828

Anal. Chem. 1982, 5 4 ,

828-830

While it was initially expected that the CN emission would also be observed for COz,this was not the case; only NO lines were observed. An oxygen atom is removed from the COz molecule and forms the intermediate species NO, which is excited and emits its strongest line at 305 nm. Just as all hydrocarbons show only CN emission, oxygen-containing molecules (e.g., 02, COz) exhibit primarily the NO p emission. Table I shows data obtained for COz. In conclusion,the MTES technique can be extended to very low detection limits and affords a relatively simple, inexpensive method of determining low-level impurities in various plant gases. Its ultimate capabilities are as yet unexplored, but

further extension of this technique provides a method for detection of not only carbon and oxygen impurities in plant gases but metal impurities as well. Metal ions in a gaseous state are detected by reactions with Nz* (3).
LITERATURE CITED
(1) (2) (3) (4)

U.S. Patent 4 150951, 1979. Capelle, G. A.; Sutton, D. G. Appl. Phys. Lett. 1977, 30, 407. Phillips, L. F. Can. J . Chern. 1963, 47, 2060. Sutton, D. G.; Westberg, K. R.; Melzer, J. E. Anal. Chem. 1979, 57, 1399.

RECEMDfor review September 18,1981. Accepted December 24, 1981.

Quickly Dissolving Amylose Indicator in Cadmium Iodide-Linear Starch Colorimetric Reagent

Jack L. Lambert," Gary T. Fina, and Earline F. Dikeman
Department of Chemistry, Kansas State Universlty, Manhattan, Kansas 66506

The starch described here is pure potato amylose and is a light, dry powder that disperses and dissolves instantly in cool water. Kept in a stoppered "salt-shaker" type dispenser such as those used for meat tenderizers, the solid amylose can be kept indefinitely and added only as needed. As the concentration of starch is not critical to titrimetric end point determination, it is necessary only to ensure the minimum required to produce the blue complex with polyiodide anionic species. Potato amylose was selected for use in the cadmium iodide-linear starch colorimetric reagent because of its greater chain length and higher content in the raw starch granules compared to other common sources. This versatile reagent was first reported in 1949 (1)and has been studied (2)or used in analytical methods for the determination of selenium (3), fluoride (4,5),chloride (6),ammonia (7),bromide (8),residual iodine in rocks (lo), peroxide in edible fats (ll), chlorine (9), total iodine and iodate in seawater (12), sulfate (13), miscellaneous anions after exchange for iodate with mercury(I1) iodate (14), and iodine from a quaternary ammonium strong base resin-triiodide disinfectant (15-1 7)as well as to monitor iodine disinfectant in stored water in space vehicles in the NASA Skylab project (18). Cadmium iodide provides iodide ion in controlled excess by release from the moderately stable iodocadmium species present in solution 2CdIz
$

breakdown of starch. The absorptivity, e, of the reagent is 1.7 X lo4per equivalent of oxidizing agent (2). The response curve is rectilinear but exhibits a small positive intercept on the concentration axis of the polyiodide species (or the oxidant that produced the polyiodide), as noted in several studies (7, 10, 11, 19-21). Hanes (22)postulated, and Rundle et al. (23-25) later confirmed, the helical structure of the starch-polyiodide complex and presented proof that it is a compound rather than an adsorption phenomenon or a simple solution of iodine in starch. Originally, it was assumed that triiodide anion, 13-, produced the chromogen, as both iodine and iodide ion were known to be necessary to the production of color. Gilbert and Marriott (26) identified the polyiodide anion in the starch helix as 312.21-, or I . : Lambert and Zitomer (7)proposed a model wherein Is- anions enter the starch helix sequentially and absorbance is produced only after the second 13-enters to produce Is2-. This model would explain the small positive intercept of the response curve on the concentration axis. Watanabe and co-workers (27) later concluded that the polyiodide species is 13-, (&I<, or IU2-. Thompson and Hamori (28) still later proposed Ill3-.
EXPERIMENTAL SECTION The method of preparation described by Lambert and Rhoads (29), which combined the selective extraction of raw starch granules reported by Krishnaswamy and Sreenivasan (30) and the 1-pentanolprecipitation procedures preferred by Schoch (31), produced a good linear starch fraction with ordinary laboratory equipment. The method described here is an improvement on that method which produces a superior linear fraction that disperses and dissolved rapidly and completely in cool water with minimal stirring. Preparation of Pure Potato Amylose. Three and one-half liters of distilled or deionized water containing 3.5 g of sodium chloride was heated to 61-64 "C in a 4-L beaker. An 80-g sample of potato starch (Fisher Scientific Co., Fair Lawn, NJ, Catalog No. S-514 or the equivalent)-but not a so-called "soluble starch"-were mixed in 400 mL of water at room temperature with rapid stirring until a smooth slurry was obtained. The slurry was added with vigorous stirring to the warm water in the large beaker, taking care that the final temperature was between 57 and 60 "C. Temperature control is important because at 65 "C the starch suspension cooks to a gelatinous material that does not filter well. Conversely, at temperatures below 57 "C, the yield
0 1982 American Chemical Society

