苑 祥，李震源，梅麗娟，張培成*

醉馬草中黃酮類化學(xué)成分研究

苑 祥1, 2，李震源1，梅麗娟2，張培成1*

1. 中國醫(yī)學(xué)科學(xué)院 北京協(xié)和醫(yī)學(xué)院藥物研究所，天然藥物活性物質(zhì)與功能國家重點(diǎn)實(shí)驗(yàn)室，北京 100050 2. 中國科學(xué)院西北高原生物研究所，青海省藏藥研究重點(diǎn)實(shí)驗(yàn)室，青海 西寧 810008

研究禾本科芨芨草屬植物醉馬草中的化學(xué)成分。綜合運(yùn)用HP-20大孔樹脂、ODS中壓色譜、Sephadex LH-20凝膠柱色譜以及制備型高效液相色譜等方法進(jìn)行系統(tǒng)的分離純化，根據(jù)化合物的理化性質(zhì)及其波譜數(shù)據(jù)，并通過對比文獻(xiàn)報道的波譜數(shù)據(jù)，鑒定化合物的化學(xué)結(jié)構(gòu)。從醉馬草乙醇提取物中分離得到15黃酮類化合物，分別鑒定為異紅草素（1）、金絲桃苷（2）、槲皮素-3--β--吡喃葡萄糖基-(3′→-3′′′)-槲皮素-3?--β--吡喃半乳糖苷（3）、3?-甲氧基槲皮素-3--α--吡喃鼠李糖基-(1→6)-β--吡喃葡萄糖苷（4）、異鼠李素-3--α--吡喃鼠李糖基-(1→2)-β--吡喃葡萄糖苷（5）、槲皮素-3--α--吡喃鼠李糖基-(1→2)-β--吡喃葡萄糖苷（6）、槲皮素-3--β--吡喃葡萄糖基-7--α--吡喃鼠李糖苷（7）、山柰酚-3--α--吡喃鼠李糖基-(1→2)-β--吡喃葡萄糖苷（8）、山柰酚-3--α--吡喃鼠李糖基-(1→4)-β--吡喃葡萄糖苷（9）、8-甲氧基槲皮素-3--β-吡喃葡萄糖苷（10）、槲皮素-3--α--吡喃鼠李糖基-(1→2)-β--吡喃半乳糖基- 7--β--吡喃葡萄糖苷（11）、5,7,3?-三羥基-8,4?,5?-三甲氧基黃酮（12）、木犀草素-7--葡萄糖醛酸苷-6??-甲酯（13）、木犀草素-7--葡萄糖醛酸苷（14）和金圣草黃素（15）。所有化合物均為首次從醉馬草中分離得到。

