New Phenolic, triterpenic and steroidal constituents from the fruits of Cuminum cyminum L.

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1 0; (): 9- ISSN 8- ISSN 9-8 JPP 0; (): 9- Received: Accepted: Nisha Chaudhary Phytochemistry Research Laboratory, Faculty of Pharmacy, Jamia Hamdard, Hamdard Nagar, New Delhi 0 0, India Shahnaz S. Husain Phytochemistry Research Laboratory, Faculty of Pharmacy, Jamia Hamdard, Hamdard Nagar, New Delhi 0 0, India Mohammed Ali Phytochemistry Research Laboratory, Faculty of Pharmacy, Jamia Hamdard, Hamdard Nagar, New Delhi 0 0, India Correspondence: Mohammed Ali Phytochemistry Research Laboratory, Faculty of Pharmacy, Jamia Hamdard, Hamdard Nagar, New Delhi 0 0, India maliphyto@gmail.com Tel: New Phenolic, triterpenic and steroidal constituents from the fruits of Cuminum cyminum L. Nisha Chaudhary, Shahnaz S. Husain, Mohammed Ali ABSTRACT Cuminum cyminum L. (Family: Apiaceae) is a small annual herb with slender, angular branched stem cultivated as a spice throughout the world. Its fruits are used to treat dyspepsia, insomnia, cold, fever, gastrointestinal, gynecological and respiratory disorders, toothache, diabetes, hypertension and epilepsy. Phytochemical analysis of the methanolic extract of the fruits of C. cyminum led to the isolation of a fatty acid ester identified as n- tricosanyl n- octadec-9- enoate () and five terpenic and steroidal constituents characterized as,,,8-tetrahydroxynaphthyl geranilan-0ʹ-al ʹ-oate (), lanost-,0 ()-dien-α-olyl n- docosanoate (), labdan-α,,0-triol--(0ʹ,ʹ- dihydroxy anthraquinone-ʹ-oate) (), stigmast--en- β--d-arabinopyranosyl-ʹ-benzoate () and lanost-,-dien-β-ol β--d- arabinopyronosyl-ʹ- n- octadec-9ʹʹ, ʹʹ-dienoate (). The presence of these phytoconstituents are reported for the first time in C. cyminum fruits and their structures have been established on the basis of spectral data analysis. Keywords: Cuminum cyminum, fruits, methanolic extract, terpenic constituents, steroidal glycoside, aliphatic ester.. Introduction Cuminum cyminum L. (Family Apiaceae), known as cumin, is a small annual herb with slender, angular branched stem. The plant is indigenous to Egypt and Syria and one of the most cultivated popular spices throughout the world. It thrives well from 9 to C and is intolerant to long periods of dry heat and frost. Cumin seeds resemble caraway seeds, but are slightly smaller having oblong shape, thicker in the middle, compressed laterally with nine ridges and yellow-brown in colour. Cumin is the second most popular spice in the world after black pepper. It is added in some Dutch cheeses, traditional bread in France, Brazilian cuisine, chili powder, sofrito, garam masala, curry powder, cuisines, baharat and other preparations of food of the Middle East, India, Cuba and Mexico. Cumin is spread on the meat along with other common seasonings. The cumin seeds are considered as carminative, analgesic, eupeptic, antispasmodic, astringent, used to treat digestive disorders, cough, diarrhea, dyspepsia, flatulence, morning sickness, colic, dyspeptic headache and bloating and to improve liver function [-]. A mixture of powdered cumin seeds, honey, salt and butter is applied to alleviate pain from scorpion bites [, ]. The seeds