There are about 240 species of the Gentiana genus grown in China, Gentiana algida Pall. is broadly distributed in the alpine zone of the world. In China, it is a little-investigated medicinal plant, that grows mainly in high cold areas such as Tibet, Sichuan, Xinjiang, Qinghai, and Jilin Provinces [1]. To the best of our knowledge, no work has been reported on the chemical constituents of Gentiana algida. As part of our continuing research on Gentiana algida Pall. [2], we have isolated six compounds from G. algida for the first time. The air-dried plants of G. algida were bought from Kumbum Monastery Tibetan Hospital in Qinghai Province of China in October of 2009 and was identified by Prof. Guo-Liang Zhang, Lanzhou University. The powdered G. algida (4.5 kg) was extracted by ethanol (30 L 4, each extraction lasting 7 days) at room temperature. The EtOH extract was concentrated under reduced pressure to give a residue (798 g), which was sequentially partitioned with petroleum ether (90–100 C) (1.0 L 4), EtOAc (1.0 L 4), and n-BuOH (1.0 L 4). The n-BuOH fraction (238 g) was chromatographed on a macroporous resin column (6.5 150 cm) and eluted with H2O–EtOH (90:10, 75:25, 50:50, 25:75, 5:95, 0:100, v/v) gradientwise and acetone to yield seven fractions (Fr. 1–Fr. 7) according to TLC analysis. Fraction 2 (3 g) was re-treated on a polyamide (60–100 mesh, 100 g) column and eluted with H2O–MeOH (5:1, v/v) to obtain 1 (25 mg), then futher eluted with H2O–MeOH (1:1, v/v) to obtain 2 (10 mg). Fraction 4 (0.5 g) was re-treated on a LH-20 (20–150 m, 30 g) column and eluted with H2O–MeOH (1:1, v/v) to obtain 3 (15 mg). Fraction 5 (1.5 g) was chromatographed on a silica gel (200–300 mesh, 50 g) column and eluted with CHCl3–MeOH (10:1, v/v) to obtain 4 (15 mg). Fraction 7 (1 g) was re-treated on a silica gel (200–300 mesh, 30 g) column and eluted with PE–EtOAc (10:1, v/v) to obtain coarse crystals of compounds 5 (30 mg) and 6 (20 mg), then recrystalled with petroleum ether to obtain 5 (15 mg) and 6 (10 mg). Isoscoparin (1), yellow powder, C22H22O11. ESI-MS m/z 461 [M – 1] +. 13C NMR (100 MHz, DMSO-d6, TMS, , ppm): 163.8 (C-2), 103.2 (C-3), 182.0 (C-4), 160.7 (C-5), 108.9 (C-6), 163.4 (C-7), 93.8 (C-8), 156.3 (C-9), 103.3 (C-10), 121.4 (C-1 ), 120.4 (C-2 ), 148.1 (C-3 ), 150.8 (C-4 ), 115.8 (C-5 ), 110.2 (C-6 ), 73.1 (C-1 ), 70.3 (C-2 ), 79.0 (C-3 ), 70.6 (C-4 ), 81.6 (C-5 ), 61.5 (C-6 ), 56.0 (OCH3-3 ) [3]. Gentiopicroside (2), white powder, C16H20O9. ESI-MS m/z 379 [M + Na] +. 1H NMR (400 MHz, DMSO-d6, TMS, , ppm, J/Hz): 7.38 (1H, s, H-3), 5.62 (1H, m, H-6), 5.60 (1H, d, J = 3.0, H-1), 5.19 (1H, m, H-10), 5.15 (1H, m, H-10), 5.04 (1H, m, H-7), 4.96 (1H, m, H-7), 4.48 (1H, d, J = 8.0, H-1 ), 3.14 (1H, t, J = 8.8, H-9). 13C NMR (100 MHz, DMSO-d6, TMS, , ppm): 96.7 (C-1), 149.1 (C-3), 103.5 (C-4), 125.1 (C-5), 116.4 (C-6), 70.1 (C-7), 134.1 (C-8), 44.5 (C-9), 116.2 (C-10), 163.1 (C-11), 98.9 (C-1 ), 73.0 (C-2 ), 77.4 (C-3 ), 69.4 (C-4 ), 76.7 (C-5 ), 61.3 (C-6 ) [4]. 3,6,4 -Trimethoxy-5,7-dihydroxyflavone (3), yellow powder, C18H16O7 [5]. -Sitosterol (4), white needles identical to an authentic sample. 10 -Hydroxy-7(11)-eremophilen-12,8 -olide (5), white needles, C15H22O3. EI-MS m/z 250 [M] +, 232, 126, 125, 109. 1H NMR (400 MHz, CDCl3, TMS, , ppm, J/Hz): 2.6 (1H, d, J = 14.0, H-6 ), 2.43 (1H, d, J = 14.0, H-6 ), 5.01 (1H, m, H-8), 2.15 (1H, dd, J = 7.2, 12.8, H-9 ), 1.96 (1H, dd, J = 11.2, 12.8, H-9 ), 1.81 (3H, s Me-13), 1.06 (s, Me-14), 0.84 (3H, d, J = 6.4, Me-15). 13C NMR (100 MHz, CDCl3, TMS, , ppm): 36.3 (C-1), 22.3 (C-2), 29.7 (C-3), 33.5 (C-4), 44.9 (C-5), 31.7 (C-6), 161.4 (C-7), 78.7 (C-8), 40.9 (C-9), 74.9 (C-10), 120.5 (C-11), 175.2 (C-12), 8.2 (C-13), 14.7 (C-14), 16.0 (C-15) [6].
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