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Antimicrobial Activity of Aroma Chemicals

and Essential Oils 1


J.A. MORRIS, A. KHETTRY, and E.W. SEITZ, Research and Development Department,
International Flavors and Fragrances, Inc., Union Beach, New Jersey 07735

ABSTRACT fragrance materials vs. these bacteriostats could be made.


Determination of the minimum inhibitory concen-
trations (MIC) of 212 common soap fragrance raw MATERIALS AND METHODS
materials demonstrated that the paper disc-petri
plate technique does not reflect the relative anti- Test Organisms
microbial activity of these materials. Commonly used The following microorganisms were used: Staphylococ-
soap bacteriostats were shown to be 100 to 1000 cus aureus, Escherichia coli and Candida albicans, obtained
times more effective than the most active fragrance from Monmouth Medical Center, Long Branch, New Jersey;
materials. Of 521 fragrance materials initially a lipophilic diphtheroid, probably a Corynebacterium
screened by the petri plate method, 44% were species, obtained from the Department of Dermatology,
inhibitory against one of the three test organisms, and University of Pennsylvania. These organisms, with the
15% were effective against all three (Staphylococcus exception of E. Coli, are normal inhabitants of human skin
aureus, Escherichia coli, Candida albicans). Of a (9,10), and two classes of organisms, Gram positive and
selected number (212) of these positive materials, diphtheroid, have been implicated in the production of
subsequently screened against a lipophilic diphtheroid body odor (9). Stock cultures were maintained on agar
organism (Corynebacterium sp.), 64 materials (30%) slants composed of tryptone (Difco), 0.3%; yeast extract
were positive against all four test organisms. However, (Difco), 0.3%; glucose, 09 K2HPO 4, 0.1%, and agar, 1%
only nine materials (4%) had a MIC as low as 50 ppm (referred to as TGY agar). Slants for the diphtheroid strain
compared to the common soap bacteriostat TCC | were supplemented with 0.5% Tween 80 (ICI America,
which had a MIC of 0.08 ppm (vs. S. aureus). In Inc.).
hand-degerming tests, no reduction of bacterial
counts was obtained with a soap containing the most Test Media
active fragrance materials. These results demonstrate
F o r the petri plate screening work, TGY agar was used
that creation of a practical antimicrobial soap
and the pH was adjusted to 5.5 with H3PO 4 to approxi-
fragrance does not appear to be possible 9
mate the pH of human skin. The medium was sterilized (15
min, 121 C) and dispensed by a New Brunswick Scientific,
INTRODUCTION Model AS-3, agar sterilizer. The diphtheroid medium was
The antimicrobial activity of aromatic substances has again supplemented with 0.5% Tween 80. For liquid
been known for more than fifty years 9 Macht and Kunkel cultures TGY medium was used without the agar, and the
(1) in 1920 and Dyche-Teague (2) in 1924 described the pH was not adjusted (pH 7.0) in order to optimize growth
antimicrobial effects of volatile oils or their vapors. More of all the test organisms. In this case the diphtheroid
recently MaruzzeUa, et al. (3-8), reported rather extensive medium was supplemented with 0.1% Tween 80.
surveys of the antirnicrobial action of many essential oils
and perfumery chemicals in both the vapor phase and by Fragrance Materials
direct contact of the liquids. Most of this previous work Fragrance raw materials, including essential oils, abso-
employed variations of the filter paper disc-petri plate lutes, essences, extracts and synthetic chemicals were
method, similar to that used for evaluating antibiotics. obtained from production inventory lots. Samples were
However, a serious disadvantage of this method is that the prepared as 10% (w/v) solutions in 95% (v/v) ethanol
data are, at best, semiquantitative and often not compar- except where solubility limitations occurred. In these cases
able from one laboratory to another because organisms are 5% solutions were prepared or alternative solvents were
exposed to unknown concentrations of test chemicals. used (e.g., diethyl phthalate, benzyl alcohol, benzyl
Based on these past results, considerable interest has benzoate) 9 In all experiments appropriate solvent controls
developed concerning the possibility of using fragrance raw were included 9 Antimicrobial chemicals included as controls
materials to create perfumes for soaps and other personal t . ,
were: TCC | (3,4,4-trlchlorocarbanilide), Monsanto; Irga-
.

care products which would have bacteriostatic activity in san DP 300


| t 9 t 9
( 2 , 4 , 4 - t n c h l o r o - 2 - h y d r o x y d l p h e n y l ether),
addition to their primary function of providing a pleasant 9 9
Clba-Gelgy; and hexachlorophene [ 2,2 -methylene his
t .

odor. This study was intended to provide a quantitative (3,4,6-trichlorophenol)]. These chemicals were also pre-
basis for critical evaluation o f this concept 9 In this paper we pared in alcoholic solutions at various concentrations
report the results of a three-stage study of the bacteriostatic depending on the final concentrations required in the
action of fragrance raw materials. First, a large number of growth media 9
materials were screened in a standardized petri plate pro-
cedure. Second, materials identified as having significant Patti Plate-Paper Disc Procedure
bacteriostatic activity were tested in liquid cultures to
The growth medium (TGY agar) was maintained at 42 C
determine their minimum inhibitory concentrations (MIC).
during dispensing into plastic petri plates (8.5 cm diameter;
Third, a soap fragrance was created from the most active
Falcon Brand). The medium was seeded at 3.3% (v/v)
fragrance materials and tested in handwashing panel studies
with 24-hour shake cultures of the appropriate organisms
to determine its degerming efficacy. In all phases of this
and dispensed at 8 ml per plate. Paper discs (0.95cm diam-
study, known antimicrobial chemicals were included as
eter; Schleicher & SchueU) were soaked with 20/A of the
controls so that direct comparison of the efficacy of the
10% test solutions, and the discs were immediately applied
1preliminary report presented at the 1976 national meeting of to the center of the solidified seeded agar, one disc per
the American Chemical Society, Miami Beach, FL, September 1978. plate. All materials were run in duplicate. It was found that

595 (SD&C 65)


5 9 6 (SD&C 6 6 ) JOURNAL OF THE AMERICAN OIL CHEMISTS' SOCIETY VOL. 56

TABLE I TABLE II
Composition of Soap Fragrance for Antimicrobial Activity of Fragrance
Hand-Degerming Test Materials in Petri Plate Cultures

