188

Use of Some Essential Oils as Natural Preservatives for Butter

R.S. Farag a,', M.N. All b and S.H. Taha b
~ Department and bFood Science and Technology Department, Faculty of Agriculture,Cairo University,Giza-Egypt

Thyme and cumin e s s e n t i a l oils w e r e u s e d in the pre- materials with a stated purity of 99% by GLC was obtained
s e n t study in an a t t e m p t to prevent butter deteriora- from Dragoco c o m p a n y (Holzminden, West Germany).
tion during storage at room temperature. Butter oxida- The s t a n d a r d materials were: cyclic t e r p e n e s (a-pinene,
tion and lipolysis w e r e f o l l o w e d by m e a s u r i n g the acid, B-pinene, camphene, limonene, ~/-terpinene, terpinolene
p e r o x i d e and TBA values. Lipolytic activity and total and phellendrene); aliphatic h y d r o c a r b o n (myrecene);
microbial and lipolytic bacterial c o u n t s w e r e also a r o m a t i c h y d r o c a r b o n (p-cymene); sesquiterpene (ca-
m e a s u r e d . During butter storage, v e r y little change in ryophyllene); phenol ethers (eugenol, thymol and methyl
the p e r o x i d e and TBA v a l u e s w e r e found while a grad- chavicol); cyclic terpene ketones (carvone, dihydrocar-
ual i n c r e a s e in the acid value w a s noticed. The addi-
vone, and thujone); aromatic aldehydes (cumin alde-
tion o f cumin and t h y m e oils at 200 ppm to butter
hyde); aliphatic alcohols (linalool and geraniol); cyclic
c a u s e d v e r y little i n c r e a s e in the acid value. The data
terpene alcohols (t-carvol, ~-terpineol and borneol) and
for lipolytic bacterial counts w e r e in general agree-
terpene esters (linalyl acetate and terpinyl acetate).
m e n t with the acid values. Thyme and cumin e s s e n t i a l
oils s h o w e d a greater anti-hydrolytic e f f e c t and act as
Identification and determination of essential oil com-
superior p r e s e r v a t i v e s c o m p a r e d to BHT. position. The essential oils were analyzed by a GCV Pye
Unicam gas c h r o m a t o g r a p h equipped with dual flame
ionization detectors. The c h r o m a t o g r a p h was fitted with
a coiled glass column (1.5 m X 4 m m ) packed with Dia-
Autooxidation and lipolysis are responsible for off-flavors tomite C (100-120 mesh) and coated with 10% PEGA. The
in lipid-containing food products. Butter deteriorates by oven t e m p e r a t u r e was p r o g r a m m e d at 4~ from
oxidative rancidity from the reaction with atmospheric 60~ to 180~ and was held at 180~ for 15 rain. Detector
oxygen and hydrolytic reactions catalyzed by lipases from and injector t e m p e r a t u r e s were 220~ and 300~ respec-
food or from microorganisms (1). Antioxidants are wide- tively. Gas flow rates for N2, H2 and air were 30, 33 and 330
ly used in m a n y foods to prevent fat rancidity. It has been ml/min, respectively. Peak identification was p e r f o r m e d
found t h a t the addition of BHT and BHA at concentra- by comparing the relative retention times of each p e a k
tions ranging from 50-500 p p m in ghee or b u t t e r r e t a r d e d with those of known compounds. Also, the essential oils
the development of both f a t t y acids and peroxides during were mixed with their major c o m p o u n d s and injected
storage (2-5). A combination of propyl gallate (100 p p m ) into GLC to verify the p e a k identity. The relative retention
and dodecyl gallate (500 p p m ) was effective as an antiox- times for thymol and cumin aldehyde are given a value of
idant in butter, as was propyl gallate at 200 p p m (6,7). 1.00, depending on essential oil origin. The p e a k areas
There has been some discussion recently of the undesira- were m e a s u r e d by triangulation, and p e r c e n t a g e of each
ble use of synthetic antioxidants, since BHA had toxic
oil c o m p o n e n t was calculated as the ratio of the p e a k
and carcinogenic effects (8). Also, BHT caused changes in
area to the total c h r o m a t o g r a p h i c area. All samples were
r a t thyroids, stimulation of DNA synthesis and induction
analyzed in triplicate and the values agreed within 2%.
of enzymes (9). Consequently, there is a need for other
Mean values are presented in the text.
types of antioxidants. The present w o r k has been conduc-
ted to s t u d y the effect of some naturally occurring essen-
Oxidation systems. Butter packaged in sterilized glass
tial oils as preservatives for butter. bottles was thoroughly mixed with BHT (200 p p m ) ,
cumin (200 p p m ) and thyme (200 p p m ) oils a n d stored at
room t e m p e r a t u r e . Samples were removed periodically
MATERIALS AND METHODS and subjected to chemical and microbiological analyses.
Source of milk. Fresh cow's milk was obtained from the Chemical analyses. Hydrolytic and oxidative rancidity
E x p e r i m e n t a l Station Herd, Faculty of Agriculture, Cairo of b u t t e r were followed by determining the acid value,
University, Giza, Egypt. peroxide n u m b e r and thiobarbituric acid (TBA) value. To
Milk processing. Milk was s e p a r a t e d into c r e a m and get the acid value, a known weight of butter fat (5 g) was
skim milk using an Alfa-Laval s e p a r a t o r (Alfa-Laval, dissolved in a neutralized alcohol (50 ml) and titrated
Sweden). Cream was c h u r n e d to obtain butter. with KOH (0.1 N) (10). For the peroxide number, a known
Butylated hydroxy toluene (BHT) and essential oils. weight of b u t t e r fat (2 g) was dissolved in a m i x t u r e of
Crystalline BHT was obtained from Sigma Chemical Com- CH3COOH:CHCI3 (3:2, v/v), a n d s a t u r a t e d solution of KI
p a n y (St. Louis, MO). The fruits and leaves of cumin (1 ml) was then added. The liberated iodine was titrated
(Cuminum cyminum, L.) and t h y m e (Thymus vu/gar/us, with sodium thiosulfate solution (0.1. N) in the presence
L.) plants were collected from the P h a r m a c y Farm, Cairo of starch as an indicator (10). The TBA test was per-
University, Giza, Egypt. The plant materials, cut into small formed by adding H20 (8 ml), TBA solution (6 ml, 0.025
pieces (ca. 100 g), were placed in a flask (2 L) together mM) and trichloroacetic acid (3 ml) to the b u t t e r fat (0.5
with double distilled w a t e r (1.5 L). A s t e a m distillation g). After heating the m i x t u r e (20 min), the interfering
continuous extraction head was a t t a c h e d to the flask. materials were e x t r a c t e d three times with ether and dis-
After s t e a m distillation the oil was isolated and dried over carded. The aqueous p h a s e was completed with distilled
anhydrous sodium sulfate. w a t e r to a known volume (25 ml) and the absorbance of
Authentic volatile compounds. A set of 24 s t a n d a r d this solution was recorded at 532 n m (11). All chemical
determinations were conducted in triplicate and the re-
*To whom correspondence should be addressed. sults are presented as average values.

