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290 J. Agric. Food Chem.

1982, 30,290-292

Isolation, Purification, and Characterization of Insect Repellents from Curcuma


longa L.

Helen C. F. Su,* Robert Horvat, and Ghulam Jilani'

Two compounds were isolated from Curcuma longa L. and identified from their spectral characteristics
as 2-methyl-6-(4-methylphenyl)-2-hepten-4-one (ar-turmerone) and 2-methyl-6-(4-methyl-l,4-cyclo-
hexadien-l-yl)-2-hepten-4-one (turmerone). ar-Turmerone and turmerone gave an average 62.9% (class
IV)and 43.1% (class 111) repellency, respectively, to TriboZium castaneum (Hbst.)after 8 weeb of study.
Turmerone was unstable thermally and also at ambient temperature in the presence of air, yielding its
dimer or the more stable ar-turmerone.

Tumeric, Curcuma longa L., is a tropical herb of the Atlanta, GA. One kilogram of the powder was extracted
Zingiberaceae family indigenous to southern Asia. The in a Soxhlet extractor with petroleum ether (bp 30-60 "C)
aromatic yellow powder from its mature rhizomes was used for 24 h. The petroleum ether solution was concentrated
in Asian countries for many centuries as a yellow vegetable under reduced pressure in a rotary evaporator to obtain
dye for silks and cottons. It is still used in foods as a the crude turmeric extract.
condiment, particularly as an essential ingredient of curry Liquid Column Chromatographic (LCC) Fraction-
powder, in medicine as a stomachic, carminative, anthel- ation of Crude Turmeric Extract. Step 1. Each 1.4-
mintic, laxative, and cure for liver ailment, and also as an 1.6-g portion of the crude extract was placed on a column
ant repellent in India (Sreenivasamurthy and Krishna- (40 X 2.0 cm i.d.) of silica gel (70-230 mesh; EM Reagent;
murthy, 1959; Watt and Breyer-Brandwijk, 1962). extra pure) and eluted with chloroform. A total of 120
The strong coloring pigment in turmeric contains cur- 2-mL fractions was collected. After the solvent was re-
cumin, 1,7-bis(4-hydroxy-3-methoxyphenyl)-1,6-heptadi- moved, every third fraction was spot tested by TLC (see
ene-3,5-dione, and two other curcuminoids (Srinivasan, below for procedure). The fractions possessing the same
1953; Lubis, 1968; Shankaracharya and Natarajan, 1974; Rr values were combined, and each combined fraction was
Krishnamurthy et al., 1976). The red-yellow essential oil bioassayed for repellency to T. castaneum. The insect
of turmeric, either from steam distillation or from an active fraction was further fractionated.
oil-selecting solvent extraction, contains d-a-phellandrene, Step 2. Each 150-160-mg sample of the active fraction
d-sabinene, zingiberene, borneol, l,&cineole, turmerone, from step 1was further separated by column chromatog-
ar-turmerone, sesquiterpene alcohols, a-and y-atlantone, raphy in a 40 X 1.2 cm i.d. column of silica gel and eluted
and bisabolene (Gildemeister and Hoffmann, 1956; Mima, with benzene. A total of 90 2-mL fractions was collected.
1959; Shankaracharya and Natarajan, 1974; Salzer, 1975 TLC analysis was made on every third fraction. The
Govindarajan, 1980). fractions with only Rf 0.25-0.28, the mixture of R 0.25-0.28
Shankar et al. (1980) conducted feeding toxicity studies and 0.30-0.33, and only R 0.30-0.33 were com6ined sep-
of turmeric and its alcohol extract in rats, guinea pigs, and arately. The two pure combined fractions were bioassayed.
monkeys. The results indicated that turmeric is nontoxic Thin-Layer Chromatographic Analysis of LCC
even at the very high level tested. There was absolutely Fractions. Brinkman EM reagent, precoated silica gel
no mortality or any morphological and histological ab- G FZu,20 X 20 cm analytical chromatoplates were used.
normalities in the experimental animals at 2.5 g of tur- A small sample of each selected fraction was applied on
meric/kg of body weight which corresponds to a maximum spots 2.5 cm above the lower edge and 1.5 cm from each
consumption of 5 g/day for an adult human with an av- other. Each plate was developed in benzene and then
erage body weight of 70 kg. It is the practice in some examined under UV at 254 nm.
southern Asian countries; i.e., India and Pakistan, to store High-Performance Liquid Chromatographic Puri-
rice or wheat by mixing it with 2% of turmeric powder fication. A Waters Associates Model ALC/GPC 244
(Chatterjee, 1980). In this paper, we report the isolation, high-pressure liquid chromatograph with a Model 6000A
purification,and identification of the repellent components pump, a U6K injector, an R401 differential refractometer,
of turmeric to Tribolium castaneum (Hbst.). and a Model 440 UV detector with a 300 X 7.8 mm i.d.
MATERIALS AND METHODS pBondapak C18column (octadecyltrichlorsilanecovalently
bonded to lo-" pPorasil packing) was used. Metha-
Extraction of Turmeric. Powdered turmeric (im- nol-water ( 8 2 by volume, degassed) was used as the eluting
ported from India) was purchased from Lex Enterprise,
system. The column effluent was monitored at 254 nm
(2.0 AUFS), and the response was recorded on a Houston
Stored-Product Insects Research and Development Omniscribe Model B-5217-1 recorder.
Laboratory, Agricultural Research, Science and Education Each of the two LCC separated components was dis-
Administration, US.Department of Agriculture, Savan- solved individually in methanol to obtain the concentration
nah, Georgia 31403 (H.C.F.S. and G.J.), and Richard B. of approximately 20 Mg/pL and filtered through a Waters
Russell Agricultural Research Center, Agricultural Re- Associates sample clarification kit with 0.5-pm Millipore
search, Science and Education Adminstration, US. De- organic filter system. For each run, a 5-pL aliquot of the
partment of Agriculture, Athens, Georgia 30604 (R.H.). stock solution was injected onto the column with the flow
lFulbright-Hays Predoctoral Research Associate, rate of the eluting solvent at 1.5 mL/min. The desired
1978-1979. Present address: Honeybee Management, peak of the compound as indicated from the recorder was
Pakistan Agricultural Research Council, Islamabad, Pak- recycled through the column once, and the effluent of the
istan. recycled peak was collected. The process was repeated
This article not subject to U S . Copyright. Published 1982 by the American Chemical Society

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