gamma diketone 2,5-heptanedione, has been shown to produce neurotoxic effects when administered to animals (Katz et al., 1980; Misumi and Nagano, 1984). 2,5-Heptanedione produced clinical signs of neuropathy and neuropathological alterations identical to those produced by the gamma diketone of n-hexane (2,5-hexanedione) at doses greater than 1000 mg/kg (ODonoghue and Krasavage, 1979). Pharmacokinetic studies have attempted to quantitate the neurotoxic risk of nheptane with that of n-hexane (Kreuzer et al., 1995). Human field studies have shown that the presence of 2,5-hexanedione in urine is a relatively specific indicator of exposure to n-hexane. Urinary 2,5-hexanedione is in fact recommended by ACGIH as a biological exposure index (BEI) to n-hexane (ACGIH, 1987). To determine the relative neurotoxic risk of n-heptane to that of n-hexane, pharmacokinetic studies have compared the urinary levels of both gamma diketones in animals and humans exposed to either n-hexane or n-heptane. These studies have shown that, when rats and human volunteers were exposed to either n-hexane (up to 300 ppm) or n-heptane (up to 500 ppm), there was a 38fold lower amount of urinary gamma-diketone in humans and rats exposed to nheptane compared with n-hexane. Since these gamma-diketones are the metabolites responsible for the neurotoxic effects, the neurotoxic risk is expected to be at least 38-times lower for n-heptane than for n-hexane. Furthermore, 2,5-heptanedione and 2,5-hexanedione also differ in neurotoxic potency. 2,5-Heptanedione appears to be approximately 2.5 to 5 times less potent in producing neurotoxicity than 2,5-hexanedione. In the rat, 2,5-heptanedione at doses greater than 1000 mg/kg/day produced signs of neuropathy and neuropathological alterations (ODonoghue and Krasavage, 1979), whereas, 2,5-hexanedione produced clinical neuropathy at doses as low as 400 mg/kg/day and altered nerve conduction at 200 mg/kg/day (Eben et al., 1979). Thus, n-heptane is unlikely to pose a neurotoxic hazard at exposures similar to that of n-hexane. Even conservatively assuming that 2,5-heptanedione and 2,5-hexanedione were equipotent as neurotoxicants, n-heptane is considered to have a neurotoxic risk that is 38 times lower than that of n-hexane. The inhalation reference concentration (RfC) for n-hexane is 0.2 mg/m3, which is based on neurotoxic effects in humans. Assuming that the inhalation rate for a 70 kg human is 20 m3/day and absorption is 100%, an oral reference dose (RfD) for n-hexane is 0.06 mg/kg/day. Since the RfD for n-hexane is based on neurotoxicity, then an oral reference dose of nheptane should also be 38 times higher than that of n-hexane or 2 mg/kg/day. An RfD of 2 mg/kg/day is a less conservative estimate of the health risks of hydrocarbons in the C5-C8 fraction than the RfD for n-hexane. With the exception of n-hexane and n-heptane, C5-C8 hydrocarbons have not been shown to cause neurotoxicity, nor can they be metabolized to the neurotoxic gamma diketone metabolites. Thus, n-heptane could be considered an appropriate surrogate for the C5-C8 hydrocarbons, with the exception of n-hexane.
16
2. Commercial Hexane
A solvent containing hexane isomers has been extensively tested for health effects as part of a USEPA mandated Test Rule under Section 4 of the Toxic Substance Control Act (see Table 3 summary). The hexane mixture, called commercial hexane, contained 53% n-hexane, 16% 3-methylpentane, 14% methylcyclopentane, 12% 2-methylpentane, 3% cyclohexane, 1% 2,3-dimethylbutane, and <1% several minor compounds. The results of the studies are summarized in Table 3. Overall, these studies show that an inhalation exposure to a hexane mixture containing 53% n-hexane produces little toxicity. There were no effects on either the peripheral or central nervous system; no reproductive or developmental toxicity; and no target organ effects. In establishing an inhalation RfC for commercial hexane mixture, the two chronic bioasssays should be considered because these studies involved lifetime exposure. The NOAEL for either the rat or mice chronic bioassay is 3000 ppm (10,307 mg/m3). Adjusting for continuous exposure (6 hours/24 hours and 5 days/7 days), the NOAEL = 1840 mg/m3. Using an uncertainty factor of 100 (animal to human extrapolation and intrahuman variability), the RfC is 18.4 mg/m3. An oral reference dose for commercial hexane can be calculated using the inhalation RfC. Assuming that the inhalation rate for a 70 kg human is 20 m3/day and absorption is 100%, then an oral RfD for commercial hexane is 5 mg/kg/day. This value is similar to the RfD derived for n-heptane. Both values are almost two orders of magnitude higher than the RfD for n-hexane demonstrating that n-hexane is uniquely toxic and is not representative of the entire C5-C8 alkane/cycloalkane fraction. Finally, these data provide further evidence that the presence of other petroleum compounds influences the toxicity of n-hexane and that mixture data should be utilized to evaluate the risk of petroleum mixtures.
3. Other C5-C8 Alkane/Cycloalkane Compounds
Other C5-C8 alkane/cycloalkane hydrocarbons have been tested for subchronic and chronic toxicity. Frontali et al (1981) compared the neurotoxicity of n-hexane with n-pentane, cyclohexane, 2-methylpentane, and 3-methylpentane. Peripheral neurotoxicity was only observed with n-hexane. This is not unexpected since only n-hexane can be metabolized to a gamma diketone (2,5-hexanedione) which is the metabolite responsible for the neurotoxic effects. n-Octane is the only other hydrocarbon in the C5-C8 alkane fraction that could potentially form the gammadiketone. However, metabolism studies were not able to detect the gamma-diketone in rats dosed with n-octane (Olson et al, 1986). Cyclohexane has been tested for subchronic toxicity. Treon et al. (1943) reported microscopic changes in the liver and kidney of rabbits exposed 6 hours/day, 5 days/week for 10 weeks to 786 ppm. No effects were observed in rabbits exposed to 434 ppm for either 10 or 26 weeks. Treon et al. (1943) also reported no treatment-related effects in a monkey exposed to 1243 ppm cyclohexane for 10 weeks. New data on cyclohexane toxicity will soon be available, as it is currently part of a USEPA mandated test rule under Section 4 of the Toxic Substances Control Act. These data may impact the RfD for this fraction and therefore should be examined upon release.
17
18
Table 3. Commercial Hexane Test Rule Studies

Study Design Increased liver weights in female rats (9000 ppm) Male rat nephropathy Increased liver weights in male and female mice at 9000 ppm No neurobehavioral or neuropathologic effects Histologic evidence of mucosal irritation in nasal turbinates and larynx at 9000 ppm Decreased severity and incidence of cystic uterine endometrial hyperplasia at 9000 ppm No neoplastic effects 9000 ppm 3000 ppm 3000 ppm Findings NOAEL Reference Duffy et al. (1991)
Toxicity Test
Species
Subchronic
Rat
0, 900, 3000, and 9000 ppm 6 hours/day, 5 days/week for 13 weeks
Subchronic
Mouse
0, 900, 3000, and 9000 ppm 6 hours/day, 5 days/week for 13 weeks
Duffy et al. (1991)
Subchronic Neurotoxicity
Rat
0, 900, 3000, and 9000 ppm 6 hours/day, 5 days/week for 2 years
Soiefer et al. (1991)
Chronic
Rat
3000 ppm
Kelly et al. (1994)
Chronic
Mouse
3000 ppm
Daughtrey et al. (1994)
Oncogenicity
Rat
9000 ppm
Kelly et al. (1994)
Oncogenicity
Mouse
Liver tumors in female mice at 9000 ppm
3000 ppm
Reproductive
Rat
0, 900, 3000, and 9000 ppm 6 hours/day, 5 days/week for 2 generations
Reduced body weight gain in offspring of both generations No adverse reproductive effects Maternal toxicity at 9000 ppm No developmental effects Maternal toxicity at 9000 ppm No developmental effects
3000 ppm
Developmental
Rat
0, 900, 3000, and 9000 ppm 6 hours/day on days 6-15 of gestation
3000 ppm (maternal) 9000 ppm (developmental) 3000 ppm (maternal) 9000 ppm (developmental)
Keenan et al. (1991)
Developmental
Mouse
0, 900, 3000, and 9000 ppm 6 hours/day on days 6-15 of gestation
Keenan et al. (1991)
In a subchronic study, rabbits were exposed by inhalation to methylcyclohexane vapor for 10 weeks. Liver and kidney effects were reported in rabbits exposed to 2880 ppm, but there were no effects at 1200 ppm (Treon et al., 1943). A monkey exposed to 370 ppm methylcyclohexane for 10 weeks showed no treatment-related effects (Treon et al., 1943). A chronic study has been conducted with methylcyclohexane (Kinkead et al., 1985). Rats, mice, hamsters, and dogs were exposed by inhalation to 0, 400, and 2000 ppm 6 hours/day, 5 days/week for 12 months. At 12 months, some of the mice, rats, and hamsters were terminated; the remaining rodents were held for an additional year and the dogs for five years. There was no increase in tumors in any of the exposed animals. The only treatment-related finding was kidney nephropathy in the 2000 ppm exposed male rats.
4. Proposed Composition-Weighted RfD for TPH Fraction Containing C5-C8 or C6-C8 Aliphatics
Based on the data given above, there are two alternatives for an RfD for this fraction: utilize the hexane RfD(0.06 mg/kg/day) for the n-hexane portion and the n-heptane RfD (2.0 mg/kg/day) for the remainder of the mass. evaluate the hexane concentration separately. If the n-hexane concentration is less than 53% as found in commercial hexane, then the RfD applied should be 5mg/kg/day. If it is greater than 53%, the RfD should be developed utilizing 0.06 for the nhexane portion and 2.0 for the remaining mass(as above). Based on the fact that only n-heptane has shown any toxicity at these levels, both of these options are very conservative. An evaluation of the composition of petroleum products was conducted to produce a single value for both the C5-C8 and the C6-C8 fractions. Both fractions were examined because the analytical group of the TPHCWG did not feel that current methodologies captured C5 reliably. It is felt that the composition of petroleum products containing n-hexane is well known and ranges from 0.05% in some gasolines to 15.7% in Sweetened Naphtha (TPHCWG Volume 2, 1997). The only products containing C5-C8 and therefore n-hexane are gasoline, crude, and the petroleum streams listed in Table 4. Gasoline and crude have low levels of n-hexane. Based on an analysis of 28 samples, n-hexane in gasoline ranges from 0.05 to 7.0% with a median content of 1.7% and an average content of 2.2% (PERF 94-05). The n-hexane content of two crude types were analyzed (TPHCWG Volume 2, 1996). The average concentration of n-hexane in crude was 1.3% with a minimum content of 0.7% and a maximum content of 1.8%. The content of nhexane in petroleum refinery streams ranges from 0.06 to 15.71%; encompassing the ranges found in gasoline and crude. Data are available from API to evaluate the n-hexane content within the fractions (C5-C8 and C6-C8) of petroleum streams and these data are shown in Table 4. Again, this is the content within the fractions, whereas the data above are for n-hexane content in the whole product. Utilizing the two recommended procedures (described above) for developing RfDs for the two fractions ( C5-C8 or C6-C8) ,the average RfDs are 2.0 and 5.0 mg/kg/day. Based on the level of conservatism inher-
19
20
Table 4. Composition-Weighted RfDs for C5 - C8 and C6 - C8 Fractions

C5 - C8 Aliphatic Fraction ________________________________ n-Hexane (wt. %) 2.4 1.8 16.9 7.8 7.8 1.7 3.3 2.0 2.0 1.8 1.8 1.7 2.0 2.7 26.4 9.7 9.2 3.0 2.3 2.0 3.7 RfD* n-Hexane (wt. %) C6 - C8 Aliphatic Fraction __________________________________ RfD* 1.9 1.9 1.5 1.8 1.8 2.0 2.0
Stream
Light catalytically cracked naphtha (API 81-03)
Light catalytically cracked naphtha (API 81-04)
Sweetened naphtha (API 81-08)
Light catalytically reformed naphtha (API 81-04)
Full range catalytically reformed naphtha (API 81-05)
Heavy catalytically cracked naphtha (API 83-18)
Heavy catalytically cracked naphtha (API 84-02)
* RfD for n-hexane is 0.06 mg/kg/day; RfD for non n-hexane hydrocarbons is 2 mg/kg/day
ent in the RfD development and the fact that the range of n-hexane in petroleum and commercial hexane are well known, 5mg/kg/day is believed to be appropriate for all situations with the exception of the rare release of high purity n-hexane(the only material believed to contain greater than 53% n-hexane). This rare situation would be easily detectable using the TPHCWG analytical methodology. Thus, the recommended oral RfD of 5 mg/kg/day and an inhalation RfC of 18.4 mg/m3 for this fraction.
B. C>8 - C10, C>10 - C12, AND C>12 - C16 ALIPHATIC FRACTION
There are minimal toxicity data available on individual components within the C9C16 aliphatic range(Appendices A and B). The data which were utilized to develop oral and inhalation criteria for this fraction were studies on JP-8 (C9-C16) and studies on dearomatized petroleum streams which together cover the entire range of the fraction (Figure 3). API data on other petroleum streams were evaluated but either the toxicity studies were not appropriate to develop criteria or the compositional information was inadequate to ascertain whether or not a match could be made to this or other fractions (Appendix C). It should also be mentioned that data are being generated (both oral and inhalation) by the USAF on n-nonane. These data should be evaluated when available; however, the data on petroleum streams are still considered preferable based on the fact that these are data on mixtures rather than on an individual compound at the low end (C9) of the fraction. MADEP had previously developed an RfD of 0.6 mg/kg/day for n-nonane based on potency vs. n-hexane; no effects were seen in a
Figure 3. Distribution of Studies in the C9 to C16 Carbon Range
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Table 5: Inhalation Studies and RfDs Developed for the C9 - C16 Fraction
Fraction C10 - C11 Subchronic (Study 1) Developmental (Study 2) C7 - C11 Subchronic (Study 3) Developmental (Study 4) JP-8 (C9 - C16) Subchronic (Study 5)
Effect No Significant Adverse Effects None
NOAEL 900 ppm (5226 mg/m3) 900 ppm (5226 mg/m3)
RfC (mg/m3) 0.9
No Significant Adverse Effects None
900 ppm (5485 mg/m3) 900 ppm (5485 mg/m3)
1.0
1.0 No Significant Adverse Effects 1000 mg/m3
subchronic inhalation study at 10x, the hexane LOAEL concentration in a similar study (MADEP,1994). Finally, in selecting appropriate RfD/RfCs, the data on JP-8 are given less weight than the petroleum stream data since the JP-8 has up to 20% aromatic content vs. the petroleum streams which have at most 1.5% aromatics and in most cases had less than 0.1% aromatics. The RfC selected for this fraction was 1.0 mg/m3 and the RfD was 0.1 mg/kg/day.
1. Summary of Inhalation Studies on Dearomatized Petroleum Streams and JP-8
All studies were conducted according to USEPA TSCA/FIFRA or OECD guidelines. All of the aliphatic streams showed male rat nephropathy, however, this endpoint was dismissed in determining the NOAELs based on a determination by the USEPA that it is not relevant to humans (Alden, 1986). The studies and inhalation RfCs developed are shown in Table 5.
a. Composition: C10-C11 Isoparaffinic solvent; aromatic content: <0.01%
Two studies were conducted on this product - a sub-chronic and a developmental study.
Study 1. Subchronic Toxicity Study (Phillips and Egan, 1984)
Sprague-Dawley rats were exposed by inhalation to 0, 300, or 900 ppm (0, 1742, or 5226 mg/m3) for 6 hours/day, 5 days/week for 12 weeks. There were questionable body weight effects at both dose levels. At study termination, increased kidney weights were observed in male rats at 300 and 900 ppm, and increased liver weights in male rats at 900 ppm. Male rat nephropathy was found at both dose levels. None of the effects observed were considered significant. NOAEL = 900 ppm (5226 mg/m3) Converting to continuous exposure and using an uncertainty factor of 1000 (10 most sensitive, 10 animal to human, 10 subchronic to chronic):
22
RfC = 0.9 mg/m3

Study 2. Developmental Toxicity Study (Mullin et al., 1990)
Rats were exposed by inhalation to 0, 300, or 900 ppm (0, 1742, or 5226 mg/m3) during gestational days 6-15. No maternal or developmental effects were observed. maternal NOEL = 900 ppm (5226 mg/m3) developmental NOEL = 900 ppm (5226 mg/m3) These data on developmental toxicity were not used in the calculation of an RfD for this fraction.
b. Composition: dearomatized white spirit; C7-C11 isoparaffins/n -alkanes/napthenes; typical aromatic content: 0.1%
Two studies were conducted on this product - a subchronic and a developmental toxicity study.
Study 3. Subchronic Toxicity Study (Phillips and Egan, 1984)
Sprague-Dawley rats were exposed by inhalation to 0, 300, or 900 ppm (0, 1828, or 5485 mg/m3) 6 hours/day, 5 days/week for 12 weeks. Decreased weight gain was observed at 900 ppm. Liver and kidney weights were increased in males at 900 ppm. Male rat nephropathy was observed at all dose levels. None of the effects observed were considered significant. NOAEL = 900 ppm (5485 mg/m3) Converting to continuous exposure and using an uncertainty factor of 1000 (10 most sensitive, 10 animal to human, 10 subchronic to chronic): RfC = 1.0 mg/m3
Study 4. Developmental Toxicity Study (unpublished data)
Sprague-Dawley rats were exposed by inhalation to 0, 300, or 900 ppm (0, 1828, or 5485 mg/m3) 6 hours/day during gestational days 6-15. No maternal or developmental toxicity were observed. maternal NOEL = 900 ppm (5485 mg/m3) developmental NOEL = 900 ppm (5485 mg/m3) These data on developmental toxicity were not used in the calculation of an RfD for this fraction.
23
c. JP-8 Jet Fuel Study 5. Subchronic Toxicity Study (Mattie et al., 1991)
Rats and mice of both sexes were exposed to JP-8 vapors at 0, 500, and 1000 mg/m3 continually for 90 days. The exposure period was followed by a 24-month recovery period. A decrease in body weight (not significant) was observed in male rats (all dose levels). A statistically significant increase in basophilic foci in the liver was observed in male rats. There was an increased splenic hematopoiesis in female rats at termination. These results were not considered to be treatment related. The only observation in mice was an increased mortality which was due to necrotizing dermatitis. The dermatitis was a result of increased fighting among the animals. Overall, there was no significant toxicological effect from exposure to JP-8 under these test conditions. NOAEL = 1000 mg/m3 Using an uncertainty factor of 1000 (10 most sensitive, 10 animal to human, 10 subchronic to chronic): RfC = 1.0 mg/m3
2. Summary of Oral Gavage Studies on Petroleum Streams and JP-8
All studies are compliant with either USEPA FIFRA or OECD guidelines. None are published, or authorized for public release in the United States. Again, all aliphatic streams showed male rat nephropathy. However, this endpoint was dismissed in determining NOAELs based on USEPA guidance (Alden, 1986). These studies and RfDs developed are shown in Table 6.
a. Composition: dearomatized aliphatic; C9-C12 isoparaffins/n-alkanes/ naphthenes; typical aromatic content: 0.1% Study 6. Subchronic Toxicity Study (unpublished data)
Sprague-Dawley rats were dosed orally with 0, 500, 2500, or 5000 mg/kg for 90 days. A high dose recovery group was also included. Decreased body weights were observed in the male rats in the 2500 and 5000 mg/kg groups. Increased food consumption in the 2500 (males) and 5000 mg/kg (males and females) was observed. Increases in platelets were observed in the 500 (males), and 2500 (males), and 5000 (males and females) dose groups. In male rats, increases in alanine aminotransferase were observed in the 2500 and 5000 mg/kg dose groups and increases in glutamyl transferase were observed in the 5000 mg/kg dose group. No changes in these parameters were observed in the female treated animals. Treatment-related microscopic changes were observed in the
24
Table 6: Oral Studies and RfDs for C9 - C16 Fraction