contained a volatile oil mainly composed of monoterpene hydrocarbons, oxygenated mono- and sesqui- terpenes, fatty acids, aldehydes, ketones and esters. The major compounds occurring in cumin are cuminaldehyde, limonene, α- and β-pinenes,,8-cineole, o- and p-cymenes, α- and γ-terpinenes, safranal and linalool [-9]. The other components present in the cumin are monoterpenic, flavone, -C-methyl-D-erythritol, sesquiterpene lactone and alkyl glycosides, amino acids, fatty acids and aromatic compounds [0-]. Several nutrients, e.g., vitamins, amino acids, protein, minerals, starch, sugars, tannins, phytic acid and dietary fiber components have also been reported in cumin seeds [8, 9]. The present paper describes the isolation and characterization of the chemical constituents from the seeds of C. cyminum of Delhi region.. Materials and Methods. General Melting points were determined on a Perfit apparatus without correction. The IR spectra were measured in KBr pellet on a Bio-Red FT-IR spectrometer. Ultraviolet (UV) spectra were obtained in methanol with a Lambda Bio 0 spectrometer. The H (00 MHz) and C (00 MHz) NMR spectra were recorded on Bruker spectrospin spectrometer using TMS as an ~ 9 ~

2 internal standard. ESI MS analyses were performed on a JEL SX 0/Da-00 instrument equipped with direct inlet probe system. Column chromatography separations were carried out on silica gel (Merck, 0 0 mesh, Mumbai, India). Precoated silica gel plates (Merck, Silica gel 0 F) were used for analytical thin layer Chromatography and visualized by exposure to iodine and UV radiations... Plant materials The dried seeds of C. cyminum were obtained from a local market of Khari Baoli, Delhi and authenticated by Dr. H.B. Singh, Scientist, Herbarium Section, National Institute of Science and Information Resources (CSIR), New Delhi. A voucher specimen (No. NISCAIR/ RHMD/00-//) is deposited in the herbarium... Preparation of extract The fruit powder of C. cyminum ( kg) was extracted exhaustively in a Soxhlet apparatus with methanol for hr. The methanolic extract was concentrated under reduced pressure to obtain dark brown viscous mass (.%). A small portion of the extract was analyzed chemically to determine the presence of different chemical constituents... Isolation of phytoconstituents The viscous dark brown mass was adsorbed on silica gel (0-0 mesh) for column chromatography, after being dissolved in a little quality of methanol for preparation of a slurry. The slurry (00 g) was air dried and subjected to chromatography over a silica gel column packed in petroleum ether. The column was eluted successively with petroleum ether, mixture of petroleum ether and chloroform (9:, :, :, and :), chloroform and the mixture of chloroform and methanol (99:, 9:, 9:, 9:8, 9:, :, :, :). Various fractions were collected separately and matched by TLC to check the homogeneity. Similar fractions having the same Rf values were combined and crystallized. The isolated compounds were recrystallized to get the following compounds:.. n-tricosanyl oleate () Elution of column with petroleum ether furnished colorless powder of, recystallized from acetone methanol (:),. mg (0.