Rapid and Simple Isolation of Zingiberene fromGinger Essential Oil

Jocel yn G. Mi l l ar*
Department of Entomology, University of California, Riverside, California 92521
Received March 4, 1998
A sesqui terpene-enri ched fracti on of gi nger oi l was treated wi th the di enophi l e 4-phenyl -1,2,4-
tri azol i ne-3,5-di one (2), whi ch sel ecti vel y formed a Di el s-Al der adduct wi th the sesqui terpene
hydrocarbon zi ngi berene (1). The adduct was puri fi ed by fl ash chromatography, then hydrol yzed
to return zi ngi berene i n good yi el d and >99% puri ty.
The sesqui terpene hydrocarbon zi ngi berene (1) [5-(1,5-
di methyl -4-hexenyl )-2-methyl -1,3-cycl ohexadi ene] has
been shown to have a consi derabl e spectrum of bi ol ogi cal
acti vi ty. For exampl e, recent studi es have determi ned
i ts anti vi ral ,
1
anti ul cer,
2
and anti ferti l i ty
3
effects. Fur-
thermore, i t has been wi del y used i n cosmeti cs and
fragrances.
4
Zi ngi berene i s a major component of com-
merci al l y avai l abl e oi l deri ved from rhi zomes of the
gi nger pl ant Zingiber officinaleRoscoe (Musal es: Zi n-
gi beraceae), but obtai ni ng pure 1 from the mi xture of
sesqui terpenes i n the oi l can be probl emati c. For
exampl e, recent publ i cati ons have descri bed the i sol a-
ti on of zi ngi berene of l ow puri ty
5
or i n l ow yi el d
1
onl y
after several tedi ous sequenti al puri fi cati on steps.
I n connecti on wi th the i denti fi cati on of sex pheromone
components of the sti nkbug Thyanta pallidovirens Stl
(Hemi ptera: Pentatomi dae), the mal es of whi ch produce
several sesqui terpene hydrocarbons,
6
we requi red 100-
mg amounts of zi ngi berene of hi gh puri ty for l aboratory
and fi el d bi oassays. A search of the l i terature reveal ed
that zi ngi berene i s general l y i sol ated from essenti al oi l s
i n preparati ve scal e by fracti onal di sti l l ati on to produce
a fracti on enri ched i n sesqui terpenes, fol l owed by one
or more l i qui d chromatographi c steps. However, zi n-
gi berene has been sel ecti vel y removed from gi nger oi l
by the formati on of Di el s-Al der adducts wi th mal ei c
anhydri de or other powerful di enophi l es duri ng the
i sol ati on of other sesqui terpene components of gi nger
oi l , such as R-curcumene and sesqui phel l andrene.
7
I t
occurred to us that thi s strategy of sel ecti ve removal
coul d be used to obtai n zi ngi berene of hi gh puri ty by
choosi ng a di enophi l e that woul d produce an adduct
amenabl e to control l ed degradati on, to return the parent
di ene. 4-Phenyl -1,2,4-tri azol i ne-3,5-di one (PTAD) (2),
whi ch was devel oped as a protecti ng group for endocycl i c
1,3-di enes i n the B-ri ng of steroi ds,
8,9
appeared to be a
l i kel y di enophi l e candi date. I n parti cul ar, PTAD reacts
wi th 1,3-di enes essenti al l y i nstantaneousl y at room
temperature to form the Di el s-Al der adducts. Fur-
thermore, the resul ti ng adducts are reported to be
readi l y degraded to the parent di enes by ei ther base
hydrol ysi s or reducti on wi th Li Al H
4
.
8,9
I n our hands, a sol uti on of PTAD i n THF was added
dropwi se to a gi nger oi l sesqui terpene fracti on i n THF
unti l the pi nk col or of unreacted PTAD persi sted. The
amount of PTAD added was not cri ti cal , because unre-
acted zi ngi berene or excess PTAD were readi l y removed
by chromatography. Thus, after concentrati on, the
mi xture was puri fi ed by fl ash chromatography on Si gel
to remove the other unreacted hydrocarbons, yi el di ng
the pure adduct 3as a col orl ess gum. Base hydrol ysi s
of 3, fol l owed by extracti on of the recovered zi ngi berene
i nto hexanes and Kugel rohr di sti l l ati on, produced 1i n
>99% puri ty by GC (Scheme 1).
Thus, the method i s amenabl e to the rapi d and
strai ghtforward preparati ve scal e i sol ati on of zi ngi b-
erene from gi nger oi l , wi thout the use of speci al i zed
equi pment such as spi nni ng band di sti l l ati on col umns
or si l i ca LC col umns i mpregnated wi th AgNO
3
. Fur-
thermore, the method shoul d be equal l y appl i cabl e to
the i sol ati on of rel ated compounds contai ni ng endocycl i c
1,3-di ene moi eti es, such as 7-epi zi ngi berene
5
or the
monoterpene R-phel l andrene.
Experimental Section
Fl ash chromatography was carri ed out wi th 0.04-
0.063-mm Si gel (Al dri ch Chemi cal Co., Mi l waukee, WI ).
1
H and
13
C NMR spectra were recorded i n CDCl
3
at 300
and 75 MHz, respecti vel y, usi ng a General El ectri c QE
300 i nstrument. GC-MS anal yses (EI , 70 eV) were
performed on a Hewl ett-Packard 5890 gas chromato-
graph i nterfaced to an Hewl ett-Packard 5970B mass
sel ecti ve detector. A DB-5 col umn was used (30 m
0.25 mm, J&W Sci enti fi c, Fol som, CA; temperature
program, 50 C/0 mi n, 10/mi n-250 C). HREI MS were
obtai ned wi th a VG7070 i nstrument.