% in Number Number %
Ingredient Fragrance MIC (ppm) Organism tested positive Positive
Aldehyde, C-I 1, undecylenic 0.4 50 (S. aureus) a S. aureus (Gram +) 521 162 31.1
Amyl cinnamyl alcohol 18.0 50 (S. aureus) E. coli (Gram -) 521 136 26.1
Anethol 10.0 100 (C. albicans) C. albicans (Yeast) 521 149 28.6
p-tert-Butyl-m-cresol 0.5 50 (S. aureus) Diphtheroid 212 101 47.6
Citronellol coeur 18.0 100 (C. albicans)
Cuminyl acetate 5.0 100 (C. albicans) All four organisms 212 a 64 30.2
Isobutyl quinoline 0.1 50 (C. albicans) Three organisms 212 89 42.0
Isocitral 0.5 100 (Diphtheroid) Two organisms 212 118 55.7
Lemma (Schiff base) 5.0 100 (S. aureus) One organism 212 144 67.9
Methyl isoeugenol 5.0 100 (Diphtheroicl) Negative 212 68 32.1
Methyl lavender ketone 7.0 100 (S. aureus)
Mousse Abs. Ess. 4.0 100 (S. aureus) aOnly those materials tested against all four organisms are in-
Musk ketone 5.0 100 (Diphtheroicl) cluded in this total.
Musk xylol 5.0 50 (Diphtheroid)
Ocmea (Schiff base) 8.0 50 (S. aureus)
Patchouli oil, dark 3.0 100 (S. aureus) i n c u b a t e d at 37 C for 18 t o 24 hr, sufficient t i m e to o b t a i n
Rosalva 2.0 100 (C. albicans) significant t u r b i d i t y . Because p o o r g r o w t h was o b t a i n e d
Sandalwood 0.5 50 (S. aureus) w i t h t h e d i p h t h e r o i d in c u l t u r e tubes, this o r g a n i s m was
Veramoss 3.0 100 (Diphtheroid)
t e s t e d in 50 m l E r l e n m e y e r flask c u l t u r e s ( 1 0 m l / f l a s k )
alndicates organism for which the MIC was determined. In some i n c u b a t e d at 37 C o n a r o t a r y s h a k e r at 2 5 0 r p m .
cases this MIC applies to more than one organism.
Handwashing Panels
20/~1 was u n i f o r m l y a b s o r b e d by t h e discs w i t h o u t exces- T h e d e g e r m i n g efficacy o f a bar soap c o n t a i n i n g 2% o f a
sive wetting. T o t a l m a t e r i a l o n t h e disc was 2 m g for m o s t fragrance c r e a t e d f r o m raw m a t e r i a l s f o u n d t o have anti-
samples. F o r t h o s e materials available in less t h a n n e a t m i c r o b i a l activity in vitro ( T a b l e I) was t e s t e d in a m o d i f i e d
f o r m , c o r r e s p o n d i n g l y less m a t e r i a l was placed o n t h e disc Cade p r o c e d u r e (11). C o m p a r i s o n was m a d e w i t h a c o n t r o l
(e.g., a 50% a b s o l u t e a c t u a l l y h a d o n l y 1 m g o n t h e disc, soap c o n t a i n i n g 2% TCC~. T h e s t a n d a r d Cade p r o c e d u r e was
t h e r e m a i n d e r b e i n g t h e s o l v e n t used for t h a t material). s h o r t e n e d to i n c l u d e o n l y a t h r e e - d a y " c o n d i t i o n i n g "
C o n t r o l discs w i t h 20/21 o f 95% e t h a n o l h a d n o effect o n p e r i o d prior to use o f t h e test soaps i n s t e a d o f t h e usual
g r o w t h o f a n y o f t h e organisms. Plates were i n c u b a t e d in an seven to t e n day period. A s t a n d a r d " s u p e r - f a t t e d " soap
u p r i g h t p o s i t i o n at 37 C for 18 t o 24 hr, d u r i n g w h i c h t i m e was used as t h e base soap in these panels. Base a n d test
a n a d e q u a t e m i c r o b i a l l a w n developed. c o u n t s were t a k e n b y t h e " f i f t h b a s i n " t e c h n i q u e described
b y Kooistra, et al. (1 2). A l i q u o t s a n d serial d i l u t i o n s o f t h e
Liquid Culture Procedure f i f t h basin were p l a t e d in T G Y agar c o n t a i n i n g 1% T w e e n
80. Panels c o n s i s t e d o f t e n people, w i t h e q u a l n u m b e r s of
M i n i m u m i n h i b i t o r y c o n c e n t r a t i o n s were d e t e r m i n e d in
males a n d females, selected f r o m r e s e a r c h personnel.
T G Y b r o t h c u l t u r e s ( 1 8 x 150 m m t u b e s ; 10.0 m l / t u b e ) .
Selected c h e m i c a l s were t e s t e d initially at 0.1, 1.0 a n d 10
p p m t o d e t e r m i n e t h e a p p r o x i m a t e range o f activity. W h e n RESU LTS AND DISCUSSION
n o positive m a t e r i a l s were f o u n d , these were r e p e a t e d at 10,
Petri Plate S c r e e n
I 0 0 , a n d 1000 p p m . Materials selected b y t h e p e t r i plate
screen were t h e n t e s t e d at 100, 500, a n d 1000 p p m . T h o s e A t o t a l o f 521 fragrance raw m a t e r i a l s were t e s t e d in this
f o u n d positive at 100 p p m were r e p e a t e d at 10, 50 a n d 100 p r e l i m i n a r y screen. Initially, chemicals were t e s t e d against S.
p p m . AU samples were r u n i n d u p l i c a t e a n d t h a t c o n c e n t r a - aureus, E. coli, a n d C. albicans. S u b s e q u e n t l y , 2 1 2 materials
t i o n at w h i c h n o g r o w t h o c c u r r e d in e i t h e r t u b e was t a k e n as were r e t e s t e d against a d i p h t h e r o i d o r g a n i s m isolated f r o m
t h e m i n i m u m inhibitory concentration. T h e effect b e i n g h u m a n skin, since this g r o u p o f organisms has b e e n impli-
m e a s u r e d was b a c t e r i o s t a s i s , since n o a t t e m p t was m a d e to c a t e d in p r o d u c t i o n o f body o d o r (9). S u m m a r i e s o f t h e
s u b s e q u e n t l y plate o u t t h e i n h i b i t e d c u l t u r e s to d e t e r m i n e p e t r i plate screen are p r e s e n t e d in T a b l e II. These d a t a
if t h e i n o c u l u m h a d b e e n killed. represent only qualitative comparison showing that the
Test materials were a d d e d to t h e t u b e s as 10% ( w / v ) G r a m positive S. aureus was o n l y slightly m o r e sensitive to
solutions. In c o n t r a s t t o t h e p e t r i plate screen, s o l u t i o n s of these fragrance materials t h a n t h e o t h e r t y p e s o f o r g a n i s m s
t h o s e m a t e r i a l s available in less t h a n n e a t f o r m were ad-
( T a b l e II). A p p r o x i m a t e l y 30% o f t h e materials were
j u s t e d a c z o r d i n g l y so t h a t t h e final s o l u t i o n s c o n t a i n e d 10% effective against a n y o n e o f t h e first t h r e e organisms. T h e
o f t h e test materials. Some materials i n c o m p l e t e l y soluble
p r o p o r t i o n of positives for t h e d i p h t h e r o i d (48%) reflects
at 10% were p r e p a r e d at 5% a n d d o u b l e aliquots were t h e biased s e l e c t i o n process for t h e c h e m i c a l s t e s t e d against
a d d e d to t h e c u l t u r e tubes. C o n t r o l a n t i m i c r o b i a l s were this organism. These m a t e r i a l s were selected f r o m t h o s e
t e s t e d in t h e range 0 . 0 2 t o 200 p p m d e p e n d i n g o n t h e f o u n d t o be positive against o n e or m o r e o f t h e first t h r e e
c h e m i c a l a n d t h e organism. F o r e x a m p l e , TCC | was t e s t e d organisms, so t h a t a h i g h e r p e r c e n t a g e o f " h i t s " was to be
at 0.02 t o 0.10 p p m against S. aureus, b u t u p to 2 0 0 p p m e x p e c t e d . F r o m t h e s e c o n d p a r t o f T a b l e II, w h i c h i n c l u d e s
against E. coli. A p p r o x i m a t e effective ranges for t h e con- d a t a o n l y f r o m t h e r e s t r i c t e d list o f materials ( d i p h t h e r o i d
trois h a d b e e n d e t e r m i n e d in p r e l i m i n a r y e x p e r i m e n t s . All test g r o u p ) we can see t h a t ca. 30% were positive for all
concentrations of ethanol added with test materials or four organisms, a n d 68% were positive for at least o n e o f
c o n t r o l s were t e s t e d a l o n e to c o r r e c t for a n y solvent t h e f o u r test organisms. Again, these data are based on a list
effects. o f materials " w e i g h t e d " t o w a r d positives. If we c o n s i d e r
T u b e s were i n o c u l a t e d w i t h 5 0 / a l o f a 1 : 10 d i l u t i o n in t h e t o t a l list o f materials ( 5 2 1 ) in t h e p e t r i plate screen a n d
sterile 0 . 8 5 % saline o f a 2 4 - h o u r s h a k e c u l t u r e ( T G Y b r o t h ) . t h e original t h r e e organisms (S. aureus, E. coli, C. albicans),
A m i n i m u m o f 3 0 0 , 0 0 0 viable o r g a n i s m s were a d d e d to we find t h a t o n l y 15% were effective against all t h r e e of
e a c h t u b e . T u b e s were m i x e d o n a V o r t e x m i x e r a n d t h e s e organisms, a n d o n l y 4 4 % were positive for at least o n e
MAY, 1979 MORRIS ET AL.: B A C T E R I O S T A S I S O F A R O M A CHEMICALS (SD&C67) 5 9 7
T A B L E Ill

