986 Spectral Assignments and Reference Data Received: 29 October 2007 Revised: 30 May 2008 Accepted: 30 May 2008 Published online in Wiley Interscience: 12 August 2008 (www.interscience.com) DOI 10.1002/mrc.2277 New terpenoids from Stachys parviflora Benth Viqar Uddin Ahmad, a∗ Saima Arshad, a Sadia Bader, a Shazia Iqbal, a Afsar Khan, a Saleha Suleman Khan, a Javid Hussain, a Rasool Bakhsh Tareen b and Amir Ahmed a,c Two new triterpenoidal saponins were isolated from the n-butanolic extract of Stachys parviflora (Lamiaceae). Their structures were elucidated on the basis of spectral data as stachyssaponin A; 3β , 15α, 19α, 21β , 22α-pentahydroxyolean-12-ene-28- oic acid 3-O-{α-L-rhamnopyranosyl-(1 → 3)-β -D-glucopyranoside}-22-O-{α-L-arabinofuranosyl-(1 → 3)-β -D-glucopyranoside} (1) and stachyssaponin B; 2β ,3β , 15α, 21β -tetrahydroxyolean-12-ene-28-oic acid 2-O-[α-L-arabinofuranoside]-3, 21-bis-O-[β - D-glucopyranoside] (2). Copyright c  2008 John Wiley & Sons, Ltd. Keywords: NMR; 1 H-NMR; 13 C-NMR; 2D NMR; triterpenoidal saponins; Stachys parviflora Introduction As part of a program to assess the chemical and biological diversity of indigenous medicinal plants, we undertook the investigation of Stachys parviflora which is well known for its curative properties, especially in the treatment of cramps, joint pains, falling sickness, and abscesses caused by guinea worm. [1] We had previously reported the isolation of saponin, flavonoid, and phenethyl glycoside from the plant. [2,3] The present article details the structural elucidation of two new saponins. Experimental Methods CC was done using silica gel, 70 – 230 mesh. Flash chromatography was carried out using silica gel 230–400 mesh. thin layer chromatography (TLC) was carried out on E. Merck silica gel plates using the indicated solvents: B:A:W = 12:3:5(n-butanol-AcOH- water); and detected at 254 nm, and by ceric sulfate reagent. The IR and UV Spectra were recorded on a Jasco-320-A and Hitachi- UV-240 spectrophotometers, respectively. Optical rotations were measured on a Jasco-DIP-360-digital polarimeter using a 10-cm cell-tube. FAB-MS were recorded on a double-focusing Varian MAT- 312 spectrometer. The 1 H- and 13 C-NMR spectra were recorded on a Bruker AMX-400 spectrometer in CD 3 OD. The 2D-NMR spectra were recorded on Bruker AMX-400 spectrometer. Chemical shifts in parts per million (δ), relative to tetramethylsilane as an internal standard, and scalar coupling were reported in Hertz. The pulse conditions were as follows: for 1 H-NMR spectra, spectrometer frequency (SF) 400.032 MHz, acquisition time (AQ) 2.281 s, number of transients (NS) 128, receiver gain (RG) 812.7, temperature (TE) 300 K, dwell time (DW) 69.6 μs, per scan delay (DE) 10 μs, dummy scans (DS) 0, F 1 2431.15 Hz, F 2 – 1.77 Hz; for the 13 C-NMR spectra SF 100.613 MHz, AQ 0.6 + 26 s, NS 25 000, RG 16 384, TE 300 K, DW 19.1 μs, DE 20 μs, DS 2, F 1 23 157.50 Hz; for the COSY 45 ◦ spectra SF 400.03 MHz, NS 32, DS 4, pulse (P1) 5.70 μs, P2 2.70 μs, TE 300 K, RG 267.4, DW 145.6 μs, DE 10 μs; for the NOESY experiments SF 300.133 MHz, NS 64, DE 305 μs, pulse width (PW) 0.0, F 1 2903.69 Hz, F 2 2903.69 Hz; for the HMQC spectra, SF 400.032 MHz, AQ 0.1491 s, NS 32, DS 16, DE 10 μs, DW 145.6 μs, RG 6502, TE 300 K, F 1 1688.52 Hz, F 2 3071.71 Hz; for the HMBC spectra SF 400.032 MHz, AQ 0.1491 s, RG 7296.2, NS 128, DW 145.6 μs, DS 16, DE 10 μs, TE 300 K, F 1 3212.30 Hz, F 2 23 236.21 Hz. Plant material The aerial parts of the plant S. parviflora Benth. (Lamiaceae) (18 kg) were collected from Quetta, Balochistan, Pakistan, in 2002 and identified by one of the authors (RBT). A voucher specimen (no. 1572) has been deposited at the herbarium of the Botany Department, Balochistan University, Quetta. Extraction and isolation The air-dried plant material (18 kg) was chopped and exhaustively extracted with n-hexane and then with methanol for a period of 30 days (3 times each) at room temperature. The extracts were evaporated on a rotary evaporator under reduced pressure to obtain crude extracts. The resulting methanol extract (250 g) was suspended in water and successively partitioned to provide n-hexane (80 g), ethyl acetate (40 g), n-butanol (85 g), and aqueous fraction (45 g). The n-butanolic extract was subjected to vacuum liquid chromatography (VLC) using CHCl 3 , CH 3 OH, and H 2 O in increasing order of polarity, and then subjected to CC using flash and column silica with a gradient of CH 3 OH in CHCl 3 and H 2 O in methanol up to 100% H 2 O obtaining four major fractions. One of the fractions eluted with 70 : 30 : : CHCl 3 : methanol was then subjected to Sephadex LH-20 and eluted with H 2 O: MeOH as a solvent gradient, and the resulting fraction was then ∗ Correspondence to: Viqar Uddin Ahmad, International Center for Chemical and Biological Sciences, H.E.J. Research Institute of Chemistry, University of Karachi, Karachi-75270, Pakistan. E-mail: vuahmad@cyber.net.pk a International Center for Chemical and Biological Sciences, H.E.J. Research Institute of Chemistry, University of Karachi, Karachi-75270, Pakistan b Department of Botany, Balochistan University, Quetta, Pakistan c Pharmaceutical Division,PCSIRLaboratoriesComplex,Karachi-75280,Pakistan Magn. Reson. Chem. 2008, 46, 986–989 Copyright c  2008 John Wiley & Sons, Ltd.