醉馬草；黃酮；異紅草素；金絲桃苷；木犀草素-7--葡萄糖醛酸苷；金圣草黃素

醉馬草(Hance) Keng.為禾本科芨芨草屬Beauv.多年生草本植物。別名醉馬芨芨、醉針茅、醉針草、馬尿掃等，是我國西北草原上有名的毒草。醉馬草在我國廣泛分布于新疆、甘肅、青海、內(nèi)蒙古等地，多生于海拔1700～4200 m的高山及亞高山草原、山坡草地和田邊。醉馬草是哈薩克醫(yī)常用的藥材，具有消腫止痛、清熱解毒等功效，并對腮腺炎和關(guān)節(jié)疼痛的治療都有很好的效果[1]?；瘜W(xué)成分研究顯示，醉馬草中含有生物堿、黃酮等多種成分[2]。醉馬草具有抗腫瘤、殺蟲、抑菌等生物活性[3-5]。為了更合理地開發(fā)利用該植物資源，充分發(fā)揮應(yīng)用價值，本實(shí)驗(yàn)對醉馬草乙醇提取物中的化學(xué)成分進(jìn)行系統(tǒng)研究，利用多種色譜分離方法共分離鑒定出15個黃酮類化合物，分別為異紅草素（isoorientin，1）、金絲桃苷（hyperoside，2）、槲皮素-3--β--吡喃葡萄糖基-(3′→-3′′′)-槲皮素-3?--β--吡喃半乳糖苷[quercetin-3--β--glucopyranoside (3′→-3′′′)- quercetin-3--β--galactopyranoside，3]、3?-甲氧基槲皮素-3--α--吡喃鼠李糖基-(1→6)-β--吡喃葡萄糖苷 [3′--methylquercetin-3--α--rhamnopyranosyl (1→6)-β--glucopyranoside，4]、異鼠李素-3--α--吡喃鼠李糖基-(1→2)-β--吡喃葡萄糖苷[isorhamnetin-3--α--rhamnopyranosyl-(1→2)-β-- glycopyranoside，5]、槲皮素-3--α--吡喃鼠李糖基-(1→2)-β--吡喃葡萄糖苷[quercetin-3--α-- rhamnopyranosyl (1→2)-β--glucopyranoside，6]、槲皮素-3--β--吡喃葡萄糖基-7--α--吡喃鼠李糖苷（quercetin-3--β--glucopyranoside-7--α-- rhamnopyranoside，7）、山柰酚-3--α--吡喃鼠李糖基-(1→2)-β--吡喃葡萄糖苷[kaempferol 3-- α--rhamnopyranosyl (1→2)-β--glucopyranoside，8]、山柰酚-3--α--吡喃鼠李糖基-(1→4)-β--吡喃葡萄糖苷[kaempferol 3--α--rhamnopyranosyl (1→4)-β--glucopyranoside，9]、8-甲氧基槲皮素-3--β-吡喃葡萄糖苷（8-methoxyquercetin-3--β- glucoside，10）、槲皮素-3--α--吡喃鼠李糖基- (1→2)-β--吡喃半乳糖基-7--β--吡喃葡萄糖苷[quercetin-3--α--rhamnopyranosyl-(1→2)-β-- galactopyranoside-7--β--glucopyranoside，11]、5,7,3?-三羥基-8,4?,5?-三甲氧基黃酮（5,7,3?- trihydroxy-8,4?,5?-trimethoxyflavone，12）、木犀草素-7--葡萄糖醛酸苷-6??-甲酯（luteolin-7--glucuronide- 6??-methyl ester，13）、木犀草素-7--葡萄糖醛酸苷（luteolin-7--glucuronide，14）和金圣草黃素（chrysoeriol，15）。所有化合物均為首次從該植物中分離得到。

1 儀器與材料

Bruker AVANCE III 500型核磁共振儀（美國Bruker-Biospin公司）；Agilent 1100 LC/MSD TOF型高分辨質(zhì)譜儀（美國Agilent公司）；Agilent 1260 型高效液相色譜儀（美國Agilent公司）；Shimadzu LC-6AD 型半制備液相色譜儀（日本Shimadzu公司）；YMC ODS-A C18制備柱（250 mm×10 mm，5 μm，日本YMC公司）；Sephadex LH-20葡聚糖凝膠色譜填料（瑞典Pharmacia Fine Chemicals公司）；MCI色譜填料（日本三菱株式會社）。

醉馬草于2018年8月采自青海省海東市平安區(qū)，經(jīng)中國科學(xué)院西北高原生物研究所梅麗娟研究員鑒定為禾本科芨芨草屬植物醉馬草(Hance) Keng.。