Effect LOAEL - 500 mg/kg/day NOAEL Oral RfD 0.1 mg/kg/day
Fraction
C9 - C12 Subchronic (Study 6)
Reversible Liver Hypertrophy and Hematological Alterations
C10 - C13 Subchronic (Study 7) Reversible Liver Hypertrophy 100 mg/kg/day
0.1 mg/kg/day
C15 - C18 Developmental (Study 8) None
> 1000 mg/kg/day
C11 - C17 Subchronic (Study 9) Reversible Liver Weight Increase
100 mg/kg/day
0.1 mg/kg/day
JP-8 (C9 - C16) Subchronic (Study 10) No Significant Adverse Effects - On Going -
750 mg/kg/day - On Going -
0.75 mg/kg/day - On Going -
n-Nonane (C9)
25
kidneys of male rats at all dose groups; the liver (hepatocellular hypertrophy) of male/female rats at all dose groups; and the stomach (edema and hyperplasia) and/or anus of males/females at the 2500 and 5000 mg/kg groups. All effects were reversible in the recovery group within the 4-week recovery period. Mechanistically, two simultaneous events seem to be occurring. (1) Male rat nephropathy. (2) The direct effect of high-dose intubation of a locally irritating substance. It is believed that the doses employed produced irritation of the gastrointestinal (GI) tract, which led to many of the other observed effects. The effects observed could not be dismissed and therefore a NOAEL could not be developed. LOAEL = 500 mg/kg/day The uncertainty factors that were used in the calculation were: 10 - animal to human 10 - most sensitive 10 - subchronic to chronic 5 - LOAEL to NOAEL A value of 5 was chosen for conversion of LOAEL to NOAEL because the effects observed in the study were all reversible within 28 days, so the adversity of the effects at the lowest dose level is questionable. RfD = 0.1 mg/kg/day
b. Composition: dearomatized aliphatic; C10-C13 isoparaffins/naphthenes/n-alkanes; typical aromatic content: 0.1% Study 7. Subchronic Toxicity Study (unpublished data)
Sprague-Dawley rats were dosed orally with 0, 100, 500, or 1000 mg/kg for 13 weeks. A high-dose recovery group was also included. Decreases in aspartate aminotransferase and glucose were observed in the 500 and 1000 mg/kg dose groups, while BUN, creatinine, alanine aminotransferase, and cholesterol were increased in the treated males. In females, liver weights were increased in the 1000 mg/kg dose groups. The liver/body weight ratio was increased in both the 500 and 1000 mg/kg dose groups (males and females). Kidney weights were increased in male rats in the 500 and 1000 mg/kg dose groups. Testicular weights were increased in the 1000 mg/kg males. Male rat nephropathy was observed in all dosed groups. Histopathology revealed centrilobular hepatocellular hypertrophy in the 500 and 1000 mg/kg dose groups (males and females). Results from the high-dose recovery
26
group revealed all treatment-related effects were reversible within a 4week period. The NOAEL was based on the liver effects observed. NOAEL = 100 mg/kg/day With an uncertainty factor of 1000 (10 most sensitive, 10 animal to human, 10 subchronic to chronic): RfD = 0.1 mg/kg/day
c. Composition: dearomatized aliphatic; C15-C18; typical aromatic content 0.6-1.5% Study 8. Developmental Toxicity Study (not published)
Sprague-Dawley rats were dosed orally with 0, 400, 800, and 1000 mg/kg during gestational days 6-15. There was no significant maternal or developmental toxicity at any of the doses tested. NOEL >1000 mg/kg These data on developmental toxicity were not used in the calculation of an RfD for this fraction.
d. Composition: C11-C17 isoparaffinic solvent; contains 22% naphthenes; typical aromatic content <0.05% Study 9. Subchronic Toxicity Study (unpublished data)
Rats were dosed orally with 0, 100, 500, or 1000 mg/kg for 90 days. A high-dose recovery group was included. Increased liver weights were observed at 500 and 1000 mg/kg for both males and females, and increased kidney weights for the 1000 mg/kg females. After the 4-week recovery period, the differences in organ weights disappeared. There were no treatment-related histopathologic effects. The NOAEL was based on liver weight changes. NOAEL = 100 mg/kg/day With an uncertainty factor of 1000 (10 most sensitive, 10 animal to human, 10 subchronic to chronic): RfD = 0.1 mg/kg/day
e. JP-8 Jet Fuel Study 10. Subchronic Toxicity Study (Mattie et al., 1995)
Male rats were exposed daily by oral gavage to 0, 750, 1500, and 3000 mg/kg of JP-8 for 90 days. Body weights were significantly decreased in both mid- and high-dose groups. Glucose, total bilirubin, Aspartate aminotransferase (AST), and Alanine aminotransferase
27
(ALT) were all significantly different than control values in all three dose groups. Dose dependent irritation of the GI tract was also observed. Neutrophil (elevation) and lymphocyte (depression) counts differed significantly from controls at all exposure levels. In the high dose group, organ/body weight ratios were significantly different for brain, liver, kidneys, spleen, and testes. However, there were no significant differences in individual organ weights. Male rat nephropathy was observed in all dose groups. The hematological and liver enzyme effects (in the absence of organ weight changes) were not considered significant. Body weight changes were dose dependent but were not significant at 750 mg/kg/day. NOAEL = 750 mg/kg/day With an uncertainty factor of 1000 (10 most sensitive, 10 animal to human, 10 subchronic to chronic): RfD = 0.75 mg/kg/day
3. Summary and Conclusions for Oral RfDs and Inhalation RfCs
It is important to remember that the mixtures data presented in this section cover the entire carbon range (see Figure 3). Therefore, it is appropriate to use the mixtures data instead of information on individual chemical compounds.
a. Inhalation
An inhalation RfC of 1.0 mg/m3 is recommended for this fraction based on inhalation studies for various petroleum streams and JP-8 (see Table 5). The inhalation RfCs range from 0.9 to 1.0 mg/m3 as shown in Table 5. An RfC of 1.0 mg/m3 is considered to be representative of this fraction. This value should be representative of the entire fraction because it is based on mixtures data rather than data on individual chemicals. The inhalation RfC of 1.0 mg/m3 for the C9 - C16 fraction is not only protective of systemic toxicity, but it also appears to be protective of developmental and reproductive endpoints.
b. Oral
Using subchronic oral gavage data for dearomatized aliphatics (C9 - C12) and dearomatized aliphatics (C10 - C13), RfDs of 0.1 mg/kg/ day were calculated. With the JP-8 jet fuel, an RfD of 0.75 mg/kg/day was calculated based on oral gavage data. Again, the use of this mixtures data is much more representative of the fraction than looking at individual surrogates. The oral RfD of 0.1 mg/kg/day for the C9 - C16 fraction is the most conservative estimate based on the experimental data for petroleum streams and JP-8. Note that not only is this RfD protective of systemic toxicity, but it also appears to be protective of developmental and reproductive endpoints.
28
C. C>16 - C21 AND C>21 - C35 ALIPHATIC FRACTION 1. Introduction
The Total Petroleum Hydrocarbon (TPH) Criteria Workgroup has recommended that the toxicity criteria developed for white mineral oils be used for developing the RfD for fractions containing aliphatic hydrocarbons C17 or higher. White mineral oils are a complex mixture of highly refined mineral hydrocarbons (MHC) consisting primarily of saturated paraffinic hydrocarbons (predominantly branched chain alkanes) and naphthenic hydrocarbons (alkanes containing one or more saturated cyclic structures). These oils are essentially pure aliphatic hydrocarbons with virtually no aromatic components or other contaminants. They are approved by the Food and Drug Administration as direct food additives and used extensively in foods, cosmetics, and pharmaceutical products. The abbreviation MHC is used throughout this report as a generic term to describe the range of aliphatic hydrocarbons present in white mineral oils. This section discusses the development of RfD values for this fraction based on the results of a toxicity study conducted by the British Industrial Biological Research Association (BIBRA) in Fischer 344 (F/344) rats as reported by Smith et al., (1996). This 90 day feeding study of several different white mineral oil samples representing different MHC sizes resulted in two distinct responses. The smaller (lower molecular weight, lower viscosity) MHC caused mesenteric lymph node histiocytosis and liver granulomas; the degree of response is inversely proportional to the molecular size of the MHC. The samples containing the larger (higher molecular weight, higher viscosity) MHC were essentially without effect (Smith et al., 1996). It should also be noted that a chronic study in Fischer 344 rats has recently been completed in Japan. Although unpublished, the preliminary results of this study support the conclusions developed below.
2. Data Summary
In the BIBRA study, male and female F/344 rats were administered a range of white mineral oils mixed in the diet at doses of 20, 200, 2,000 and 20,000 ppm for 90 days. The data will be expressed as the composite average daily intakes (approximately 2, 20, 200, and 2,000 mg/kg/day, respectively). This study utilized seven samples which represented a full range of white mineral oils. The refining history and physical properties of the oils are shown in Table 7. Rats exposed to the lower molecular weight oils (average molecular weight 320 - 420) had histological changes in the liver and the mesenteric lymph nodes. Mesenteric lymph node histiocytosis was noted at doses of 20 mg/kg/day or higher, whereas liver granulomas were only noted at the 2,000 mg/kg/day dose in rats exposed to lower molecular weight oils (N10A, N15H, P15H, N70A and N70H). The incidence and severity of the effects were inversely related to the molecular weight of the MHC. Females were more sensitive than males. Rats fed up to 2,000 mg/kg/day of the higher molecular weight white mineral oils (BIBRA samples P70H and P100H) showed no effect on body weight, clinical signs, or mortality No treatment related toxicity or histopathological effects were seen in these animals.
29
30
Refining Method Acid Treatment Hydrogenation Hydrogenation Acid Treatment Hydrogenation Hydrogenation Hydrogenation 99.8 69.5 68.0 76.4 15.0 16.6 13.3 320 330 350 410 420 485 510 Viscosity (cSt) 40 0C Average Molecular Weight Average Carbon Number Distribution C15-30 C17-30 C18-30 C21-35 C22-37 C27-43 C28-45
Table 7. Test Materials and Physical Properties of White Mineral Oils Used in BIBRA Studya
Sample
Crude Type
N10A
Naphthenic
N15H
Naphthenic
P15H
Paraffinic
N70A
Naphthenic
N70H
Naphthenic
P70H
Paraffinic
P100H
Paraffinic
Sample abbreviations: N=Naphthenic, P = Paraffinic, A = Acid Treated, H = Hydrogenated, Number (10,15,70, 100) = Approximate viscosity (cSt) at 40oC. a Data were obtained from Smith et al. (1996).
Table 8. Liver Granuloma Response in the BIBRA Studya

Category Low Molecular Weight (C17-34) High Molecular Weight (C>34) Sample ID N10A, N15H, P15H, N70A, N70H P70H, P100H NOAEL (mg/kg/day) 200 LOAEL (mg/kg/day) 2,000
2,000
ndb
NOAEL = No observed adverse effect level LOAEL = Lowest observed adverse effect level
a b
Data obtained from Smith et al., (Smith et al., 1996). No response was noted at the highest concentration used in the study (2,000 mg/kg/day).
3. Rationale for RfD Values a. Determination of Two Distinct Responses
The effects of MHC in F/344 rats appear to be inversely related to molecular weight (MW). The average MWs of the white mineral oils used in the BIBRA study are shown in Table 7. Administration of the low molecular weight oils resulted in granulomatous effects in F/344 rats, with greater responses seen with the lowest MW oils. Essentially no effects were noted with the high molecular weight oils. Based on these differences in toxicological responses, two separate RfD values are warranted. Based on the low molecular weight oil data, a lower RfD is recommended for the lower molecular weight TPH aliphatic fractions (C17-34, average MW 240-480), whereas a higher RfD value is recommended for the higher molecular weight TPH aliphatic fraction (C>34, average MW >480) based on the high molecular weight oils which showed no effect. The lack of effects seen with the high molecular weight MHC is consistent with studies showing essentially no absorption for alkanes above C32 (Albro and Fishbein, 1970). In this study, the retention of selected aliphatic hydrocarbons was measured in rats after intragastric administration. An inverse relationship with a high correlation coefficient (0.96) was found between molecular size and retention of the aliphatic hydrocarbons. The largest molecule used in this study was squalane (C30), 96-100% of the oral dose was recovered unchanged in the feces, indicating essentially no absorption.
b. Determination of No Observed Adverse Effect Level (NOAEL)
The critical effect used to determine RfDs for TPH aliphatic fractions C>17 is liver granuloma formation based on the MHC data. As shown in Table 8, the lowest observed adverse effect level (LOAEL) for the low molecular weight oils was 2,000 mg/kg/day; the no observed adverse effect level (NOAEL) for these oils was 200 mg/kg/day. The NOAEL for the high molecular weight oils was 2,000 mg/kg/day. Although mesenteric lymph node histiocytosis was noted at lower doses, this finding was not considered to be an adverse effect because it is a normal adaptive response to the ingestion of foreign material (Schuurman et al., 1994). The normal physiological function of the mesenteric lymph node is to filter foreign material from the lymph. Previous work by Albro and Fishbein (1970) showed that
31
aliphatic hydrocarbons are absorbed by the small intestine and enter the lymph. Mesenteric lymph nodes draining the gut- associated lymphoid tissue normally show sinus-histiocytosis (Shuurman et al., 1994). Mesenteric lymph node histiocytosis is, therefore, not considered an adverse effect in the F/344 rat.
c. Justification of Safety Factors
The RfDs for TPH aliphatic fractions C>17 were calculated using a safety factor of 100. This factor takes into account animal to human extrapolation (a factor of 3), individual susceptibility (a factor of 10), and subchronic to chronic extrapolation (a factor of 3). Normally a safety factor of 1000 would be used; however, the overall weight of evidence on MHC justifies a lower uncertainty factor in this case for the reasons described below: i. Extensive human exposure to both natural dietary oils and MHC does not indicate any clinical effects. Although MHCinduced lipid granulomas have been observed in human tissues, these findings are considered to be clinically unimportant and are pathologically distinct from the lesions noted in F/344 rats. Human lipid granulomas are characterized as benign, circumscribed lesions containing mineral oil in the center, without evidence of inflammation, fibrosis, or significant liver dysfunction (Wanless and Geddie, 1985). In contrast, the liver granulomas in F/344 rats are characterized as reactive with associated inflammation and occasional parenchymal cell necrosis (Smith et al., 1996). This evidence supports a lower uncertainty factor for animal to human extrapolation F/344 rats appear to be a uniquely sensitive strain of rats. The inflammatory effects noted in F/344 rats were not seen in dogs, mice or different strains of rats (Long-Evans or Sprague Dawley) fed comparable doses of similar MHC (Firriolo et al., 1995; Smith et al., 1995). In addition, historical chronic studies of MHC at doses of up to 10% in the diet of Sprague Dawley rats were without clinical or histological effects (Shubik et al., 1962). Thus, the F/344 rat appears to have a predisposition for granulomatous effects. When compared to other rat strains (Wistar, Sprague Dawley), the F/344 rat had a higher incidence of spontaneous granulomas in the mesenteric lymph nodes (Ward et al., 1993). A review of the National Toxicology Program (NTP) chronic and subchronic studies also revealed a highly variable incidence of spontaneous liver granulomas and mesenteric lymph node histiocytosis in untreated control F/344 rats with the incidence higher in females than in males. These studies suggest that the F/344 rat is predisposed to the development of inflammatory granulomatous lesions (Miller et al., 1996), thereby providing further support for a lower uncertainty factor for animal to human extrapolation.
ii.
32
iii
Although F/344 rats have a predisposition for inflammatory granulomatous responses, these effects do not appear to progress to tumors nor alter lifetime, body weight or health status of the rats. For example, in a recent Japanese study, F/344 rats administered 5% MHC in the diet for 24 months displayed mesenteric lymph node histiocytosis, but no differences in body weight, survival, or tumor incidence were noted in any of these animals compared to controls (Takahashi, 1996). NTP conducted studies on chlorinated paraffins (C23, 43% Cl), comprised of linear saturated hydrocarbons with a molecular size similar to the low molecular weight oils used in the BIBRA study (Bucher et al., 1987). F/344 rats received chlorinated paraffin by gavage at doses up to 900 mg/kg (females) and 3,750 mg/kg (males) 5 days a week for up to 2 years. F/344 rats had effects similar to those seen with MHC, i.e. mesenteric lymph node histiocytosis and liver granulomas. Interestingly, there were no treatment related effects on F/344 rat survival, body weight, tumor incidence, or health status. These results suggest that the granulomatous lesions noted in F/344 rats exposed for 90 days to MHC, will not result in any adverse clinical effects following chronic exposure, and justify a lower uncertainty factor for subchronic to chronic extrapolation.
d. Calculation of RfD
The RfD for TPH aliphatic fractions containing C>17 was calculated according to USEPA guidelines in which the NOAEL, based on the critical effect, is divided by an appropriate safety factor. i. RfD for C17-34 MHC The RfD for TPH fractions containing aliphatic fractions of C17-34 is 2 mg/kg/day based on the NOAEL for low molecular weight oils for liver granulomas (200 mg/kg/day) and a safety factor of 100. ii. RfD for C>34 MHC The RfD for TPH fractions containing aliphatic fractions C>34 is 20 mg/kg/day based on the NOAEL for high molecular weight oils for liver granulomas (2,000 mg/kg/day) and a safety factor of 100.
33
Table 9. Development of Oral RfDs for TPH Aliphatic Fractions C 17

