Professional Documents
Culture Documents
Essential Oil and Glycosidically Bound Volatiles Of: Origanum Vulgare L. Ssp. Hirtum (Link) Ietswaart
Essential Oil and Glycosidically Bound Volatiles Of: Origanum Vulgare L. Ssp. Hirtum (Link) Ietswaart
ABSTRACT] The glycosidically bound volatiles were isolated from dried plant material by percolation with
ethyl acetate and by extraction with water during hydrodistillation of essential oil[ Fifteen volatile aglycones
were identi_ed by gas chromatography!mass spectrometry "GC!MS# on two columns with di}erent polarity of
the stationary phases[ The main aglycones were] thymoquinone\ benzyl alcohol\ thymol\ 1!phenylethanol\
carvacrol and 0!octen!2!ol[ The content of aglycones was 08 and 10 mg kg−0 with respect to the method of
isolation[ The chemical composition of aglycones was compared with the chemical composition of essential oil\
which consisted mainly of thymol and carvacrol[
After the hydrolysis of glycosides\ D!"¦#!glucose and unknown disaccharide were identi_ed[ The hydrolysis
of disaccharide\ D!"¦#!glucose and D!"¦#!galactose were detected by TLC and GC!MS as trimethylsilylethers[
Copyright Þ 1999 John Wiley + Sons\ Ltd[
KEY WORDS] Ori`anum vul`are ssp[ hirtum^ Lamiaceae^ glycosidically bound volatiles^ sugar moiety^ essential
oil\ gas chromatography!mass spectrometry
Copyright Þ 1999 John Wiley + Sons\ Ltd[ Flavour Fra`r[ J[ 1999^ 04] 089Ð083
081 J[ MASTELIC
AND I[ JERKOVIC
ł \ M[ MILOS ł
residue was dissolved in 9[4 ml pentane ] ether 0 ]0 v:v Table 1. Constituents and percentage composition of
and this solution was used for GC!MS analyses of essential oil from Origanum vulgare L. ssp. hirtum (Link)
carbohydrates[ Ietswaart isolated by hydrodistillation
Peak Compound I0 Peak area Methods of
No[ "HP!19M# ) identi_cation
Gas Chromatography-Mass Spectrometry (GC-MS)
Hydrocarbons
The volatile aglycones and the essential oil were 0[ a!Thujene 0923 9[4 I0\ I1\ MS
1[ b!Pinene 0986 9[2 I0\ I1\ MS
analyzed by gas chromatography!mass spectrometry 2[ Sabinene 0001 9[1 I0\ I1\ MS
"Hewlett!Packard\ model 4789\ with a mass selective 3[ d!2!Carene 0020 9[2 I0\ I1\ MS
detector\ model 4860A#[ Two columns with di}erent 4[ b!Phelandrene 0101 9[0 I0\ I1\ MS
5[ g!Terpinene 0120 1[9 I0\ I1\ MS
polarity of stationary phases were used[ GC operating 6[ Terpinolene 0159 9[2 I0\ Ð\ MS
conditions were]22\23 column HP!19M "Carbowax 19M#\ 7[ p!Cymene 0162 00[7 I0\ Ð\ MS
49 m×9[1 mm i[d[\ _lm thickness 9[1 mm\ column tem! 8[ Caryophyllene 0467 9[6 I0\ I1\ MS
09[ a!Humulene 0526 9[0 I0\ I1\ MS
perature programmed from 69>C isothermal for 3 min\ 01[ a!Murolene 0582 9[2 I0\ I1\ MS
then increased to 079>C at a rate of 3>C min−0^ column 02[ b!Farnesene 0588 9[2 I0\ I1\ MS
HP!090 "Methylsilicone#\ 14 m×9[1 mm i[d[\ _lm thick! 03[ b!Bisabolene 0690 9[6 I0\ I1\ MS
04[ d!Cadinene 0618 9[1 I0\ I1\ MS
ness 9[1 mm\ column temperature programmed from
69>C isothermal for 1 min\ then increased to 199>C at Oxygene containing compounds
a rate of 2>C mm−0^ carrier gas helium\ ~ow rate 0 ml 05[ 0!Octen!2!ol 0304 1[9 I0\ I1\ MS
06[ Bornyl acetate 0431 9[0 I0\ I1\ MS
min−0\ injector temperature 149>C\ volume injected 0 07[ Terpinen!3!ol 0448 0[9 I0\ I1\ MS
m\ split ratio 0 ] 49[ MS conditions] ionization voltage 08[ Methylthymyl ether 0454 9[4 I0\ Ð\ MS
69 eV\ ion source temperature 179>C\ mass range 29Ð 19[ 1!Isopropyl!0!methoxy!