F r a g r a n c e Materials w i t h A n t i m i c r o b i a l A c t i v i t y

S'taph. aureus E. r C. albicans Diphtheroid


Zone Zone Zone Zone
diam. MIC diam. MIC diam. MIC diam. MIC
Chemical a mm ppm mm ppm mm ppm mm ppm

Acetanilide 0b NT c 13 NT 11 NT NT NT
N - A c e t yl m e t h y l a n t h r a n i l a t e 0 > 1000 0 > 1000 0 > 1000 0 >1000
A l d e h y d e , C-8 0 500 12 500 12 500 12P d 1000
A l d e h y d e , C-9 0 NT 0 NT 13 NT NT NT
A l d e h y d e , C- 11 U n d e c y l e n i c 0 50 0 > 1000 22 50 17 500
A l d e h y d e , C-16 0 NT 15 NT 0 NT NT NT
A l d e h y d e , C- 18 0 > 1000 15 1000 20 500 15 1000
Allyl a m y l g l y c o l a t e 0 NT 0 NT 11P NT NT NT
Amaryllide 14 > 1000 13 1000 19 1000 17 1000
Ambrarome Absolute 13 NT 0 NT 0 NT NT NT
Amyt cinnamic aldehyde coeur 0 > 1000 0 > 1000 0 > 1000 0 500
A m y l cinnamyl alcohol 0 50 0 >1000 0 500 12 500
A m y l salicylate 0 > 1000 0 > 1000 0 > 1000 0 500
A m y r i s Oil 13 NT 0 NT 0 NT NT NT
A n e t h o l , USP 0 500 0 500 0 100 0 500
Anisyl acetate 0 > 1000 13 > 1000 12P 1000 0 1000
Armoise Essence 0 1000 0 >1000 0 1000 0 >1000
A r r a s a l d e h y d e , 50% 20 500 14 1000 17 500 26 500
Aubepine 0 > 1000 18 > 1000 11 1000 0 1000
A u r a l v a ( S c h i f f base) 11 NT 12 NT 12 NT NT NT
Balsam C o p a i b a , USP 0 >1000 0 >1000 0 >1000 0 >1000
B a l s a m Peru Oil 18 >1000 15 >1000 11 >1000 14 500
Basil Oil 0 500 0 500 0 500 0 1000
Bay Oil 18 500 13 1000 20 500 14 1000
Benzaldehyde 0 1000 0 1000 0 1000 0 >1000
Benzoic Acid 21 1000 28 1000 21 1000 28 1000
Benzoin coeur 18 > 1000 16 1000 10 > 1000 12 1000
Benzophenone 0 NT 0 NT 12 NP NT NT
Benzyl acetate 0 > 1000 0 > 1000 0 1000 0 1000
Benzyl alcohol 0 > 1000 12 P > 1000 0 > 1000 0 500
Benzyl benzoate 0 > 1000 0 > 1000 0 > 1000 0 500
Benzyl propionate 0 >1000 0 1000 0 500 0 1000
B e n z y l salicylate 0 >1000 0 >1000 0 >1000 0 1000
Bergamot MPF 0 1000 0 >1000 0 500 0 1000
Beta g a m m a h e x e n y l f o r m a t e 13 NT 17 NT 0 NT NT NT
Beta n a p h t h y l a n t h r a n i l a t e 16 1000 20 1000 22 500 15 1000
Beta p i n e n e c o e u r 0 500 0 >1000 0 1000 0 >1000
Birch T a r r e c t i f i e d 14 500 14 > 1000 17 1000 14 500
Boise de Rose filtered 0 >1000 17 1000 0 500 0 1000
Borneol 0 1000 0 1000 0 500 12 500
C a m p h e n e 46 0 1000 0 > 1000 0 1000 0 >1000
C a m p h o r Oil W h i t e 12 500 *e > 1000 0 500 12 >1000
C a r a w a y Oil 0 5 O0 0 > 1000 0 500 0 500
C a r d a m o m Oil, G u a t e m a l a 0 > 1000 0 > 1000 0 500 0 500
Carvone, dextro 10 1000 11 1000 0 500 0 1000
C a r v o n e , laevo 0 > 1000 11 1000 11 500 0 1000
Cashmeran 0 500 0 > 1000 0 > 1000 14 500
C a s t o r e u m Abs. 50% 11 NT 0 NT 0 NT NT NT
C e d a r l e a f Oil 0 1000 0 1000 0 500 0 >1000
Cedarwood, White 0 1000 0 >1000 0 >1000 0 500
Cedrone S 12 NT 0 NT 0 NT NT NT
Cedrus Atlantica coeur 11 500 0 >1000 0 >1000 11 1000
Celery seed oil 0 NT 0 NT 12 NT NT NT
C h a m o m i l e oil 0 1000 0 > 1000 0 500 0 >1000
Cinnamalva 0 > 1000 19 1000 13 500 0 1000
Cinnamic Alcohol 14 >1000 19 >1000 27 500 16 1000
C i n n a m o n leaf oil, C e y l o n 18 500 17 1000 14 500 12 500
Ciste, colorless 12 NT 0 NT 0 NT NT NT
Citral, d i m e t h y l a c e t a l 0 500 0 >1000 33 500 14 500
Citral, r e f i n e d 15 500 14 500 46 500 16 500
C i t r o n a m a ( S c h i f f base) 0 NT 0 NT 11 NT NT NT
Citronella, F o r m o s a , J a v a 11 NT 0 NT 17 NT NT NT
Citronellal 0 500 0 > 1000 12 5 O0 0 500
Citronellol coeur 12 1000 10 1000 48 I O0 t8 500
Citronellyl acetate 0 > 1000 0 > 1000 0 > 1000 0 500
Citronellyl e t h y l ethel" 11 NT 0 NT 0 NT NT NT
Citronellyi isobutyrate 0 > 1000 0 > 1000 0 > 1000 0 >1000
Citrus oil, distilled 0 1000 0 > 1000 0 500 0 >1000
Clove l e a f oil 14 500 19 1000 19 500 15 500
Clove b u d oil 16 500 16 1000 18 500 14 S00
Cecal 15 500 14 > 1000 11 1000 15 S00
Coniferan 0 1000 0 > 1000 0 > 1000 0 SO0
C o r i a n d e r oil 0 1000 11 1000 0 500 0 1000
C o r n m i n t oil 10 NT 0 NT 0 NT NT NT
Coronal, beta 12 NT 0 NT 0 NT NT NT
C o r t e x A l d e h y d e , 50% 17 1000 21 500 16 > 1000 16 500
Coumarin 13 > 1000 15 > 1000 16 1000 12P 1000
C u m i n oil 0 500 0 1000 0 500 0 1000
Cuminyl alcohol 15P 1000 14 500 23 500 16 1000
598 (SD&C68) JOURNAL OF THE AMERICAN OIL CHEMISTS' SOCIETY VOL. 56