TPH Aliphatic Fraction C17-34 C>34 NOAELa (mg/kg/day) 200 2,000 LOAELa (mg/kg/day) 2,000 ndb r 100 100 RfD (mg/kg/day) 2 20
RfD = Reference Dose TPH = Total Petroleum Hydrocarbons NOAEL = No observed adverse effect level LOAEL = Lowest observed adverse effect level
a b
Based on the liver granuloma response noted in Smith et al., (1996). No response was noted at the highest concentration used in the study (2,000 mg/kg/day).
iii. Conclusion As shown in Table 9, the RfD for TPH fractions containing an aliphatic carbon range of C17 - C34 MHC is 2 mg/kg/day; for fractions containing aliphatic fractions C>34 , the RfD is 20 mg/kg/day. The RfDs were based on the results of the BIBRA study (Smith et al., 1996) using liver granulomas as the critical effect. The use of these RfD values for determining cleanup levels for TPH aliphatic fractions C>17 should provide an adequate margin of safety to protect human health.
34
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Ibbotsun, B.G. et al. (1989). A Site-Specific Approach for the Development of Soil Cleanup Guidelines for Trace Organic Compounds in Petroleum Contaminated Soils, Vol. I, Chapter 24, p. 321, Lewis Pub. IRIS CD-Rom, Vol. 27, Expiration: January 31, 1996. Koblis, K. et al. (1993). Impact of Surrogate Selection on Risk Assessment for Total Petroleum Hydrocarbons, J. Soil Cont. 2, p. 125. Kostecki, P et al. (1993). Review of Present Risk Assessment Models for Petroleum Contaminated .T. Soils. Chapter 24, p. 553. Principles and Practices for Petroleum Contaminated Soils, Lewis Pub. Krewski, D. and R.D. Thomas. (1992). Carcinogenic mixtures. Risk Analysis. 12(1):105-113. LaGoy, P and Quirke, T.C. (1994). Establishing Generic Remediation Goals for the Polycyclic .K. Aromatic Hydrocarbons: Critical Issues, Environ. Health Perspect Vol. 102, pp. 348-352. Lewtas, J. (1993). Complex Mixtures of Air Pollutants: Characterizing the Cancer Risk of Polycyclic Organic Matter. Environmental Health Perspectives 100, p. 211. Magee, B.H. et al. (1993). Risk-Based Target Cleanup Levels for TPH in Soils. Chapter 17 p. 303 in Hydrocarbon Contaminated Soils, Vol. III, Lewis Pub. Massachusettes Department of Environmental Protection. Interim Final Petroleum Policy: Development of Health-Based Alternative to the Total Petroleum Hydrocarbon (TPH) Parameter, Boston, MA, June 1994. Metcalf & Eddy. (1993). Extending Luft Manual Methods to Crude Oil to Determine Soil Cleanup Levels. Report submitted to Western States Petroleum Association, Glendale, CA. September 17. Michelson, T.C. and Boyce, C.P (1993). Cleanup Standards for Petroleum Hydrocarbons. Part 1. . Review of methods and recent developments. J. Soil Cont. 2(2):109-124. Michelson, T.C. and Boyce, C.P (1993). Case Study Comparisons of Site-Specific Cleanup Standards . with Generic TPH Standards. Part II J. Soil Cont. 2(2): 265. Millner, G.C., James, R.C. and Nye, R.C. (1992). Human Health-Based Soil Cleanup Guidelines for Diesel Fuel No. 2. J. Soil Cont. 1(2):103-157. Odencrantz, J.E. et al. (1993). A Better Approach to Soil Cleanup Level Determinations. Levine Fricke, Inc. Pepelko, W.E. and Withey, J.R. (1985). Methods for route-to-route extrapolation of dose. Tox. Ind. Health 1:153-175. Plotkin, B. and Bruya, J. (1994). Suggestions for Alternatives to TPH as a Criteria for Cleanup. Friedman & Bruya, Inc. PRC Environmental Management, Inc. (1993). Site Inspection at the Underground Storage Tank Site, U.S. Army Reserve Center, Fort Shafter Flats, Hawaii, Vol. II. Report submitted to the U.S. Army Corps of Engineers. August 24. Rhodes, I., and Albertson, B. (1997). Review of TPH Analytical Methodologies. TPHCWG, Volume 2. Ryer-Powder, J.E. and Sullivan, M.J. (1993). Update on the Derivation of an Oral Reference Dose for Diesel Fuel No.2, in Principles and Practices for Diesel Contaminated Soils, Vol. III, pp. 49-56. Ryer-Powder, J.E., and Sullivan, M.J. 1994. Chapter 2: Update on the Derivation of an Oral Reference Dose for Diesel Fuel No. 2. in Principles and Practices for Diesel Contaminated Soils, Vol. III, Eds. P Kostecki; E.J. Calabrese and C.P Barkan, Amherst Sc. Publ. .T. .L. Rosenblatt, D.H. et al. (1994). Evaluation of Health Risks from a Buried Mass of Diesel Fuel Before and After Bioremediation. J. Soil Cont. 3, p. 1. Schoeny, R.S. (1993). Creative Approaches in the Study of Complex Mixtures: Evaluating Comparative Potencies. Chapter 25, p. 591. Principles and Practices for Petroleum Contaminated Soils, Lewis Pub.
Shull, L. R. et al. (1994). Health Risk Assessment of Petroleum Hydrocarbons in Environmental Media; Proceedings of Federal Environmental Restoration III & Waste Minimization II Conference (Apr. 27-29, 1994) Vol. I, pp. 1-9. Sullivan, M. et al. (1991). A Method for Incorporating Biodegradation Rates of Benzene into the Risk Assessment Process in Hydrocarbon Contaminated Soils, Vol. 1., pp. 605-613. Lewis Pub. Sullivan, M.J. et al. (1991). A Risk Assessment for Crude Oil in Residential Surface Soils in Hydrocarbon Contaminated Soils, Vol. 1, pp. 615-645. Lewis Pub. Takahashi, M. (1996). Toxicological Studies of White Mineral Oils. The Toxicology Forum 1996 Annual European Meeting, Oxford, United Kingdom. Pp. 573-583. USEPA. (1986). Guidelines for Health Risk Assessment of Chemical Mixtures. 51 FR 34014-34025. September 24. USEPA. (1987). The Risk Assessment Guidelines of 1986, EPA600/08-87-045, August. USEPA. (1989). Risk Assessment Guidance for Superfund, Volume 1 Human Health Evaluation Manual (Part A), Interim Final, EPA/540/1-89/002, December. USEPA. (1990). Review Draft, Interim Methods for Development of Inhalation Reference Concentrations, EPA/600/8-90/006A, August. USEPA. (1991). Guidelines for Developmental Toxicity Risk Assessment, Federal Register Vol. 56 No. 234, December 5. USEPA. (1992). Master List Responses for 2QTR 1992. Memo from Joan Ann-Marie Burke. USEPA, ORD, ECAO, Cincinnati, Ohio. June 19. S. Dollarhide to
USEPA. (1994). Review Draft Guidelines for Reproductive Toxicity Risk Assessment, EPA600.AP .94.001, February. USEPA. (1995). Proposed Guidelines for Neurotoxicity, Federal Register Vol. 60, No. 192, October 4. USEPA. (1995). The Use of the Benchmark Dose Approach in Health Risk Assessment, EPA/630/R94/007. February. Warshawksy, D. et al. (1993). Factors Affecting Carcinogenic Potential of Mixtures. Fund. Appl. Toxicol. 20, pp. 376-382. Washington State Department of Ecology. (1994). Total Petroleum Hydrocarbon (TPH) Cleanup, Focus (May). Welsh, R.J. et al. (1993). Estimation of the Migration Potential of Diesel Fuel Constituents from Soil to Groundwater: A Case Study. J. Soil Cont. 2, p. 343.
AROMATIC SURROGATES
Ambrose, A.M., Booth, A.N., DeEds, F. and Cox, Jr, A.J. (1960). A toxicological study of biphenyl, a citrus fungistat. Food Res. 25: 328-336. Clark, D.G., Butterworth, S.T., Martin, J.G., Roderick, H.R., and Bird, M.G. (1989). Inhalation toxicity of high flash aromatic naphtha. Toxic. Ind. Health. 5(3):415-428. Douglas, J.F., McKee, R.H., Cagen, S.Z., Schmitt, S.L., Beatty, P .W., Swanson, M.S., Schreiner, C.A., Ulrich, C.E., and Cockrell, B.Y. (1993). A neurotoxicity study assessment of high flash aromatic naphtha. Toxic. Ind. Health. 9(6):1047-1058. McKee, R.H., Wong, Z.A., Schmitt, S.L., Beatty, P Swanson, M., Schreiner, C.A., and Schardein, J.L. ., (1990). The reproductive and developmental toxicity of high flash aromatic naphtha. Toxic. Ind. Health. 6(3/4):441-460. NTP (National Toxicology Program). (1980). Unpublished Subchronic Toxicity study: Naphthalene (C52904), Fisher 344 rats. Prepared by Batelles Columbus Laboratories Under Subcontract No. 76-34-106002.
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NTP (1986). NTP Technical Report on the Toxicology and Carcinogenesis of Xylenes (mixed) (60.2% . m-xylene, 13,6% pxylene, 17.0 ethylbenzene and 9.1% 0-xylene) (CAS No. 1330-20-7) in F344/N rats and B6C3F1 mice (gavage studies). U.S. DHHS, PHS, NIH, NTP Research Triangle Park, NC. , NTP TR 327, NIH Publ. No. 6-2583. NTP (1989). Toxicology and Carcinogenesis Studies of Toluene in F344/N Rats and B6C3F1 Mice. . Technical report Series No. 371. Research Triangle Park, NC. Quast, J.F., Humiston, C.G., Kalnins, R.Y.et al. (1979). Results of a toxicity study of monomeric styrene administered to beagle dogs by oral intubation for 19 months. Toxicology Research Laboratory, Health and Environmental Sciences, DOW Chemical Co., Midland, MI. Final Report. USEPA. (1988). 13-Week Mouse Oral Subchronic Toxicity Study. Prepared by Toxicity Research Laboratories, Ltd., Muskegon, MI for the Office of Solid Waste, Washington, DC. USEPA. (1989a). Mouse Oral Subchronic Study with Acenaphthene. Study conducted by Hazelton Laboratories, Inc., for the Office of Solid Waste, Washington, DC. USEPA. (1989b). Mouse Oral Subchronic Toxicity Study of Fluorene. Prepared by Toxicity Research Laboratories, Ltd., Mukegon, MI for the Office of Solid Waste, Washington, DC. USEPA. (1989c). Subchronic Toxicity in Mice with Anthracene. Final Report. Hazelton Laboratories America, Inc. Prepared for the Office of Solid Waste, Washington, DC. USEPA. (1989d). Mouse Oral Subchronic Toxicity Study of Pyrene. Prepared by Toxicity Research Laboratories, Ltd., Muskegon, MI for the Office of Solid Waste, Washington, DC. Wolf, M.A., Rowe, V.K., McCollister, D.D., Hollingsworth R.L. and Oyen F. 1956. Toxicological studies of certain alkylated benzenes and benzene. Arch. Ind. Health. 14:387-398.
C5 - C8 ALIPHATIC FRACTION
ACGIH. (1987). Documentation on the recommended Biological Exposure Index for n-hexane. API. (1980). 26 Week Inhalation Toxicity Study of Heptane in the Rat. Conducted by BioDynamics Inc. for the American Petroleum Institute, Biodynamics Project No. 78-7233. BP America, Inc. Constituent-Based Toxicity Assessment of Complex Petrolem-Derived Mixtures: Consideration of Gasoline Constituent Date in an Additive Risk Model. Cleveland, Ohio, October 1993. Daughtrey, W.C., Putman, D.L., Duffy, J., Soiefer, A.I., Kirwin, C.J, Kirwin, L.N., and Keenan, T.H. (1991). Cytogenetic studies on commercial hexane solvent. J. Appl. Toxicol. 14: 161-165. Daughtrey, W.C., Neeper-Bradley, T., Duffy, J., Haddock, L., Keenan, T., Kirwin, C., and Soiefer, A. (1994a). Two-generation reproduction study on commercial hexane solvent. J. Appl. Toxicol. 14: 387-393. Daughtrey, W.C., Duffy, J.S., Haddock, L.S., Kelly, D.W., Keenan, T.H., Richter, W.R., and Rhoden, R.A. (1994b). Chronic inhalation study of commercial hexane in mice. Toxicologist 14: 317 (Abstract #1234). Duffy, J., Newton, P Cockrell, B., Soiefer, A., Kirwin, C., and Daughtrey, W.C. (1991). A thirteen week ., inhalation toxicity study of commercial hexane in the rat and mouse. Toxicologist 11: 315 (Abstract #1219) Eben, A., Flucke, W., Mihail, F., Thyseen, J., and Kimmerle, G. (1979). Toxicological and metabolic studies of methyl n-butylketone, 2,5-hexanedione, and 2,5-hexanediol in male rats. Ecotoxicol. Environ. Saf. 3: 204. Frontali, N., Amantini, M.C., Spagnolo, A., Guarcini, A.M., and Saltari, M.C. (1981). Experimental neurotoxicity and urinary metabolites of the C5-C7 aliphatic hydrocarbons used as glue solvents in shoe manufacture. Clin. Toxicol. 18: 1357-1367.
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Katz, G.V., ODonoghue, J.L., DiVincenzo, G.D., and Terhaar, D.J. (1980). The relative neurotoxicity of methyl-n-butyl ketone, n-hexane, and their metabolites. Toxicol. Appl. Pharmacol. 52: 433-441. Keenan, T., Neeper-Bradley, T., Dodd, D., Kirwin, C., Duffy, J., and Soiefer, A. (1991). Developmental toxicity study of commercial hexane vapor in rats and mice. Toxicologist 11: 315 (Abstract #1218). Kelly, D.W., Duffy, J.S., Haddock, L.S., Daughtrey, W.D., Keenan, T.H., Newton, P .E., and Rhoden, R.A. (1994). Chronic inhalation study of commercial hexane in rats. Toxicologist 14: 317 (Abstract #1233). Kinkead, E.R., Haun, C.C., Schneider, M.G., Vernot, E.H., and Macewen, J.D. (1985). Chronic Inhalation Exposure of Experimental Animals to Methylcyclohexane. Air Force Aerospace Medical Research Laboratory report #TR-85-032. Kirwin, C., San, R., Harbell, J., Soiefer, A., Daughtrey, W., and Keenan, T. (1991). Salmonella and CHO/HGPRT mutation assays of commercial hexane. Toxicologist 14: 315 (Abstract #1220). Kreuzer, P .E., Stmer, A., Richter, M., Kessler, W., and Filser, J.G. (1995). Neurotoxic risk of n-heptane in rats and humans compared with that of n-hexane. International Toxicologist 7(1): 18-P-5 (Abstract). Misumi, J., and Nagano, M. (1984). Neurophysiological studies on the relation between the structural properties and neurotoxicity of aliphatic hydrocarbon compounds in rats. Brit. J. Indus. Med. 41: 526-532. ODonoghue, J.L., and Krasavage, W.K. (1979). The structural-activity relationship of aliphatic diketones and their potential neurotoxicity. Toxicol. Appl. Pharmacol. 48:A55. Olson, C.T., Yu, K.O., Hobson, D.W., and Serve, M.P (1986). The metabolism of n-octane in Fischer . 344 rats. Toxicol. Lett. 31: 147-150. Petroleum Environmental Research Forum (PERF)-CAP 94-05. Refinery Stream Speciation Project. Soiefer, A., Robinson, K., Broxup, B., Duffy, J., Keenan, T., and Daughtrey, W.D. (1991). A subchronic neurotoxicity study of commercial hexane vapor in the rat. Toxicologist 11: 315 (Abstract #1217). Takeuchi, Y., Ono, Y., Hisanaga, N., Kitoh, J., and Sugiura, Y. (1980). A comparative study on the neurotoxicity of n-pentane, n-hexane, and n-heptane in the rat. Brit. J. Ind. Med. 37: 241-247. Takeuchi, Y., Ono, Y., Hisanaga, N., Kitoh, J., and Sugiura, Y. (1981). A comparative study of the toxicity of n-pentane, n-hexane, and n-heptane to the peripheral nerve of the rat. Clin. Toxicol. 18: 13951402. TPHCWG - Volume No. 1, Petroleum Component Tables. Treon, J.F., Crutchfield, W.E., Jr., and Kitzmiller, K.V. (1943). The physiological response of animals to cyclohexane, methylcyclohexane, and certain derivatives of these compounds. J. Ind. Hyg. Toxicol. 25: 323-347.

Mattie, D.R., Alden, C.L., Newell, T.K., Gaworski, C.L., and Flemming, C.D. (1991). A 90-day continuous vapor inhalation toxicity study of JP-8 jet fuel followed by 20 or 21 months of recovery in Fischer 344 rats and C57BL/6 mice. Toxic. Pathol 19(2):77-87. Mattie, D.R., Marit, G.B., Flemming, C.D., and Cooper, J.R. (1995). The effects of JP-8 jet fuel on male Sprague-Dawley rats after a 90-day exposure by oral gavage. Toxic. Ind. Health. 11(4):423435. Mullin, L.S., Ader, A.W., Daughtrey, W.C., Frost, D.Z., and Greenwood, M.R. (1990). Toxicology update - Isoparaffinic hydrocarbons: A summary of physical properties, toxicity studies and human exposure data. J. Appl. Toxic. 10(2):135-142.
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Phillips, R.D. and Egan, G.F. (1981). Teratogenic and dominant lethal investigation of two hydrocarbon solvents. The Toxicologist Vol.1 (1):15 (Abstract 53). Phillips, R.D. and Egan, G.F. (1984). Subchronic inhalation exposure of dearomatized white spirit and C10-C11 isoparaffinic hydrocarbon in Sprague-Dawley rats. Fund. Appl. Toxic. 4:808-818.

Albro P and Fishbein, L. 1970. Absorption of aliphatic hydrocarbons by rats. Biochem. Biophys. Acta. .W. 219:437-446. Bucher, J.R., Alison, H., Montgomery, C.A., Huff, J., Haseman, J.K., Farnell S., Thompson, R., and Prejean, J.D. 1987. Comparative toxicity and carcinogenicity of two chlorinated paraffins in F344/N rats and B6C3F1 mice. Fund. App. Tox. 9:454-468. Firriolo, J.M., Morris, C.F., Trimmer, G.W., Twitty, L.D., Smith, J.H., and Freeman, J.J. 1995. Comparative 90-day feeding study with low viscosity white mineral oils in Fischer 344 and Sprague Dawley derived CRL:CD rats. Toxicol. Pathol. 23:26-33. Miller, M.J., Lonardo E.C., Greer, R.D., Bevan, C., Edwards, D.A., Smith, J.H., and Freeman JJ. 1996. Variable responses of species and strains to white mineral oils and paraffin waxes. Reg. Tox. Pharm. 23:55-68 National Toxicology Program (NTP). 1986. Toxicology and carcinogenesis studies of chlorinated paraffins (C23, 43% Chlorine, CAS No 63449-39-8) in F/344N rats and B6C3F1 mice (gavage studies). NTP TR 305, NIH publication No. 86-2561. Schuurman, H.J., Kuper, C.F., and Vos, J.G.. 1994. Histopathology of the immune system as a tool to assess immunotoxicity. Toxicology 86:187-212. Shubik, P Saffiotto, U., Lijinsky, G., Pietra, G., Rappoport, H., Toth, B., Raha, C.R., Feldman, R., and ., Ramahi, H. 1962. Studies of the toxicity of petroleum axes. Toxicol. Appl. Pharmacol. 4(Supp):1-62. Smith, J.H., Bird, M.G., Lewis, S.C., Freeman, J.J., Hogan, C.K., and Scala, R.A. 1995. Subchronic feeding study of four white mineral oils in dogs and rats. Drug and Chem. Tox. 18:83-103. Smith, J.H., Mallett, A.K., Priston, R.A.J., Brantom, P .G., Worrell, N.R., Sexsmith, C., and Simpson, B.J. 1996. Ninety-day feeding study in Fischer 344 rats of highly refined petroleum-derived food-grade white oils and waxes. Toxicol. Pathol. 24:214-230. Stefanski, S.A., Elwell, M.R., and Stomberg, P .C. 1990. Ch. 22 Spleen, lymph nodes, and thymus. In: Pathology of the Rat (eds. Boorman, G.A., Eustis, S.L., Elwell, M.R., Montgomery, C.A., and MacKenzie, W.F.) Academic Press, San Diego p. 385. Takahashi M. 1996. Carcinogenicity study of white mineral oil. Presented at the European Toxicology Forum March 28, 1996. Oxford U.K. p. 573. Wanless, I.R. and Geddie, W.R. 1985. Mineral oil lipogranulomata in liver and spleen. Arch. Pathol. Lab Med. 109:283-286. Ward, J.M., Uno, H., and Frith, C.H. 1993. Immunohistochemistry and morphology of reactive lesions in lymph nodes and spleen of rats and mice. Toxicol. Pathol. 21:199-205
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APPENDIX A
Literature Review (Individual Compounds)
OBJECTIVE
A literature review was conducted by EA Engineering, Science, and Technologies, Inc. (EA) in order to identify toxicity data and toxicity factors for approximately 250 chemical constituents of petroleum products. This information will be used to select surrogates for specific petroleum hydrocarbon fractions. Exxon Biomedical Sciences, Inc. (EBSI) reviewed the EA work product and identified surrogates for which additional toxicity data were available. A second literature review, conducted at EBSI, captures these data.
SEARCH STRATEGY - EA
EA identified USEPA toxicity factors including reference dose and slope factors at national and regional levels, and state-assigned toxicity values. An on-line search of the National Library of Medicine (NLM) electronic bibliography files, Hazardous Substances Data Base (HSDB) and Registry of Toxic Effects of Chemical Substances (RTECS) was subsequently conducted to identify toxicity data for materials that were not assigned a federal or state toxicity factor. EA supplied a detailed explanation of this process in a document entitled Summary of Work for Project 6a of the TPH Criteria Work Group. The EA deliverable is a compendium of toxicity factors and toxicity data, presented in spreadsheet format, accompanied with a list of references.
SEARCH STRATEGY - EBSI