3!methyl benzene 0465 9[3 I0\ I1\ MS
299 mass units[ The analysis of carbohydrates as tri! 10[ Borneol 0544 0[0 I0\ I1\ MS
methylsilylethers was performed on column HP!090 11[ p!Cymen!7!ol 0676 9[0 I0\ I1\ MS
with the same GC!MS conditions[ In this case mass 12[ Thymol 1012 38[2 I0\ I1\ MS
13[ Carvacrol 1038 13[5 I0\ I1\ MS
range was 29Ð599 mass units[
Total 85[2)
Identification and Quantitative Determination I0 retention indices on HP!19M^ I1 retention indices on HP!090^
MS mass spectra^ Ð not detected[
Individual peaks were identi_ed by comparison of their
retention indices with those of authentic samples\ as
well as by comparison of their mass spectra with those Glycosidically Bound Volatiles
stored in data base "Wiley library#[ The percentage com!
position of the samples was computed from the GC The content of glycosidically bound volatile compounds
peak areas without using correction factors[ The con! in dried plant material was 08 mg kg−0 "Method A#
tents of aglycones were calculated from the GC!peak and 10 mg kg−0 "Method B#[ Fifteen aglycones were
areas related to the GC!peak area of 0!octanol "lib! identi_ed[ Among the aglycones were identi_ed ali!
erated from octyl!b!D!glucoside#[ Preliminary GC!MS phatic alcohols\ terpene compounds and derivatives of
analysis showed the absence of 0!octanol as potential phenylpropanes[ The results are shown in Table 1[ The
aglycone[ main aglycones for both methods were] thymoquinone
"36[9 and 39[1)#\ benzyl alcohol "1[9 and 7[8)#\ thy!
mol "5[7 and 6[4)#\ 1!phenylethanol "9[7 and 4[5)#\ 0!
octen!2!ol "4[3 and 0[2)# and carvacrol "4[4 and 0[8)#[
Results and Discussion Minor di}erences among the content and the com!
Free Volatile Compounds position of aglycones depend on the isolation methods[
By comparison of the chemical composition of the
The yield of essential oil was 1[8)[ The content and essential oil "Table 0# and aglycones "Table 1# _ve com!
chemical composition of the essential oil is given in pounds were established to be identical] 0!octen!2!ol\
Table 0[ Sixteen compounds were identi_ed rep! terpinen!3!ol\ p!cymen!7!ol\ thymol and carvacrol[ Our
resenting 85[2) of the total oil[ The main components results show only moderate correlation in the chemical
were thymol "38[2)#\ carvacrol "13[5)# and p!cymene composition of free and glycosidically bound volatiles[
"00[7)#[ The essential oil also contained smaller quan! The aglycones\ such as aliphatic alcohols\ 1!phenyl!