T A B L E III ( c o n t d . )

Fragrance Materials with Antimicrobial Activity

Staph. aureus E. coli C. albicans Diphtheroid


Zone Zone Zone Zone
diam. MIC diam. MIC diam. MIC diam. MIC
Chemical a mm ppm mm ppm mm ppm mm ppm

Cuminyl acetate 0 >1000 * >1000 0 100 0 500


Cyclamal extra 0 NT 0 NT 23P NT NT NT
2-Cyclohexyl eyelohexanone 0 NT 0 NT 10 NT NT NT
C y c l o s i a base 14 >1000 16 >1000 14 >1000 13 1000
Cymene coeur 0 1000 0 >1000 0 500 0 >1000
C y p r e s s oil, F r e n c h 0 1000 0 >1000 0 1000 0 >1000
Decalaetone 15 >1000 0 >1000 12 1000 14 1000
n-Decanol 20 NT 0 NT 13 NT NT NT
Dibenzyl ether 0 >1000 0 >1000 0 >1000 0 500
D i b u t y l sulfide, 1 0 % 0 NT 16P NT 0 NT NT NT
Diethyl phthalate 0 >1000 0 >1000 14P 500 12P 1000
Dihydro cuminyl alcohol 14 1000 16 500 23 500 19 500
Dimethyl anthranilate 0 1000 0 1000 14P 500 0 500
Dimethyl benzyl carbinol 0 >1000 16P >1000 10 1000 0 >1000
Dimethyl octanol 0 NT 0 NT 12 NT NT NT
Dimethyl phenyl acetaldehyde 14 NT 0 NT 0 NT NT NT
Dimethyl phenyl ethyl carbinol 0 >1000 14 1000 21 1000 15 1000
Dimethyl phthalate 0 NT 11 NT 12 NT NT NT
Dimethyl sulfide 0 1000 0 >1000 0 500 0 1000
Diphenyl oxide 0 >1000 0 >1000 0 500 0 500
Dipropylene glycol 0 >1000 0 >1000 0 >1000 0 >1000
p-Ethyl acetophenone 0 NT 0 NT 13P NT 0 NT
Ethyl benzaldehyde 0 500 11 500 11 500 0 500
Ethyl benzoate 0 1000 0 1000 0 500 0 1000
Ethyl- 3-hydroxy-3-phenyl propionate 0 NT 13 NT 12 NT NT NT
Ethyl linalool 18 500 14P 1000 11 500 20 500
Ethyl methacrylate 0 >1000 0 >1000 0 >1000 0 >1000
Ethyl phenyl glyeidate 0 NT 0 NT 11 NT NT NT
Ethyl vanillin 14 >1000 18 1000 19 1000 15 1000
E u c a l y p t u s oil, 7 0 - 7 5 % 0 >1000 0 >1000 0 1000 0 >1000
Eugenol, USP 16 500 21 500 22 500 15 500
F e n n e l oil, S w e e t 0 500 0 500 0 500 0 500
Fir b a l s a m , A b s . 12 500 15P 1000 0 >1000 0 1000
Fraistone 0 NT 0 NT 26P NT NT NT
Furfural 12 >1000 11 >1000 0 1000 0 >1000
Galaxolide 0 1000 0 >1000 0 >1000 0 500
Galbanum coeur 0 tO00 0 >1000 0 >1000 0 500
Geraniol coeur 14 1000 16 500 38 500 20 500
G e r a n i o l e n e , light 13 NT 0 NT lIP NT NT NT
Geranium, African 12 NT 0 NT 19 NT NT NT
Geranoxy acetaldehyde, 50% 0 NT 0 NT 13 NT NT NT
Geranyl benzoate 0 >1000 0 >1000 0 >1000 0 >1000
Geranyl methyl tiglate 0 NT 0 NT 14 NT NT NT
Geranyl propionate 0 >1000 0 >1000 0 >I000 0 500
G r a p e f r u i t oil 0 500 0 >1000 0 500 0 >1000
Guaiene 0 500 0 >1000 0 >I000 12 500
G u a i c w o o d oil 13 500 0 >1000 0 >1000 0 500
Hay Abs. 12 NT 0 NT 0 NT NT NT
Hedione 0 >1000 0 >1000 0 1000 12 500
Helional 0 500 14 1000 14 500 16 500
Heliotropyl acetate 0 NT 13P NT 12P NT NT NT
n-Hexanol 0 NT 11 NT 0 NT NT NT
Hexyl cinnamic aldehyde 0 >1000 0 >1000 0 >1000 0 500
Hydratropal acetone 18 NT 0 NT 21 NT NT NT
Hydratropic alcohol, white 12 >1000 14 1000 13 1000 11 >1000
Hydroxy citronellal dimethyl acetal 0 NT 0 NT 11 NT NT NT
Hydroxy citronellal 20 >1000 16 >1000 13 1000 14 >1000
H y s s o p oil 11 NT 0 NT 0 NT NT NT
Indisan 12 500 0 >1000 0 >1000 10 500
Indole 18 1000 23 500 17 500 22 500
Iralia 11 NT 0 NT 0 NT NT NT
Iritone 10 NT 0 NT 0 NT NT NT
Isoamyl pentenoate 12 NT 0 NT 0 NT NT NT
Iso b e t a g a m m a h e x e n y l a c e t a t e 18 >1000 18 1000 12 1000 12 >1000
lsoborneol 0 >1000 0 500 0 500 0 1000
Isobutyl benzyl carbinol 11 500 12 1000 15 500 14 500
Isobutyl cinnamate 11 NT 0 NT 0 NT NT NT
Isobutyl furyl propionate 0 NT 0 NT 10 NT NT NT
Isobutyl quinoline 29 100 13 >1000 26P 50 17 100
Isocitral 19 500 11 1000 20 500 18 100
Isoeugenol 23 500 18 500 14 500 14 500
Isoeugenyl benzoate 0 >1000 0 >1000 0 >1000 0 >1000
Isojasmone 12 NT 0 NT 12 NT NT NT
lsomuguet aldehyde, 50% 25 NT 0 NT 11 NT NT NT
isopropyl cyclohexyl propanol 15 NT 0 NT 18 NT NT NT
Isopropyl quinoline 29 500 13 500 16 100 20 500
Isopulegol M Extra 10 1000 12 1000 0 1000 0 1000
Jasmonate 0 NT 0 NT 11 NT NT NT
MAY, 1979 MORRIS ET AL.: BACTERIOSTASIS OF AROMA CHEMICALS (SD&C 69) 599
T A B L E III ( c o n t d . )