[CdI]+

+ [CdIJ-

F!

Cd2+

+ [CdI,I2-

Retrogradion (precipitation due to intermolecular hydrogen bonding) of starch is retarded in cadmium iodide solution, probably due to complexation of cadmium ion by the hydroxyl groups on the starch molecule. Toxicity of the cadmium retards the growth of microorganisms. The response of the reagent to oxidizing agents is nearly independent of pH between 7.0 and 0.5. At pH 7.5, a precipitate of cadmium hydroxide appears, and a t pH values lower than 0.5, dissolved oxygen slowly oxidizes the iodide. The absorbance produced at 614 nm is temperature dependent, so for precise measurements the temperature should be controlled. The reagent is essentially nonselective toward strong oxidants. The pH of the reagent prepared with pure potato amylose is approximately 6.0, which is within the 5.9-6.3 pH range recommended for prevention of hydrolytic
0003-2700/82/0354-0828$0 1.25/0

ANALYTICAL CHEMISTRY, VOL. 54, NO. 4, APRIL 1982

829

Table I. Determination of Visual Limit of Sensitivity of Linear Starch-Polyiotsde Complex by Method o f Known Addition with Iodide-Selective Electrode ppm of Isample, mL a stock solution 26.6 22.SC 20.9 19.oc before reduction 0.76 0.12 O.l!Z 0.10
0.0!3

after reduction 13.91 0.47

0.40

difference, ppm I, 13.15 0.35 0.28 0.25 0.23

color obsd

0.35 0.32

It. blue ft. blue f t . blue no color

a Measurements done in triplicate except for the stock solution. b Color observed after 5 min of development time; depth of solution 3 cm in a 1 3 X 100 mm test tube Milliliters of stock solution against a white background. diluted to 500 mL.

is reduced due to incomplete extraction of the linear fraction from the starch granules. After 2 h at 57-60 "C, the beaker with the starch suspension was covered and allowed to settle overnight (7 h minimum). A large capacity centrifuge ?withspeeds of 8000-10 000 rpm and a radius of at least 10 cm compacted the residual starch granules in 20-30 min for easy decantation. Filtration of the supernatant solution containing the extracted amylose was done in two steps: first through fast filter paper (such as Whatman No. 4 or No. 113 or No. 114 if a wet strengthened paper is preferred) with suction on a Buchner funnel, followed by filtration through a 5-mm mat of washed diatomaceous earth on fast or medium filter paper. As some water was retained by the starch granules that were removed by centrifugation, it was necessary to add water which contains 1g of sodium chloride/L to bring the total volume back to 3.9 L. The solution was heated to 75-80 "C to form a clear solution. The stirring was increased until cavitation occurred so that when 200 mL of 1-pentanol (Fisher Scientific Co., Fair Lawn, NJ, Catalog No. A-394 or the equivalent) was added, the solution was thoroughly saturatedl. A 1- to 1.5-cm layer of 1-pentanol covered the aqueous solution after stirring was stopped. The beaker was covered and tlhe contents were allowed to cool overnight (although periods up to 2 days do no harm to the product). As much of the supernatant liquid as possible was removed by slow siphoning, with care taken to recover the undissolved 1-pentanol for reuse. It was often necessary to dislodge some starch-pentanol complex from the water-pentanol interface with a stirring rod. The 1-pentanol precipitation was repeated twice, using 3.5 L of water containing 3.5 g of sodium chloride and 200 mL of 1-pentanol in each step as before. After removal of the supernatant liquid from the third precipitation, the remaining starch-pentanol slurry was stirred vigorously as an equal volume of methanol was added. Care was taken to produce a dense, filterable precipitate that had no lumps. The precipitate was filtered with suction through a fast filter paper on a Buchner funnel until a firm cake was obtained. The filter cake was transferred to a medium-size beaker and sufficient 1-pentanol was added to cover the starch. Stirring for 3 h with a magnetic stirring bar produced a fine dispersion of the precipitated amylose. After the solids were allowed to settle, the 1-pentanol was decanted and a volume of 1-pentanol equal to the previous volume was added. After the mixture was stirred gently for 6 more h and the solids were allowed to settle, the precipitated amylose was filtered with suction through fast filter paper on a Biichner funnel until the cake began to crack. A 200-mL portion of methanol was added with stirring to resuspend the amylose and the suction was continued. The moist cake was then dried at room temperature under vacuum. The typical yield was 3-4 g* The final treatments with 1-pentanol and methanol, followed by drying under vacuum, are crucial to the quality of the final product. Properly prepared, the product should smell faintly of 1-pentanol,if at all. Stored in a sealed bottle, it is stable for years. It dissolves rapidly and completely in cool water with little or no