% yield), R f 0. (petroleum ether); m.p. - C; UV λmax: nm; IR ν max (KBr): 9, 8, 8,, 08,,, 0, 00, 80, 9 cm - ; H NMR (CDCl ): δ. (H, m, H-9),.0 (H, m, H-0),. (H, t, J=.0 Hz, H-ʹ),. (H, t, J=, Hz, H -),. (H, m, H-8),.0 (H, m, H -),.0 (H, m, CH ),. (H, m, CH ),. (H, m, CH ),. (H, m, CH),.0 (0 H, brs, 0 x CH),.8 (H, brs, 8 x CH), 0.89 (H, t, J=. Hz, CH -8), 0.8 (H, t, J=. Hz, CH -ʹ); C NMR (CDCl ): δ. (C-), 9.8 (C-9),.0 (C-0),.0 (C-ʹ),. (CH ), 9. ( x CH ), 9. ( x CH ), 9.0 ( x CH ),.8 (CH ),. (CH -8),.0 (CH -ʹ); +ve TF MS m/z (rel.int.): 0 [M] + (C H 80 ) (.), 8 (.8), (.), 9 (8.)... Tetrahydroxynaphthyl geranilan-0ʹ-al ʹ-oate () Elution of the column with petroleum ether-chloroform (:) yielded pale yellow crystals of, recystallized from acetonemethanol (:),.8 mg (0.0 % yield); R f 0. (petroleum etherchloroform, :); m.p C; UV- λmax (MeH): 0, 9, 9 nm (log ε.,.,.); IR ν max (KBr): 8,, 98, 8,, 80,,,, 0,, 9, 0, 89 cm -. ; H NMR (CDCl ): δ 9.9 (H, d, J=.8 Hz, H-0ʹ),.8 (H, d, ~ 0 ~ J=8.0 Hz, H-),. (H, d, J=8. Hz, H-),. (H, d, J=8.0 Hz, H-),. (H, d, J=8. Hz, H-),.8 (H, m, H-ʹ),.0 (H, d, J=. Hz, H -ʹ),.8 (H, m, H-ʹ),. (H, m, H -ʹ),. (H, m, H-ʹ),.8 (H, m, H -),. (H, d, J=.0 Hz, Me-8ʹ),. (H, d, J=.0 Hz, Me-9ʹ); C NMR (CDCl): δ. (C-),.8 (C-),.8 (C-),. (C-), 9. (C-),. (C-), 8.08 (C-),.09 (C-8),.8 (C-9),.8 (C-0), 9.8 (C-ʹ),.0 (C-ʹ),.9 (C-ʹ),. (C-ʹ), 9. (C- ʹ),.8 (C-ʹ),.8 (C-ʹ),. (C-8ʹ),.8 (C-9ʹ), 9.8 (C-0ʹ); +ve FAB MS m/z (rel.int.): 0 [M] + (C 0H ) (.), 9 (.8), 9 (.)... Lanostdienyl docasanoate () Elution of the column with chloroform gave colorless crystals of, recystallized from acetone- methanol (:),. mg (0.09% yield); R f : 0. (chloroform); m.p. - C; UV- λmax: 0 nm (log ε.8); IR ν max (KBr): 9, 8,,,, 8, 0, 088, cm - ; H NMR (CDCl): δ. (H, m, H-),.0 (H, m, H-),.0 (H, dd, J =.,.0 Hz, H-β),.98 (H, brs, Me- ),.0 (H, m, H ʹ),. (0H, m, 8 x CH, x CH),. (H, brs, Me-9),. (H, brs, Me-8),.9 (H, brs, Me-0),. ( H, brs, Me-9), 0.9 (H, d, J =. Hz, Me-), 0.9 (H, brs, Me-8), 0.9 (H, d, J =. Hz, Me-), C NMR (CDCl ): δ. (C-), 9.0 (C-), 8. (C-), 8. (C-),.8 (C- ),. (C-), 9. (C-),. (C-8), 0.8 (C-9),. (C- 0),. (C-),. (C-),.08 (C-), 0. (C-),. (C-), 0.9 (C-),.8 (C-),. (C-8), 8. (C-9),. (C-0),.8 (C-),.9 (C-),.8 (C-), 9. (C-), 8. (C-),. (C-),.9 (C-),. (C- 8), 8. (C-9),.8 (C-0), 9.9 (C-ʹ),.0 (C-ʹ),. (C-ʹ), 9. to.8 (C-ʹ to C-0ʹ),. (C-ʹ),.8 (C-ʹ). +ve FAB MS m/z: 8 [M] + (C H 9 ) (.8), (.), 9 (8.), (.)..8. Trihydroxy labdanyl anthraquinoate () Elution of the column with chloroform-methanol (9:) afforded brown colored mass of, recystallized from chloroform-methanol (9:),.9 mg (.0% yield); R f value: 0. (chloroform- methanol, 9:); m.p. 8-8 C; UV λmax (MeH): 09, 8, 0 nm (log.,.,.); IR νmax (KBr):, 8, 0, 9, 8,, 0,,, 9,,, 0 cm - ; H NMR (DMSd ):.8 (H, d, J=8. Hz, H-9ʹ),.8 (H, d, J=.0 Hz, H-ʹ),. (H, dd, J=.0, 8. Hz, H-ʹ),. (H, d, J=8. Hz, H-ʹ),.9 (H, d, J=8. Hz, H-8ʹ),. (H, brs, H =),.8 (H, ddd, J=.,.,. Hz, H-β),. (H, d, J=8. Hz, H -0),.8 (H, d, J=. Hz, H-α),.8 (H, m, H -a),.0 (H, m, H -b),.0 (H, m, H -a),. (H, m, H-8, H-),. (H, m, H-9),. (H, m, H-b, H-a),. (H, m, H-a),. (H, m, H-a, H -a),.8 (H, m, H -a, H -b),. (H, m, H -b),. (H, m, H -b, H -b),. (H, brs, Me-9),.9 (H, m, H - b),. (H, brs, Me-),. (H, d, J=.0 Hz, Me-8), 0.9 (H, t, J=.0 Hz, Me-); C NMR (DMS-d ): δ. (C-), 9. (C-), 8. (C-),.08 (C-), 8. (C-),. (C-),. (C-), 0. (C-8), 0.8 (C-9),. (C-0), 9.8 (C-), 8.9 (C-),. (C-),.0 (C-),.9 (C-),. (C-),. (C-),.0 (C-8),.9 (C-9),. (C-0), 8.0 (C-ʹ),. (C-ʹ),. (C-ʹ),.89 (C-ʹ), 9. (C-ʹ),. (C-ʹ),.8 (C-ʹ),.0 (C-8ʹ),.0 (C-9ʹ),.0 (C-0ʹ),. (C-ʹ),.8 (C-ʹ), 9. (C-ʹ), 9. (C-ʹ),. (C-ʹ); +ve TF MS m/z (rel.int.): 9 [M] + (C H 8) (.), (9.8), 8 (.), (.)..9. β-sitosterol --arabinobenzoate ()