JAOCS, Vol. 68, no. 3 (March 1990)

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USE OF SOME ESSENTIAL OILS AS NATURALPRESERVATIVES FOR BUTTER
Determination of lipolytic activity. T h e lipolytic activ- (1). A n e x p e r i m e n t w a s c o n d u c t e d u s i n g BHT a t 200 p p m
ity o f b u t t e r w a s d e t e r m i n e d a n d is d e f i n e d as t h e n u m b e r a l o n g w i t h o t h e r e x p e r i m e n t s in o r d e r to c o m p a r e t h e
o f m o l e s o f free f a t t y a c i d s ( a s oleic a c i d ) n e u t r a l i z e d b y a n t i o x i d a t i v e a b i l i t y o f t h e e s s e n t i a l oils t o w a r d s b u t t e r
NaOH (12). r a n c i d i t y . T h e e x p e r i m e n t a l p e r i o d w a s t e r m i n a t e d (18
Microbiological analysis. T h e c o u n t s o f t o t a l b a c t e r i a days) when an objectionable odor was obviously noticed
a n d lipolytic b a c t e r i a w e r e c a r r i e d o u t w i t h m e l t e d b u t t e r with the control sample.
(1 ml) d i l u t e d w i t h a p p r o p r i a t e v o l u m e s o f s t e r i l i z e d sa- Table 1 s h o w s t h e p e r o x i d e a n d TBA v a l u e s for t h e
line s o l u t i o n a n d t h o r o u g h l y m i x e d w i t h a s u i t a b l e m e d i - s y s t e m s c o n s i s t i n g o f b u t t e r c o n t a i n i n g BHT, c u m i n a n d
um. T r e p t o n e - g l u c o s e - y e a s t - a g a r m e d i u m w a s u s e d for t h y m e oils. The r e s u l t s s h o w t h a t t h e c h a n g e s in t h e
t o t a l b a c t e r i a l c o u n t (13). T r e p t o n e - g l u c o s e - y e a s t - p e r o x i d e v a l u e s d u r i n g t h e first 15 d a y s w e r e v e r y low in
s t e r i l i z e d ghee m e d i u m w a s u s e d for lipolytic b a c t e r i a all cases. This m e a n s t h a t v e r y little a u t o o x i d a t i o n h a d
(14). The p l a t e s c o n t a i n i n g t h e m e d i a a n d b u t t e r w e r e t a k e n place. On t h e e i g h t e e n t h d a y o f t h e s t o r a g e , t h e
i n c u b a t e d a t 32~ for f o u r d a y s in all cases. peroxide values were remarkably increased, and the au-
Statistical analysis. T h e c a l c u l a t e d A-values for all t o o x i d a t i o n p r o c e s s c o m m m e n c e d . C o n c e r n i n g TBA
c h e m i c a l values, lipolytic a c t i v i t y a n d c o u n t s o f t o t a l b a c - values, n o obvious c h a n g e o c c u r r e d in a n y c a s e s t h r o u g h -
t e r i a a n d lipolytie b a c t e r i a w e r e s t a t i s t i c a l l y a n a l y z e d us- o u t t h e e n t i r e e x p e r i m e n t a l p e r i o d . This w o u l d i n d i c a t e
ing a s p l i t d e s i g n (15). t h a t t h e s e c o n d a r y p r o d u c t s , s u c h as a l d e h y d e s a n d ke-
t o n e s , h a d n o t y e t been f o r m e d .
It is w o r t h m e n t i o n i n g t h a t t h e s t a t i s t i c a l a n a l y s i s indi-
cated that there were significant differences between
RESULTS AND DISCUSSION
TBA v a l u e s a n d v a r i o u s s t o r a g e p e r i o d s for e a c h s y s t e m
C o n s u m e r s all o v e r t h e w o r l d a r e b e c o m i n g i n c r e a s i n g l y u n d e r s t u d y . However, t h e d i f f e r e n c e s in TBA v a l u e s oc-