stirring. All previous preparations of potato amylose dissolved completely but only after warming and stirring. No precipitate nor turbidity, either from growth of microorganisms or from retrogradation, was observed after periods exceeding a year in the cadmium iodide-linear starch colorimetric reagent containing this pure potato amylose, prepared with 11.0 g of cadmium iodide and 2.5 g of the amylose as previously described (2). The reagent is usable for periods exceeding 4 years after preparation even if a few flakes of precipitate appear. Sensitivity Studies on the Cadmium IodideLinear Starch Reagent. The detection limit of the cadmium iodide-linear starch reagent was specified as the concentration below which a properly prepared quaternary ammonium strong base resin-triiodide water disinfectant released iodine into distilled water, while at the same time maintaining its capability to serve as a demand-type bactericide and virucide. The use of handheld comparators for determining residual iodine disinfectant in the stored water aboard the Skylab space vehicles likewise prompted interest in the detection limit of the reagent. A stock solution of iodine was prepared by permitting elemental iodine to dissolve for 2 days in the dark in approximately 2 L of doubly distilled water. Doubly distilled water was used instead of chlorine demand-free water to avoid chloride ion and possible interhalogen species. A 99-mL sample of the stock solution was analyzed for iodide ion concentration with an iodide-selective electrode. One milliliter of iodide solution of known concentration was added and the iodide concentration again determined. A 300-mL sample of the solution was then passed through a 20-mesh zinc metal column and the last 99-mL portion analyzed again by the known addition method. The difference between the total reduced iodide Concentrationand the initial iodide concentration was the concentration of iodine in the stock solution. Similar measurements were made on several dilutions of the stock solution with doubly distilled water at or near the concentration corresponding to the previously estimated lower limit of visual detection of the amylose-polyiodide blue complex. The measurements were done in triplicate by the method of known addition, with the results shown in Table I. The iodine demand of the doubly distilled water resulted in concentrations of iodine lower than that calculakd from the dilutions alone. Independent determinations of the iodine demand at similar dilutions corresponded with the observed reductions in iodine concentration. The visual lower limit of sensitivity was found to be 0.25 ppm of iodine when viewed through 3 cm of solution in a 13 X 100 mm test tube against a white background. This compared favorably with previous observations that the blue color can be seen at 0.20 ppm of iodine with the reagent in larger quantities of solution (32), although a spectrophotometer could not distinguish the absorbance compared to a water blank in 1-cm cuvettes.

RESULTS AND DISCUSSION

Linear potato starch (amylose) fraction, prepared by an improved precipitation and dehydration procedure, is stable indefinitely in the solid state. It disperses and dissolves rapidly and completely in cool water and can be added as needed in place of unstable starch indicator solutions in iodimetric titrations. A sealed, "salt-shaker" type of dispenser would be convenient for this type of use. In cadmium iodide solution, where both the growth of microorganisms and retrogradation are suppressed, it provides a sensitive and reproducible colorimetric reagent that is stable on storage in a sealed bottle for years. While temperature dependent, its nonselective response to oxidants is essentially constant over the p H range 0.5-7.0. The lower visual limit for the detection of iodine is 0.25 ppm.