3 Elution of the column with petroleum ether chloroform (:) gave colorless crystal of ; recystallized from acetone-methanol,.mg 0.0 % yield); R f value: 0. (petroleum ether- chloroform, :); m.p. - C; UV- λmax: 0, nm (log ε.,.8); IR νmax (KBr): 8, 0, 9, 8,, 8, 0, 8, 9, 0, 88, cm - ; H NMR (CDCl ):.8 (H, dd, J=., 8.0 Hz, H-ʹʹ),. (H, J=.,. Hz, H-ʹʹ),. (H, m, H-ʹʹ),. (H, m, H-ʹʹ),.0 (H, m, H-ʹʹ),. (H, dd, J=.0,. Hz, H-),.0 (H, d, J=. Hz, H-ʹ),.0 (H, brm, w /=8. Hz, H-α),. (H, brs, H -ʹ),. (H, dd, J=.,. Hz, H-ʹ),.9 (H, m, H-ʹ),. (H, m, H-ʹ),.0 (H, brs, Me-9), 0.9 (H, d, J=. Hz, Me-), 0.89 (H, d, J=. Hz, Me-), 0.8 (H, d, J=. Hz, Me-), 0.8 (H, d, J=. Hz, Me-9), 0. (H, brs, Me-8), (9H, m, x CH, x CH); C NMR (CDCl ): δ. (C-),.8 (C-),.0 (C-),. (C-),.8 (C-),.0 (C-), 0.9 (C-),.8 (C-8),. (C-9),.0 (C-0),.09 (C-), 9. (C-),. (C-),.80 (C- ),. (C-), 8. (C-),. (C-),.8 (C-8), 9. (C-9),.0 (C-0), 8. (C-),.9 (C-),.80 (C-),.9 (C-), 9. (C-), 9. (C-), 9. (C-), 8. (C- 8),.0 (C-9), 0.8 (C-ʹ),.8 (C-ʹ), 8.8 (C-ʹ),. (C-ʹ),.9 (C-ʹ), 8. (C-ʹʹ), 9. (C-ʹʹ),. (C-ʹʹ),. (C-ʹʹ),.9 (C-ʹʹ), 9. (C-ʹʹ), 9.0(C-ʹʹ); 0 [M] + (C H ) (.), (.8)..0. Lanostdienyl arabinolinoleiate () Elution of the column with chloroform-methanol (99:) yielded colorless crystals of, recystallized from acetone-methanol (:),.9 mg (0.0% yield); R f 0.9 (chloroform-methanol, :99); m.p. -8 C; UV λmax (MeH): 0, nm (log ε.,.); IR ν max (KBr):, 8, 9, 8,,,, 8, 9, 9, 0, cm - ; H NMR (DMS-d ): δ. (H, m, H-),. (H, m, H-),. (H, dd, J =., 9. Hz, H-α),.8 (H, brs, Me-),. (H, brs, Me-),. (H, brs, Me-9),. (H, brs, Me-8),.8 (H, brs, Me-0),. (H, brs, Me-9), 0.98 ( H, d, J=.8 Hz, Me-), 0.9 (H, brs, Me-8),. (H, d, J=. Hz, H-ʹ),. (H, dd, J=.,.8 Hz, H-ʹ),.0 (H, d, J=. Hz, H -ʹ),.98 (H, m, H-ʹ),. (H, m, H-ʹ),.09 (H, t, J=. Hz, H -ʹʹ),.0 (H, m, H-9ʹʹ),. (H, m, H-0ʹʹ),.08 (H, m, H-ʹʹ),.98 (H, m, H-ʹʹ),.80. (H, m, 0 x CH, x CH), 0.89 (H, t, J =. Hz, Me- 8ʹʹ); C NMR (CDCl ): δ. (C-), 8. (C-),.0 (C-), 8.0 (C-),.8 (C-), 8.8 (C-), 9. (C-),. (C-8), 0. (C- 9),. (C-0),. (C-),. (C-),.8 (C-), 9.8 (C-),.8 (C-),.0 (C-),. (C-),. (C-8), 8. (C-9),. (C-0), 8. (C-),.9 (C-),. (C- ),. (C-), 9.8 (C-),. (C-),.89 (C-),.0 (C-8), 8. (C-9),. (C-0), 0.8 (C-ʹ), 8. (C-ʹ),.8 (C-ʹ),.0 (C-ʹ),.80 (C-ʹ),.0 (C-ʹʹ),.8 (C-ʹʹ),.8 (C-ʹʹ), 9. (C-ʹʹ), 9.8 (C-ʹʹ), 9. (C- ʹʹ), 9. (C-ʹʹ),. (C-8ʹʹ), 8. (C-9ʹʹ), 0.0 (C-0ʹʹ),.8 (C-ʹʹ),.8 (C-ʹʹ),.8 (C-ʹʹ),. (C-ʹʹ), 9.0 (C-ʹʹ), 9.8 C-ʹʹ),.89 (C-ʹʹ),. (C-8ʹʹ); +ve FAB MS m/z (rel.int.): 8 [M + H] + (C H 89 ) (.).. Results and discussion Compound, named n-tricosanyl oleate, was obtained as a colorless powder from petroleum ether eluants. It exhibited characteristic IR absorption bands for ester ( cm - ), unstauration (08 cm - ) and long aliphatic chain (9 cm - ). Its mass spectrum showed a molecular ion peak at m/z 0 corresponding to the molecular formula of a fatty acid ester CH80. The ion peaks arising at m/z [CH (CH)CH=CH ~ ~ (CH ) -C] +, 9 [M-] and 8 [CH (CH ) CH=CH(CH ) - C] + indicated that oleic acid was esterified with a C aliphatic alcohol. The H NMR spectrum of showed two one-proton multiplets at δ. and.0 assigned to vinyl H-9 and H-0 proton, respectively. A two-proton triplet at δ. (J=.0 Hz) was ascribed to oxygenated methylene H-ʹ proton. The other methylene proton resonated from δ. to δ.8. Two threeproton triplets at δ 0.89 (J=. Hz) and 0.8 (J=. Hz) were accounted to the terminal C-8 and C-ʹ primary methyl protons, respectively. The C NMR spectrum of compound displayed signals for ester carbon at δ. (C-), vinylic carbons at δ 9.8 (C-9) and.0(c-0), oxygenated methylene carbon at δ.0 (C-ʹ), other methylene carbons in the range of δ.-.8, and methyl carbons at δ. (Me-8) and.0(me-ʹ). n the basis of the above account the structure of has been characterized as n- tricosanyl n- octadec-9-enoate ' CH (CH ) CH=CH (CH ) C-CH (CH ) CH n-tricosanyl oleate () Compound, designated as Tetrahydroxynaphthyl geranilan-0'-al- '-oate, was obtained as pale yellow crystals from petroleum etherchloroform (:) eluants. Its UV absorption maxima at 9 and 9 nm indicated aromatic nature of the molecule. It showed IR absorption bands for hydroxyl groups (8, cm - ), ester function (0 cm - ), aldehydic group (80 cm - ) and aromatic ring (,, 0 cm - ). n the basis of mass and C NMR spectra, the molecular ion peak of was determined at m/z 0 consistent with the molecular formula of a naphthyl ester, C 0H. The ion peaks arising at m/z 9, 9 [C fission] + suggested that a C0 unit was linked to a tetrahydroxy naphthyl ring. The H NMR spectrum of exhibited four one-proton doublets at δ.8 (J= 8.0 Hz),. (J = 8.),. (J= 8.0 Hz) and. (J = 8. Hz) assigned correspondingly to ortho-coupled aromatic H-, H-, H- and H- protons. A two-proton doublet at δ.0 (J =. Hz) and two one-proton multiplets at δ.8 and.8 were ascribed to methylene H -ʹ attached to the ester group and to methine H-ʹ and H-ʹ protons, respectively. The other methylene protons appeared as two-proton multiplets at δ.,. and.8. Two three-proton doublets at δ. (=.0 Hz) and. (J =.0 Hz) and a one-proton doublet at δ 9.9 (J=.8 Hz) were due to secondary C-8ʹ and C-9ʹ methyl and C-0ʹ aldehydic protons, respectively. The C NMR spectrum of displayed signals for ester carbon at δ 9.8 (C-ʹ), aldehydic carbon at δ 9.8 (C- 0ʹ), aromatic carbons between δ..8, methyl carbons at δ. (C-8ʹ) and.88 (C-9ʹ) and methylene and methine carbons between δ.0.8. n the bans of the foregoing discussion, the structure of has been elucidated as,,,8- tetrahydroxynaphthyl geranilan-0ʹ-al ʹ-oate. This is a new naphthyl ester. 0' CH ' ' H ' H H Tetrahydroxynaphthyl geranilan-0'-al-'-oate () C ' ' ' 8' 9'

4 Compound, designated as lanostdienyl docosanoate, was obtained as a colourless crystalline mass from chloroform eluants. It responded positively to Liebermann-Burehardt test for triterpenoids and showed distinctive IR absorption bands for ester function ( cm - ), unsaturation ( cm - ) and long aliphatic chain ( cm - ). n the basis of mass and C NMR spectral data, the molecular ion peak of was determined at m/z 8 consistent with the molecular formula of a triterpenic ester C H 9. The ion peaks arising at m/z [M ] +, [CH (CH ) 0C] + and 9 [CH (CH ) 0C] + suggested that a C fatty