2 提取與分離

將陰干的醉馬草全草粉碎（45 kg），然后經(jīng)80%乙醇回流提取（3×2 h），料液比1∶8，得浸膏（約3.2 kg）。浸膏經(jīng)水分散后，依次用石油醚和醋酸乙酯萃取。所得水部位（1.5 kg）加水分散，上樣HP-20大孔樹脂，使用乙醇溶液梯度洗脫（0、20%、50%、70%、95%），得到20個流分Fr. A～T。Fr. H（12.6 g）使用ODS中壓柱色譜分離，甲醇-水（0∶10～10∶0）梯度洗脫，經(jīng)HPLC分析后合并，共獲得12個亞流分（Fr. H1～H12）。Fr. H9（200 mg）經(jīng)Sephadex LH-20凝膠柱色譜（0～50%乙醇溶液梯度洗脫）、pre-HPLC（25%甲醇溶液）分離得到化合物1（12 mg）、3（16 mg）、10（9 mg）和12（7 mg）。Fr. H5（79 mg）經(jīng)Sephadex LH-20凝膠柱色譜（10%～50%乙醇溶液梯度洗脫）得到化合物2（25 mg）和14（30 mg）。Fr. H2（153 mg）經(jīng)Sephadex LH-20凝膠柱色譜（0～50%乙醇溶液梯度洗脫）、pre-HPLC（18%甲醇）分離得到化合物4（5 mg）、5（17 mg）、6（6 mg）、8（8 mg）和9（19 mg）。Fr. K（7.8 g）經(jīng)Sephadex LH-20凝膠柱色譜（0～90%乙醇溶液梯度洗脫）分離得獲得30個亞流分（Fr. K1～K30），F(xiàn)r. K5（18 mg）經(jīng)pre-HPLC（21%甲醇）分離得到化合物7（6 mg）。Fr. K9（156 mg）經(jīng)Sephadex LH-20凝膠柱色譜（25%乙醇等度洗脫）得到化合物11（26 mg）和13（15 mg）。Fr. K17（60 mg）經(jīng)pre-HPLC（28%甲醇）分離得到化合物15（11 mg）。

3 結(jié)構(gòu)鑒定

化合物1：黃色無定形粉末；ESI-MS/: 449 [M＋H]+，分子式為C21H20O11；1H-NMR (500 MHz, methanol-4): 7.34 (2H, overlapped, H-6?, 2?), 6.86 (1H, d,= 8.0 Hz, H-5?), 6.51 (1H, s, H-3), 6.45 (1H, s, H-8), 4.84 (1H, overlapped, H-1?), 4.13 (1H, t,= 9.0 Hz, H-2?), 3.83 (1H, d,= 12.0 Hz, H-6?b), 3.69 (1H, dd,= 12.0, 5.0 Hz, H-6?a), 3.43 (2H, overlapped, H-4?, 5?), 3.37 (1H, m, H-3?)；13C-NMR (125 MHz, methanol-4): 184.1 (C-4), 166.3 (C-2), 164.9 (C-7), 162.1 (C-5), 158.7 (C-9), 151.1 (C-4?), 147.1 (C-3?), 123.5 (C-1?), 120.3 (C-6?), 116.8 (C-2?), 114.1 (C-5?), 109.2 (C-6), 105.2 (C-3), 103.9 (C-10), 95.1 (C-8), 82.7 (C-5?), 80.1 (C-3?), 75.3 (C-1?), 72.5 (C-2?), 71.8 (C-4?), 62.9 (C-6?)。以上數(shù)據(jù)與文獻(xiàn)報道的基本一致[6]，故鑒定化合物1為異紅草素。

化合物2：黃色晶針[氯仿-甲醇（1∶1）]；ESI-MS/: 465 [M＋ H]+，分子式為C21H20O12；1H-NMR (500 MHz, DMSO-6): 12.66 (1H, s, 5-OH), 10.86 (1H, s, 7-OH), 9.77 (1H, s, 4?-OH), 9.19 (1H, s, 3?-OH), 7.68 (1H, dd,= 8.5, 2.0 Hz, H-6?), 7.53 (1H, d,= 2.0 Hz, H-2?), 6.82 (1H, d,= 8.5 Hz, H-5?), 6.41 (1H, d,= 2.0 Hz, H-6), 6.21 (1H, d,= 2.0 Hz, H-8), 5.39 (1H, d,= 8.0 Hz, H-1?), 5.17 (1H, d,= 4.5 Hz, 4?-OH), 4.90 (1H, d,= 5.5 Hz, 2?-OH), 4.47 (2H, m, 3?, 6?-OH), 3.65 (1H, m, H-6?b), 3.57 (1H, m, H-6?a), 3.46 (1H, m, H-5?), 3.36 (1H, m, H-4?), 3.33 (1H, m, H-2?), 3.28 (1H, m, H-3?)；13C-NMR (125 MHz, DMSO-6): 177.5 (C-4), 164.2 (C-7), 161.3 (C-5), 156.3 (C-9), 156.3 (C-2), 148.5 (C-4?), 144.9 (C-3?), 133.5 (C-3), 122.1 (C-6?), 121.1 (C-1?), 116.0 (C-5?), 115.2 (C-2?), 104.0 (C-10), 101.8 (C-1?), 98.7 (C-6), 93.6 (C-8), 75.9 (C-5?), 73.2 (C-3?), 71.2 (C-2?), 68.0 (C-4?), 60.2 (C-6?)。以上數(shù)據(jù)與文獻(xiàn)報道基本一致[7]，故鑒定化合物2為金絲桃苷。