conscious of the nutritional value and the safety of their c u r r e d w i t h i n t h e s e c o n d d e c i m a l p o i n t s . In o u r e x p e r i -
f o o d a n d its i n g r e d i e n t s . A t t h e s a m e time, t h e r e is a n ence these changes are meaningless.
i n c r e a s e d p r e f e r e n c e for n a t u r a l f o o d s a n d f o o d ingre- Table 2 a n d F i g u r e 1 s h o w t h e a c i d v a l u e s a n d A - a c i d
d i e n t s w h i c h a r e g e n e r a l l y b e l i e v e d to be safer, h e a l t h i e r v a l u e s for t h e s y s t e m s u n d e r s t u d y . W i t h o u t a d d i t i o n of
a n d less s u b j e c t to h a z a r d s t h a n f o o d s c o n t a i n i n g artifi- BHT o r e s s e n t i a l oils ( c o n t r o l ) , t h e a c i d v a l u e of b u t t e r
cial f o o d a d d i t i v e s . C u m i n a n d t h y m e oils w e r e d e m o n - g r a d u a l l y i n c r e a s e d w i t h t h e s t o r a g e time. T h e a c i d v a l u e s
s t r a t e d to h a v e a positive a n d effective i n h i b i t o r y effect on for b u t t e r c o n t a i n i n g BHT w e r e s i g n i f i c a n t l y l o w e r t h a n
s y n t h e t i c m e d i a c o n t a i n i n g b a c t e r i a , y e a s t a n d fungi (16). t h a t o f t h e control. The a d d i t i o n o f c u m i n a n d t h y m e oils
T h e s e m i c r o o r g a n i s m s a r e k n o w n to be r e s p o n s i b l e for a t 200 p p m t o b u t t e r p o s s e s s e d a m o r e s i g n i f i c a n t lower-
f o o d d e t e r i o r a t i o n . C o n s e q u e n t l y , t h e e s s e n t i a l oils u n d e r ing effect on t h e a c i d v a l u e s t h a n d i d BHT. T h e r e f o r e , b o t h
s t u d y w e r e a d d e d t o b u t t e r in a n a t t e m p t to s t u d y t h e i r s y s t e m s w e r e m o r e effective t h a n BHT in p r e v e n t i n g b u t -
effect on p r e v e n t i n g lipid o x i d a t i o n a n d hydrolysis. It h a s t e r hydrolysis. We also o b s e r v e d t h a t b u t t e r c o n t a i n i n g
been r e p o r t e d t h a t t h e 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 h y m e oil h a d a l o w e r A - a c i d v a l u e t h a n b u t t e r c o n t a i n i n g
t i o n (MIC) r e q u i r e d to p r e v e n t c e r t a i n t y p e s o f m i c r o o r - c u m i n oil. The effectiveness o f a d d e d m a t e r i a l s in p r o t e c t -
g a n i s m s w a s 200 p p m of t h e s e e s s e n t i a l oils (16). Hence, ing b u t t e r , b a s e d on t h e i r abilities to slow d o w n t h e a c i d
t h e e s s e n t i a l oils w e r e a d d e d to t h e n a t u r a l m e d i u m ( b u t - v a l u e rise, followed t h i s s e q u e n c e : t h y m e ~ c u m i n ~ BHT
t e r ) a t 200 p p m , w h i c h is s i m i l a r to MIC. T h e levels o f control.
e s s e n t i a l oils a d d e d t o b u t t e r a r e b e y o n d t h e c o n c e n t r a - Table 3 i l l u s t r a t e s t h e l i p a s e a c t i v i t y o f all s y s t e m s
tion o f a n t i o x i d a n t s a d d e d in i n d u s t r y to f o o d p r o d u c t s under study determined at the end of the experiment.