LITERATURE CITED
Arthur, P.; Moore, T E.; Lambert, J. L. J . Am. Chem. Soc., 1949, 77,3260. Lambert, J. L. Anal. Chem. 1951, 23,1247. Lambert, J. L.; Arthur, P.; Moore, T. E. Anal. Chem. 1951, 23,1101. Lambert, J. L. Anal. Chem. 1953, 25,271. Hinze, W. L.; Humphrey, R. E. Microchem. J . 1975, 20,246. Lambert, J. L.; Yasuda, S. K. Anal. Chem. 1955, 27,444. Zltomer, F.; Lambert, J. L. Anal. Chem. 1962, 3 4 , 1738. Zitomer, F.; Lambert, J. L. Anal. Chem. 1963, 35,1731. Lambert, J. L.; Olguin, J. Anat. Chem. 1969. 4 1 , 838.

830
(10) (11) (12) (13) (14) (15) (16) (17) (18) (19) (20) (21) (22) (23) (24)

Anal. Chem. 1982, 5 4 , 830-832

Crouch, W. H., Jr. Anal. Chem. 1982, 34,1698. Swoboda, P. A. T.; Lea, C. H. Chem. Ind. (London) 1958, 1090. Schnepfe, M. M. Anal. Chlm. Acta 1972. 58, 83. Hlnze, W. L.; Humphrey, R. E. Anal. Chem. 1973, 45. 814. Hlnze, W. L.; Humphrey, R. E. Anal. Chem. 1973, 45, 385. Taylor, S. L.; Flna, L. R.; Lambert, J. L. Appl. Mlcroblol. 1970, 20, 720. Lambert, J. L.; Flna. G. T.; Flna. L. R. Ind. Eng. Chem., Prod. Res. Dev. 1980. 79, 258. Hatch, G. L.; Lambert, J. L.; Flna, L. R. Ind. Eng. Chem., Prod. Res. Dev. 1980, 79, 259. Smith, C. A.; Spleth, H. H. Anal. Chem. 1973, 45, 422. Sugawara, K.; Koyama, T.; Terada, K. Bull. Chem. SOC.Jpn. 1955, 28,494. Novick. R. P. Blochem. J . 1982, 8 3 , 236. Lambert, J. L. Anal. Chem. 1951, 23, 1251. Hanes, C. S. New Phytol. 1937, 36,189. Rundle, R. E.; Baldwin, R. R. J . Am. Chem. SOC. 1943, 6 5 , 544. Rundie, R. E.; French, D. J . Am. Chem. Soc. 1943, 6 5 , 558. (25) Rundle, R. E.; Foster, J. F.; Ealdwln, R. R. J . Am. Chem. Soc. 1944, 66. 2116. (28) Gilbert, G. A.; Marriott, J. V. R. Trans. Faraday Soc. 1948, 4 4 , 84. (27) Watanabe, T.; Ogawa, K.; Ono, S. Bull Chem. Soc. Jpn. 1970, 43, 950. (28) Thompson, J. C.; Hamorl, E. J . Phys. Chem. 1971, 75, 272. (29) Lambert, J. L.; Rhoads, S. C. Anal. Chem. 1958, 28, 1629. (30) Krishnaswamy, K. 0.; Sreenlvasan, A. J . Blol. Chem. 1948, 778, 1253. (31) Schoch, T. J. "Advances In Carbohydrate Chemlstry"; Pigman, W. W., Wolfram, M. L., Eds.; Academic Press: New York, 1945; Vol. 1. p 259. (32) Dlkeman, E. A. M.S. Thesis, Kansas State University, 1972.

RECEIVED for review January 16,1981.

Accepted December 28,1981. This work was supported in part by National Science Foundation Grant CHE79-15217.

Indirect Determination of Cyanide by Single-Column Ion Chromatography

D.

L. DuVal, J. S. Fritz," and D. T. Gjerde'

USDOE,Iowa
State lJnivers/?y, Ames, Iowa 5001 1

Ames Laboratory

Many different methods for determining cyanide have been developed over the years. Most colorimetric methods are based on the Konig reaction of cyanogen halide with an aromatic amine and pyridine and are sensitive from about 0.01 to 2 ppm cyanide (I). Two common titrimetric methods are Liebig's and Volhard's methods (I). Both use the formation of a very stable silver cyanide complex as the basis for their methods and are used above 20 ppm cyanide. The sample must also be free from interferences. The chromatographic method described in this article was developed by modifying a pneumatoamperometric method developed by Beran and Bruckenstein (2) that used the reaction of cyanide with iodine listed below.