化合物3：棕色無定形粉末，ESI-MS/: 911 [M＋H]+，分子式為C42H38O23；1H-NMR (500 MHz, methanol-4): 7.81 (1H, d,= 2.0 Hz, H-2???), 7.67 (1H, d,= 2.0 Hz, H-2?), 7.55 (1H, dd,= 8.5, 2.0 Hz, H-5?), 7.53 (1H, dd,= 8.5, 2.0 Hz, H-5???), 6.83 (1H, d,= 8.5 Hz, H-6???), 6.81 (1H, d,= 8.5 Hz, H-6?), 6.35 (2H, m, H-8, 8??), 6.15 (2H, s, H-6, 6??), 5.22 (1H, d,= 7.5 Hz, H-1????), 5.13 (1H, d,= 7.5 Hz, H-1?????), 3.81 (1H, d,= 3.0 Hz, H-4?????), 3.78 (1H, t,= 9.0 Hz, H-2?????), 3.68 (1H, dd,= 12.0, 2.0 Hz, H-6b?????), 3.60 (1H, dd,= 11.0, 6.0 Hz, H-6a?????), 3.55～3.50 (3H, m, H-6b????, 2?????, 5?????), 3.45 (2H, m, H-6a????, 2????), 3.39 (1H, t,= 9.0 Hz, H-4????), 3.31 (1H, t,= 9.0 Hz, H-3????), 3.19 (1H, m, H-5????)；13C- NMR (125 MHz, methanol-4): 179.5 (C-4′′), 179.5 (C-4), 166.0 (C-7), 166.0 (C-7′′), 163.1 (C-5′′), 163.0 (C-5), 159.0 (C-2), 158.7 (C-2′′), 158.5 (C-9′′), 158.4 (C-9), 150.0 (C-4′′′), 149.8 (C-4′), 145.9 (C-3′), 145.8 (C-3′′′), 135.7 (C-3′′), 135.6 (C-3), 123.2 (C-6′′′), 123.0 (C-6′), 122.9 (C-1′), 122.8 (C-1′′′), 117.8 (C-2′′′), 117.5 (C-2′), 116.1 (C-5′), 116.0 (C-5′′′), 105.7 (C-10), 105.6 (C-10′′), 105.4 (C-1′′′′′), 104.2 (C-1′′′′), 99.9 (C-6, 6′′), 94.7 (C-8, 8′′), 78.4 (C-5′′′′), 78.1 (C-3′′′′), 77.2 (C-5′′′′′), 75.7 (C-2′′′′), 75.1 (C-3′′′′′), 73.2 (C-2′′′′′), 71.2 (C-4′′′′), 70.0 (C-4′′′′′), 62.5 (C-6′′′′), 61.9 (C-6′′′′′)。以上數(shù)據(jù)與文獻(xiàn)報道基本一致[8]，故鑒定化合物3為槲皮素-3--β--吡喃葡萄糖基-(3′→-3′′′)-槲皮素-3?--β--吡喃半乳糖苷。