TABLE 1

Influence of BHT and Some E s s e n t i a l Oils on the Peroxide (PV) and TBA Values of Butter Stored at Room Temperature

Storage Butter + BHT Butter + Cumin oil Butter + Thyme oil

period Butter (control) (200 ppm) (200 ppm) (200 ppm)

(day) PV A-pv TBA ATBA PV 5-PV TBA ATBA PV A - P V TBA ATBA PV A - P V TBA ATBA

0 2.3 0.0 0.01 0.00 2.2 0.0 0.02 0.00 2.2 0.0 0.02 0.00 2.5 0.0 0.01 0.00
3 2.5 0.2 0.02 0.01 2.3 0.1 0.03 0.01 2.2 0.0 0.03 0.01 2.7 0.2 0.03 0.02
6 2.5 0.2 0.02 0.01 2.6 0.4 0.03 0.01 2.4 0.2 0.03 0.01 2.7 0.2 0.03 0.02
9 2.4 0.1 0.02 0.01 2.8 0.6 0.03 0.01 2.8 0.6 0.04 0.02 2.8 0.3 0.05 0.04
12 2.5 0.2 0.06 0.05 2.9 0.7 0.06 0.04 3.2 1.0 0.06 0.04 3.1 0.6 0.05 0.04
15 3.4 1.1 0.06 0.05 3.1 0.9 0.07 0.05 3.8 1.6 0.04 0.02 3.4 0.9 0.07 0.06
18 5.9 3.6 0.07 0.06 6.0 3.8 0.08 0.06 7.1 4.9 0.05 0.03 5.5 3.0 0.05 0.04

Peroxide value is expressed as milliequiv, peroxide/kg fat.

