Year : 2009 | Volume
: 1 | Issue : 4 | Page : 231--233
Lipid Constituents from Cissus quadrangularis Leaves
Vandana Jain1, Achal Thakur1, Lal Hingorani2, KS Laddha1,
1 Medicinal Natural Product Research Laboratory, Pharmaceutical Division, Institute of Chemical Technology, Matunga (E), Mumbai 400 019, India
2 Pharmanza Herbal Pvt. Ltd, Kaniya 388 435, Gujarat, India
Medicinal Natural Product Research Laboratory, Pharmaceutical Division, Institute of Chemical Technology, Matunga (E), Mumbai 400 019
A phytochemical investigation of Cissus quadrangularis leaves yielded five additional known compounds including eicosyl eicosanoate, tetratriacontanol, tetratriacontanoic acid, α-amyrin and β-sitosterol. The characterization of isolated compounds was achieved by chemical and spectral studies (IR, 1H NMR and Mass Spectroscopy).
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Jain V, Thakur A, Hingorani L, Laddha K S. Lipid Constituents from Cissus quadrangularis Leaves.Phcog Res 2009;1:231-233
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Jain V, Thakur A, Hingorani L, Laddha K S. Lipid Constituents from Cissus quadrangularis Leaves. Phcog Res [serial online] 2009 [cited 2021 Apr 19 ];1:231-233
Available from: http://www.phcogres.com/text.asp?2009/1/4/231/58104
Cissus quadrangularis Linn. (Family: Vitaceae) is commonly distributed thorough out the hotter parts of India and Sri Lanka and is known as asthisanhara in Sanskrit , . The plant is useful for treatment of bone fracture, diarrhoea, skin disorders and scurvy  . The plant is reported to contain high amount of dietary antioxidants that includes vitamin C, carotenoids and polyphenols. Ketosteroids, tetracyclic triterpenoid (7-oxo, onocer-8-ene-3β, 21α-diol), pentacyclic triterpenoids (δ-amyrin and β-amyrone), stillbene derivatives, β-sitosterol and lipids have been reported from aerial parts, specifically from stems ,,, . Literature survey reveals that majority of studies have been carried out on aerial parts or specifically on stems of the plant. Leaves that constitute only 5-8 % of the aerial plant parts  have not been explored for its chemical constituents, hence it was thought worthwhile to emphasize on the phytochemical characterization of C. quadrangularis leaves. The present investigation deals with the extraction, isolation and characterization of marker constituents from the hexane extract of leaves of C. quadrangularis.
Materials and Methods
Melting point is uncorrected, IR spectra were recorded in KBr and 1 H NMR spectra (300 MHz) were measured in CDCl 3 with TMS as internal standard. Column chromatography and TLC were carried out using silica gel G (60-120) at room temperature and spots were visualized by exposure to iodine vapors or spraying with Anisaldehyde sulphuric acid reagent.
Plant material was collected locally and authenticated by Prof. H. M. Pandit, Botany Dept., Khalasa College, Mumbai, a voucher specimen is deposited in Medicinal Natural Product Research Laboratory, ICT, Mumbai.
Extraction and isolation
Dried and powdered leaves (1.0 kg) were extracted using hexane by Soxhlet extraction. Wax fraction was obtained by adding acetone to the hexane extract. The precipitated wax was further purified by giving washing with cold acetone. Compounds I-III were obtained by subjecting the wax fraction to column chromatography using varying percentage of chloroform in hexane. Fractions (250 ml) were collected and monitored by TLC. Subjecting the remaining acetone soluble fraction of hexane extract to column chromatography using varying percentage of ethyl acetate in hexane resulted in isolation of compound IV and V.
Eicosyl eicosanoate (I): Fraction 25-30 of the hexane-chloroform (9:1) eluate was purified by preparative TLC (hexane-chloroform 7:3) to give I (15 mg), IR υmax cm -1 : 2916, 2850, 1735, 1460, 1180, 730. 1 H NMR δ: 0.88 (6H, t), 1.25 [(CH 2 )n, br s], 2.3 (2H, t), 4.05 (2H, t). Mass: M + 592 (C 40 H 80 O 2 ) and m/z 313, 280.
Tetratriacontanol (II): Fraction 58-66 of the hexane-chloroform (7:3) eluate afforded compound II (200 mg). IR νmax cm -1 : 3500-3200, 2920, 2850, 1465, 1180, 1050, 725. 1H NMR δ: 0.88 (6H, t), 1.25 [(CH2 ) n , br s], 3.64 (2H, t). MS m/z: 476 (M-18).
