Identification of new compounds from Fumaria parviflora Lam .

Article history: Received on: 21/02/2017 Accepted on: 27/03/2017 Available online: 30/04/2017 Fumaria parviflora Lam. (Fumariaceae) is a traditional medicinal herb chiefly used as a blood, skin and liver related disorder along with several human ailments like abdominal cramps, diarrhea, fever, antidyspeptic, cholagogue, diaphoretic, diuretic, laxative, sedative, tonic and syphilis. Phytochemical investigation of a methanolic extract of the plant of the plant led isolation of six new compounds characterized as 2hexadecanoxybenzyl alcohol (2), 2-O-β-D-arabinopyranosyloxy-2'-O-β-D-arabinopyranosyl-(8'''-geranilanyl) benzoate (3), n-tetracosanoylO-β-D-arabinopyranoside (4), n-tetracosanoyl O-β-D-arabinopyranosyl-(2ʹ→1′′)O-β-Darabinopyranoside (5), 2,6,7trihydroxynaphthyl-2-O-β-D-glucopyranosyl-(6a→1b)-O-β-Dglucopyranosyl-(6b→1c)-O-β-D-glucuranopyranosyl-(2c→1d)-O-β-D-glucuranopyranosyl-2d-n-octadecanoate (6) and 2-hydroxy benzoic acid-2-O-β-D-arabinopyranosyl-(2a→1b)-O-β-D-arabinopyranosyl-(2b→1c)-O-β-Darabinopyranosyl-(2c→1d)-O-β-D-glucuranopyranosyl-2d-(2',6',10',14'-tetramethyl)-n-hexadecan-1'-oate (7) along with a known fatty acid ntriacontanoic acid (1). The structural elucidations of the compounds were established on the basis of spectral data analysis and chemical means.


General
UV spectra were measured with a Lambda Bio 20 Spectrophotometer (Perkin Elmer, Rotkreuz, Switzerland) in methanol.The 1 H (400 MHz) and 13 C (100 MHz) NMR spectra were recorded on a Bruker ARX-Spectrometer (Rheinstetten, Baden-Wurttemberg, Germany), using CDCl 3 and DMSO-d 6 as solvents and TMS (Fluka analytical, Sigma-Aldrich, Netherland) as an internal standard.Melting points were determined by a thermoelectrically heated Perfit melting point apparatus (Ambala, India) without correction.Infra Red (IR) spectra were recorded using KBr pellets with a Jasco FT-IR-5000 Spectrometer (FTS 135, Kawloon, Hong Kong).Mass-spectrometric detection was carried out on ESI MS (Q-TOF-ESI) (Waters Corp., Manchester, UK), an electrospray-ionisation (ESI) technique with positive ionization mode.Column chromatography was performed on silica gel (Qualigens, Mumbai, India), 60-120 mesh and solvents taken were purchased from Merck Specialties (E.Merck, Pvt. Ltd.New Delhi, India).Pre-coated Aluminum TLC plates of silica gel 60 F 254 (Merck, Darmstadt, Germany) were used to run and spots were visualized by exposure to iodine vapors, and UV radiations and spraying with anisaldehyde-sulphuric acid solution.

Plant material
The F. parviflora whole plant was collected from the herbal garden of Jamia Hamdard, New Delhi and identified by Prof. Javed Ahmad, In-charge of the herbal garden.A specimen voucher of the drug was deposited in the herbarium of the Faculty of Pharmacy with a reference number PRL-JH/2011/05.

Preparation of extract and isolation
The dried F. parviflora whole plant (2.5 kg) was coarsely powdered and extracted with methanol for 48 h using a Soxhlet extractor in hot and cold cycle of interval of 5-6 hrs.The extract was dried under reduced pressure to obtain a dark brown residue (380 g).The extract was partioned with hexane (750 ml X 3) and chloroform (500 ml X 3) and excluded.The remaining portion of extract (80 g) was dissolved in minimum amount of methanol and adsorbed mechanically by heating on water bath with column grade silica gel (60-120 mesh) to obtain a slurry that was chromatographed over silica gel column loaded in chloroform and the eluants of each fraction were examined by precoated Aluminum TLC plate.The column was eluted with gradient mixtures of chloroform-methanol (99:1, 97:3, 19:1, 93:7, 9:1, 3:1, v/v) to isolate the compounds 1-7.
Further a spot of TLC chromatogram of methanolic extract was scraped out, sonicated to 2-3 min at 30 ºC in MS grade methanol and filtered first whatman's filter paper and then 0.2 sized µ filter for Mass spectrometer.Its molecular weight and m/z fragments were analyzed and compared with the same R f of eluants of column which is showed in Fig. 1.

RESULTS AND DISCUSSION
Compound 1 was a known fatty acid identified as ntriacontanoic acid (melissic acid) (Chibnall et al., 1933).

13C
NMR spectral data, the molecular ion peak of 7 was established at m/z 960 consistent with the molecular formula of a salicyl tetraglycosidic ester (C 47 H 76 O 20 ).An ion peak generating at m/z 137 [O-1a fission, C 6 H 4 (O)COOH] + indicated the location of salicylic unit at one of the terminal of the molecule.The ion fragments produced at m/z 295 [O-C 1 fission, C 20 H 39 O] + , 427 [O-1d fission, C 20 H 39 O-C 5 H 8 O 4 ] + , and 401 [O-1c fission, C 6 H 5 (COOH)-C 5 H 8 O 5 -C 5 H 8 O 4 ] + , suggested the diterpenic unit at another terminal and presence of four C 5 sugar units in the molecule.