EBSI modified the EA deliverable to include additional toxicity data and omitted studies that were less than 4 weeks in duration. Studies of this length are not appropriate for determining a No Observable Effect Level (NOAEL). One exception, however, was the addition of developmental studies to the EA deliverable. The USEPAs approach to determining a reference dose (RfD) or reference concentration (RfC) is based on the NOAEL. Ideally, a NOAEL is derived from a chronic study. Modifying factors are then applied to the NOAEL to account for variability and uncertainties in the data. When no chronic study is available, an RfD or RfC may be calculated from a suitable subchronic study. To identify additional toxicology studies, an on-line search of the following bibliographic and summary databases was conducted: TOXLINE (1965+); MEDLINE (1963+); EMBASE (1974+); American Petroleum Institute Literature (APILIT) (1963+); RTECS; HSDB; IRIS; and the Chemical Carcinogenesis Research Information Service (CCRIS) database. A detailed explanation of EBSIs search strategy is provided in Attachment I. Fourteen constituents in the C3 to C15 range were identified as having potential data and were subject to an on-line search. These constituents are listed in Table 1 of Attachment I. The literature search focused on subchronic, chronic and developmental studies from oral or inhalation routes of exposure. Acute studies were excluded. A second search for studies for surrogates in the C13 to C26 range followed. Forty-eight chemicals (Table 2 of Attachment I) were nominated. However, no significant new studies were found for this carbon range.
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DELIVERABLE
The modified EA deliverable is provided in Attachment II. In addition, a toxicity summary for the added studies can be found in Appendix B, entitled Toxicity Summaries for Aromatic and Aliphatic Constituents in the C4 to C22 Carbon Range.
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ATTACHMENT I
Results of Total Petroleum Hydrocarbons (TPH) Literature Search
INTEROFFICE CORRESPONDENCE TO: FROM: REFERENCE: SUBJECT:
DATE: February 28, 1996
Results of Total Petroleum Hydrocarbons (TPH) Literature Search
LITERATURE SEARCH CONDUCTED TO IDENTIFY TPH SURROGATES
This literature search supported a project to revise methods for determining safe levels of Total Petroleum Hydrocarbons (TPH). There is much variation on what specific hydrocarbons should be used as surrogates. This project was to determine surrogate substances for carbon ranges, e.g., n-hexane for C6, and calculate reference doses for these surrogates. The TPH Criteria Workgroup had a search done on 250 chemicals identified as being possible surrogates and provided a list with dose, species, route, duration, estimated tox factor, critical effect and a citation. Info Services was asked to locate additional tox studies, especially chronic and subchronic, that could be used to calculate the reference doses. We also did a search to verify chemical and physical constants provided by the Workgroup.
ON-LINE AND MANUAL SOURCES SEARCHED On-line Sources
Toxline (1965 - present) Medline (1963 - present) Excerpta Medica (1974 - present) American Petroleum Institute Literature - APILIT (1963 - present) Registry of Toxic Effects of Chemical Substances (RTECS) Hazardous Substances Data Bank (HSDB) EPAs Integrated Risk Information Service (IRIS) Chemical Carcinogenesis Research Info. Service (CCRIS) Chemical Information System (CIS) Databases: ENVIROFATE (Environmental Fate)
MANUAL SOURCES
Verschueren, K. (1983). Handbook of Environmental Data on Organic Chemicals, 2nd Edition. New York: Van Nostrand Reinhold. Mackay, D., Shiu, W.Y. and Ma, K.C. (1992). Illustrated Handbook of PhysicalChemical Properties and Environmental Fate for Organic Chemicals. Boca Raton: Lewis Publishers.
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LITERATURE SEARCH FOR TOX DATA FOCUSED ON C3-C15 CHEMICALS AND PNAS
The original TPH Criteria Workgroup list had identified 254 chemicals (C3-C26) as possible TPH surrogates and found useful tox studies on about 65 materials. We scanned the C3-C15 compounds and identified 14 materials on which it was believed there were additional tox studies. Table 1 lists the chemicals we tagged. The search focused on oral and dermal chronic and subchronic studies and, any inhalation, genetox, reprotox and carcinogenicity studies were included also. Acute studies were excluded. The printouts were arranged by chemical. There was too much information on biphenyl to do a reasonable search in the time allotted. We provided the summary printouts (RTECS, HSDB, etc.) and would fill in missing endpoint gaps later. For trimethylbenzenes, the search was limited to the past 5 years. However, it was noticed that the API C9 aromatic hydrocarbons studies were omitted from the Workgroup list. These studies were done in the mid to late 1980s and would not have been captured by my search. Therefore, for trimethylbenzenes only, I expanded the time range on the search to include the API studies. In the set of printouts for methylcyclopentane (MCP), you will also find references on commercial hexane. Since MCP is a component of commercial hexane, I included the published studies and TSCA submissions on commercial hexane in the search for this material since they may be relevant. No materials were identified in the C15-C26 range. Most of those materials are PNAs with a few long chain alkanes and cyclic compounds. The ATSDR on PNAs issued in 1994 is a good review of PNA tox. However, I was requested to search for any studies in the 1994 to present time range, and the search was broadened to include C13-C26. Table 2 lists the 48 chemicals searched for this part. Note that I did not search for references on benz(a)pyrene (BAP). A scan of the hits (over 2,000) turned up primarily BAP as a positive control in skin-painting studies of non-PNA materials. I found very few significant studies on these compounds from 1994 to present. This is not surprising. The carcinogenicity of PNAs is fairly well established and since there is no commercial use for these materials, there is little justification for their study. I found very few studies on the alkanes and cyclics on the list. The available tox studies on long chain alkanes are old (pre 1980) and are mostly acute studies since many of these substances were evaluated as possible cosmetic ingredients.
ADDITIONAL SOURCES FOR CHEMICAL/PHYSICAL CONSTANTS WERE LOCATED
Joan Tell provided a list of about 125 chemicals with chemical/physical constants such as solubility, Henrys law constant, etc. The request was to look for published values for the materials to verify the values provided by the Workgroup and to indicate if the values found were experimental (measured) or theoretical (calculated). Beth Meriwether, who did the bulk of this part, did not use the references cited by the Workgroup in their list since this request was for additional sources. However, I do recommend that you view the references used by the Workgroup as an additional source.
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The main source used for this search was Mackay, et.al. (Illustrated Handbook of Physical-Chemical Properties and Environmental Fate for Organic Chemicals). Beth found a good portion of the values in this book. The highlight of this source is that it is a compendium of published values and gives parameters and references. Beth used Verschueren, HSDB, and ENVIROFATE to fill in the data gaps. She did not find all the values, especially diffusion coefficients, and I went online to look for other possible, mainly on-line, sources. The numeric property databases on STN are very expensive and a search could cost a considerable sum. Joan Tell and I discussed this and she decided to use what she had. Variability in diffusion coefficients is not significant and she felt reasonably confident with the values given by the Workgroup.
SEARCH RESULTS WERE ORGANIZED BY CHEMICAL
This writeup covers all the searches done in support of this project. Most of the tox studies printouts were given to you although Deb did direct me to give a portion of the C15-C26 printouts to Michelle. Joan has the chemical and physical constants portion of the search. Please feel free to contact me if you have any questions or need additional information. /lah cc: M.D. Andriot D.A. Edwards A.C. Holladay E.J. Meriwether J.G. Tell
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Table A-1. List of 14 Chemicals (C3-C15)

isopropylbenzene n-butylbenzene p-cymene 1-pentene methylcyclohexane n-propylbenzene t-butylbenzene tetralin cyclohexene biphenyl trimethylbenzene (all isomers) diethylbenzene (all isomers) durene methylcyclopentane
Table A-2. Additional Materials (C13-C26)