化合物4：黃色無定形粉末，ESI-MS/: 625 [M＋H]+，分子式為C28H32O16；1H-NMR (500 MHz, methanol-4): 8.05 (1H, d,= 2.0 Hz, H-2?), 7.46 (1H, dd,= 8.5, 2.0 Hz, H-6?), 6.85 (1H, d,= 8.5 Hz, H-5?), 6.33 (1H, s, H-8), 6.11 (1H, d,= 2.0 Hz, H-6), 5.83 (1H, d,= 8.0 Hz, H-1?), 5.10 (1H, d,= 1.0 Hz, H-1?), 0.79 (3H, d,= 6.5 Hz, H-6???)；13C-NMR (125 MHz, methanol-4): 179.3 (C-4), 165.6 (C-7), 163.2 (C-5), 158.3 (C-2), 158.1 (C-9), 150.5 (C-3?), 148.4 (C-4?), 134.4 (C-3), 123.3 (C-6?), 123.1 (C-1?), 115.9 (C-5?), 114.6 (C-2?), 106.0 (C-10), 102.5 (C-1?), 100.1 (C-1???), 99.6 (C-6), 94.5 (C-8), 77.9 (C-3?), 77.1 (C-5?), 75.6 (C-2?), 73.9 (C-4???), 72.4 (C-4?), 72.3 (C-3???), 70.6 (C-2???), 69.8 (C-5???), 68.3 (C-6?), 57.1 (-OCH3), 17.4 (C-6???)。以上數(shù)據(jù)與文獻(xiàn)報道基本一致[9]，故鑒定化合物4為3?-甲氧基槲皮素-3--α--吡喃鼠李糖基-(1→6)-β--吡喃葡萄糖苷。

化合物5：黃色無定形粉末，ESI-MS/: 625 [M＋H]+，分子式為C28H32O16；1H-NMR (500 MHz, methanol-4): 7.93 (1H, d,= 2.0 Hz, H-2?), 7.49 (1H, dd,= 8.5, 2.0 Hz, H-6?), 6.85 (1H, d,= 8.5 Hz, H-5?), 6.32 (1H, s, H-8), 6.12 (1H, d,= 2.0 Hz, H-6), 5.85 (1H, d,= 8.0 Hz, H-1?), 5.11 (1H, d,= 1.0 Hz, H-1?), 0.81 (3H, d,= 6.5 Hz, H-6???)；13C-NMR (125 MHz, methanol-4): 179.3 (C-4), 165.7 (C-7), 163.2 (C-5), 158.4 (C-2), 158.2 (C-9), 150.6 (C-3?), 148.3 (C-4?), 134.3 (C-3), 123.4 (C-6?), 123.3 (C-1?), 117.7 (C-5?), 116.0 (C-2?), 114.4 (C-10), 102.8 (C-1???), 100.1 (C-1?), 99.7 (C-6), 94.5 (C-8), 80.3 (C-2?), 78.8 (C-3?), 78.4 (C-5?), 73.9 (C-4???), 72.4 (C-3???), 72.3 (C-2???), 71.8 (C-4?), 69.9 (C-5???), 62.4 (C-6?), 56.9 (-OCH3), 17.4 (C-6???)。以上數(shù)據(jù)與文獻(xiàn)報道基本一致[10]，故鑒定化合物5為異鼠李素- 3--α--吡喃鼠李糖基-(1→2)-β--吡喃葡萄糖苷。