Fragrance Materials with Antimicrobial Activity

Staph. aureus E. c e l l C. albicans Diphtheroid


Zone Zone Zone Zone
diam. MIC diam. MIC diam. MIC diam. MIC
Chemical a mm ppm mm ppm mm ppm mm ppm

J a s m o n e , cis 19 1000 13 1000 21 500 15 500


Jasmutone 11 500 12 >1000 13 500 14 500
Labdanax 0 NT 13 NT 12 NT NT NT
Labdanol 11 NT 0 NT 0 NT NT NT
L a b d a n u m resin, A b s . 14 500 0 >1000 0 >1000 12 1000
Lactone HB 17 1000 12 500 16 500 14 500
Lauryl alcohol 0 1000 0 >1000 0 >1000 10 500
Laurine extra 17 >1000 12 >1000 13 1000 0 1000
L a v a n d i n abrialis 0 NT 12 NT 0 NT NT NT
Lavandulol 11 1000 12 1000 13 500 0 500
L a v e n d e r A b s . , Camilli 0 >1000 0 >1000 0 1000 0 1000
Lemma (Schiff base) 18 100 11 >1000 12 500 12 500
L e m o n oil, Cal. 0 500 0 >1000 0 500 0 >1000
Lemongrass 13 500 14 500 15 500 17 500
L i m e oil, w a s h e d 0 1000 0 >1000 0 500 0 1000
Limonene 0 >1000 0 >1000 0 500 0 >1000
Linalool oxide 0 NT 10 NT 0 NT NT NT
Linalool 0 1000 18 1000 0 500 0 500
Linalyl acetate 0 >1000 0 >1000 0 >1000 0 500
Linalyl cinnamate 0 >1000 0 >1000 0 >1000 0 >1000
L o v a g e oil 14 NT 0 NT 14 NT NT NT
L R G N o . 182 ( E t h o x y c y c l o h e x a n o n e ) 11 NT 0 NT 0 NT NT NT
L R G N o . 1 1 8 1 (Nee-, i s o m e n t h o n e s ) 13 NT 12 NT 0 NT NT NT
Lyral 16 1000 13 >1000 12 1000 14 1000
Mace, w h o l e e x t r a c t 12 >1000 0 >1000 0 >1000 0 500
Maltol 0 >1000 16 >1000 16 1000 14 >1000
M a n d a r i n oil 0 1000 0 >I000 0 500 0 >1000
Menthol, USP 11 500 11 500 10 500 0 500
Methaltyl pentenoate 14 NT 0 NT 0 NT NT NT
p-Methoxy hydrotropic aldehyde 18 1000 26 1000 15 500 16 500
Methyl anthranilate 12 >1000 14 >1000 16 1000 0 1000
p-Methoxy phenoxy acetaldehyde 18 1000 20 >1000 16 1000 19 500
Methyl benzoate 0 >1000 0 >1000 0 1000 0 1000
p-Methoxy phenoxy acetaldehyde dimethyl acetal 0 >1000 0 >1000 11 1000 0 1000
Methyl cinnamate 11 1000 12 >1000 13 500 0 500
s-Methyl cinnamic aldehyde 20 1000 17 500 18 500 0 500
Methyl cyclocitrone 10 NT 0 NT 0 NT NT NT
Methyl eugenol 12 1000 11 1000 16 500 12 500
Methyl heptenol 12 NT 13 NT 0 NT NT NT
Methyl hexyl acetaldehyde 34 >1000 22 >1000 15 1000 16 500
Methyl isoeugenol 0 >1000 0 >1000 0 >1000 0 100
Methyl lavender ketone 13 100 13 1000 18 500 15 500
Methyl/3-naphthyl ketone 0 NT 0 NT 15 NT NT NT
Methyl octin carbonate 0 >1000 0 >1000 0 500 0 500
Methyl octyl acetaldehyde 18 NT 0 NT 0 NT NT NT
2 - M e t h y l - 2 - p e n t e n o i c acid 19P 1000 35 1000 18 1000 25 1000
Methyl-p-cresol 0 >1000 0 1000 0 500 0 >1000
Methyl phenyl ethyl alcohol 13 >1000 19 >1000 16 1000 0 >1000
Methyl p-toluate 0 1000 0 1000 0 500 0 1000
Miel B l a n c , D e l a i r e 11 NT 17P NT 0 NT NT NT
Mousse Abs., Verte Maroc 21 50 15 1000 18 500 18 500
Muguet aldehyde 40 NT 0 NT 14 NT NT NT
Muscagene 12 NT 0 NT 0 NT NT NT
Musk ambrette 0 >1000 0 >1000 0 >1000 13P 500
Musk ketone 0 >1000 0 >1000 0 >1000 12P 100
Musk xylol 0 >I000 0 >1000 0 >1000 11P 50
Myrac aldehyde 17 NT 0 NT 33 NT NT NT
Myrrh coeur 13 1000 0 >1000 0 >1000 12 1000,
Myrtenal 13 NT 10 NT 0 NT NT NT
M y r t l e oil, C h a r a b o t 12 NT 11 NT 0 NT NT NT
Naame (Schiff base) 12 NT 0 NT 0 NT NT NT
Nareisse k e t o n e 0 NT 0 NT 13 NT NT NT
Narcitol 14 NT 18 NT 13 NT NT NT
Nerol 14 500 13 1000 41 500 12 500
Nerolidol 15 NT 0 NT 0 NT NT NT
Neroly blanc 13 NT 10 NT 0 NT NT NT
Nortonkalactone 29 >1000 13 >1000 17 >1000 0 >1000
N u t m e g oil 0 500 0 >1000 0 500 0 1000
O a k m o s s essence 22 50 0 >1000 12 1000 14 500
Ocimene 22 500 14 1000 13 500 16 500
O c m e a ( S c h i f f base) 13 50 10 500 13 50 13 100
O p o p o n a x oil 10 NT 0 NT 0 NT NT NT
O r a n g e oil, Fla. 0 500 20 >1000 0 500 22 500
Orange, terpeneless Abs. 11 NT 0 NT 0 NT NT NT
Orange terpenes 12 NT 16 NT 0 NT NT NT
Orenyle 14 NT 0 NT 0 NT NT NT
O r i g a n u m oil, S p a n i s h 33 500 24 500 13 500 16 500
Orivone 11 NT 0 NT 39 NT NT NT
600 (SD&C 70) JOURNAL OF THE AMERICAN OIL CHEMISTS' SOCIETY VOL. 56

T A B L E lIl (contd.)