1-t-butyl-3,4,5-trimethylbenzene 2,6-dimethylundecane fluorene n-heptylbenzene heptylcyclohexane 4-methylbiphenyl n-tridecane tridecene 1,4,5-trimethylnaphthalene anthracene 4,4-dimethylbiphenyl 1-methylfluorene n-octylbenzene phenanthrene n-tetradecane 9-methylanthracene 2-methylanthracene 1-methylphenanthrene n-pentadecane 9,10-dimethylanthracene 2-ethylanthracene fluroanthene n-hexadecane 1-phenylnaphthalene pyrene 1,2-benzofluorene 2,3-benzofluorene n-heptadecane 1-methylpyrene benz(a)anthracene chrysene n-octadecane triphenylene 5-methylchrysene n-nonadecane benz(b)fluroanthene benz(k)fluoranthene benz(a)pyrene (not searched) benz(e)pyrene n-eicosane perylene benz(ghi)perylene n-heneicosane 3-methylcholanthrene 1,2,5,6-dibenzanthracene picene coronene n-hexacosane
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ATTACHMENT II
EBSI MODIFIED DELIVERABLE
APPENDIX B
Toxicity Summaries for Both Aromatic and Aliphatic Constituents in the C4 to C22 Carbon Range
2-BUTENE (C4)
Male and female Wistar rats were exposed to 2-butene (42.4% cis-2-butene; 55.3% trans-2-butene) in a combined repeat dose and reproductive/developmental toxicity study. Animals were exposed at nominal concentrations of 0, 2500 and 5000 ppm 2-butene, 6 hours/day, 7 days/week. Actual concentrations were 0, 2476 and 5009 ppm or 0, 5.7 and 11.5 g/m3, respectively. Exposure of mated females ended after treatment on day 19 of gestation. A significant decrease in body weight was noted in the high dose females during premating weeks 0 to 2, and one day after parturition. Food consumption was decreased for this group in the first pre-mate week. In males, total white blood cell count and lymphocyte number were significantly increased. However, this increase did not follow a dose relationship and was within historical control values. Plasma Ca-levels were significantly decreased in males at 11.5 g/m3. No reproductive effects were observed in the parental animals. No effects were observed on the number of pups born, sex ratio or viability index. The NOAEL was 5.7 g/m3 for the P generation and > 11.5 g/m3 for the F1 generation.
Koten-Vermeulen, J.E.M.v., Plassche, E.J. v.d. 1992. SIDS Dossier on the HPV P1 Chemical: 2-Butene. RIVM, Rijksinstituut Voor Volksgezondheid en Milieuhygiene National Inst.
CYCLOPENTENE (C5) - 99.8%
Wistar II rats (10/sex/group) were exposed to 0, 870 or 8110 ppm cyclopentene vapor, 6 hours/day, 5 days/week for 3 weeks. Body weight gain was decreased in females at 8110 ppm. Appearance, behavior and gross evaluations were unremarkable. Wistar II rats (10/sex/group) exposed to 0, 112, 317 or 1139 ppm cyclopentene vapor, 6 hours/day, 5 days/week for 12 weeks tolerated test concentrations without any detectable effects. Animals were observed daily and weighed weekly. Hematology, clinical chemistry and urine analyses were unremarkable, as were animal appearance and behavior. No macroscopic or histological changes were observed. The NOEL was 1139 ppm for this study.
Kimmerle G., Thyssen, J. 1975. Acute, subacute and subchronic inhalation toxicity of cyclopentene. Int. Arch. Arbeitsmed. 34:177-184.
TOLUENE (C7)
An oral RfD of 0.2 mg/kg/day for toluene is currently on IRIS. This value is based on a subchronic oral gavage study in rats (NTP, 1989). Groups of 10 rats/sex/group were administered toluene in corn oil at levels of 0, 312, 625, 1250, 2500, or 5000 mg/kg for 5 days/week for 13 weeks. All animals in the 5000 mg/kg dose group died within the first week. At the 2500 mg/kg dose level, one female and 8 males died; however, two of these deaths were attributed to gavage errors. No significant changes in hematology or urinalysis were observed in the treated animals at any dose level. In females, liver, kidney and brain weights were all significantly increased at doses of 1250 mg/kg or greater. In males, liver and kidney weights were significantly increased at the 625 mg/kg dose level and above. Lesions in the liver and nephrosis were observed in animals at 2500 and 5000 mg/kg. Histopathological changes were also observed in the brain and urinary
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bladder at 1250, 2500, and 5000 mg/kg dose levels. The NOAEL for this study is 312 mg/kg based on liver and kidney weight changes in the male rats at 625 mg/kg. The RfD of 0.2 mg/kg/day was calculated using the NOAEL of 312 mg/kg, which was converted to 223 mg/kg/day based on the gavage schedule of 5 days/week. An uncertainty factor of 1000 (10 for animal to human; 10 for most sensitive; and 10 for subchronic) was applied to the NOAEL (223 mg/kg/day) to obtain 0.2 mg/kg/day.
CYCLOHEXANE (C6)
Under TSCA Section 4, the EPA and cyclohexane producers entered into an Enforceable Consent Agreement in November 1994 to conduct the following studies: 2-generation reproduction study (in progress, report to CMA 2/97); 90-day inhalation study in mice (report to CMA 6/96); 90-day neurotoxicity study in rats (report to CMA 6/96); 90-day inhalation study in rats (in progress, report to CMA 1/97); and a developmental study in rats (pilot completed, study start 3/96). In the inhalation developmental pilot study conducted under TSCA Section 4, rats were exposed to 0, 3000, 6000 or 9000 ppm cyclohexane. At 6000 and 9000 ppm, maternal weight gain and overall food consumption was reduced. There was an increased incidence of stain chin and stain face, and generally diminished response of the animals to a sound stimulus while being exposed. No statistically significant differences were noted between control and treated groups in fertility, number of implants, number of resorptions, number of live fetuses, sex ratio, or mean fetal weight. There were no external fetal alterations noted.
Bevan, C. J. (Draft Document). 1995. Cyclohexane Testing Program Update.
Rabbits exposed to 786 ppm cyclohexane, 6 hours/day, 5 days/week for 10 weeks showed microscopic changes in the liver and kidney. No effects occurred in rabbits exposed to 434 ppm for either 10 or 26 weeks. No treatment related effects occurred in monkeys exposed at 1243 ppm cyclohexane for 10 weeks.
Treon, J.F., Crutchfield, W.E., Jr., and Kitzmiller, K.V. 1943. The physiological response of animals to cyclohexane, methylcyclohexane, and certain derivatives of these compounds. J. Ind. Hyg. Toxicol. 25:323-347.
In a study to assess the neurotoxic potential of cyclohexane, rats were exposed to a vapor of 1500 or 2500 ppm, 3 to 10 hours/day, 5 to 6 days/week, for periods up to 30 weeks. No histopathologic effects were detected in the peripheral nervous system; however, the central nervous system was not evaluated.
Frontali, N., Amantini, M.C., Spagnoto, A., Guarcini, A.M., Saltari, M.C., Burgnone, F., and Perbillini, L. Experimental neurotoxicity and urinary metabolites of C5-C7 aliphatic hydrocarbons used as glue solvents in shoe manufacture. Clinical Toxicology, 18(12):1357-1367, 1981.
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1-HEXENE (C6)
In a reproduction/developmental screening study (OECD guideline 421), rats (12/sex/group) were administered 1-hexene in corn oil, by oral gavage, at levels of 0, 100, 500 or 1000 mg/kg/day at a volume of 5 mL/kg. The F0 males were treated for 28 days prior to mating and until sacrifice (44 days of dosing). The F0 females were treated 14 days prior to mating, during mating, throughout gestation and lactation, and until sacrifice (41-55 total days of dosing). F1 pups were sacrificed on day 4 of lactation. No mortality or clinical signs of toxicity were observed during the study. Body weight, body weight gain, food consumption and organ weights were unremarkable. Histopathologic evaluations of ovaries, testes, epididymides, liver, and peripheral nerve were unremarkable. Kidney effects, indicative of hydrocarbon nephropathy, were evident in males at all dose levels. No evidence of impaired reproductive capabilities was observed in the F0 parents, and no evidence of developmental toxicity was observed in the F1 pups. A NOAEL for reproductive toxicity was considered to be 1000 mg/kg/day.
Springborn Laboratories (SLS). March 24, 1995. Reproduction/Developmental Toxicity Screening Test in Rats with 1-Hexene. Submitted to Chemical Manufacturers Association. SLS Study No. 3325.1.
Rats (40/sex/group) were exposed to a vapor concentration of 1-hexene at 0, 300, 1000, or 3000 ppm, 6 hours/day, 5 days/week, for 90-days. No mortalities occurred. Female rats exposed at 3000 ppm had significantly reduced body weights. Male rats exposed to 3000 had slightly lower body weights compared to controls. Exposure to 1-hexene had no effect on neuromuscular coordination in females or on sperm count in males. At termination, serum phosphorous levels were significantly elevated in males at 300 ppm, and in male and female rats at 1000 and 3000 ppm. In addition, hematocrit and RBC levels were elevated for males and females at 3000 ppm and in females at 1000 ppm. Mean corpuscular hemoglobin and mean corpuscular hemoglobin concentrations were depressed in females exposed at 1000 and 3000 ppm. Both absolute and relative testes weights increased in males at 3000 ppm. However, no gross or histopathologic lesions were observed in any tissue at interim or terminal sacrifice. The NOAEL was determined at 1000 ppm. However, based on the effects observed at 1000 ppm, a NOAEL of 300 ppm may be more appropriate.
APME Monomer Dossier Re: SCF PMN/Reference No. 18820 (1-Hexene), 1995.
Rats (10/sex/group) were administered undiluted 1-hexene by oral gavage, daily for 13 weeks at 101, 350, 700 and 1010 mg/kg body weight/day. Some animals died (3 at 350 mg/kg; 7 at 700 mg/kg and 5 at 1010 mg/kg) due to hydrocarbon induced chemical pneumonitis. Food consumption was reduced in rats exposed at 700 and 1010 mg/kg 1-hexene, and body weight gain was reduced in males. Hematology, clinical chemistry and urinalysis were generally unremarkable except for some minor changes seen in these values in males exposed at 700 and/or 1010 mg/kg. Neurotoxicity evaluations showed some non-dose related differences between control and treated animals. Histologic evaluations were performed only with animals that died during the study, to avoid a statistical distortion resulting from the aspiration-related deaths. Most treated animals showed effects indicative
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of forestomach irritation from undiluted 1-hexene. The NOAEL for this study was determined to be 350 mg/kg body weight/day.
APME Monomer Dossier Re: SCF PMN/Reference No. 18820 (1-Hexene), 1995.
METHYLCYCLOHEXANE (C7)
Rats, mice, hamsters and dogs were exposed to a vapor of methylcyclohexane at 0, 400 or 2000 ppm, 6 hours/day, 5 days per week for 12 months. At 12 months, some of the rats, mice, and hamsters were terminated. The remaining rodents were held an additional year and the dogs for five years. There was no increase in tumors in any of the exposed animals. The only treatment related finding was kidney nephropathy in the 2000 ppm exposed rats. Hemolysis of blood samples prohibited clinical chemistry evaluations for the female rats.
Kinkead, E.R., Haun, C.C., Schneider, M.G., Vernot, E.H., and Macewen, J.D. (1985) Chronic inhalation exposure of experimental animals to methylcyclohexane. Air Force Aerospace Medical Research Report AFAMRL-TR-85-03.
Rabbits were exposed to a vapor of methylcyclohexane for 10 weeks. Liver and kidney effects were reported in rabbits exposed to 2880 ppm; however, there were no effects at 1200 ppm. No treatment related effects were reported in a monkey exposed to 370 ppm methylcyclohexane for 10 weeks.
Treon, J.F., Crutchfield, W.E., Jr., and Kitzmiller, K.V. (1943). The physiological response of animals to cyclohexane, methylcyclohexane, and certain derivatives of these compounds. J. Ind. Hyg. Toxicol. 25:323-347.
ETHYLBENZENE (C8)
The chosen study is a rat 182-day oral bioassay in which ethylbenzene was given 5 days/week at doses of 13.6, 136, 408, or 680 mg/kg/day in olive oil gavage (Wolf et al., 1956). There were 10 albino female rats/dose group and 20 controls. The criteria considered in judging the toxic effects on the test animals were growth, mortality, appearance and behavior, hematologic findings, terminal concentration of urea nitrogen in the blood, final average organ and body weights, histopathologic findings, and bone marrow counts. The LOAEL of 408 mg/kg/day is associated with histopathologic changes in liver and kidney. The RfD of 0.1 mg/kg/day was calculated using the NOAEL of 136 mg/kg, which was converted to 97.1 mg/kg/day based on the gavage schedule of 5 days/week. An uncertainty factor of 1000 (10 for animal to human; 10 for most sensitive; and 10 for subchronic) was applied to the NOAEL (97.1 mg/kg/day) to obtain 0.1 mg/kg/day.
STYRENE (C8)
Four beagle dogs/sex were gavaged with doses of 0, 200, 400, or 600 mg styrene/kg bw/day in peanut oil for 560 days (Quast et al., 1979). No adverse effects were observed for dogs administered styrene at 200 mg/kg-day. In the higher dose groups, increased numbers of Heinz bodies in the RBCs, decreased packed cell volume, and sporadic decreases in hemoglobin and RBC counts were observed. In
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addition, increased iron deposits and elevated numbers of Heinz bodies were found in the livers. Marked individual variations in blood cell parameters were noted for animals at the same dose level. Other parameters examined were body weight, organ weights, urinalyses, and clinical chemistry. The NOAEL in this study is 200 mg/kg-day and the LOAEL is 400 mg/kg-day. The RfD of 0.2 mg/kg/day was calculated using the NOAEL of 200 mg/kg/day. An uncertainty factor of 1000 (10 for animal to human; 10 for most sensitive; and 10 for subchronic) was applied to the NOAEL (200 mg/kg/day) to obtain 0.2 mg/kg/day.
XYLENES (C8)
Groups of 50 male and 50 female Fischer 344 rats and 50 male and 50 female B6C3F1 mice were given gavage doses of 0, 250, or 500 mg/kg/day (rats) and 0, 500, or 1000 mg/kg/day (mice) for 5 days/week for 103 weeks (NTP, 1986). The animals were observed for clinical signs of toxicity, body weight gain, and mortality. All animals that died or were killed at sacrifice were given gross necropsy and comprehensive histologic examinations. There was a dose-related increased mortality in male rats, and the increase was significantly greater in the high-dose group compared with controls. Although increased mortality was observed at 250 mg/kg/day, the increase was not significant. Although many of the early deaths were caused by gavage error, NTP (1986) did not rule out the possibility that the rats were resisting gavage dosing because of the behavioral effects of xylene. Mice given the high dose exhibited hyperactivity, a manifestation of CNS toxicity. There were no compound related histopathologic lesions in any of the treated rats or mice. Therefore, the high dose is a FEL and the low dose a NOAEL. The RfD of 2 mg/kg/day was calculated using the NOAEL of 250 mg/kg, which was converted to 179 mg/kg/day based on the gavage schedule of 5 days/week. An uncertainty factor of 100 (10 for animal to human and 10 for most sensitive) was applied to the NOAEL (179 mg/kg/day) to obtain 2 mg/kg/day.
ISOPROPYLBENZENE (CUMENE) (C9)
Rats were exposed to cumene vapor at concentrations of 0, 100, 500 and 1200 ppm (0, 0.50, 2.48 and 6.01 mg/L), 6 hours/day, 5 days/week for 13 weeks. A satellite group received a single 6-hour exposure, in order to evaluate neurobehavior. Alterations in functional observational battery (FOB) were observed in the satellite group at 500 and 1200 ppm, at 1 and 6 hours post exposure, but not at 24 hours post exposure. Effects included abnormal gaits, increased activity, decreased rectal temperature, and decreased toe pinch withdrawal reflexes. Necropsies were not performed in the single exposure study. In the 13 week inhalation study, no exposure related deaths occurred. No differences were observed in mean body weight; however, decreased food consumption was noted Week 1 for females exposed at 500 and 1200 ppm. A consistent increase in water consumption was noted in males exposed at 500 and 1200 ppm from Week 2 onward. These groups also demonstrated changes in several hematologic and clinical chemistry parameters. No exposure-related changes were seen in brain measurements, functional observational battery, or nervous system
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histopathology. Motor activity decreased in males exposed to 500 and 1200 ppm. This effect was not observed in a subsequent 13 week inhalation study, reported by the same author. There were no exposure-related effects on spermatogenesis. Liver, kidney and adrenal gland weights were increased in the 500 and 1200 ppm groups. Renal proximal tubular cell hypertrophy, hyperplasia, and hyaline droplet formation was evident in males exposed to 500 and 1200 ppm cumene. Cataracts were observed, however, in a non-dose dependent manner and in both exposed and control animals. Cumene was not considered neurotoxic. The NOAEL for this study was determined at 100 ppm.
Cushman, J.R., Norris, J.C., Dodd, D.E., Darmer, K.I., and Morris, C.R. 1995. Subchronic inhalation toxicity and neurotoxicity assessment of cumene in Fischer 344 rats. J. Am. Coll. Tox. 14(2): 129-147.
In a second 13 week inhalation study, conducted to assess the high incidence of cataracts observed in the first study, rats were exposed to cumene vapor, 6 hours/day, 5 days/week at concentrations of 0, 50 (permissible exposure limit), 100, 500 and 1200 ppm (0, 0.25, 0.50, 2.50 and 6.00 mg/L), with a 4 week recovery period. No animals died during the study. Body weights were unremarkable. Although some relative and absolute liver, kidney and adrenal gland weights were increased in rats exposed at 500 or 1200 ppm, no histopathological evaluations were conducted. The eyes were the only tissue evaluated histopathologically. No treatment related ophthalmic effects were observed. No serum chemistry or hematological evaluations were conducted. No changes in functional observational battery, auditory brain stem response, or motor activity were observed in any dose group. No treatment related neurotoxic or ototoxic effects were noted. The NOAEL for this study is 100 ppm, and is in agreement with the initial 13 week study conducted by Cushman et al. (1995).
Cushman, J.R., Norris, J.C., Dodd., D.E., Darmer, K.I., and Morris, C.R. 1995. Subchronic inhalation toxicity and neurotoxicity assessment of cumene in Fischer 344 rats. J. Am. Coll. Tox. 14(2): 129-147.
Rats were exposed to cumene vapor at concentrations of 0, 105, 300, or 599 ppm (0, 0.53, 1.5 and 3.0 mg/L), 6 hours/day, 5 days/week for approximately 28 days. No animals died during the study. Hypoactivity and irritation effects were noted during exposure. Absolute and relative liver and/or kidney weights were increased. No changes were reported in mean body weight, clinical, gross or microscopic pathology findings. The NOAEL was > 3 mg/L.
EUCLID Data Sheet: Cumene. 1995. Section 5.4 Repeated Dose Toxicity. ICI Chemicals & Polymers. EBSI Document No. 96MRR 54.
Female rats were exposed to 0, 100, 500 or 1200 ppm cumene vapor, 6 hours/day, on days 6 - 15 of gestation. No dams died, aborted or delivered early. However, body weight gain was significantly reduced throughout the exposure period in dams in the 1200 ppm group, and maternal food consumption was reduced at 1200 and 500 ppm. Gross observations, body weight, and organ weights were unremarkable except for a significant increase in relative liver weight at 1200 ppm. No significant changes were noted in gestational parameters and no increased incidence of either malformations or variations were noted. The NOEL for developmental toxicity was greater than 1200 ppm.
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EUCLID Data Sheet: Cumene. 1995. Section 5.9 Developmental Toxicity/Teratogenicity. ICI Chemicals & Polymers. EBSI Document No. 96MRR 54.
Female rabbits were exposed to 0, 500, 1200 or 2300 ppm cumene vapor, 6 hours/day, on days 6 - 18 of gestation. Maternal toxicity occurred in all three treatment groups as evidenced by maternal deaths, reduced relative liver weight (2300 ppm), and reduced maternal weight gain and food consumption during the exposure period. There were no significant changes in gestational parameters and no increased incidence of malformations or variations. However, one significant variation, ecchymosis of the head, was observed at 500 ppm but was within range of historical control values. The NOEL for developmental toxicity was greater than 2300 ppm.
EUCLID Data Sheet: Cumene. 1995. Section 5.9 Developmental Toxicity/Teratogenicity. ICI Chemicals & Polymers. EBSI Document No. 96MRR 54.
Groups of 10 female Wistar rats were administered 139 doses of cumene by gavage in olive oil at 154, 462, or 769 mg/kg/day over a 194-day period; 20 rats given olive oil served as controls (Wolf et al., 1956). Body weights were measured throughout the study. Most hematological evaluations were conducted after the 20, 40, 80, and 130th doses, and blood urea nitrogen determinations, and gross and histological examinations (lungs, heart, liver, kidneys, testes, spleen, adrenals, pancreas, femoral bone marrow) were conducted at the end of the study. Effects were not observed at 154 mg/kg/day but a slight but significant increase in average kidney weight occurred at 462 mg/kg/day. A moderate increase in average kidney weight occurred at 769 mg/kg/day. Therefore, 154 mg/kg/day is the NOAEL and 462 mg/kg/day is the LOAEL based on increased kidney weight. The RfD of 0.04 mg/kg/day was calculated using the NOAEL of 154 mg/kg, which was converted to a 110 mg/kg/day based dosing schedule of 139 doses in 194 days. An uncertainty factor of 3000 (10 for animal to human; 10 for most sensitive; 10 for subchronic; and an additional 3 for inadequate database) was applied to the NOAEL (110 mg/kg/day) to obtain 0.04 mg/kg/day.
N-NONANE(C9)
Harlan-Wistar rats were exposed by inhalation to 0, 1900, 3100 or 8400 mg/m3 (0, 360, 590, or 1600 ppm) n-nonane 6 hours/day, 5 days/week for 13 weeks. Two deaths resulted at 1600 ppm. Exposure to 1600 ppm produced excessive salivation, mild coordination loss, and fine tremors throughout the first 4 days of exposure. Salivation and lacrimation continued throughout the study. Mean body weights or mean body weight changes were significantly lower in the 1600 ppm group. There were no hematological, serum chemistry or histopathologic changes that were considered treatment-related. No effects were observed at 360 or 590 ppm.
Carpenter et al. 1975. Petroleum hydrocarbon toxicity studies XVII. Animal response to n-nonane vapor. Toxicol. Appl. Pharmacol. 44: 53-61.
N-PROPYLBENZENE (C9)
Rabbits (15/group) were fed n-propylbenzene in the diet at 0, 2.5 and 25 mg/kg/day for a 6 month period. Appearance, body weight, organ weights and protein function
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of the liver were unremarkable. Some animals showed mild protein dystrophy of the liver and kidneys. At 25 mg/kg, a non-significant decrease in RBC count was noted with deposition of hemosiderin in the spleen, indicating red-cell destruction. Leukocyte counts increased (nonsignificant) in both dose groups.
National Research Council. 1977. Drinking Water and Health. p. 763.
1,3,5-TRIMETHYLBENZENE (C9)
Sprague Dawley rats (10/sex/dose group) were administered 1,3,5-trimethylbenzene in corn oil by oral gavage for a 14 day period at concentrations of 0, 60, 150 and 600 mg/kg/day at a constant volume of 5mL/kg/day. A high dose recovery group was retained an additional 14 days. All animals survived treatment. No adverse clinical signs or treatment-related effects were observed in body weight, body weight gain or food consumption. Ophthalmic and necropsy findings were unremarkable. An increase in cholesterol levels was noted in mid- and high-dose females. An increase in white blood cell counts with corresponding increases in neutrophils and lymphocytes was noted in high dose males. At treatment termination, relative liver weights were significantly increased for mid- and high dose females and high dose males. In addition, relative adrenal weight was significantly increased in high dose males. All high dose animals exhibited centrilobular hepatic hypertrophy following treatment. All noted effects reversed by the end of the 14-day recovery period. The NOEL for this study was determined at 60 mg/kg, based on increased cholesterol levels and liver weight at 150 and 600 mg/kg.
IIT Research Institute. 14-Day Oral Gavage Toxicity Study of 1,3,5-Trimethylbenzene in Rats with a Recovery Group. IITRI Project No. L08512. Study 1. February 1995.
Sprague Dawley rats (10/sex/dose group) were administered 1,3,5-trimethylbenzene in corn oil by oral gavage, 5 days per week for a 90 day period at concentrations of 0, 50, 200 and 600 mg/kg/day at a constant volume of 5mL/kg/day. A high dose recovery group was retained an additional 28 days without treatment. All tissues from the control and high dose groups underwent microscopic examination. Lesions and limited tissues were evaluated in the low and mid-dose groups. No histologic evaluations were conducted for the recovery group. All animals survived treatment. No statistically significant effects were reported for body weight, body weight gain or food consumption. However, cumulative body weight gain decreased by 11% in high dose males. Ophthalmic exams were unremarkable. Phosphorus levels increased for high dose females. Also, a significant increase in absolute and relative liver weight was reported for high dose females at treatment termination. In males, relative liver and kidney weights were significantly increased at treatment termination. No treatment-related microscopic lesions were observed in any animal. Any treatment-related effect was absent by the end of the 28-day recovery period. A NOEL was established at 200 mg/kg based on increased phosphorous levels, liver and kidney weight reported at 600 mg/kg/day.
IIT Research Institute. 90-Day Oral Gavage Toxicity Study of 1,3,5-Trimethylbenzene in Rats with a Recovery Group. IITRI Project No. L0851. Study 2. May 1995.
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T-BUTYLBENZENE (C10)
t-Butylbenzene did not induce morphological transformation of Syrian hamster embryo cells.
Rivedal, E., Mikalsen, S.-O., Roseng, L.E., Sanner, T., and Eide, I. 1992, Effects of hydrocarbons on transformation and intercellular communication in Syrian hamster embryo cells. Pharm. Tox. 71: 57-61.
t-Butylbenzene was negative in bacterial mutation assays, a mitotic gene conversion assay, and in a cultured rat-liver cell line for structural chromosome damage.
Dean, B.J., Brooks, T.M., Hodson-Walker, G., and Hutson, D.H. 1985. Genetic toxicology testing of 41 industrial chemicals. Mutat. Res. 153:57-77.
N-DECANE (C10)
Rats were exposed to 540 ppm n-decane vapor 18 hours/day, 7 days/week for a total of 123 days. There was a significant weight gain and increase in total leukocyte count compared to controls. No changes were noted in polymorphonuclearlymphocyte ratios, in bone marrow composition, and no significant gross or microscopic organ changes were noted. No information was given as to whether the hematological changes were within normal biological variation. Some rats held for one month without additional exposure did not differ from the controls.
Nau, C.A., Neal, J., and Thornton, M. 1966. C9-C12 fractions obtained from petroleum distillates. Arch Environ. Health 12: 382-393.
DIETHYLBENZENE (C10)
Rats (25 females/group) were administered 0, 20, 100 or 200 mg/kg/day diethylbenzene in corn oil at 5 mL/kg, on days 6 through 15 of gestation. No treatmentrelated mortalities or clinical signs of toxicity were observed. Mean maternal body weight gain and food consumption were reduced at 100 and 200 mg/kg. Mean fetal body weight gain was reduced at 200 mg/kg, a level which was maternally toxic. A greenish-blue discoloration of the amniotic sac was evident at 100 and 200 mg/kg, and increased in intensity in a dose-dependent manner. No treatmentrelated malformations or developmental variations were observed. The NOEL for maternal toxicity was considered 20 mg/kg/day and the NOEL for fetal toxicity was considered 100 mg/kg/day.
Submission from Monsanto Chemical Company to U.S. EPA. TSCA 8(e) Reporting. May 27, 1992. EPA Document No. 88-920003153.
NAPHTHALENE (C10)
Rabbits exposed to naphthalene by oral route at doses up to 400 mg/kg/day on gestation days 6 to 18 showed no apparent adverse reproductive effects (or signs of developmental toxicity).
Pharmakon Research International (PRI), Inc. 1986. Developmental toxicity study in rabbits: Naphthalene. Report to Texaco, Inc. Beacon, NY. PH 329-TX-001-85.
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Mice exposed to naphthalene (in corn oil) at a dose of 300 mg/kg/day on days 7 to 14 of gestation had a decreased number of live pups per litter. No congenital abnormalities were observed.
Plasterer, M.R., Bradshaw, W.S., Booth, G.M., et al. 1985. Developmental toxicity of nine selected compounds following prenatal exposure in the mouse: naphthalene, p-nitrophenol, sodium selenite, dimethyl phthalate, ethylene thiourea and four glycol ether derivatives. Toxicol. Environ. Health 15:25-38.
In a 90 day oral gavage study, mice were administered 5.3, 53 or 133 mg/kg naphthalene. No treatment-related mortalities or body weight changes were reported in either sex, and no organ weight changes were observed in males. A significant decrease in absolute brain, liver and spleen weight was noted for females at the highest dose; however, organ to body weight ratios were significantly different only for the spleen. Although spleen weight decreased, there was no evidence of immunotoxicity in any treatment group for either sex. No histopathologic evaluations were performed in this study. Exposed mice showed no alterations in hematology. Several serum chemistry parameters including BUN levels in females (all doses) and total serum protein in both sexes (53 and 133 mg/kg), showed significant dose-related changes. A corresponding increase in albumin levels was noted in males, and an increase in globulin levels was noted in both males and females. Electrolyte values were generally unaffected by treatment, except for decreased calcium levels in males administered 53 or 133 mg/kg naphthalene. Although there were some changes, serum chemistry parameters gave little evidence of significant toxicity at any dose level.
Shopp, G.M., White, K.L., Jr., Holsapple, M.P et al., 1984. Naphthalene toxicity in CD-1 mice: General ., toxicology and immunotoxicology. Fund. App. Toxicol. 4:406-419.
Naphthalene was not teratogenic to pregnant rats administered up to 450 mg/kg/day, by gavage, on gestation days 6 to 15. However, there was a trend toward a dose-related increase in malformations.
National Toxicology Program (NTP). 1991a. Developmental toxicity of naphthalene (CAS No. 91-20-3) administered by gavage to Sprague-Dawley (CD) rats on gestational days 6 through 15. Research Triangle Park, NC: National Toxicology Program, National Institute of Environmental Health Sciences, U.S. Department of Health and Human Services, Public Health Service, National Institutes of Health. TER-91006.
In a 13 week subchronic oral study, rats and mice exposed to naphthalene at doses up to 400 and 200 mg/kg/day, respectively, showed no evidence of cardiovascular, gastrointestinal, respiratory, neurologic, renal or hepatic effects. No histopathological lesions of the testes were noted in mice or rats at any dose level.
Battelles Columbus Laboratories (Battelle). 1980a. Subchronic toxicity study: Naphthalene (C52904) B6C3F1 mice. Report to U.S. Department of Health and Human Services, National Toxicology Program, Research Triangle Park, NC. Battelles Columbus Laboratories (Battelle). 1980b. Subchronic toxicity study: Naphthalene (C52904), Fischer 344 rats. Report to U.S. Department of Health and Human Services, National Toxicology Program, Research Triangle Park, NC.
B6C3F1 mice were exposed to naphthalene vapors at 10 or 30 ppm, 6 hours/day, 5 days/week for a 2 year period. Both sexes displayed chronic inflammation and metaplasia of the olfactory epithelium, hyperplasia of the respiratory epithelium, and a dose-related increase in inflammatory lesions of the lungs. No treatment-
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related effects were observed for gastrointestinal, hematological, renal, hepatic, immunological or neurological systems. Female (but not male) mice exposed to 30 ppm naphthalene for a lifetime exhibited a significant increase in pulmonary alveolar/bronchiolar adenomas. NTP concluded no incidence of carcinogenicity in males and limited evidence in female mice based on increased incidence of pulmonary alveolar/bronchiolar adenomas.
National Toxicology Program (NTP). 1992a. Technical report series No. 410. Toxicology and carcinogenesis studies of naphthalene (CAS No. 91-20-3) in B6C3F1 mice (inhalation studies). Research Triangle Park, NC: U.S. Department of Health and Human Services, Public Health Service, National Institutes of Health. NIH Publication No. 92-3141.
In a 13 week subchronic dermal study, rats treated with up to 1000 mg/kg/day naphthalene, 6 hours/day, 5 days/week, showed an increased incidence of excoriated skin lesions and papules. Similar lesions were seen in the control and low dose groups. At the high dose, naphthalene exacerbated the severity of the lesions. No reported respiratory, cardiovascular, gastrointestinal, hematological, hepatic or renal effects.
Frantz, S.W., VanMiller, J.P and Jengler, W.C. 1986. Ninety-day (subchronic) study with naphthalene in ., albino rats. Report to Texaco, Inc., Beacon, NY, by Bush Run Research Center Union Carbide, Export, PA. Project No. 49-539 revised (unpublished).
A provisional RfD for naphthalene of 0.04 mg/kg/day was developed by the USEPA. This RfD was based on an oral subchronic NTP unpublished study (NTP, 1980). In this study, rats were administered naphthalene by gavage 5 days/week for 13 weeks. The dose levels used in this study were not published in any of the available summaries. However, the NOEL was identified to be 50 mg/kg/day. The critical effect was decreased body weight. Using the gavage schedule of 5 days/week, the 50 mg/kg/day is converted to 35.7 mg/kg/day. An uncertainty factor of 1000 (10 for animal to human; 10 for most sensitive; and 10 for subchronic) is used to calculate the RfD of 0.04 mg/kg/day. This provisional RfD is not on IRIS nor is it in HEAST. This value was on IRIS but was pulled pending further review. The value was also removed from HEAST due to the uncertainty in the calculation of the RfD.
TETRALIN (C10)
Cataracts resulted in guinea pigs (2/2) after 6 days of inhalation exposure, 30 minutes/day at a concentration of 71 mg/kg/day tetralin. No details were provided regarding dose, animal weight or test material purity. Saturated vapor (658 ppm) was assumed. There data are of questionable reliability due to limitations in study design.
Badinand, A., Paufique. L., and Rodier, J. 1947. Experimental intoxication with Tetraline. Arch. Mal. Prof. Med. Trav. Secur. Soc., 8:124.
Cataracts were produced by Day 11 in rabbits exposed by oral route to 118 to 235 mg/kg/day tetralin. However, this study lacked controls, used immature animals and provided no details on test material purity. Therefore, these data are of questionable reliability due to limitations in study design.
G. Basile, 1939. Experiments on the action of some hydrogenation products of naphthalene (Tetralin and Decalin) on the lens and its posterior capsule of the rabbit. Boll. Ocul. 18:951-957.
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METHYLNAPHTHALENE (C11)
Exposure to methylnaphthalene in the diet of mice at concentrations of 0, 0.075 or 0.15% for 81 weeks resulted in a high incidence of pulmonary alveolar proteinosis, increased sera lipid and phospholipid levels, and peripheral blood monocytes for both sexes. The incidence of bronchiolar/alveolar adenomas was significantly increased in males in both treatment groups but not in females. The increase was not dose-dependant, and was not accompanied by an increase in incidence of bronchiolar/alveolar carcinomas. The LOAEL for this study was 0.075%.
Murata, Y., Denda, A., Maruyama, H., and Konishi, Y. (1993) Chronic toxicity and carcinogenicity studies of 1-methylnaphthalene in B6C3F1 mice. Fund. Appl. Tox. 21: 44-51.
Four groups of CD-1 mice (20/sex/group) were gavaged daily with 0, 175, 350, or 700 mg/kg/day acenaphthene for 90 days (USEPA, 1989a). The toxicological evaluations of this study included body weight changes, food consumption, mortality, clinical pathological evaluations (including hematology and clinical chemistry), organ weights and histopathological evaluations of target organs. The results of this study indicated no treatment-related effects on survival, clinical signs, body weight changes, total food intake, and ophthalmological alterations. Liver weight changes accompanied by microscopic alterations (cellular hypertrophy) were noted in both mid- and high-dose animals and seemed to be dose-dependent. Additionally, high-dose males and mid- and high-dose females showed significant increases in cholesterol levels. Although increased liver weights, without accompanying microscopic alterations or increased cholesterol levels, were also observed at the low dose, this change was considered to be adaptive and was not considered adverse. The LOAEL is 350 mg/kg/day based on hepatotoxicity; the NOAEL is 175 mg/kg/day. The RfD of 0.06 mg/kg/day was calculated using the NOAEL of 175 mg/kg/day. An uncertainty factor of 3000 (10 for animal to human; 10 for most sensitive; 10 for subchronic; and an additional 3 for inadequate database) was applied to the NOAEL (175 mg/kg/day) to obtain 0.06 mg/kg/day.
BIPHENYL (C12)
Fifteen weanling albino rats of each sex were placed in each of eight experimental groups: 0.0, 0.001, 0.005, 0.01, 0.05, 0.10, 0.50, and 1.0% biphenyl in the diet (Ambrose et al., 1960). Dietary levels of 0.5% biphenyl and greater were associated with kidney damage, reduced hemoglobin levels, decreased food intake, and decreased longevity. One animal in each of the lower dose groups and control group had detectable blood in the renal pelvis. A NOAEL of 0.1% of diet is chosen because of the uncertainty of the significance of the effects observed at lower doses as compared to the more certain AEL of 0.5% of diet. The RfD of 0.05 mg/kg/day was calculated using the NOAEL of 0.1%, which was converted to 50 mg/kg/day. An uncertainty factor of 100 (10 for animal to human and 10 for most sensitive) and a modifying factor of 10 were applied to the NOAEL (50 mg/kg/day) to obtain 0.05 mg/kg/day.
Ambrose, A.M., Booth, A.N., DeEds, F., and A.J. Cox, Jr. 1960. A toxicological study of biphenyl, a citrus fungistat. Food. Res. 25: 328-336.
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Pregnant rats were administered biphenyl by oral gavage on days 6 to 15 of gestation, at doses of 125, 250, 500 or 1000 mg/kg. No fetal or maternal toxicity resulted from exposure at doses < 500 mg/kg. Evidence (nonsignificant) of fetotoxicity was noted at 1000 mg/kg and consisted of reduced number of live fetuses, reduced fetal weight, and increased resorptions. However, this dose was maternally toxic and produced mortality in five dams. Neither teratogenicity nor maternal toxicity was evident at doses ranging from 125 to 500 mg/kg biphenyl.
Khera, K.S., Whalen, C., Angers, G., and Trivett, G., 1979. Assessment of teratogenic potential of piperonyl butoxide, biphenyl and phosalone in the rat. Toxicol. Appl. Pharmacol. 47(2): 353-358.
FLUORENE (C13)
Fluorene (C13) has an RfD of 0.04 mg/kg/day that is on IRIS. This value is based on an oral 13-week study in mice. Mice (25/sex/group) were exposed to 0, 125, 250, or 500 mg/kg/day of fluorene suspended in corn oil by gavage for 13 weeks (USEPA, 1989b). A significant decrease in the red blood cell count and packed cell volume were observed in females in the 250 mg/kg/day group and in males and females at the 500 mg/kg/day dose level. In both high dose males and females, there was a significant decrease in BUN and a significant increase in total serum bilirubin. At 250 and 500 mg/kg/day, there was a significant increase in liver weight. A significant increase in spleen and kidney weight was observed in males and females at 500 mg/kg/day and males at 250 mg/kg/day. Increases in liver and spleen weights in high dose animals were accompanied by histopathological increases in the amounts of hemosiderin in the spleen and Kupffer cells of the liver. The LOAEL is 250 mg/kg/day based on hematological effects and the NOAEL is 125 mg/kg/day. The RfD for fluorene was calculated by taking the NOAEL of 125 mg/kg/day and applying an uncertainty factor of 1000 (10 for animal to human; 10 for most sensitive; and 10 for subchronic) and a modifying factor of 3 for lack of adequate toxicity data in a second species and reproductive/developmental data.
US EPA. 1989. Mouse oral subchronic toxicity study. Prepared by Toxicity Research Laboratories, LTD., Muskegon, MI for the Office of Solid Waste, Washington, DC.
ANTHRACENE (C14)
Anthracene was administered to groups of 20 male and female CD-1 (ICR)BR mice by oral gavage at doses of 0, 250, 500, and 1000 mg/kg/day for at least 90 days (USEPA, 1989c). Mortality, clinical signs, body weights, food consumption, opthalmology findings, hematology and clinical chemistry results, organ weights, organto-body weight ratios, gross pathology, and histopathology findings were evaluated. No treatment-related effects were noted. The no observed-effect level (NOEL) is the highest dose tested (1000 mg/kg/day). The RfD of 0.3 mg/kg/day was calculated using the NOAEL of 1000 mg/kg/day. An uncertainty factor of 3000 (10 for animal to human; 10 for most sensitive; 10 for subchronic; and an additional 3 for inadequate database) was applied to the NOAEL (1000 mg/kg/day) to obtain 0.3 mg/kg/day.
US EPA. 1989. Subchronic Toxicity in Mice with Anthracene. Final Report. Hazelton Laboratories America, Inc. Prepared for the Office of Solid Waste, Washington, DC.
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FLUORANTHENE (C16)
Male and female CD-1 mice (20/sex/group) were gavaged for 13 weeks with 0, 125, 250, or 500 mg/kg/day fluoranthene (USEPA, 1988). A fifth group of mice (30/sex) was established in the study for baseline blood evaluations. Body weight, food consumption, and hematological and serum parameter values were recorded at regular intervals during the experiment. At the end of 13 weeks, the animals were sacrificed and autopsied, which included organ weight measurement and histological evaluation. All treated mice exhibited nephropathy, increased salivation, and increased liver enzyme levels in a dose-dependent manner. However, these effects were either not significant, not dose-related, or not considered adverse at 125 mg/kg/day. Mice exposed to 500 mg/kg/day had increased food consumption and increased body weight. Mice exposed to 250 and 500 mg/kg/day had statistically increased SGPT values and increased absolute and relative liver weights. Compound-related microscopic liver lesions (indicated by pigmentation) were observed in 65 and 87.5% of the mid- and high-dose mice, respectively. Based on increased SGPT levels, kidney and liver pathology, and clinical and hematological changes, the LOAEL is considered to be 250 mg/kg/day, and the NOAEL is 125 mg/kg/day. The RfD of 0.04 mg/kg/day was calculated using the NOAEL of 125 mg/kg/day. An uncertainty factor of 3000 (10 for animal to human; 10 for most sensitive; 10 for subchronic; and an additional 3 for inadequate database) was applied to the NOAEL (125 mg/kg/day) to obtain 0.04 mg/kg/day.
US EPA. 1988. 13-Week mouse oral subchronic toxicity study. Prepared by Toxicity Research Laboratories, Ltd., Muskegon, MI for the Office of Solid Waste, Washington, DC.
PYRENE (C16)
An oral RfD of 0.03 mg/kg/day for pyrene is currently on IRIS. This value was based on a subchronic oral gavage study in mice (USEPA, 1989d). Groups of 20 mice/sex/group were administered pyrene in corn oil at levels of 0, 75, 125, or 250 mg/kg for 13 weeks. Nephropathy was present in 4 (control), 1 (75 mg/kg/day), 1 (125 mg/kg/day), and 9 (250 mg/kg/day) male mice. Similar lesions were seen in female mice: 2 (control), 3 (75 mg/kg/day), 7 (125 mg/kg/day), and 10 (250 mg/kg/day). Decreased kidney weights were observed in the 125 and 250 mg/kg/day dose groups. The NOAEL was determined to be 75 mg/kg/day and the LOAEL was 125 mg/kg/day for nephropathy and decreased kidney weights. The RfD for pyrene was calculated by taking the NOAEL of 75 mg/kg/day and applying an uncertainty factor of 1000 (10 for animal to human; 10 for most sensitive; and 10 for subchronic) and a modifying factor of 3 for lack of adequate toxicity data in a second species and reproductive/developmental data.
US EPA. 1989. Mouse Oral Subchronic Toxicity of Pyrene. Study conducted by Toxicity Research Laboratories, Muskegon, MI for the Office of Solid Waste, Washington, DC.
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BENZ(A)ANTHRACENE (C18)
Classified as a B2 carcinogen - use B(a)P slope factor and a potency factor. Mice (20 male - B6AF1/J) were administered two doses of benz(a)anthracene at a concentration of 3% by oral gavage (Klein, 1963). Treated animals exhibited an increased incidence of hepatomas (80%) and pulmonary adenomas (85%) over the control incidence values of 10% and 30%, respectively. In the same study, 40 male B6AF1/J mice were administered by gavage benz(a)anthracene 15 times at a concentration of 3%. Animals exhibited elevated incidences of hepatomas (46%) and lung adenomas (95%) over control values of 0% and 30%, respectively (Klein, 1963). Benzo(a)pyrene and benz(a)anthracene (a range of concentrations) were applied to the backs of C3H/He mice (sex unspecified) three times a week for 50 weeks (20 to 40 mice/dose) (Bingham and Falk, 1969). Benzo(a)pyrene was dissolved in decalin and benz(a)anthracene was dissolved in toluene for application. At 50 weeks, animals were sacrificed and tumors were quantitated. Tumors were classified as either malignant or benign, but no further details were provided. No solvent controls were included in the study. Both compounds produced malignant and benign skin tumors. Benz(a)anthracene appeared to be less potent than benzo(a)pyrene; however, the use of different solvents could be a confounding factor.
Bingham, E. and Falk, H.L. (1969). The modifying effect of carcinogens on the threshold response. Arch. Environ. Health 19:779-783. Klein, M. (1963). Susceptibility of strain B6AF/J hybrid infant mice to tumorigenesis with 1,2-benzanthracene, deoxycholic acid, and 3-methylcholanthrene. Cancer Res. 23:1701-1707.
CHRYSENE (C18)
Classified as a B2 carcinogen - use B(a)P slope factor and a potency factor.