化合物6：黃色無定形粉末，ESI-MS/: 611 [M＋H]+，分子式為C27H30O16；1H-NMR (500 MHz, methanol-4): 7.62 (1H, d,= 2.0 Hz, H-2?), 7.59 (1H, dd,= 8.5, 2.0 Hz, H-6?), 6.83 (1H, d,= 8.5 Hz, H-5?), 6.36 (1H, s, H-8), 6.17 (1H, d,= 2.0 Hz, H-6), 5.06 (1H, d,= 7.5 Hz, H-1?), 4.47 (1H, d,= 1.0 Hz, H-1???), 3.76 (1H, dd,= 11.0, 1.0 Hz, H-6?b), 3.59 (1H, m, H-2???), 3.50 (1H, dd,= 9.5, 3.0 Hz, H-3???), 1.08 (3H, d,= 6.5 Hz, H-6???)；13C-NMR (125 MHz, methanol-4): 179.4 (C-4), 166.1 (C-7), 163.0 (C-5), 159.3 (C-9), 158.5 (C-2), 149.8 (C-4?), 145.8 (C-3?), 135.6 (C-3), 123.5 (C-6?), 123.1 (C-1?), 117.7 (C-2?), 116.0 (C-5?), 105.7 (C-10), 104.7 (C-1?), 102.4 (C-1???), 99.9 (C-6), 94.8 (C-8), 78.1 (C-3?), 77.2 (C-5?), 75.7 (C-2?), 73.9 (C-4???), 72.2 (C-3???), 72.1 (C-2???), 71.4 (C-4?), 69.7 (C-5???), 68.5 (C-6?), 17.9 (C-6???)。以上數(shù)據(jù)與文獻(xiàn)報道基本一致[11]，故鑒定化合物6為槲皮素-3--α--吡喃鼠李糖基-(1→2)- β--吡喃葡萄糖苷。

化合物7：黃色無定形粉末，ESI-MS/: 611 [M＋H]+，分子式為C27H30O16；1H-NMR (500 MHz, methanol-4): 7.66 (1H, d,= 2.0 Hz, H-2?), 7.53 (1H, dd,= 8.5, 2.0 Hz, H-6?), 6.82 (1H, d,= 8.5 Hz, H-5?), 6.31 (1H, s, H-8), 6.10 (1H, d,= 2.0 Hz, H-6), 5.72 (1H, d,= 7.5 Hz, H-1?), 5.16 (1H, d,= 1.0 Hz, H-1???), 0.87 (3H, d,= 6.5 Hz, H-6???)；13C-NMR (125 MHz, methanol-4): 179.4 (C-4), 165.7 (C-7), 163.1 (C-5), 158.3 (C-9), 158.1 (C-2), 149.6 (C-4?), 145.9 (C-3?), 134.6 (C-3), 123.3 (C-6?), 123.0 (C-1?), 117.3 (C-2?), 116.1 (C-5?), 105.9 (C-10), 102.6 (C-1???), 100.8 (C-1?), 99.6 (C-6), 94.4 (C-8), 77.5 (C-3?), 77.1 (C-5?), 75.7 (C-2?), 74.0 (C-4???), 72.4 (C-3???), 72.3 (C-2???), 70.9 (C-5???), 69.8 (C-4?), 62.0 (C-6?), 17.3 (C-6???)。以上數(shù)據(jù)與文獻(xiàn)報道的基本一致[12]，故鑒定化合物7為槲皮素-3--β--吡喃葡萄糖基-7--α--吡喃鼠李糖苷。

化合物8：黃色無定形粉末，ESI-MS/: 595 [M＋H]+，分子式為C27H30O15；1H-NMR (500 MHz, methanol-4): 8.03 (2H, d,= 8.5 Hz, H-2?, 6?), 6.84 (2H, d,= 8.5 Hz, H-3?, 5?), 6.33 (1H, d,= 2.0 Hz, H-8), 6.12 (1H, d,= 2.0 Hz, H-6), 5.67 (1H, d,= 7.5 Hz, H-1?), 5.16 (1H, d,= 1.0 Hz, H-1???), 3.97 (1H, m, H-5???), 3.94 (1H, m, H-2???), 3.90 (1H, dd,= 9.5, 7.5 Hz, H-3???), 3.78 (1H, m, H-6?a), 3.72 (1H, dd,= 9.5, 3.0 Hz, H-3?), 3.66 (1H, dd,= 9.5, 3.0 Hz, H-2?), 3.59 (1H, m, H-4?), 3.54 (1H, m, H-5?), 3.45 (1H, t,= 6.0 Hz, H-4???), 3.29 (1H, m, H-6?b), 0.88 (1H, d,= 6.5 Hz, H-6???)；13C-NMR (125 MHz, methanol-4): 179.5 (C-4), 165.6 (C-7), 163.2 (C-5), 161.3 (C-4?), 158.4 (C-9), 158.4 (C-2), 134.4 (C-3), 132.2 (C-2?, 6?), 123.0 (C-1?), 116.1 (C-3?, 5?), 105.9 (C-10), 102.6 (C-1???), 100.6 (C-1?), 99.6 (C-6), 94.5 (C-8), 79.6 (C-2?), 77.0 (C-3?), 75.8 (C-5?), 74.0 (C-4???), 72.4 (C-2???), 72.3 (C-3???), 70.8 (C-4?), 69.8 (C-5???), 62.2 (C-6?), 17.2 (C-6???)。以上數(shù)據(jù)與文獻(xiàn)報道的基本一致[13]，故鑒定化合物8為山柰酚-3--α--吡喃鼠李糖基-(1→2)-β--吡喃葡萄糖苷。

