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