acid was esterified with a triterpenol. The H NMR spectrum of showed two one-proton multiplets in the deshielded region at δ. and.0 assigned to vinylic H- and H- protons, respectively. A oneproton double doublet at δ.0 with coupling interactions of. and.0 Hz was ascribed to β-oriented oxygenated methine H- proton. A three-proton broad singlet at δ.98 was due to C- methyl protons located on the vinylic C-0 carbon. Five threeproton broad singlets at δ.,.,.9,. and 0.9 and two three-proton doublets at δ 0.9 (J =. Hz) and 0.9 (J =. Hz) were associated correspondingly to tertiary C-9, C-8, C-0, C- 9, C-8 and secondary C- and C- methyl protons. A threeproton triplet at δ 0. (J =. Hz) was due to primary C-ʹ methyl protons. The C NMR spectrum exhibited signals for ester carbon at δ 9.9 (C- ), vinylic carbons at δ.8 (C-),. (C-),. (C-0) and.9 (C-), oxygenated methine carbon at δ 8. (C-) and methyl carbons from δ 8. to.8. The H and C NMR spectral data of the triterpenic skeleton of were compared with the reported data of lanostenes [0-]. n the basis of spectral data analysis, the structure of has been established as lanost-,0 ()-dien-α-olyl n-docosanoate. This is a new triterpenic ester Hz),.9 (J = 8. Hz) and.8 (J = 8. Hz), assigned to aromatic meta-coupled H-ʹ, and ortho-coupled H-ʹ, H-8ʹ and H- 9ʹ protons, respectively. A one-proton double doublet at δ. with coupling interactions of.0 and 8. Hz was ascribed to ortho-, meta-coupled H-ʹ proton. A one-proton triple doublet at at δ.8 (J=.,.,. Hz), a broad singlet at δ.0 and a doublet at δ. integrating for two protons each were attributed correspondingly to β-oriented carbinol H-, oxygenated methylene H - and hydroxymethylene H -0 protons. Two three-proton broad singlets at δ. and., a three-proton doublet at δ. (J =.0 Hz) and a three-proton triplet at δ 0.9 (J =.0 Hz) were associated with the tertiary C- and C-9, secondary C-8 and primary C- methyl protons, respectively of labdane-type diterpenoid. The other methine and methylene protons resonated in the range of δ The C NMR spectrum of exhibited signals for keto carbons at δ 9. (C-ʹ). (C-ʹ), ester carbon at δ. (C-ʹ), aromatic carbons from δ.0 to.0 oxygenated methylene carbons at δ. (C-) and. (C-0), carbinol carbon at δ. (C-) and methyl carbons at δ.9 (C-),. (C-),.0 (C-8) and.9 (C- 9). The H and C NMR values of labdanyl unit of were compared with those of the related diterpenes [, ]. n the basis of the foregoing account the structure of has been established as labdan-α,,0- triol--(0ʹ,ʹ- dihydroxy anthraquinone -ʹ-oate). This is a new labdane type diterpene linked with an anthraquinone moiety. 0 8 ' 8' 0 Trihydroxy labdanyl anthraquinoate () 9 8 ' ' Compound, named β-sitosterol --arabinobenzoate, was C (CH ) 0 - CH obtained as a colorless crystalline mass from petroleum etherchloroform (:) eluents. It gave positive tests for glycosides and showed characteristic absorption bands for hydroxyl groups (8, Lanostdienyl-n-docosanoate () 0 cm - ), ester function ( cm - ), unsaturation (8 cm - ) and aromatocity (0, 0 cm - ). n the basis of mass and C Compound, named trihydroxy labdanyl anthraquinoate, was NMR spectra, the molecular weight of was established as m/z 0 obtained as a brown coloured mass from chloroform-methanol