TBA value refers to the absorbance of the coloured product formed at 532 nm.
L.S.D. (0.05) values for A-P.V. between treatments and various experimental periods within each treatment were 0.081 and 0.192,
respectively.
L.S.D. (0.05) values for A-TBA-test between treatments and various experimental periods within each treatment were 0.0033 and 0.007,
respectively.

JAOCS, Vol. 68, no. 3 (March 1990)

190

R.S. FARAG ETAL.

TABLE 2

Effect of B H T a n d Some Essential Oils on t h e A c i d v a l u e s (A.V.) of B u t t e r S t o r e d at R o o m T e m p e r a t u r e

Storage Butter (control) Butter + BHT Butter + Cumin oil Butter + Thyme oil
period (200 ppm) (200 ppm) (200 ppm)

(day) A.V. A-A.V. A.V. A-AN. A.V. A-AN. A.V. A-AN.

0 0.5 0.0 0.7 0.0 0.5 0.0 0.9 0.0

3 1.4 0.9 1.5 0.8 1.3 0.8 1.4 0.5
6 2.5 2.0 2.4 1.7 1.6 1.1 1.5 0.6
9 4.0 3.5 3.2 2.5 2.2 1.7 2.0 1.1
12 4.9 4.4 3.9 3.2 3.0 2.5 3.0 2.1
15 6.3 5.8 4.5 3.8 3.8 3.3 4.0 3.1
18 7.4 6.9 5.6 4.9 4.1 3.6 4.6 3.7

Acid value is expressed as milligrams of KOH required to neutralize 1 g fat.

L.S.D. (0.05) values for A-AN. between treatments and various experimental periods within each treatment were 0.131 and 0.351,
respectively.

8-
O Butter (Control)
o Butter * BHT (200 ppm) . ~
6" -F I~utter * Cumin oll (200 ppm)
Q x I~utter * Thyme oll 1200 ppm) ~ ~ " ~ .io
>

4
!

I l I ( I I

0 3 6 9 12 15 18
Storage period (day)
F I G . 1. C h a n g e s in t h e a c i d v a l u e s o f b u t t e r s t o r e d at r o o m t e m p e r a t u r e w i t h o u t a n d
w i t h a d d e d B H T a n d e s s e n t i a l oils.