BENZO(B)FLUORANTHENE (C20)
Classified as a B2 carcinogen - use B(a)P slope factor and a potency factor. Seven PAHs (benzo(a)pyrene, benzo(b)fluoranthene, benzo(j)fluoranthene, benzo(k)fluoranthene, indeno(1,2,3-cd)pyrene, cyclopentadieno(cd)pyrene, and coronene) were tested at varying concentrations to determine their dose-response relationships as carcinogens when applied topically to the backs of female NMRI mice two times a week for the lifetime of the animal (40 mice/dose) (Habs et al., 1980). At death, all animals were dissected and their dorsal skin examined histologically for tumor formation. A clear dose-response relationship was observed at the site of application for benzo(a)pyrene. Benzo(b)fluoranthene showed a clear carcinogenic effect. Benzo(j)fluoranthene exhibited weak carcinogenic effects, while benzo(k)fluoranthene and indeno(1,2,3-cd)pyrene showed no carcinogenic effect. In this study, the results were reported as tumors and no other distinction was defined. However, it is assumed that the tumors were all carcinomas based on this statement from the study, Animals at an advanced state of macroscopically clearly infiltrative growth were killed. Benzo(a)pyrene, benzo(b)fluoranthene, benzo(j)fluoranthene, benzo(k)fluoranthene at concentrations between 0.01% and 0.5% dissolved in acetone were applied to the clipped backs of female Swiss mice (20/dose/chemical) three times
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per week for the lifetime of the animals (Wynder and Hoffmann, 1959). Results show that benzo(a)pyrene, benzo(b)fluoranthene, and benzo(j)fluoranthene produced high incidences of skin papillomas and carcinomas at all dose levels. Benzo(k)fluoranthene produced a limited number of papillomas only at the high dose level (0.5%). There were no control groups in the study.
Habs, M., Schmahl, D., and Misfeld, J. (1980). Local carcinogenicity of some environmentally relevant polycyclic aromatic hydrocarbons after lifelong topical application to mouse skin. Arch. Geschwulstforsch. 50:266-274. Wynder, E.L. and Hoffmann, D. (1959). The carcinogenicity of benzo(b)fluoranthene. Cancer. 12:1194.
BENZO(K)FLUORANTHEN (C20)

BENZO(GHI)PERYLENE (C20)
Female Ha/ICR/mil Swiss albino mice (20/dose level) received three weekly topical applications of B(a)P, benzo(ghi)perylene, or indeno(1,2,3-cd)pyrene for one year at various concentrations (Hoffmann and Wynder, 1966). B(a)P and benzo(ghi)perylene were dissolved in dioxane and indeno(1,2,3-cd)pyrene was dissolved in acetone. There were dioxane controls but no acetone controls. Tumors were observed at 30 and 35% in the higher dose groups (0.1 and 0.5%, respectively) of indeno(1,2,3-cd)pyrene but not in the two lower dose groups (0.01 and 0.05%). B(a)P produced tumors in 85 and 95% of the animals at concentrations of 0.05 and 0.1%, respectively.
Hoffmann, D. and Wynder, E.L. Krebsforsch. 68:137-149. (1966). Beitrag zur carcinogen Wirkung von Dibenzopyrenen. Z.
DIBENZ(AH)ANTHRACENE (C22)

BENZO(A)PYRENE (C20)
Classified as a B2 carcinogen- slope factor 7.3 (mg/kg/day)-1. Male and female CFW mice were administered dietary doses of B(a)P at concentrations up to 1000 ppm for varying lengths of time (23 to 238 days) (Rigdon and Neal 1966, 1969). Treated mice exhibited an increased incidence of forestomach tumors. At the 250 ppm level, 64% of the animals developed papillomas or carcinomas of the forestomach, while 100% of the mice fed 1000 ppm exhibited tumors of the forestomach. Male and female CFW mice were fed B(a)P at concentrations of 0, 1, 10, 20, 30, 40, 45, 50, 100, and 250 ppm in the diet (Neal and Rigdon, 1967). The food was given ad libitum and the amount of food ingested per day was not measured and was assumed to be 4 g per day. The number of mice per exposure level ranged from 23 to 73 and the length of exposure varied from 70 8 to 197 days. There were also variations in the age of the animals at the beginning of the experiment (17 to 101 days) and the interval between the end of exposure and death (2 to 101 days).
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There were 289 mice in the control group. At autopsy, the stomachs were removed and washed with tap water and macroscopically examined. Select specimens were fixed for histological analysis. No forestomach tumors were reported in the 0, 1, and 10 ppm dose groups. The incidence of forestomach tumors in the 20, 30, 40, 45, 50, 100, and 250 ppm dose groups were 1/23, 0/37, 1/40, 4/40, 24/34, 19/23, and 66/73, respectively. There is no indication of any other organ being evaluated either macro- or microscopically. Brune et al. (1981) exposed five groups of Sprague-Dawley rats (32/sex/group) to B(a)P at a concentration of 0.15 mg/kg. Three of the groups were exposed by gavage with different exposure regimens: 5 times/week, every 3rd day, or every 9th day. The remaining two groups were exposed through the diet either 5 times/week or every 9th day. Exposures lasted for the lifetime of the animals which varied from 87 to 131 weeks. This study focused on the development of tumors in the gastrointestinal tract. The combined incidence of tumors of the forestomach, esophagus, and larynx was 3/64, 3/64, and 10/64 for the control group, the group fed B(a)P in the diet every 9th day, and the group fed B(a)P in the diet 5 times/week, respectively. Tumors at other sites (i.e., mammary gland, kidney, pancreas, lung, urinary bladder, testes) were determined to be spontaneous because they were observed at similar incidence levels in control animals. However, the data on the number of tumors in treated and control animals at these other locations were not reported in the study. Thyssen et al. (1981) exposed (nose only) Syrian golden hamsters to an average concentration of 2.2, 9.5, and 46.5 mg/m3 of B(a)P for 4.5 hours/day, 7 days/week for the first 10 weeks and 3 hours/day for the remainder of the treatment time. Animals exposed to 0, 2.2, 9.5 and 46.5 mg/m3 were exposed for a total 96.4, 95.2, 96.4, and 59.5 weeks, respectively. The numbers of animals per group were 27, 27, 26, and 25 for controls, low-, mid-, and high-dose groups, respectively. Animals were sacrificed and organs were fixed and sections for histology were prepared. Tumor data were presented for respiratory tract, digestive tract, and other sites. In the other sites category, tumors from the pituitary gland, harderian gland, thyroid gland, parathyroid gland, liver, pancreatic islets, pancreatic ducts, kidneys, adrenal gland, and colon were combined. The incidences of respiratory tract tumors were 0/27 for controls, 0/27 for the low-dose group, 9/26 for the mid-dose group, and 13/25 for the high-dose group. The highest exposure level did have an impact on survival time when compared with controls (59.5 weeks for high-dose vs. 96.4 weeks for control).
Brune, H., Deutsch-Wenzel, R.P Habs, M., Ivankovic, S., and Schmahl, D. (1981). Investigation of the ., tumorigenic response to benzo(a)pyrene in aqueous caffeine solution applied orally to Sprague-Dawley rats. J. Cancer Res. Clin. Oncol. 102:153-157. Neal, J. and Rigdon, R.H. (1967). Gastric tumors in mice fed benzo(a)pyrene: A quantitative study. Tex. Rep. Biol. Med. 25: 553-557. Rigdon, R.H. and Neal, J. (1966). Gastric carcinomas and pulmonary adenomas in mice fed benzo(a)pyrene. Tex. Rep. Biol. Med. 24:195-207. Rigdon, R.H. and Neal, J. (1969). Relationship of leukemia to lung and stomach tumors in mice fed benzo(a)pyrene. Proc. Soc. Exp. Biol. Med. 130:146-148. Thyssen, J., Althoff, J.K.G., and Mohr, U. (1981). Inhalation studies with benzo(a)pyrene in Syrian golden hamsters. J. Natl. Cancer Inst. 66:575-577.
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APPENDIX C
Review of American Petroleum Institutes (APIs) Toxicity Data on Selected Refinery Streams
SUMMARY
REVIEW OF AMERICAN PETROLEUM INSTITUTES (APIS) TOXICITY DATA ON SELECTED REFINERY STREAMS
The objective of this section was to review the APIs toxicity data on selected refinery streams in order to: 1. determine whether any of the refinery streams were similar in overall composition to the equivalent carbon ranges of the surrogate fractions 2. determine whether any of the toxicity studies on the refinery streams could be used to develop oral reference doses (RfDs). Analytical data were available for each of the streams which were tested. However, the compositional analysis varied depending on the stream. Detailed GCMS data were available for each of the nine naphtha streams. From this data, the weight percent of the individual hydrocarbons with differing carbon numbers was determined. However, for the remaining 28 streams, the only analytical data available were information from the distillation curve for the material (i.e., the volume percent distilled off at differing temperatures) and the total percent of saturates, olefins, and aromatics in the stream. Using the distillation data, the carbon numbers of the compounds distilling off at different temperatures was estimated. It was then assumed that each boiling range had the same percentage of aliphatics and aromatics. In this way, the percent of aliphatics and aromatics in the various fractions was estimated. However, it must be noted that since the analytical data were limited, the estimates of the percent aliphatics and aromatics in the carbon ranges of interest remain highly uncertain. During the next phase of the review, the toxicity data on the refinery streams were evaluated in order to determine whether the data could be used to develop oral reference doses (RfDs). The data reviewed included 26 chronic dermal carcinogenicity studies, 5 subchronic inhalation studies, and 30 subchronic dermal studies. There were no chronic or subchronic oral studies available for any of the refinery streams. Based on the review, it was determined that only two subchronic inhalation studies would be considered appropriate by the US EPA for the development of oral RfDs. The chronic data were deemed unacceptable for the development of RfDs because in these studies only a single dose was tested and systemic effects were not evaluated. The subchronic dermal studies could not be used because no absorption data were available. In addition, only two of the five subchronic inhalation studies were done using multiple doses and could be used to develop oral RfDs. However, in order to develop oral RfDs from the subchronic inhalation studies, route-to-route extrapolation was required. It should be noted that recently, the EPA has expressed concern regarding the use of route-to route extrapolation due to the uncertainties associated with this methodology. In conclusion, based on the review it was determined that the refinery streams were not similar in composition to any of the surrogate fractions. Furthermore, of the available data, only two subchronic inhalation studies would be considered appropriate by the US EPA for the development of oral RfDs.
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I. INTRODUCTION
A. PURPOSE OF THE DOCUMENT
The review of the American Petroleum Institutes data on selected refinery streams was undertaken in order to determine: whether, based on composition, any of the refinery streams are similar in overall composition to the carbon ranges of the surrogate fractions whether any of the toxicity studies on the refinery streams can be used to develop oral reference doses (RfDs)
II. EPA METHODOLOGY FOR THE DEVELOPMENT OF ORAL REFERENCE DOSES (RfDs)
The US EPA issued The Risk Assessment Guidelines of 1986 in which methods for developing reference dose (RfD) values were given. Since then, Proposed Guidelines for Neurotoxicity; Guidelines for Developmental Toxicity Risk Assessment; and Guidelines for Reproductive Toxicity Risk Assessment have been published, which also discuss the development of RfDs. In addition, the IRIS database provides guidance for developing RfD values in Background Document 1 (Reference Dose: Use in Health Risk Assessment), last revised March 15, 1993. Information on the development and use of RfDs is also provided in the Risk Assessment Guidance for Superfund Volume 1 Human Health Evaluation. A review of the available information shows that US EPAs methods for determining the RfDs remains unchanged.
A. DEFINITION OF THE REFERENCE DOSE (RfD)
The RfD is an estimate (with uncertainty spanning perhaps an order of magnitude) of daily exposure to the human population, including sensitive subgroups, that is likely to be without appreciable risk of deleterious effects during a lifetime (USEPA, 1989). The RfD is used to evaluate potential noncarcinogenic effects of exposure to a given compound. It is not used to evaluate carcinogenic endpoints. The RfD is operationally derived from a no-observed-adverse-effect-level (NOAEL) by application of uncertainty factors (UFs) that reflect various types of data sets used to estimate a reference value and application of a modifying factor (MF), which reflects the completeness of the database.
B. APPROPRIATE DATA FOR DEVELOPMENT OF AN ORAL RfD
The first step in developing an RfD is to choose a critical study and determine the NOAEL. The NOAEL is the highest dose at which no adverse effects are observed. If a NOAEL is not available, a lowest-observed-adverse-effect-level (LOAEL) can be used; however, this adds an additional uncertainty factor into the equation. The most appropriate source of the NOAEL for the oral RfD (or LOAEL) is from a
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chronic oral study. If there is no chronic oral data, subchronic oral data can also be used, but this also adds another level of uncertainty into the derivation of the RfD. In some cases, chronic and subchronic inhalation studies can be utilized in the development of oral RfDs when there are no oral data available. However, this is not common practice because there is a great deal of uncertainty surrounding the use of inhalation data in this respect. Most other toxicity data (i.e., dermal, acute, or genotoxic) are not recommended for use in the development of oral RfDs. Acute oral LD50s have been used to develop tentative RfDs, which represent a dose that the true RfD will fall below. These RfDs are calculated by taking the oral LD50 and dividing it by 100,000 or 1,000,000. The use of dermal data to develop oral RfDs is an area currently being reviewed by the US EPA. The biggest concern centers around the lack of data on the amount of the compound absorbed through the skin which causes the toxic endpoint. Because this is an ongoing issue with US EPA, there is no regulatory guidance on the methodology to be used in the calculation process. In a published document by Ryer-Powder and Sullivan (1994), an oral RfD was derived from a dermal carcinogenicity study. In the development of the RfD, an absorption factor of 1% was incorporated into the equation. However, no data were provided that support the use of this extremely conservative value. Information on absorption of various compounds through the skin is available in the literature. In a report on the bioavailability of petroleum constituents, default dermal absorption values from soil were recommended for benzo(a)pyrene, benzene, toluene, and xylene of 3 - 30%, 10%, 10%, and 75%, respectively (Brainard and Beck, 1992). Another report investigated the absorption of radiolabeled benzo(a)pyrene from lubricants of different viscosities and found a range of 18 to 23% for dermal absorption (CONCAWE, 1990). More reasonable estimates of absorption can be obtained from these studies.
C. DERIVATION OF AN ORAL RfD
The RfD is calculated using the following equation: RfD = NOAEL (or LOAEL)/(UF x MF) where the RfD is expressed in mg/kg/day; the NOAEL (or LOAEL) represents a critical effect; the uncertainty factor (UF) can range from 1 to 10,000; and the modifying factor (MF) can range from 1 to 10 based on the completeness of the data set.
D. UNCERTAINTY AND MODIFYING FACTORS
Following are explanations of the different uncertainty factors used in the derivation of RfDs: Use a 10-fold factor when extrapolating from valid experimental results in studies using prolonged exposure to average healthy humans. This factor is intended to account for the variation in sensitivity among the members of the human population and is referenced by the EPA as 10H.
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Use an additional 10-fold factor when extrapolating from valid results of long-term animal studies when results of humans studies are either not available or inadequate. This factor accounts for the uncertainty involved in extrapolating from animal data to humans and is referenced by the EPA as 10A. Use an additional 10-fold factor when extrapolating from less than chronic results on experimental animals when there are no useful long-term human data. This factor is intended to account for the uncertainty involved in extrapolating from less than chronic NOAELs to chronic NOAELs and is referenced by the EPA as 10S. Use an additional 10-fold factor when deriving an RfD from a LOAEL, instead of a NOAEL. This factor is intended to account for the uncertainty involved in extrapolating from LOAELs to NOAELs and is referenced by the EPA as 10L. The modifying factor is an additional uncertainty factor and is applied based on the strength of the database and professional judgment. The MF can range from 1 to 10.
III. DESCRIPTION OF THE API DATA ON REFINERY STREAMS