consistent to the molecular formula of a steroidal glycosidic ester, (9:) eluents. It gave positive tests for phenols and had UV C H. The ion peak arrising at m/z due to removal of the absorption maxima at 8 and 0 nm for anthraquinones. Its IR glycosidic chain indicated that β-sitosterol unit was linked as an spectrum exhibited characteristic absorption bands for hydroxyl aglycone to the sugar unit. The H NMR spectrum of displayed groups (, 8, 0 cm - ), ester function ( cm - ) and five deshielded signals as one-proton double doublets at δ.8 (J = aromatic ring (0,, 0 cm - ). n the basis of mass and 8.0,. Hz) and. (J =.,. Hz) and three one-proton C NMR spectra, the molecular ion peak of has been established multiplets at δ.,. and.0 assigned to five aromatic at m/z 9 consistent with the molecular formula of an protons. A one- proton double doublet at δ. with coupling anthraquinone unit linked to C 0 compound, C H 8. The ion interactions of.0 and. Hz was ascribed to vinylic H- proton. peaks arising at m/z [C ʹ - fission] +, [M ] + and 8 A one-proton broad multiplet at δ.0 with half width of 8. Hz [C fission] + indicated that dihydroxy anthraquinonoate unit was accounted to oxygenated methine H-α proton. Two threeproton broad singlets at δ 0. and.0, four three-proton doublets was linked with the diterpenoid. The H NMR spectrum of exhibited four one-proton doublets at δ.8 (J =.0 Hz),. (J = at δ 0.9 (J=. Hz), 0.89 (J=. Hz), 0.8 (J=. Hz) and 0.8 ~ ~ H H C H ' ' ' ' ' ' ' H ' 0' 9' H

5 (J=. Hz) were associated correspondingly to tertiary C-8 and C- 9, secondary C-, C- and C- and primary C-9 methyl protons, all attached to saturated carbons. A one-proton doublet at δ.0 (J =. Hz) was due to anomeric H-ʹ proton. The other sugar protons appeared as a one-proton double doublet at δ. (J =.,. Hz), two one-proton multiplets at δ.9 and. and a twoproton broad singlet at δ.. The other methine and methylene protons resonated from δ.9 to.09. The C NMR spectrum of exhibited signals for aromatic carbons from δ 8. to.9, ester carbon at δ 9.0 (C- ), vinylic carbons at δ.8 (C-) and.0 (C-), oxygenated methine carbon at δ.0 (C-), anomeric carbon at δ 0.8 (C- ), other sugar carbons from δ.8 to.9 and steroidal carbons between δ The H NMR and C NMR spectral data of the steroidal nucleus were compared with other stigmastene- type molecules [-]. The presence of H- signal in the deshielded region at δ. and C-ʹ signal at δ.8 suggested location of the phenyl ester at C ʹ. n the basis of these evidences the structure has been formulated as stigmast--en-β--d-arabinopyranosyl-ʹ-benzoate. This is a new steroidal glycosidic ester. H ' ' H ' ' '' C '' '' '' ' '' '' β-sitosteryl--β-d-arabino-'-benzoate () Compound, named lanostdienyl arabinolinoleiate, was obtained as a colorless crystalline mass from chloroform- methanol (99:) Eluants. It gave positive tests for glycosides and had IR absorption bands for hydroxyl groups (, 8 cm - ), ester function ( cm - ), unsaturation ( cm - ) and long aliphatic chain ( cm - ). n the basis of mass and C NMR spectra, the molecular ion peak of was determined at m/z 8 [M + H] + consistent with the molecular formula of a tetracyclic triterpenic glycosidic ester CH89. The H NMR spectrum of exhibited six one-proton