T h e i r e f f e c t i v e n e s s t o i n h i b i t t h e lipolytic a c t i v i t y in t h e t h e r e is a r e l a t i o n s h i p b e t w e e n i n h i b i t o r y effect on t h e
b u t t e r c a n be r a n k e d in t h e following o r d e r : c u m i n = growth of microorganisms and the chemical composition
t h y m e _ BHT > c o n t r o l . S t a t i s t i c a l a n a l y s i s r e v e a l e d t h a t of t h e t e s t e d e s s e n t i a l oils. Generally, t h e e x t e n t of t h e
BHT a n d t h y m e oil a d d e d t o b u t t e r h a d t h e s a m e l o w e r i n g i n h i b i t o r y effect of t h e oils c a n be a t t r i b u t e d t o t h e p r e s -
effect o n l i p a s e activity. However, t h e v a l u e s for l i p a s e ence of an aromatic nucleus containing a polar functional
a c t i v i t y s h o w t h a t c u m i n a n d t h y m e oils a c t a s a m o r e g r o u p . T h e w i d e s p r e a d u s e of p h e n o l s a n d r e l a t e d c o m -
s u p e r i o r p r e s e r v a t i v e for b u t t e r t h a n BHT. Table 3 lists p o u n d s as d i s i n f e c t a n t s is well e s t a b l i s h e d . T h y m o l oil
t h e t o t a l b a c t e r i a l a n d lipolytic b a c t e r i a l c o u n t s m e a s - h a d a h i g h e r i n h i b i t o r y a c t i o n t h a n c u m i n oil, w h i c h
u r e d a t t h e e n d o f t h e s t o r a g e p e r i o d . It is obvious t h a t t h e m i g h t be d u e t o t h e p r e s e n c e of p h e n o l i c OH g r o u p s . It is
a d d e d m a t e r i a l s to b u t t e r s i g n i f i c a n t l y l o w e r e d b o t h t o t a l well k n o w n t h a t t h e OH g r o u p is m u c h m o r e r e a c t i v e a n d
b a c t e r i a a n d lipolytic b a c t e r i a l c o u n t s . However, t h e de- easily f o r m s h y d r o g e n b o n d s w i t h t h e a c t i v e sites of t h e
gree of effectiveness was largely dependent upon added h y d r o l y t i c enzymes.
materials. For instance, the systems containing cumin T h e c h a n g e s in p e r o x i d e v a l u e s a n d TBA v a l u e s for all
a n d t h y m e oils w e r e m o r e effective t h a n BHT in l o w e r i n g systems during the various experimental periods were
t h e t o t a l a n d lipolytic b a c t e r i a l c o u n t s . S t a t i s t i c a l a n a l y - s m a l l c o m p a r e d to t h e c h a n g e s in t h e a c i d v a l u e s . Hence,
sis r e v e a l e d t h a t t h e t r e n d t o w a r d s d e c r e a s i n g t h e lipolyt- t h e s e r e s u l t s i n d i c a t e t h a t t h e m a i n c a u s e o f b u t t e r spoil-
ic b a c t e r i a l c o u n t s w a s in a c c o r d a n c e w i t h t h e t r e n d for age is h y d r o l y t i c r a n c i d i t y a n d n o t o x i d a t i v e r a n c i d i t y .
l i p a s e activity. T h e e s s e n t i a l oils s t u d i e d s h o w e d a m o r e p o w e r f u l a n t i -
T h e v o l a t i l e s u b s t a n c e s o f t h e s e e s s e n t i a l oils w e r e qual- h y d r o l y t i c effect t h a n BHT. T h e e s s e n t i a l oils c a n be ob-
i t a t i v e l y a n d q u a n t i t a t i v e l y d e t e r m i n e d b y gas-liquid tained from materials which are widely cultivated, inex-
c h r o m a t o g r a p h y (Table 4). T h e m o s t a b u n d a n t c o m - p e n s i v e a n d safe. T h e r e f o r e , t h e a u t h o r s r e c o m m e n d e d
p o u n d s in c u m i n a n d t h y m e oils w e r e c u m i n a l d e h y d e t h e e s s e n t i a l oils d e r i v e d f r o m t h y m e a n d c u m i n s h o u l d
(55.7%) a n d t h y m o l (42.7%), respectively. It s e e m s t h a t be u s e d t o e x t e n d t h e s h e l f life of b u t t e r .

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USE OF SOME ESSENTIAL OILS AS NATURAL PRESERVATIVES FOR B U T r E R

TABLE 3

I n f l u e n c e o f B I t T a n d Some Essential Oils on the L i p a s e A c t i v i t y a n d B a c t e r i a l C o u n t s i n B u t t e r a t the Eighteenth Day o f S t o r g e

Lipid s y s t e m Lipase activity Total b a c t e r i a l c o u n t X 104 Lipolytic b a c t e r i a l c o u n t X 103

Butter (control) 0.61 c 280 a 418.3 r

Butter + BHT (200 p p m ) 0.45 b 180.5 b 207.0 b
Butter + C u m i n oil (200 p p m ) 0.32 a 57.0 r 153.6 a
Butter + T h y m e oil (200 p p m ) 0.39~ b 104.5 d 156.0 a

LS.D. (0.05) v a l u e s for lipase activity, t o t a l bacterial c o u n t s a n d lipolytic b a c t e r i a l counts w e r e 0.11, 37.53 a n d 45.41, respectively.
a,b,r v a l u e s w i t h i n a c o l u m n followed by t h e s a m e letter a r e n o t significantly different by L.S.D. test.