Gi nger oi l (Spectrum Chemi cal Co., Gardena, CA) was
Kugel rohr di sti l l ed at 0.1 mmHg. The fi rst fracti on
(oven temperature <60 C) contai ni ng monoterpenoi ds
was di scarded. A second fracti on (oven temperature
* To whom correspondence shoul d be addressed. Tel .: 909 787 5821.
Fax: 909 787 3086. E-mai l : jocel yn.mi l l ar@ucr.edu.
Scheme 1
1025 J . Nat. Prod. 1998, 61, 1025-1026
S0163-3864(98)00069-X CCC: $15.00 1998 Ameri can Chemi cal Soci ety and Ameri can Soci ety of Pharmacognosy
Publ i shed on Web 07/11/1998
60-100 C) consi sted al most excl usi vel y of sesqui ter-
pene hydrocarbons, i ncl udi ng R-curcumene (9.2%), zi n-
gi berene (46%), sesqui phel l andrene (17.4%), and bi sab-
ol ene (10.1%). A porti on of the sesqui terpene fracti on
(2.0 g, ca. 9.8 mmol of mi xed sesqui terpenes) i n 20 mL
of dry THF was sti rred at room temperature, whi l e
addi ng PTAD (0.88 g, 5 mmol ; Al dri ch) i n THF (10 mL)
dropwi se. The mi xture was concentrated and puri fi ed
by fl ash chromatography, el uti ng wi th 15% Me
2
CO i n
hexanes. The adduct 3(1.18 g, ca. 69% yi el d based on
percentage of zi ngi berene i n the starti ng materi al ) was
recovered as a col orl ess gum:
1
H NMR (CDCl
3
, 300
MHz) 7.3-7.5 (5H, m, Ph), 5.98 (1H, br d, J ) 5.7 Hz,
H-3), 5.09 (1H, br t, J ) 7 Hz, H-10), 4.97 (1H, dd, J )
5.7, 2.6 Hz, H-4), 4.71 (1H, m, H-6), 2.31 (1H, ddd, J )
13.1, 8.8, 3.5 Hz, H-6), 2.08 (3H, m, H-5, H-9, H-9), 1.93
(3H, d, J ) 1.6 Hz, H-15), 1.70 (3H, s, H-12), 1.63 (3H,
s, H-13), 1.5 (1H, m, H-8), 1.25 (2H, m, H-6, H-8), 1.10
(1H, m, H-7), 0.88 (3H, d, J ) 6.3 Hz, H-14);
13
C NMR
(75.48 MHz) 156.43, 156.10, 140.83, 131.84, 131.58,
129.04, 128.05, 125.37, 124.07, 120.59, 55.10, 53.60,
41.34, 36.51, 34.30, 28.57, 25.71, 24.88, 19.40, 17.70,
16.31; EI MS m/z 379 [M]
+
(92), 268 (6), 241 (100), 204
(28), 178 (33), 119 (63), 91 (27); HREI MS m/z 379.2258
(cal cd for C
23
H
29
N
3
O
2
, 379.2260).
The adduct 3(0.33 g, 0.87 mmol ) was refl uxed under
Ar i n 10 mL of 2.1 M KOH i n 95% EtOH for 3 h. The
cool ed mi xture was di l uted wi th H
2
O and extracted wi th
hexanes (3 25 mL). The hexane extracts were washed
wi th saturated aqueous NaCl , dri ed over Na
2
SO
4
, and
passed through a short col umn of Si gel (ca. 4 cm 1.5
cm di am), el uti ng wi th hexanes, to remove traces of
ani l i ne. The sol uti on was concentrated, then Kugel rohr
di sti l l ed, yi el di ng 143 mg (81%) of zi ngi berene (1), >
99% pure by GC-MS. The mass spectrum and reten-
ti on ti mes exactl y matched those of zi ngi berene i n the
crude gi nger oi l , and the
1
H NMR spectrum cor-
responded cl osel y to l i terature data.
5
Acknowledgment. We thank the Cal i forni a Pi sta-
chi o Commi ssi on for fi nanci al support of thi s work, and
the Anal yti cal Servi ces Laboratory, UC Ri versi de, for
obtai ni ng the HRMS.
References and Notes
(1) Denyer, C. V.; Jackson, P.; Loakes, D. M. J . Nat. Prod. 1994,
57, 658-662.
(2) Yamahara, J.; Mochi zuki , M.; Rong, H. Q.; Matsuda, H.;
Fuji mura, H. J . Ethnopharmacol. 1988, 23, 299-304.
(3) Ni , M.; Chen, Z.; Yan, B. Huadong Huagong Xueyuan Xuebao
1988, 14, 675-679; ChemAbstr. 1989, 111, 195149w.
(4) Handa, K. L.; Sharma, M. L.; Ni gam, N. C. Perf. Cosmetics 1963,
44, 233-236.
(5) Breeden, D. C.; Coates, R. M. Tetrahedron 1994, 50, 11123-
11132.
(6) Mi l l ar, J. G. Tetrahedron Lett. 1997, 38, 7971-7972.
(7) Connel l , D. W.; Sutherl and, M. D. Aust. J . Chem. 1965, 19, 283-
288.
(8) Barton, D. H. R.; Shi oi ri , T.; Wi ddowson, D. A. J . Chem. Soc.
(C) 1971, 1968-1973.
(9) Barton, D. H. R.; Lusi nchi , X.; Rami rez, J. S. Tetrahedron Lett.
1983, 24, 2995-2998.
NP9800699
1026 J ournal of Natural Products, 1998, Vol. 61, No. 8 Notes