I2

+ HCN e H++ I- + ICN

K,, = 0.73

They describe the equilibrium as being well-known, pH dependent, and quantitative from a pH of 2 to 7. Since the final concentration of iodide is proportional to the initial concentration of cyanide, the new chromatographic method analyzes iodide using a single-column ion chromatograph and a conductivity meter to determine cyanide concentrations. Excess iodine in the solution is removed by adsorption onto a short precolumn containing unfunctionalized XAD-4 resin (Rohm and Haas). After a series of injections the precolumn is removed from the chromatograph and the iodine is desorbed by flushing with a nitric acid-acetone mixture. The precolumn is then placed back in the injection line of the chromatograph.

EXPERIMENTAL SECTION
Chemicals. All chemicals were of reagent grade quality from Fisher Chemical Co. and were used as received. A 0,100 M iodine solution in 95% ethanol was prepared. The container used to store the iodine solution was wrapped in aluminum foil to slow the decomposition by W and visible radiation. Cyanide samples for calibration and for testing the procedure under various conditions were prepared from a stock solution of potassium cyanide that was prepared fresh weekly and contained approximately 100 ppm of cyanide. The stock solution was made up in approximately 0.001 M sodium hydroxide. Cyanide samples of varying concentration were prepared by dilution of the stock solution just before analysis.

'Present address: Exxon Research and Engineering Co., P.O. Box 121, Linden, NJ 07036.
0003-2700/82/0354-0630$01.25/0

A 0.05 M acetate buffer solution with a pH of 4.75 was made with glacial acetic acid and 0.1 M sodium hydroxide. A 0.05 M nitric acid solution was prepared in acetone and the 100 ppm iodide stock solution used in these experiments was made from potassium iodide. Interferences were made from stock solutions of nitrate salts for the cations and sodium or potassium salts for the anions. Apparatus. A single-column ion chromatograph described by Gjerde et. al. (3) was used with a Model 213A Wescan conductivity detector and cell. The chromatograph was modified by adding a glass precolumn as described below. The precolumn was fitted with a female leur adapter on one end so that samples could be injected by hand directly into the precolumn using a syringe. The other end of the precolumn was connected to the injector valve, containing the sample loop, with a short length of Teflon tubing. This arrangement allows the operator to load the sample loop and remove the excess iodine from the sample at the same time. The precolumn should be glass so that the operator can tell when the precolumn needs to be flushed. When the resin in the precolumn at the end closest to the injector valve starts to turn yellow, the iodine needs to be flushed from the precolumn. The precolumn, 75 X 2 mm i.d., was packed with unfunctionalized Rohm and Haas XAD-4 resin, 100-150 mesh. The analytical column, 500 X 2 mm i.d., was packed with anion exchange resin prepared from 325 to 400 mesh Rohm and Haas XAD-1 (3). The resin had an anion exchange capacity of 0.025 mequiv/g. Other specific conditions were as follows: eluent pumping rate, 1.5 mL/min; sample loop size, 100 pL; detector full scale, 1 pmho; detector output, 10 mV; recorder full scale, 10 mV. Procedure. If necessary, dilute the sample so that the expected cyanide concentration will be in the range of 0.5-10 ppm. Pipet accurately 40 mL of sample into a 50-mL volumetric flask and add 1 mL of 5 x low2 M acetate buffer (pH 4.75). Mix well and then add 0.3 mL of 0.1 M iodine in 95% ethanol. A yellow color should remain after mixing, indicating that an excess of iodine has been added. The excess iodine is necessary for the reaction of cyanide and iodine to be quantitative. Dilute the solution to 50 mL, mix, and allow the solution to sit at least 5 min. Then slowly inject a 2-3 mL aliquot of the sample into the ion chromatograph through the XAD-4 precolumn. The slow injection time, approximately 5-10 s, through the precolumn allows the resin more time to adsorb the excess iodine. Separate the iodide ion, resulting from reaction of cyanide with iodine, chromatographically by elution with 2 X M sodium or potassium phthalate, pH 6.25. The retention time for iodide on the column
0 1982 American Chemical Soclety