deshielded multiplets at δ.,.,.0,.,.08 and.98 assigned to vinylic H-, H-, H-9ʹʹ, H-0ʹʹ, H-ʹʹ and H-ʹʹ protons, respectively. A one-proton double doublet at δ. with coupling interactions of. and 9. Hz was ascribed to α-oriented oxygenated H- methine proton. Seven three-proton broad singlets at δ.8,.,.,.,.8,. and 0.9 were ascribed to C- and C- methyl protons located on the vinylic C- carbon and to tertiary C-9, C-8, C-0, C-9 and C-8 methyl protons, respectively. A three- proton doublet at δ 0.98 (J=.8 Hz) and a three-proton triplet at δ 0.89 (J=. Hz) were associated with correspondingly to secondary C- and primary C-8ʹʹ methyl protons. A two-proton triplet at δ.09 (J =. Hz) was due to C- ʹʹmethylene protons adjacent to the ester group. The other methine and methylene protons appeared from δ.80 to.. A one-proton doublet at δ. (J =. Hz) was accounted to anomeric H-ʹ proton. The other sugar protons appeared in the range of δ... The C NMR spectrum of showed signals for vinylic 8 ~ ~ carbons from δ.8 to.8, ester carbon at δ.0 (C ʹʹ), oxygenated methine carbon at δ.0 (C-), anomeric carbon at δ 0.8 (C-ʹ), other sugar carbons between δ and methyl carbons in the range of δ The shifting of H-ʹ signal as a double doublet at δ. (J =.,.8 Hz) in the deshielded region in the H NMR spectrum and C-ʹ at 8. in the C NMR spectrum suggested the location of the ester linkage at C. The H and C NMR spectral data of the triterpenic skeleton of were compared with the reported data of lanostenes [0-]. n the basis of spectral data analyses, the structure of has been elucidated as lanost-, -dien-β-ol β--d- arabinopyranosyl-ʹn octadec -9ʹʹ, ʹʹ-dienoate. This is a new lanostenyl glycosyl ester. H ' H ' ' ' '' 9'' 0'' '' ' 8'' C (CH ) CH=CH - CH - CH=CH - (CH ) -CH Lanostdienyl arabinolinoleiate (). Conclusion Phytochemical investigation of a methanolic extract of the fruits of Cuminum cyminum on subjection to silica gel column chromatography led to the isolation of a variety of chemical constituents including acyl and phenolic esters, triterpenoids and a steroidal glycosidic ester. All the compounds are isolated from the fruits for the first time.. Acknowledgement The authors are thankful to the Head, SAIF, Central Drug Research Institute, Lucknow for recording mass spectral data of the compounds.. References. Joshi SG. Medicinal plants: Family Apiaceae. xford and IBH Publishing Co. Pvt. Ltd, Delhi, Mhaskar KS, Blatter E, Caius JF. editors. In Kiritikar and Basu's Illustrated Indian Medicinal Plants. New Delhi: Satguru Publications 000,.. Perry LM. Medicinal Plants of East and Southeast Asia. The MIT Press, Massachusetts and London, Al Yahya M, Collpharm A. Phytochemical studies of plant used in traditional medicine in Saudi Arabia. Fitoterapia 98; ():9-8.. Agrawal S. Volatile oil constituents and wilt resistance in cumin (Cuminum cyminum L.). Curr Sci Ind 99; : 8.. Bettaieb I, Bourgou S, Wannes WA, Hamrouni I, Limam F, Marzouk B. Essential oils, phenolics and antioxidant activities of different parts of cumin (Cuminum cyminum L.). J Agric Food Chem 00; 8: Eikani MH, Goodarznia I, Mirza M. Supercritical carbon dioxide extraction of cumin seeds (Cuminum cyminum L.). Flav Frag J 999; :9.

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