F r a g r a n c e Materials w i t h A n t i m i c r o b i a l A c t i v i t y

Staph. a u r e u s E. coli C. albicans Diphtheroid


Zone Zone Zone Zone
diam. MIC diam. MIC diam. MIC diam. MIC
Chemical a mm ppm mm ppm mm ppm mm ppm

Oxyphenylon 0 >1000 12 >1000 13 >1000 12 >1000


Para-cresol 20 >1000 29 1000 16 1000 22 1000
Para-cresyl a c e t a t e c o e u r 0 >1000 0 >1000 12P 1000 0 >1000
Para-isopropyl hydratropic aldehyde 13 NT 0 NT 12 NT NT NT
Para-methyl benzyl acetate 11 NT 0 NT 0 NT NT NT
Para-methyl dimethyl benzyl carbinol 14 >1000 14 1000 13 1000 14 1000
Para-tert b u t y l c y c l o h e x a n o n e 11 1000 0 >1000 27P 500 0 500
Para-tert b u t y l - m e t a - c r e s o l 85 50 26 500 85P 50 22 100
Para-tolyl alcohol 12 >1000 16 >1000 17 >1000 12 >1000
P a t c h o u l i oil, d a r k 12 100 0 >1000 0 >1000 13 500
P a t c h o u l i oil, light 0 500 0 >1000 0 >1000 13 500
Peach a l d e h y d e c o e u r 12 NT 0 NT 14 NT NT NT
P e p p e r oil, b l a c k 0 1000 0 >1000 0 >1000 0 >1000
Peppermint 0 NT 0 NT 10 NT NT NT
Persicol ("/-undecalact o n e ) 0 NT 0 NT 11 NT NT NT
Petinerol 0 1000 12 1000 14 500 0 1000
P e t i t g r a i n S.A. 0 >1000 0 >1000 0 500 0 1000
Petitgrain terpenes 14 500 11 >1000 10 500 12 >1000
Phellandrene 18 NT 18 NT 0 NT NT NT
Phenoxy ethyl propionate 0 >1000 0 >1000 12 500 0 1000
Phenyl acetaldehyde 21 100 40 1000 33 500 18 100
Phenyl ethyl acetate 0 NT 25P NT 0 NT NT NT
Phenyl ethyl alcohol 0 >t000 16 >1000 0 >1000 0 >1000
Phenyl ethyl cinnamate 0 >1000 0 >1000 0 >1000 0 >1000
Phenyl ethyl phenyl acetate 13 >1000 0 >1000 0 >1000 0 500
Phenyl propyl alcohol 12 >1000 1.6 >1000 16 1000 14 1000
Phenyl propyl aldehyde 12 500 27 500 14 1000 0 500
Phixia 16 >1000 15 >1000 12 1000 14 1000
Piconia 13 NT 0 NT 0 NT NT NT
P i m e n t o b e r r y oil 16 500 1"7 1000 18 500 14 500
Pine n e e d l e oil, Siberian 0 500 0 >1000 0 1000 0 1000
Pine oil 12 1000 14 >1000 11 500 12P 1000
P r o p y l e n e glycol, USP 0 >1000 0 >1000 0 >1000 0 >1000
Rosacene 14 1000 12 1000 29 500 0 1000
Rosalva 16 1000 0 >1000 16 100 14 500
R o s e m a r y oil, Span. Tunis. 0 1000 0 >1000 0 1000 0 >1000
Rosetone NT >1000 0 >1000 0 >1000 0 >1000
Rosin g u m 12 NT 0 NT 0 NT NT NT
Sandalwood 11 50 0 >1000 * >1000 11 500
Santalol 13 500 0 >1000 0 >1000 13 500
Sauge sclaree Abs. 12 500 0 >1000 0 >1000 11 500
S e s q u i t e r p e n e s PC 13 500 12P >1000 0 >1000 12 500
S p e a r m i n t oil 0 1000 0 >1000 0 500 0 1000
S p r u c e oil 0 500 0 >1000 0 1000 0 1000
St. G u a i o l 13 500 0 >1000 0 >1000 12 500
St. J o h n ' s b r e a d c o n c . 10% 0 NT 15P NT 0 NT NT NT
S t y r a x alva essence 21 NT 0 NT 0 NT NT NT
S t y r a x clarified, e x t r a 11 >1000 0 >1000 0 500 11 >1000
Surfleurs H a y 11 NT 0 NT 0 NT NT NT
Tabac absolute 10 NT 0 NT 0 NT NT NT
T a n g e r i n e oil, Fla. 0 500 0 >1000 0 500 0 >1000
Terpineol 12 1000 19 1000 20P 1000 0 1000
T h u j a oil 0 500 0 >1000 0 500 0 1000
Thyme, white 25 500 27 500 14 500 16 500
Tiglyl p i p e r i d i d e 0 NT 13 NT l 1 NT NT NT
T o l u resin abs. 5 0 % 17 >1000 17 >1000 11 1000 13 1000
Tonalid 0 >1000 0 >1000 0 >1000 0 500
T o n k a abs. 11P >1000 14 >1000 14P >1000 0 >1000
Trans-decahydro beta naphthol 15 1000 15 1000 22P 500 16 500
Trans-3-pentenyl acetone 0 NT 18 NT 0 NT NT NT
T r e e m o s s abs., F r e n c h , 5 0 % 18 100 0 >1000 0 1000 14 500
Trimethyl cyclohexanol 0 NT 11 NT 0 NT NT NT
Trimethyl cyclohexenone 0 > 1000 12 1000 10 1000 0 >1000
Trimethyl cyelohexenol 15 NT 13 NT 0 NT NT NT
U n d e c y l e n i c acid 21 NT 0 NT 13 NT NT NT
Vanillin 16 >1000 22 >1000 19 1000 12P >1000
V e l t o l plus 0 >1000 20 >1000 12 >1000 0 >1000
Veramoss 0 500 0 >1000 12 500 12 100
Verdural extra 13 NT 13 NT 0 NT NT NT
V i o l e t t o n e A, colorless 16 NT 12 NT 0 NT NT NT
W i n t e r g r e e n oil 0 >1000 * >1000 0 500 0 1000
Yaracetal 0 NT 13 NT 0 NT NT NT
Ylang concrete 0 >1000 0 >1000 0 1000 0 1000
Zingerone 11 >1000 12 >1000 11 >1000 0 >1000

CONTROLS
Hexachlorophene 13 0.10 12 50 0 50 0 100
MAY, 1979 MORRIS ET AL.: BACTERIOSTASIS OF AROMA CHEMICALS (SD&C 71)601

TABLE III (contd.)