Acetylation of II: To 20 mg of II, pyridine and acetic anhydride (2 ml each) were added and the mixture was left overnight at room temperature. It was then diluted with water, extracted with diethyl ether, washed successively with dil HCl, H2O, NaHCO3 solution and water (each 2 Χ 50 ml) and then dried (Na2SO4). Removal of solvent gave a residue, 15 mg, IR υmax cm-1: 2920, 2850, 1720, 1460, 1180, 730. Mass: M + 536.
Tetratriacontanoic acid (III): Fraction (75-80), eluted from hexane-chloroform (1:1) yielded compound III, 25 mg. IR υmax cm -1 : 3600-3200, 2920, 2850, 1695, 1460, 1180, 730. 1H NMR δ: 0.88 (6H, t), 1.25 [(CH 2 ) n , br s], 2.35 (2H, t). Mass: M + 508 (C 34 H 68 O 2 ).
α-Amyrin (IV): Fraction (70-85) eluted from hexane-ethyl acetate (9:1) resulted in compound IV (100 mg), further purified by recrystallization with acetone. IR νmax cm -1 : 3450-3200, 2916, 2846, 1461, 1360, 1035, 725. 1 H NMR δ: 0.770 (3H, d), 0.793 (3H, s) 0.830 (3 H, s), 0.872 (3H, br s), 0.940 (3H, s), 0.969 (3H, s), 1.000 (3H, s), 1.072 (3H, s), 1.90 (2H, m), 3.218 (1H, q) 5.128 (1 H, t). Mass: M + 426 (C 30 H 50 O).
β-Sitosterol (V): Eluted in hexane-ethyl acetate (8:2) fractions 100-110 afforded compound V, 120 mg, identified by comparison with an authentic sample (melting point, co-TLC).
Results and Discussion
Compounds I-V were isolated by silica gel chromatography followed by preparative TLC of wax fraction and acetone soluble fraction obtained from hexane extract of C. quadrangularis leaves.
Compound I, melting point 67-69 °C, had IR absorption bands at 2916 and 2850 (long chain aliphatic group), 1735 (ester carbonyl group) and 730 cm-1 (long chain). 1 H NMR spectrum of the compound showed a triplet at δ 0.88 for terminal methyl group, a broad singlet at δ 1.25 for - (CH 2)n, a triplet at δ 2.3 for -CH2 -O-C(=O)- and a triplet at δ 4.05 for -CH 2 -C(=O)-O- proton, suggesting a long chain aliphatic ester. A [M]+ at m/z 592 suggested the molecular formula as C40H80O2 . Further fragment ions at m/z 313 and 280 suggest the acid and alcohol moiety of C 20 chain length. Thus on the basis of above data and comparing the literature values  , compound I is characterized as eicosyl eicosanoate [I]. [INLINE:1]
Compound II, melting point 90-92 °C, possessed IR bands at 3500-3200, 2920, 2850, 1050, 730 cm -1 for a long chain aliphatic alcohol. 1 H NMR displayed a triplet δ 0.88 for terminal methyl group, a broad singlet at δ 1.25 for -(CH 2 )n and methoxyl protons adjacent to hydroxyl group resonated at δ 3.64 as a triplet. A [M] + was absent in its mass spectrum instead it has showed [M-H 2 O] at m/z 476 (characteristic of long chain alcohols which gives M-18 peak), suggesting the molecular formula as C 34 H 70 O. On acetylation it afforded a mono acetate, (M) + at m/z 536(C 36 H 72 O 2 ). The compound II is characterized as tetratriacontanol. [INLINE:2]
Compound III, melting point 96-98 °C possessed IR bands at 3620-3200, 1695, 730 cm-1 for long chain aliphatic carboxylic acid. The 1 H NMR displayed a triplet at δ 0.88 for terminal methyl group and the methoxyl protons adjacent to carbonyl group resonated at δ 2.35 as a triplet. Presence of [M] + at m/z 508 (C 34 H 68 O 2 ), indicates the acid to be tetratriacontanoic acid. [INLINE:3]
Compound IV, melting point 178-182 °C, has been isolated from acetone soluble fraction. The compound was confirmed as α-amyrin by comparing the spectral data ( 1 H NMR and MS) with the literature values ,. [INLINE:4]
Compound V (β-sitosterol) melting point 130-135 °C, has been isolated from acetone soluble fraction and characterized by comparison with an authentic sample. [INLINE:5]
In conclusion, from the leaves of C. quadrangularis five additional marker constituents namely eicosyl eicosanoate, tetratriacontanol, tetratriacontanoic acid, α-amyrin and β-sitosterol have been isolated and characterized by spectral analysis and by comparison with the authentic samples. The isolated compounds can serve as important marker constituents for standardization of C.quadrangularis extract and its formulations.
The author thanks to the UGC, Delhi for providing the financial assistance.
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