A. CONTENT OF API DATA ON REFINERY STREAMS
During the past 15 years, the API has conducted a series of toxicological studies on selected refinery streams. The streams which were tested represent high volume refinery processes. These streams vary widely in composition and range from materials in the naphtha range having carbon numbers predominantly in the C4 to C11 range (light ends) to heavy vacuum residuums which have carbon numbers predominantly greater than C34. The names of the refinery streams, their API code numbers, and description of their composition (i.e., predominant carbon number range, percent aliphatics and aromatics) are provided in Table C-1. In order to determine their toxicity, a battery of toxicity tests was carried out on each stream. In addition to acute toxicity testing, some of the streams were tested to evaluate their genotoxic potential (see Table C-2). In some cases, subchronic dermal and inhalation studies as well as chronic dermal studies were also conducted (see Table C-3). However, it should be noted that no subchronic or chronic oral studies were carried out on any of the streams.
B. EVALUATION OF GENETOX DATA
Genotoxicity assays are used to assess the potential of a test article to cause mutagenicity, clastogenicity (chromosome damage), or DNA damage in either in vitro or in vivo test systems. The results of these assays can assist in the assessment of the carcinogenicity potential of a compound. Typically, compounds that are carcinogenic are also genotoxic. So, conversely, compounds that are genotoxic could potentially go on to produce cancer.
104
TABLE C-1. Description of API Refinery Streams

API Refinery Stream Code 78-02 78-03 78-04 78-05 78-09 78-10 79-01 79-03 79-04 79-05 81-03 81-04 81-07 81-08 81-09 81-10 81-13 81-14 81-15 83-01 83-02 83-03 83-04 83-05 83-06 83-07 83-08 83-09 83-11 83-12 83-15 83-16 83-18 83-19 84-01 84-02 85-01
a
Name Home heating oil Medium catalytic cracked stock (30%) Home heating oil Low catalytic cracked stock (10%) Home heating oil High catalytic cracked stock (50%) Naphthenic base stock Paraffinic base stock Paraffinic base stock Naphthenic base stock Paraffinic base stock Paraffinic base stock Paraffinic base stock Light catalytically cracked naphtha Light catalytically cracked naphtha Hydrodesulfurized kerosine Sweetened naphtha Hydrodesulfurized middle distillate Hydrodesulfurized middle distillate Vacuum residuum Vacuum residuum Catalytically cracked clarified oil Home heating oil Light catalytic cracked stock (10%) Home heating oil Light catalytic cracked stock (30%) Home heating oil Light catalytic cracked stock (50%) Light catalytically reformed naphtha Full range catalytically reformed naphtha Heavy catalytically reformed naphtha Light catalytically cracked distillate Light catalytically cracked distillate Straight run kerosine Straight run middle distillate Hydrotreated light naphthenic distillate Hydrotreated heavy naphthenic distillate Light paraffinic distillate solvent extract Heavy catalytically cracked naphtha Light alkylate naphtha Light paraffinic distillate Heavy thermally cracked naphtha Stoddard solvent
Predominant Carbon Number Range C10 - C24a C10 - C24a C10 - C22a C15 - C30 C15 - C25a > C19a C20 - C50 C20 - C50 C20 - C50 C20 - C50 C4 - C11 C4 - C11 C9 - C16 C4 - C12 C11 - C25 C11 - C25 > C34 > C34 > C20 C10 - C19a C10 - C19a C10 - C20a C5 - C11 C4 - C12 C7 - C12 C9 - C25 C9 - C25 C9 - C16 C11 - C20 C15 - C30 C20 - C50 C15 - C30 C6 - C12 C7 - C10 C15 - C30 C6 - C12 C9 - C11
Percent Aliphatic NAb NAb NAb 76.2 89.8 86.2 62.3 71.9 72.2 68.1 91.4 79.7 82 96 75 69.1 NAb 29.5 8 75.6 70.5 72.7 60.5 37.5 8.7 27.6 31.7 82 78.8 67.3 53.1 39.1 36.2 99.9 79.1 84.3 85.5
Percent Aromatic NAb NAb NAb 23.8 10.2 13.8 37.7 28.1 27.8 51.9 8.6 20.3 18 4 25 30.9 NAb 34.7 58.3 24.4 29.5 27.3 39.5 62.5 91.3 72.4 68.3 18 21.2 31.9 46.9 60.9 63.8 <0.1 20.9 15.7 14.5
Value is estimated because data are not provided. b No data are available for this parameter.
105
TABLE C-2. Genotoxicity Data Available for Selected Refinery Streamsa

API Refinery Streamsb 78-02 78-03 78-04 78-05 78-09 78-10 79-01 79-03 79-04 79-05 81-03 81-04 In vitro Mouse Lymphoma In vivo Rat Bone Marrow Modified Ames In vivo SCEc In vitro SCEc
Ames
+ Ed E E E E 1) E (w A)e - (w/o A) 2) -
+f + -
E -
81-07 81-08 81-09 81-10 81-14 81-15 83-01 83-02 83-03 83-04 83-05
+ (w A) - (w/o A)
E (w A) - (w/o A) + (w A) - (w/o A)
+ (w A) - (w/o A) 83-06 1)+ (w A) - (w/o A) 2) E 83-07 + (w A) - (w/o A) + 83-08 + 83-09 E (w A) + (w/o A) 83-11 1) + (w A) - (w/o A) 2) + 83-12 + 83-15 83-16 + 83-18 + 83-19 84-01 + (w A) - (w/o A) 84-02 + 85-01 + a Descriptions of the genotoxicity assays can be found in Table C-4. b Data on the composition of the refinery streams can be found in Table C-1. c Sister chromatid exchange d Equivocal results e Activation f Results questionable
106
TABLE C-3. Subchronic and Chronic Toxicity Data Available for Selected Refinery Streams
Subchronic ___________________________________ API Refinery Streamsa 78-02 78-03 78-04 78-05 78-09 78-10 79-01 79-03 79-04 79-05 81-03 81-04 81-07 81-08 81-09 81-10 81-13 81-14 81-15 83-01 83-02 83-03 83-04 83-05 83-06 83-07 83-08 83-09 83-11 83-12 83-15 83-16 83-18 83-19 84-01 84-02 85-01
a
Chronic ___________________________________ Oral Inhalation Dermal X X
Oral
Inhalation
Dermal
X X X X X X X X X X X X X X X X X X X
X X X X X X X X X X X X
X X X X X X X X X X X X X X X X X X X X X X X X X X X
Data on the composition of refinery streams can be found in Table C-1.
107
With the API refinery streams, there have been several different genotoxicity assays performed to assess the potential for these streams to cause mutagenicity or clastogenicity (chromosomal damage) in both in vivo and in vitro test systems (see Table C-4). However, there have been no assays conducted which look at the potential for these streams to cause DNA damage. In looking at the results of these assays (see Table C-2), the majority of the results were either negative or equivocal. Equivocal results mean that the results of the assay were uncertain and a conclusion on genotoxicity could not be made. There were some positive results for several streams, but these results were not conclusive because either only one genotoxicity assay was performed or there were mixed results on several genotoxicity assays. For example, API refinery stream 83-12 (hydrotreated light naphthenic distillate) had positive results in an in vitro mouse lymphoma assay. However, this was the only assay performed on this stream and there was no other supporting evidence. In the OECD guidelines for genetic toxicology testing, it is recommended that a battery of tests be performed to assess the genotoxic potential of a compound because the results from one test are inconclusive and have low confidence. For several of the API refinery streams, there were multiple genotoxicity assays and there were both positive and negative results obtained. For example, with API refinery stream 79-03 (paraffinic base stock), two of the assays were negative, one was equivocal, and one assay had positive results. The interpretation of these results would be equivocal because of the mixed results. Further testing would be needed to ascertain the genotoxic potential of this refinery stream. Genotoxicty tests are not typically used by the US EPA in the development of RfDs. As mentioned previously, data from these types assays are typically used to assess the potential of compound to be carcinogenic. Because the majority of the genotoxicity data for the API refinery streams was negative, this indicates a low potential of the streams to cause genotoxicity and a low concern for carcinogenic potential of the streams.
C. EVALUATION OF SUBCHRONIC AND CHRONIC DATA
As stated above, the API conducted a number of subchronic inhalation and dermal studies as well as some chronic dermal carcinogenicity studies on selected refinery streams (Table C-3). In order to develop oral RfDs, the EPA currently recommends that these values be developed from chronic oral or inhalation studies. Alternatively, subchronic oral or subchronic inhalation studies can be used. However, to date, dermal studies (either chronic or subchronic) have not been used by the EPA to develop oral RfDs for petroleum hydrocarbons due to the uncertainty associated with the amount test material absorbed and problems with the protocols (e.g., studies do not follow TSCA or OECD guidelines). Furthermore, in the majority of cases the only effects observed in these dermal studies are local effects such as erythema and/or edema. Repeated dermal exposure protocols with severely irritating materials are not representative of human exposure scenarios. In those studies where systemic effects were evaluated, they were typically not observed. Thus, the use of dermal studies for the development of oral reference doses remains questionable.
108
TABLE C-4. Description of Genotoxicity Tests Conducted on API Refinery Streams

Genotoxicity Test Mutagenic Assays In vitro mouse lymphoma Evaluate test article for its ability to induce forward mutation in the mouse lymphoma cell line. A forward mutation alters the gene and inactivates it. With this type of mutation, there is a detectable change in appearance or structure. Evaluate the test article for mutagenic activity in a bacterial mutation system , Salmonella, with and without a mammalian S9 activation component. Objective
Ames and modified Ames
Clastogenic (chromosome damage) Assays In vivo rat bone marrow Evaluate test article for its ability to induce chromosome aberrations, which are changes in chromosome structure, in somatic (body) cells. First, animals are treated with the test article and then the cells are removed and analyzed. Evaluate the ability of the test article to induce SCEs in rodent bone marrow, spleen, or spermatagonia cells. An SCE is when segments of two chromatids of a chromosome are exchanged. First, animals are treated with the test article and then the cells are removed and analyzed. Evaluate the ability of the test article to induce SCEs in Chinese hamster ovary (CHO) cells, with and without metabolic activation.
In vivo sister chromatid exchange (SCE)
In vitro SCE
IV. ANALYTICAL COMPOSITION OF REFINERY STREAMS

Although analytical data are available for each of the streams that were tested, the extent of the compositional analysis varies depending on the specific stream. For example, detailed GC-MS data are available for each of the nine naphtha streams. From this data, the weight percent of individual hydrocarbons with differing carbon numbers can be determined (e.g., API 81-03, API 84-01, Attachment I). However, in other cases, the only analytical data available on the stream are the information from the distillation curve for the material (i.e., the volume percent distilled off at differing temperatures) and the total percent of saturates, olefins and aromatics in the stream. Using the distillation data, the carbon numbers of the compounds distilling off at different temperatures can be estimated. If one then assumes that each boiling range has the same percentage of aliphatics and aromatics, the percent of aliphatics and aromatics in the various fractions can be estimated (e.g., API 81-07, Attachment I). However, it must be recognized that these assumptions may, at times, result in either significant overestimates or underestimates of these fractions.
109
For the purpose of illustration, the composition of API Stream 81-03 (Light Catalytic Cracked Naphtha) which was determined by GC-MS and API 81-07 (Hydrodesulfurized Kerosine) whose composition was determined based on distillation curve data are presented in tabular form in Tables C-5 and C-6, respectively. In order to better compare these data to the surrogate fractions, simpler tables were prepared from the information presented in Tables C-5 and C-6. As shown in Tables C-7 and C-8, the total weight percent of compounds with carbon numbers in the various ranges (both aliphatic and aromatic) were added together. For example, as shown in Table C-3, API 81-03 contains 30.1% C5 aliphatics and 29.1% C6 aliphatics for a total of 59.2% aliphatics in the C5 - C6 range (Table C-5), etc.
V. DEVELOPMENT OF RfDs FOR SELECTED REFINERY STREAMS BASED ON THE BEST AVAILABLE CHRONIC OR SUBCHRONIC DATA
A. CHRONIC DATA
For certain streams, the only data available are from 2-year chronic dermal carcinogenicity studies. Although one can attempt to develop oral RfDs from such studies, it is doubtful that this is appropriate for the following reasons: Only a single dose level is used in these studies. Thus, they do not provide a dose response assessment and therefore an accurate determination of toxicity is not possible. When tumors are observed, they occur only at the site of application (i.e., skin tumors). Systemic effects are typically not evaluated. Protocols not consistent with TSCA and OECD guidelines. Based on the reasons stated above, such data should not be used to develop an oral RfD, which will protect against systemic effects. Therefore, oral RfDs were not developed from these studies.
B. DEVELOPMENT OF RFDs FROM SUBCHRONIC INHALATION AND SUBCHRONIC DERMAL STUDIES 1. Subchronic Inhalation Studies.
As shown in Table C-3, subchronic inhalation studies were conducted on five refinery streams. Of these five studies, only two were appropriate for the development of an oral RfD. These two studies were API 81-03 (Light Catalytic Cracked Naphtha) and 85-01 (Stoddard Solvent); both were 13-week inhalation studies with multiple dose levels. The remaining three subchronic inhalation studies (API 8107 Hydrodesulfurized Kerosene, 81-09 and 81-10 Hydrodesulfurized Middle Distillate) were inappropriate because they were conducted using a single dose level and hence no dose response information was available. The examples of the development of oral RfDs from API 81-03 (Light Catalytic Cracked Naphtha) and 85-01 (Stoddard Solvent) are shown on the following pages.
110
TABLE C-5. Light Catalytic Cracked Naphtha (API 81-03)

BP Range >nC3-</=nC4 >nC4-</=nC5 >nC5-</=nC6 >nC6-</=nC7 >nC7-</=nC8 >nC8-</=nC9 >nC9-</=nC10 >nC10-</=nC11 >nC11-</=nC12 86 13 <0.0001 ~0.1 0.18 0.24 0.25 0.23 1.3 2.2 3.8 0.09 0.1 10.2 4.7 0.67 3.17 22.3 2.9 4.66 0.69 28.2 2.87 1.9 21.6 1.2 wt % Aliphatic BP Range wt % Toluene wt % Et.benzene wt % Xylenes wt % nC6 wt % nC7 wt % nC9 wt % Naphthalene wt % Pyrene(C16) wt % Aromatic wt % BP Range Benzene wt % PNAs
GC/MS ID
wt % Aliphatic C#(GC/MS)
wt % Aromatic C#(GC/MS)
C4s
2.4
C5s
30.1
C6s
29.1
2.9
C7s
16.8
4.7
C8s
5.8
3.8
C9s
1.3
1.4
C10s
0.21
0.26
C11s
0.14
0.1
C12s
86
13
HC Type/Summed
99
99
HC Type/MS HC Type/D1319
90 91
10 9
111
112
TABLE C-5. Continued

D-1319 ~~~C# nC5 46 27 12 5 56 35 9 4.5 2.7 1 0.5 Vol %
D-86 DISTILLATION DATA:
vol % Distilled off: IBP 5% 10% 20% 30% 40% 50% 60% 70% 80% 90% 95% EP(99%) degF 93 106 113 123 133 143 153 168 185 208 244 280 350 nC6 nC7 nC8 nC8 nC10 degC 34 41 45 51 56 62 67 76 85 98 118 138 177
Saturates Olefins Aromatics -</=nC6 >nC6-</=nC7 >nC7-</=nC8 >nC8-</=nC9 >nC9-</=nC10 >nC10-</=nC11 >nC11-</=nC12
D-1319 56 35 9
Vol %
Saturates Olefins Aromatics
API Gravity: 70 Density: 0.7005
TABLE C-6. Hydrodesulfurized Kerosene (API 81-07)
Detailed hydrocarbon data not available, thus the table with C# distribution and BP range cannot be generated
GC/MS ID >nC3-</=nC4 >nC4-</=nC5 >nC5-</=nC6 >nC6-</=nC7 >nC7-</=nC8 >nC8-</=nC9 >nC9-</=nC10 >nC10-</=nC11 >nC11-</=nC12 0.8 0.13 0.1
wt % Aliphatic C#(GC/MS) BP Range wt % Toluene wt % Et.benzene wt % Xylenes wt % nC6 wt % nC7 wt % nC9 wt % Naphthalene wt % Pyrene(C16)
wt % Aliphatic BP Range
wt % Aromatic wt % BP Range Benzene
wt % PNAs
C4s
C5s
C6s
C7s
C8s
C9s
C10s
C11s
C12s
HC Type/Summed
0.0
0.0
0.0
0.0 0.0002 ~0.9
HC Type/MS HC Type/D1319
71 78
29 22
113
114
TABLE C-6. Continued

Assume that each boiling range has the same % of aliphatics and % of aromatics: degC nC9 nC11 BP Range 0 0 ~~~C# (78 vol%) vol % Aliphatic BP Range (22 vol%) vol % Aromatic BP Range
D-86 Distillation Data:
vol % Distilled off:
degF
nC12
IBP 5% 10% 20% 30% 40% 50% 60% 70% 80% 90% 95% EP(99.5%) nC13 >nC8-</=nC9 >nC9-</=nC10 >nC10-</=nC11 >nC11-</=nC12 >nC12-</=nC13 >nC13-</=nC14 >nC14-</=nC15 3.9 19.5 31.2 19.5 3.5 1.1 5.5 8.8 5.5 1 nC14 nC15
310 382 395 409 420 429 436 443 452 464 480 493 533
154 194 202 209 215 221 224 228 233 240 249 256 278
D-1319
Vol %
77.3 0.5 22.2
API Gravity: 41.8 Density: 0.8157
TABLE C-7. API SAMPLE 81-03 (Light Catalytic Cracked NAPHTHA)

Wt% Aliphatic C4 C5-C6 C7-C8 C9-C10 C11-C12 C13-C16 C17+ 2.4 59.2 22.6 1.5 0.14 2.9 8.5 1.7 0.1 Wt% Aromatic
TABLE C-8. API SAMPLE 81-07 (Hydrodesulfurized Kerosine)