Fragrance Materials with Antimicrobial Activity
Staph. aureus E. cell C. albicans Diphtheroid
Zone Zone Zone Zone
diam. MIC diam. MIC diam. MIC diam. MIC
Chemicala mm ppm mm ppm mm ppm mm ppm
Trichlorocarbanilide (TCC| 11 0.08 0 >200 0 100 0 50
Trichlorohydroxy diphenyl ether (Irgasan DP300 | 25 0.04 22 0.06 12 6.0 0 50

aThe majority of materials in this table are identified by trivial or trade names commonly used in the fragrance industry. Chemical names for
all materials (where applicable) can be obtained from fragrance handbooks.
bo indicates no inhibition of growth was detected.
CNT indicates chemical was not tested against this organism.
dp indicates the zone of inhibition around the paper disc was only partially cleared.
e. Indicates a general reduction of growth was evident, but no measureable zone of inhibition was observed.

organism. least one organism in the petri plate assay. For com-
While these petri plate data are poor indicators of the pleteness, all other materials tested, but for which no
relative antimicrobial activity of the test materials, the antimicrobial activity was found, are listed in Table IV.
method is a practical way of screening large numbers of Twenty-three materials were found to be effective at 50 or
materials. The zones of inhibition ranged from 10 mm 100 ppm (none at 10 ppm) against at least one organism.
diameter (just barely larger than the paper disc) to a clear- These include a wide variety of structural types: phenolics,
ing of the entire plate (85 mm), but these zone sizes do not terpenoids, heterocyclics, esters, alcohols, etc. Five of these
accurately reflect relative antimicrobial effectiveness. For materials are essential oils or absolutes including two of the
example, TCC | produced a cleared zone of 11 mm against oakmoss type which are complex mixtures of phenolics,
S. aureus but was subsequently shown to be much more depsides, resinoids, and other compounds. E. coli (Gram
bacteriostatic than any of the fragrance materials. The size negative) was least senstivie in the liquid cultures, with C.
of the cleared zone in this method is dependent on the albicans(yeast) and the diphtheroid being somewhat more
solubility and rate of diffusion of the sample in the aqueous sensitive than S. aureus (Gram positive). This is somewhat
medium. The very low solubility of TCC| responsible for surprising since previous work had indicated that the Gram
its apparently poor bacteriostatic activity in this type of positive organisms are usually more sensitive than other
assay. Some materials produced no definite cleared zones, types to bacteriostatic action of fragrance materials (3,5).
but obvious reduction of growth over the entire plate This result is also in contrast to the findings of our own
compared to control plates indicated either rapid diffusion petri plate screen in which S. aureus was most sensitive
of the material or an antimicrobial effect of the vapor. An (Table II). This again emphasizes that qualitative petri
attempt to minimize vapor effects and evaporative loss of plate screening methods and quantitative minimum inhibi-
volatile materials by using a double agar layer technique tory concentration methods are not necessarily comparable.
proved to be unworkable for this large number of samples. T h e results for antimicrobial activity against the diphthe-
reid organism may have been affected by the inclusion of
Minimum Inhibitory Concentration
Tween 80 in the growth medium. This surfactant may have
The petri plate screen identified 309 materials with increased the solubility of some of the fragrance materials
significant antimicrobial activity against at least one of the or aided in their penetration of the bacterial cells wails and
test organisms. Because of the large numbers of tubes membranes. This question can only be answered by retest-
involved in determining a minimum inhibitory concentra- ing the other organisms in the presence of Tween 80.
tion (minimum 24 tubes per sample in our procedure), the The results for all three antimicrobial compounds
number o f materials to be tested was reduced to 212. This (TCC | Irgasan DP 300 | and hexachlorophene) are in close
list of materials (the same materials as those tested against agreement with accepted industry values (~<0.1 ppm)
the diphtheroid organism in the petri plate assay)included against S. aureus. However, E. coli, C. albicans and the
those that showed relatively strong antimicrobial activity in diphtheroid were all somewhat resistant to these anti-
the petri plate screen or were suspected of having signifi- microbials with the exception of Irgasan DP 300 | against
cant antimicrobial activity because of structural considera- E. eoli. No MIC for TCC | vs E. cell was obtained. At the
tions (e.g., phenolics). highest level tested (200 ppm), growth still occurred, and
The original range of concentrations to be tested (0.1 to testing of higher concentrations was not attempted since
10 ppm) was selected to be 100-fold higher than the the practical use limit of T C C ~ had already been far
approximate MIC for TCC | (0.1 ppm). When no positives exceeded. The apparent resistance of the diphtheroid may
were found among 40 of the antimicrobial fragrance have been due to a neutralizing effect of the Tween 80 in
materials producing large zones of inhibition, even at 100 the growth medium. Use of Tween 80-containing-medium
ppm, further preliminary experiments were conducted on for the other test organisms increased the apparent MIC of
12 selected materials to determine the appropriate range of these antimicrobials in each case. No correlation between
concentrations. Since most of the materials tested were type of organisms or chemical structure of test materials
insoluble o r v e r y poorly soluble above 1000 ppm (0.1%), and bacteriostatic activity was evident from these data.
this was selected as the maximum concentration to be Some materials were effective at relatively low concentra-
tested even though many of the test materials were ineffec- tion against one organism and negative against one or more
tive at this concentration. The final levels tested were 100, of the other organisms (e.g., amyl cinnamyl alcohol, 50
500 and 1000 ppm, with 10 and 50 ppm tested for all those ppm for S. aureus; 1000 ppm for E. cell). Furthermore,
found positive at 100 ppm. This broad range was necessary chemical with related structures were not always equally
to encompass the wide variety of materials tested. effective against the same organisms (e.g., amyl cinnamyl
Petri plate and MIC results are summarized in Table III alcohol, 50 ppm for S. aureus; amyl cinnamic aldehyde,
which includes all materials found to be positive against at 1000 ppm for the same organism). Several compounds
602 (SD&C72) JOURNAL OF THE AMERICAN OIL CHEMISTS' SOCIETY VOL. 56