tities of g!terpinene "1[9)#\ 0!octen!2!ol "1[9)#\ bor! ethanol\ benzyl alcohol\ eugenol\ linalool\ geraniol\
neol "0[0)# and terpinen!3!ol "0[9)#[ Similar results neral\ a!terpineol and terpinen!3!ol can\ more or less\
were reported for the Green origin of Ori`anum vul`are be considered as ubiquitous in aglycone fractions of
ssp[ hirtum[20 Lamiaceae0 family[ From our results it can be seen that
Copyright Þ 1999 John Wiley + Sons\ Ltd[ Flavour Fra`r[ J[ 1999^ 04] 089Ð083
ESSENTIAL OIL OF ORIGANUM VULGARE 082
Table 2. Identified constituents and percentage com- 080"39#\ 036"13#\ 017"7#\ 092"5#\ 62"78#[ b!anomer of
position of volatile aglycones from Origanum vulgare L. glucopyranose] M¦ 324"1#\ 294"1#\ 180"1#\ 106"06#\
ssp. hirtum (Link) Ietswaart isolated by two different 193"099#\ 080"27#\ 078"3#\ 036"10#\ 017"5#\ 092"5#\
methodsa 62"66#\ a!anomer of galactopyranose] M¦ 324"0#\
Peak Compound Peak area ")# Methods of 294"2#\ 154"1#\ 106"16#\ 194"10#\ 193"099#\ 080"37#\
No[ identi_cation 036"11#\ 017"00#\ 092"5#\ 62"67#\ b!anomer of gal!
A B
actopyranose] M¦ 324"2#\ 250"1#\ 208"3#\ 107"00#\
0[ 2!Hexen!0!ol tr 2[3 I0\ I1\ MS 106"49#\ 194"11#\ 193"099#\ 081"07#\ 080"82#\ 036"23#[ In
1[ 0!Octen!2!ol 4[3 0[2 I0\ I1\ MS
2[ Benzaldehyde 9[7 9[8 I0\ I1\ MS
general\ galactose is rarely found as the glycone moiety
3[ Terpinen!3!ol 9[5 Ð I0\ I1\ MS of glycosidically bound volatiles[ This monosaccharide
4[ Thymoquinone 36[9 39[1 I0\ I1\ MS unity was identi_ed as glycone part in glycosides of
5[ Methyl salicylate Ð 9[5 I0\ I1\ MS
Thymus vul`aris "Lamiaceae#\08 as well as the disac!
6[ Benzyl alcohol 1[9 7[8 I0\ I1\ MS
7[ 1!Phenyl ethanol 9[7 4[5 I0\ I1\ MS charide part in glycosides of b!D!glucopyranosyl!"0!1#!
8[ p!Cymen!7!ol 9[7 0[1 I0\ I1\ MS b!D!galactopyranose in Coleus forskohlii "Lamiaceae#[14
09[ Eugenol 9[3 Ð I0\ I1\ MS
00[ Thymol 5[7 6[4 I0\ I1\ MS Acknowled`ements * This work was supported by Croatian National
01[ Carvacrol 4[4 0[8 I0\ I1\ MS Grant\ Project 900!992[
02[ 0!H!Indole 1[1 9[7 I0\ I1\ MS
03[ 1!"p!Metoxyphenyl# ethanol Ð 0[2 Ð\ Ð\ MS
04[ Butyl phtalate monoester 2[9 tr Ð\ Ð\ MS
Copyright Þ 1999 John Wiley + Sons\ Ltd[ Flavour Fra`r[ J[ 1999^ 04] 089Ð083
083 J[ MASTELIC
AND I[ JERKOVIC
ł \ M[ MILOS ł
22[ Mastelic J\ Milos² M\ Kus²trak D\ Radonic A[ Croat[ Chem[ Acta[ 25[ Soczewinski E\ Wojciah M\ Pachowicz K[ Chem[ Anal[ "Warsaw#
0887^ 60] 036[ 0887^ 32] 712[
23[ Mastelic J\ Kus²trak D[ Acta Pharm[ 0886^ 36] 022[ 26[ Franken!Luykx JMM\ Klopper WJ[ Brauwissenschaft 0856^ 19]
24[ Bieganowska ML\ Petruzynik A[ Chem[ Anal[ "Warsaw# 0886^ 062[
31] 235[
Copyright Þ 1999 John Wiley + Sons\ Ltd[ Flavour Fra`r[ J[ 1999^ 04] 089Ð083