Wt% Aliphatic C4 C5-C6 C7-C8 C9-C10 C11-C12 C13-C16 C17+ 23.4 54.2 6.6 14.4 Wt% Aromatic
115
2. Subchronic Dermal Studies
As shown in Table C-3, subchronic dermal studies were conducted on 30 refinery streams. However, absorption data were not available for any of the these studies. Thus, a default assumption would be required to estimate the amount of material absorbed. Because of the uncertainties inherent in such default assumptions, the use of dermal data to develop oral RfDs remains highly controversial and is currently not accepted by the USEPA. Therfore, oral RfDs were not developed from the dermal data on these refinery streams.
VI. CONCLUSIONS
The API Refinery Stream Data cannot be used to develop oral RfDs for the following reasons: The composition of the refinery streams does not adequately match the equivalent carbon ranges of the surrogate fractions. The chronic dermal carcinogenicity studies were conducted with a single dose and systemic effects were not evaluated. The development of oral RfDs from the two multidose subchronic inhalation studies required route-to-route extrapolation; although currently acceptable, route-to-route extrapolation is not recommended by the USEPA. Absorption data were not available for any subchronic dermal study so a default value (50%) was used; due to the uncertainties inherent in such default assumptions, the use of dermal data to develop oral RfDs remains controversial and is currently not accepted by the USEPA.
116
EXAMPLE 1. API 81-03 Light Catalytical Cracked Naptha In this study, rats were exposed to 1510, 2610 or 4520 ppm for five days/week for 13 weeks. There was a dose-related increase in liver weights for both males and females in the high-level group. These liver weight changes were associated with a trace severity of cellular hypertrophy. Thus, 2610 ppm was considered to be the NOAEL in this study. In order to calculate an oral RfD from this study, the inhalation NOAEL must first be converted to an equivalent oral dose.
DEVELOPMENT OF AN ORAL RFD FROM SUBCHRONIC INHALATION STUDY: 1. Convert inhalation NOAEL to Oral NOAEL
Inhalation NOAEL = 2610 ppm = 8753 mg/m3 (MW = 82) Daily exposure period = 6 hr/24 hr Assumed daily respiratory volume for a 0.31 kg rat = 0.2 m3 /day Conversion of 5 day/week dosing regimen to 7 day/week continuous exposure = 5/7 Estimated ratio of inhaled dose systemically absorbed = 0.5 (Pepelko and Withey 1985)
Equivalent Oral Dose =
8753 mg/m3 x 6hr/24 hr x 0.2 m3/day x 5d/7d x 0.5 = 520 mg/kg/day 0.3 kg
Calculation of RfD:
Equivalent Oral Dose Uncertainty Factor Uncertainty Factor = 1000 (to account for use of subchronic study, variation within species, variation between species). 520 mg/kg/day = 0.521 mg/kg/day 1000
117
EXAMPLE 2 API 85-01 Stoddard Solvent Development of an oral RfD from a 13-week inhalation toxicity study in the rat (animals dosed 6 hr/day, 5 days/week x 13 weeks - Tox. Appl. Pharmacol. 32:282297, 1975).
1. Convert inhalation NOAEL to oral NOAEL
Inhalation NOAEL = 1.9 mg/L = 1900 mg/m3 Daily exposure period = 6 hr/24 hr Assumed daily respiratory volume for a 0.3 kg rat = 0.2 m3/day Conversion of 5 day/week dosing regimen to 7 day/week continuous exposure = 5/7 Estimation of ratio of inhaled dose systemically absorbed = 0.5
Equivalent Oral Dose:
1900 mg/m3 x 6 hr/24hr x 0.2m3/day x 5 d/7 d x 0.5 = 113 mg/kg/day 0.3 kg

Calculation of Oral RfD: (uncertainty factor = 1000)
113 mg/kg/day = 0.11 mg/kg/day 1000
118
REFERENCES
API Health Environ. Sci. Dep. Rep. Brainard, J. and Beck, B.D. (1992). A review of bioavailability of petroleum constituents. Submitted to the Association of the Environmental Health of Soils. Presentation at 1992 West Coast Soils and Groundwater Conference. Carpenter, C.P Kincead, E.R., Geary, D.L., Sullivan, L.J., and King, J.M. (1975). Petroleum hydrocarbon ., toxicity studies III. Animal and human response to vapors of Stoddard solvent. Tox. Appl. Pharmacol. 32:282-297. CONCAWE (1990). Factors Affecting the Skin Penetration and Carcinogenic Potency of Petroleum Products Containing Polycyclic Aromatic Hydrocarbons. Report No. 90/55. IRIS CD-Rom, Vol. 27. Expiration: January 31, 1996. Pepelko, W.E. and Withey, J.R. (1985). Methods for route-to-route extrapolation of dose. Tox. Ind. Health 1:153-175. Ryer-Powder, J.E., and Sullivan, M.J. (1994). Chapter 2: Update on the Derivation of an Oral Reference Dose for Diesel Fuel No. 2. In Principles and Practices for Diesel Contaminated Soils, Vol III, (P .T. Kostecki; E.J. Calabrese and C.P Barkan, Eds.). Amherst, MA, Amherst Sc. Publ. .L USEPA (1987). The Risk Assessment Guidelines of 1986 (EPA600/08-87-045), August. USEPA (1989). Risk Assessment Guidance for Superfund Volume 1 Human Health Evaluation Manual. (EPA/540/1-89/002), December. USEPA (1990) Review Draft, Interim Methods for Development of Inhalation Reference Concentrations (EPA/600/8-90/006A), August. USEPA (1991). Guidelines for Developmental Toxicity Risk Assessment, Federal Register Vol. 56 No. 234, December 5. USEPA (1994). Review Draft Guidelines for Reproductive Toxicity Risk Assessment (EPA600.AP .94.001), February. USEPA (1995). Proposed Guidelines for Neurotoxicity, Federal Register Vol. 60, No. 192, October 4, 1995.
119
ATTACHMENT I
Composition Data on Selected API Refinery Streams
Table I-1. Light Catalytic Cracked Naphtha (API 81-03)

wt % BP Range >nC3-</=nC4 >nC4-</=nC5 >nC5-</=nC6 >nC6-</=nC7 >nC7-</=nC8 >nC8-</=nC9 >nC9-</=nC10 >nC10-</=nC11 >nC11-</=nC12 86 13 <0.0001 ~0.1 0.18 0.24 0.25 0.23 1.3 2.2 3.8 0.09 0.1 10.2 4.7 0.67 3.17 22.3 2.9 4.66 0.69 28.2 2.87 1.9 21.6 1.2 Aliphatic BP Range Aromatic wt % BP Range Benzene wt % Toluene wt % Et.benzene wt % Xylenes wt % nC6 wt % nC7 wt % nC9 wt % Naphthalene wt % Pyrene(C16) wt % wt % PNAs
wt %
wt %
GC/MS ID
Aliphatic C#(GC/MS)
Aromatic C#(GC/MS)
C4s
2.4
C5s
30.1
C6s
29.1
2.9
C7s
16.8
4.7
C8s
5.8
3.8
C9s
1.3
1.4
C10s
0.21
0.26
C11s
0.14
0.1
C12s
86
13
HC Type/Summed
99
99
HC Type/MS
90
10
HC Type/D1319
91
123
124
Table I-1. Continued

Assume that each boiling range has the same % of aliphatics and % of aromatics: degC nC5 BP Range ~~~C# (91 vol%) vol % Aliphatic BP Range (9 vol%) vol % Aromatic BP Range
degF
nC6 5
0 0 46 27 12 0.5
IBP 5% 10% 20% 30% 40% 50% 60% 70% 80% 90% 95% EP(99%) nC7 nC8 nC8 nC10 >nC3-</=nC4 >nC4-</=nC5 >nC5-</=nC6 >nC6-</=nC7 >nC7-</=nC8 >nC8-</=nC9 >nC9-</=nC10 >nC10-</=nC11 >nC11-</=nC12 0 0 4.5 2.7 1
93 106 113 123 133 143 153 168 185 208 244 280 350
34 41 45 51 56 62 67 76 85 98 118 138 177
D-1319
Vol %
56 35 9
Table I-2. API Sample 81-03 (Light Catalytic Cracked Naphtha)

Wt% Aliphatic C4 C5-C6 C7-C8 C9-C10 C11-C12 C13-C16 C17+ 2.4 59.2 22.6 1.5 0.14 2.9 8.5 1.7 0.1 Wt% Aromatic
125
126
Table I-3. Stoddard Solvent, Mineral Spirits (API 85-01)

BP Range >nC3-</=nC4 >nC4-</=nC5 >nC5-</=nC6 >nC6-</=nC7 >nC7-</=nC8 >nC8-</=nC9 >nC9-</=nC10 >nC10-</=nC11 >nC11-</=nC12 >nC12-</=nC13 83 16 <0.0001 ~0.7 0.52 0.27 9.7 3.7 28.5 5.5 35.6 6.0 9.1 0.52 0.13 ND ND 0.03 0.49 ND ND 5.3 0.61 wt % Aliphatic BP Range wt % Toluene wt % Et.benzene wt % Xylenes wt % nC6 wt % nC7 wt % nC9 wt % Naphthalene wt % Aromatic wt % BP Range Benzene wt % Pyrene(C16) wt % PNAs
GC/MS ID
C4s
C5s
C6s
C7s
C8s
0.21
0.52
C9s
10.4
6.4
C10s
36.8
6.5
C11s
27.4
2.3
C12s
8.1
0.49
C13s
0.44
HC Type/Summed
83
16
99
99
HC Type/MS
83.5
16.5
HC Type/D1319
83.6
16.4

Assume that each boiling range has the same % of aliphatics and % of aromatics:
vol % degC >nC9 BP Range 0 0 ~~~C# (83.5 vol%) vol % Aliphatic BP Range (16.5 vol%) vol % Aromatic BP Range
Distilled off:
degF
nC10 42.0 37.6 4.2 8.3 7.4 0.83
IBP 5% 10% 20% 30% 40% 50% 60% 70% 80% 90% 95% EP(98.5%) >nC3-</=nC4 >nC4-</=nC5 >nC5-</=nC6 >nC6-</=nC7 >nC7-</=nC8 >nC8-</=nC9 >nC9-</=nC10 >nC10-</=nC11 >nC11-</=nC12 nC11 <nC12
321 327 329 332 337 342 345 349 354 361 374 385 404
161 164 165 167 169 172 174 176 179 183 190 196 207
D-1319
Vol %
83.5 <0.1 16.5
127
Table I-4. API Sample 85-01 (Stoddard Solvent)

Wt% Aliphatic C4 C5-C6 C7-C8 C9-C10 C11-C12 C13-C16 C17+ 0.21 47.2 35.5 0.44 0.52 12.9 2.8 Wt% Aromatic
TABLE I-5. API Sample 83-15 (Hydrotreated Heavy Naphthenic Distillate)

Wt% Aliphatic C4 C5-C6 C7-C8 C9-C10 C11-C12 C13-C16 C17+ (c20+) 58.7 37.2 Wt% Aromatic
128
Table I-6. Light Parrafinic Distillate (API 84-01)

BP Range >nC3-</=nC4 >nC4-</=nC5 >nC5-</=nC6 >nC6-</=nC7 >nC7-</=nC8 >nC8-</=nC9 >nC9-</=nC10 >nC10-</=nC11 >nC11-</=nC12 0.9 2.4 2.4 8.3 7.6 8.9 0.39 0.04 11.8 5.0 1.64 7.29 18.7 0.9 4.97 0.6 18.4 0.94 1.18 13.9 0.72 wt % Aliphatic BP Range wt % Toluene wt % Et.benzene wt % Xylenes wt % nC6 wt % nC7 wt % nC9 wt % Naphthalene wt % Pyrene(C16) wt % Aromatic wt % BP Range Benzene wt % PNAs
GC/MS ID
C4s
2.2
C5s
19.6
C6s
19.8
0.9
C7s
14.3
5.0
C8s
9.4
8.9
C9s
5.7
9.1
C10s
1.9
1.7
C11s
0.9
C12s
HC Type/Summed
74
26
74
26 nd ~0.02
99
100
HC Type/MS
78
22
HC Type/D1319
79
21
129
130
D-86 Distillation Data: degC nC5 BP Range 0 0 nC6 ~~~C# (79 vol%) vol % Aliphatic BP Range (21 vol%) vol % Aromatic BP Range
degF
nC7 nC8 ~nC9 ~nC10
IBP 5% 10% 20% 30% 40% 50% 60% 70% 80% 90% 95% EP(97.5%) >nC3-</=nC4 >nC4-</=nC5 >nC5-</=nC6 >nC6-</=nC7 >nC7-</=nC8 >nC8-</=nC9 >nC9-</=nC10 >nC10-</=nC11 >nC11-</=nC12 16 24 16 16 4.0 2.0 4.2 6.3 4.2 4.2 1.1 0.5
100 122 133 152 170 187 211 236 260 283 316 340 359
38 50 56 67 77 86 99 113 127 139 158 171 182
D-1319
Vol %
48.4 30.6 21
TABLE I-7. API Sample 84-01 (Light Parrafinic Distillate)

Wt% Aliphatic C4 C5-C6 C7-C8 C9-C10 C11-C12 C13-C16 C17+ 2.2 39.4 23.7 7.6 0.9 0.9 13.9 10.8 Wt% Aromatic
131
132
BP Range >nC3-</=nC4 >nC4-</=nC5 >nC5-</=nC6 >nC6-</=nC7 >nC7-</=nC8 >nC8-</=nC9 >nC9-</=nC10 >nC10-</=nC11 >nC11-</=nC12 5.7 0.11 16.8 1.4 27.3 5.1 19.6 4.45 22.84 0.24 0.2 10.3 3.0 19.6 2.16 6.7 3.0 2.0 3.9 0.01 wt % Aliphatic BP Range wt % Toluene wt % Et.benzene wt % Xylenes wt % nC6 wt % nC7 wt % nC9 wt % Naphthalene wt % Aromatic wt % BP Range Benzene wt % Pyrene(C16) 27.5 72.4 0.0005
Table I-8. Full Range Catalytically Reformed Naphtha (API 83-05)

wt % PNAs
GC/MS ID
C4s
0.01
C5s
4.0
C6s
7.1
3.0
C7s
10
19.6
C8s
4.9
27.3
C9s
1.1
18.4
C10s
0.11
4.2
C11s
C12s
HC Type/Summed
27.2
72.5
100
~0.4
100
HC Type/MS
36
64
HC Type/D1319
37
63
TABLE I-8. Continued
D-86 DISTILLATION DATA: degC ~~nC6 0 0 nC7 BP Range ~~~C# (37 vol%) vol % Aliphatic BP Range (63 vol%) vol % Aromatic BP Range
degF
nC8
IBP 5% 10% 20% 30% 40% 50% 60% 70% 80% 90% 95% EP(97.5%) nC9 nC10 nC11 >nC3-</=nC4 >nC4-</=nC5 >nC5-</=nC6 >nC6-</=nC7 >nC7-</=nC8 >nC8-</=nC9 >nC9-</=nC10 >nC10-</=nC11 >nC11-</=nC12 1.9 5.6 11.1 11.1 5.6 1.9 3.2 9.5 18.9 18.9 9.5 3.2
136 168 182 213 230 245 259 271 286 306 326 351 392
58 76 83 101 110 118 126 133 141 152 163 177 200
D-1319
Vol %
36 0.5 63
133
TABLE I-9. API Sample 83-05 (Catalytic Reformed Naphtha)

Wt% Aliphatic C4 C5-C6 C7-C8 C9-C10 C11-C12 C13-C16 C17+ 0.01 11.1 14.9 1.2 Wt% Aromatic 3.0 46.9 22.6 -
TABLE I-10. API Sample 81-15 (Catalytic Cracked Clarified Oil)

Wt% Aliphatic C4 C5-C6 C7-C8 C9-C10 C11-C12 C13-C16 C17+ (C20+) 8.0 58.3 Wt% Aromatic
134
Table I-11. Hydrodesulfurized Kerosene (API 81-07)
Detailed hydrocarbon data not available, thus the table with C# distribution and BP range cannot be generated
GC/MS ID >nC3-</=nC4 >nC4-</=nC5 >nC5-</=nC6 >nC6-</=nC7 >nC7-</=nC8 >nC8-</=nC9 >nC9-</=nC10 >nC10-</=nC11 >nC11-</=nC12 0.8 0.13 0.1
wt % Aliphatic C#(GC/MS) BP Range wt % Toluene wt % Et.benzene wt % Xylenes wt % nC6 wt % nC7 wt % nC9 wt % Naphthalene wt % Pyrene(C16)
wt % Aliphatic BP Range
wt % Aromatic wt % BP Range Benzene
wt % PNAs
C4s
C5s
C6s
C7s
C8s
C9s
C10s
C11s
C12s
HC Type/Summed
0.0
0.0
0.0
0.0 0.0002 ~0.9
HC Type/MS
71
29
HC Type/D1319
78
22
135
136
TABLE I-11. Continued
D-86 DISTILLATION DATA: degC nC9 nC11 BP Range 0 0 ~~~C# (78 vol%) vol % Aliphatic BP Range (22 vol%) vol % Aromatic BP Range
degF
nC12
IBP 5% 10% 20% 30% 40% 50% 60% 70% 80% 90% 95% EP(99.5%) nC13 >nC8-</=nC9 >nC9-</=nC10 >nC10-</=nC11 >nC11-</=nC12 >nC12-</=nC13 >nC13-</=nC14 >nC14-</=nC15 3.9 19.5 31.2 19.5 3.5 1.1 5.5 8.8 5.5 1 nC14 nC15
310 382 395 409 420 429 436 443 452 464 480 493 533
154 194 202 209 215 221 224 228 233 240 249 256 278
D-1319
Vol %
77.3 0.5 22.2
TABLE I-12. API Sample 81-07 (Hydrodesulfurized Kerosine)

Wt% Aliphatic C4 C5-C6 C7-C8 C9-C10 C11-C12 C13-C16 C17+ 23.4 54.2 6.6 14.4 Wt% Aromatic
137

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Total Petroleum Hydrocarbon Criteria Working Group Series

Total Petroleum Hydrocarbon Criteria Working Group Series

Figure 1. Risk Assessment for Petroleum Contaminated Media

Figure 2. Toxicity Assessment Methods

b. Appropriate Data for Development of an RfD/RfC

d. Uncertainty and Modifying Factors

Carbon Range a Critical Effect Hepatoxicity, Nephrotoxicity 5.0 18.4 0.4

Aromatic Oral RfD (mg/kg/day) Neurotoxicity

Aromatic Inhalation RfC (mg/m3)

Aliphatic Oral RfD (mg/kg/day)

Aliphatic Inhalation RfC (mg/m3)

C>8 - C10 C>10 - C12 C>12 - C16 NA Nephrotoxicity 2.0 NA

Hepatic and hematological changes Hepatic (foreign body reaction) granuloma

C>16- C21 C>21 - C35

III. EVALUATION OF AROMATIC FRACTIONS

Table 2. RfDs/RfCs Associated with Compounds in the Equivalent Carbon Ranges

Equivalent Carbon Ranges

Compounds w/Toxicity Data

0.2 0.1 0.2 2.0

0.4 1.0 1.0 NA

C>8 - C10 C>10 - C12 C>12 - C16 0.04

C9 Aromatics Naphthalenes/ Methylnaphthalenes

C>16 - C21 C>21 - C35 NA NA NA

IV. EVALUATION OF ALIPHATIC FRACTIONS

Table 3. Commercial Hexane Test Rule Studies

0, 900, 3000, and 9000 ppm 6 hours/day, 5 days/week for 13 weeks

0, 900, 3000, and 9000 ppm 6 hours/day, 5 days/week for 13 weeks

Duffy et al. (1991)

0, 900, 3000, and 9000 ppm 6 hours/day, 5 days/week for 2 years

Soiefer et al. (1991)

0, 900, 3000, and 9000 ppm 6 hours/day, 5 days/week for 2 years

Kelly et al. (1994)

0, 900, 3000, and 9000 ppm 6 hours/day, 5 days/week for 2 years

Daughtrey et al. (1994)

0, 900, 3000, and 9000 ppm 6 hours/day, 5 days/week for 2 years

Kelly et al. (1994)

0, 900, 3000, and 9000 ppm 6 hours/day, 5 days/week for 2 years

Liver tumors in female mice at 9000 ppm

Daughtrey et al. (1994)

0, 900, 3000, and 9000 ppm 6 hours/day, 5 days/week for 2 generations

Daughtrey et al. (1994)

0, 900, 3000, and 9000 ppm 6 hours/day on days 6-15 of gestation

Keenan et al. (1991)

0, 900, 3000, and 9000 ppm 6 hours/day on days 6-15 of gestation

Keenan et al. (1991)

Table 4. Composition-Weighted RfDs for C5 - C8 and C6 - C8 Fractions

Light catalytically cracked naphtha (API 81-03)

Light catalytically cracked naphtha (API 81-04)

Sweetened naphtha (API 81-08)

Light catalytically reformed naphtha (API 81-04)

Full range catalytically reformed naphtha (API 81-05)

Heavy catalytically cracked naphtha (API 83-18)

Heavy catalytically cracked naphtha (API 84-02)

Figure 3. Distribution of Studies in the C9 to C16 Carbon Range

Effect No Significant Adverse Effects None

NOAEL 900 ppm (5226 mg/m3) 900 ppm (5226 mg/m3)

RfC (mg/m3) 0.9

No Significant Adverse Effects None

900 ppm (5485 mg/m3) 900 ppm (5485 mg/m3)

1.0 No Significant Adverse Effects 1000 mg/m3

RfC = 0.9 mg/m3

Table 6: Oral Studies and RfDs for C9 - C16 Fraction