TABLE 4 REFERENCES
Chemical Composition o f Cumin and Thyme E s s e n t i a l Oils 1. Allen, J.C., a n d R.J. Hamilton, Rancidity in Foods, Applied
Science Publishers, L o n d o n a n d New York, 1983, pp. 85 a n d 173.
C u m i n oil T h y m e oil 2. Axistova, V.P., G.S. Poyarkova, Z.P. C h u z h o v a , E.a. Sidorova a n d
H.V. M a r k a r i m a , Dairy Sci. (Abstr.) 33:3867 (1971).
3. E1-Emary, M., A~A. EI-Nimr a n d N.S. A h m e d , Egyptian Dairy
Component RRT~ % RRT~ %
So/. 2:149 (1974).
4. P r a s a d , S., a n d S.K. Gupta~ Asian J. Dairy Res. 2:45 (1984).
a-Pinene 0.07 0.3 0.08 1,1 5. Rao, C.N., B.V. Rao, T.J. Rao a n d G.R.R. Rao, Ibid. 3:127 (1985).
f~-Pinene 0.11 20.6 0.10 0.3
6. Riel, RR., a n d V.T.X. Vovon, Diary Sci. (Abstr.) 37:3701 (1975).
Camap hene 0.17 0.4 -- --
7. Sokolov, F.S., Ibid. 37-.2320 (1975).
Limonene 0.24 5.4 0.20 0.3
8. J o h s o n , LE., a n d W.M Cort, B e v e r a g e s No. 148, 10, No. 149, 10,
T-Terpinene 0.28 0.2 0.13 0.1
14 (1985).
Terpinolene 0.37 12.0 -- --
9. W u r t z e n , G., P. Olsen a n d E. Poulsen, Food Sc/. Technol. (Abstr.)
Phelleridrene -- -- 0.17 1.5
18:2T18 (1986).
p-Cymene 0.34 4.0 0.27 36.0
10. Official M e t h o d s of Analysis o f t h e A s s o c i a t i o n of Official A n a -
Caryophyllene 0.78 0.6 -- --
lytical Chemist, 15th edn., e d i t e d by W. Horowitz, AOAC, W a s h -
Thymol -- -- 1.00 42.7
ington, D.C., 1975.
Thujone -- -- 0.41 0.2
11. Keeney, P.G., Candy and Snack Industry 136:68 ( t 971 ).
Cumin aldehyde 1.00 55.7 -- --
12. Longeneck, H.E., a n d D.E. Haley, J. Am. Oil Chem. Sac. 57:19
Borneol -- -- 0.52 0.7
(1935).
Linalyl a c e t a t e -- -- 0.19 1.0
13. A m e r i c a n Public Health Association, Standard Methods for the
Terpinyl a c e t a t e -- -- 0.13 0.1
Examination of Dairy P r o d u c t s , 1 l t h edn., New York, 1960.
Unidentified
14. E1-Sadek, M.G., a n d S,a..Z. M a h m o u d , Practical Applied Micro-
compounds -- 0.8 -- 16.0 b/o/(x.Ty, EI-Saada Publishers, Cairo, Egypt, 1967. p. 105 a n d 106.
~RRT refers to t h e relative r e t e n t i o n t i m e for t h e ma~or c o m p o u n d 15. T h o m a s , M.L, and F.H. Jackson, Statistical Methods in Agricul-
for e a c h e s s e n t i a l oil w h i c h is given a v a l u e o f 1.00. tural Research, A g r i c u l t u r a l E x t e n s i o n , University of Califor-
nia, 1972.
16. Farag, R.S., Z.Y. D a w a n d S.H. Abo-Raya, J. Food Sc/. 54:74
(1989).

[Received N o v e m b e r 20, 1988; a c c e p t e d O c t o b e r 14, 1989]

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JAOCS, Vol. 68, no. 3 (March '1990)