TABLE IV

Fragrance Materials Showing No Antimicrobial


Activity in Petri Plate Screen a

Abalyn Dodecyl nitrile Mentha citrat a


Abitol Elemi oil Menthanyl acetate
Acetate, C-q Estragon oil Menthone
Acetophenone Menthyl pentenoate
Ethyl acetate
Agrumea (Schiff base) Ethyl acetoacetate Methyl a c e t o p h e n o n e
Alcohol, C-12 Ethyl amyl ketone Methyl chavicol
Aldehyde, C- I 0 Ethyl p e n t e n o a t e Methyl diphenyl ether
Aldehyde, C-I 2, Laurie Methyl h e p t e n o n e
Allyl caproate Ethyl hutanol
Methyl-n-hexyl ether
Allyl cyclohexyl propionate Ethyl butyl ketone
Methyl hexyl ketone
Allyl ionone Ethyl butyrate
Methyl i o n a n t h e m e
Ambrain ex gum labdanum Ethyl geranate
Methyl ionone, gamma
Amyl acetate Ethyl isovalerate Methyl isohexyl carbinyl acetate
Amyl vinyl carbinyl acetate Farnesol Methyl nonyl acetaldehyde
Amyl vinyl carbinol l.arnesyl acetate 3-Methyl pentanol
Aprol 100 l'leuramone Moskene
Astratone Floralozone Mugyl acetone
Badiane Oil, l:ringhian Flouve oil Musk 36A
Benzyl isoeugenol Fructone Myrcenyl acetate
Benzyl phenyl acetate Galbanol Neoindisan
Bergamal Gamma terpinene coeur Nerolin
Besabolene Gelsone Neryl acetate
Beta g a m m a Hexenyl acetate Geranyl acetate Octea c y m b a r u m
Beta gamma Hexenol Geranyl acetone Oenanthic ether
Borneol Geranyl phenyl acetate Olibanol
n-Butyl pentenoate Geranyl tiglate Olibanum Olearome
Butyl benzoate Glycolierral Orange, bitter
Butyl methacrylate Grisalva Para cresyl caprylate
Butyl undecylenate Grisavan Para cresyl isobutyrate
Cabreuva oil Hay oil Parsley seed oil
Carbitol Pennyroyal
Hay oil, High Alps
Carrot oil Phenyl acetaldehyde, dimethyl acetal
Helycrisum oil
Caryophyllene Phenyl ethyl chloride
Herbac
Caryophyllene acetate Phenyl ethyl isobutyrate
Hercolyn D
Cassie Essence Abs. Phenyl ethyl salicylate
Hexylene glycol
Castor Oil Picol formate
cis-3-Hexenyl salicylate
Cedrenyl acetate Hexyl pentenoate Pinocarvyl acetate
Cedramber n-Hexyl isopentenoate Proflora
Celestolide Hexyl methacrylate Pseudo linalyl acetate coeur
Citralva Raldeine Omega
Hexyl salicylate
Citrindol Reseda body
Hyacinth body
Citroflex No. 2 Rhodinol
Hydratropic aldehyde, dimethyl acetal
Citron, CI Chauvet Rhodinol residue
Indotene
Citronellyl crotonate Rhodinyl formate
Isoborneol
Citronellyl formate Rose oxide
Isobornyl acetate
Citronellyl propionate Shimus Oil
Isobutyl isobutyrate
Citroviol lsobutyl p e n t e n o a t e Sinpine
Civet, Artificial lsobutyl phenyl acetate Styralyl acetate
Cognac oil Isobutyl salicylate Talia
Copaiba oil Isohexyl p e n t e n o a t e Terpinolene
Cubeb oil Isolongifolene Terpinyl acetate
Cyclacet Isomenthone Tetrahydro linalool
Cycloctal Isopropyl myristate T e t r a h y d r o muguol
Cyclohexyl ethyl acetate Isopropyl palmitate Tolpine
Cyclotene Jasmal Triethylene glycol
Cyclotropal Jessemal Trimethyl n o n a n o n e
4-Damascol Labdalva Trimethyl undecyl aldehyde
Decanyl acetate Lavandulyl acetate Trimofix R
Dimethyl malonate Leaf acetal Triplal
Dihydro floralate Lemon terpenes Turfurol acetate
Dihydro cyclacet Lilial Turpentine
Dih3~dro pseudo ionone Linalyl benzoate Vanilla concentrate (20%)
Dihydro terpinyl acetate Linalyl propionate Vanitrope
Diisobutyl ketone Linseed oil, abs. Vanoris
Dimethyl benzyl carbinyl acetate Lolitol Verdox coeur
Dimethyl benzyl carbinyl b u t y r a t e Longifolene Vertenex
Dimethyl octanyl acetate Lyrame (Schiff base) Vertofix coeur
Dimetol Maraniol Vetiveryl acetate
Dimyrcetol Marjolaine Essence Vionex acetate
Dipentene Mate Abs. Wormwood Abs., terpeneless
Dodecalide Melonal Ylang concrete

aThe majority of materials in this table are identified by trivial or trade names commonly used in the fragrance industry. Chemical names
for all materials (where applicable) can be obtained from fragrance handbooks.
MAY, 1979 MORRIS ET AL.: BACTERIOSTASIS O F AROMA CHEMICALS (SD&C73) 6 0 3
were inadvertantly tested more than once because of TABLE V
reliance on trade names in selection of test materials. For
Hand-Degerming Efficacy of Test Fragrance Soap
example, hydroxycitronellal was also tested under the
names cyclosia base, laurine, and phixia. Results were %
comparable for all four materials (see Table III), emphasiz- Soap Base count a Test c ount a Difference b
ing the reproducibility of these methods.
Experimental fragrance 3.03 x 106 3.90 x 106 + 19.2 c
Using the data accumulated in vitro, those materials
identified as having the strongest antimicrobial activity TCC | (Control) 2.23 x 106 3.66 x 105 - 72.0 d
were tested in hand-degerming experiments. The two soaps
aMean of 10 subjects, 5 male, 5 female.
tested as described in METHODS contained, respectively, 2%
bMean of % difference for each of ten subjects.
(w/w) of TCC@ (control) or 2% (w/w) of a fragrance whose
CNot statistically significant.
composition is given in Table I. This fragrance was created
dStatistically significant reduction at a 99.5% confidence level.
to maximize antimicrobia] efficacy within the limits of a
reasonably pleasant soap aroma. No skin-degerming was
achieved with the test soap (Table V), whereas the control ACKNOWI.EDGMENTS
soap (TCC | showed significant reduction of bacterial The authors express appreciation to Mrs. Mary Lou Lang (Mon-
counts. The failure to achieve the usual count reduction mout h Medical Center) for cultures of test organisms; to Dr. John
Labows (Monell Chemical Senses Center) and Dr. Ken McGinley
with TCC~, i.e. 90-99%, was probably due to the somewhat
(D e pa rt me nt of Dermatology, University of Pennsylvania), for the
shortened test period in this modified Cade procedure. diphtheroid culture; to l)r. Ronald Leight for the statistical analysis
Considerable individual variation was encountered, due, in of the hand-degerming panel data; and to Mrs. Gall Harris for
part, to insufficient "conditioning" with a blank soap to valuable technical assistance.
allow skin flora to "normalize" prior to the start of the
actual test period. Nevertheless, the reduction observed REFERENCES
with the control soap (TCC| was found to be statistically I. Macht, D.I., and W.M. Kunkel, Proc. Soc. Exp. Biol. Med.
18:68 (1920).
significant at a 99.5% confidence level, indicating that this 2. Dyehe-Teague, l".C., Perf. Es.sent. Oil Record 1:6 (1924).
modified procedure would have shown degerming if it had 3. Maruzzella, J.C., and P.A. Henry, J. Am.Pharnl. Assoc. 47(4):
occurred with the fragranced soap. 294 (1958).
4. Maruzzella, J.C.,and L. Liguori, Ibid. 4 7 : 2 5 0 (1958).
5. Maruzzella, J.C., and N.A. Sicurella, Ibid. Sei. Ed. 4 9 :6 9 2
CONCLUSIONS (1960).
6. Maruzzella, J.C., Am. Perfum, 77(1):67 (1962),
The purpose of this study was to determine if fragrance 7. Maruzzella, J.C., Am, Perfum Cosmet. 78 : 19 (1963).
raw materials could be demonstrated to have antimicrobial 8. Maruzzella, J.C., and N. Kirsch, Perf. Essent. Oil Record
5 4 ( 1 2 ) : 8 2 3 (1963).
activity comparable to well known bacteriostatic agents. It 9. Shehadeh, N.H., and A.M. Kligman, J. Invest. Dermatol.
is apparent from the data presented here that in terms of 4 0 ( I ) : 6 1 (1963).
bacteriostasis, the best fragrance material is 100 to 1000 10. Somerville, D.A., J. Med. Mierobiol. 6:215 (1973).
times less effective than common soap antimicrobials 11. Cade, A . R . , J . Soc. Cosmet. Chem. 2:281 (1951).
12. Kooistra, J.A., E.A. Bannan, and R.O. Carter, J. Soe. Cosmet.
against one of the major types of skin organism. Thus, the Chem. 17:343 (1966).
creation of a practical fragrance with significant antimicro-
bial activity appears highly unlikely. [Received August 18, 1978]