化合物9：黃色無定形粉末，ESI-MS/: 595 [M＋H]+，分子式為C27H30O15；1H-NMR (500 MHz, methanol-4): 8.01 (2H, d,= 8.5 Hz, H-2?, 6?), 6.85 (2H, d,= 8.5 Hz, H-3?, 5?), 6.34 (1H, d,= 2.0 Hz, H-8), 6.14 (1H, d,= 2.0 Hz, H-6), 5.71 (1H, d,= 7.5 Hz, H-1?), 5.18 (1H, d,= 1.0 Hz, H-1???), 0.90 (1H, d,= 6.5 Hz, H-6???)；13C-NMR (125 MHz, methanol-4): 179.4 (C-4), 165.6 (C-7), 163.2 (C-5), 161.3 (C-4?), 158.5 (C-9), 158.4 (C-2), 134.4 (C-3), 132.1 (C-2?, 6?), 123.1 (C-1?), 116.1 (C-3?, 5?), 106.0 (C-10), 102.6 (C-1???), 100.2 (C-1?), 99.7 (C-6), 94.5 (C-8), 80.1 (C-4?), 78.9 (C-5?), 78.4 (C-3?), 74.0 (C-4???), 72.4 (C-2???), 72.3 (C-3???), 71.8 (C-2?), 69.9 (C-5???), 62.6 (C-6?), 17.5 (C-6???)。以上數(shù)據(jù)與文獻(xiàn)報道基本一致[14]，故鑒定化合物9為山柰酚-3--α--吡喃鼠李糖基-(1→4)-β--吡喃葡萄糖苷。

化合物10：黃色無定形粉末，ESI-MS/: 495 [M＋H]+，分子式為C22H22O13；1H-NMR (500 MHz, methanol-4): 7.87 (1H, d,= 2.0 Hz, H-2?), 7.64 (1H, dd,= 8.5, 2.0 Hz, H-6?), 6.84 (1H, d,= 8.5 Hz, H-5?), 6.20 (1H, s, H-6), 5.16 (1H, d,= 7.5 Hz, H-1?), 3.86 (3H, s, -OCH3), 4.17 (1H, m, H-6?b), 3.78 (1H, m, H-6?a), 3.40～3.75 (5H, m, sugar-H)；13C-NMR (125 MHz, methanol-4): 178.4 (C-4), 164.7 (C-7), 158.6 (C-2), 158.0 (C-5), 150.3 (C-9), 148.5 (C-4?), 145.9 (C-3?), 135.7 (C-3), 129.1 (C-8), 123.0 (C-6?), 122.9 (C-1?), 117.7 (C-5?), 116.2 (C-2?), 105.3 (C-10), 100.1 (C-1?), 97.5 (C-6), 77.2 (C-3?), 75.1 (C-5?), 73.2 (C-2?), 70.0 (C-4?), 62.0 (-OCH3), 61.9 (C-6?)。以上數(shù)據(jù)與文獻(xiàn)報道基本一致[15]，故鑒定化合物10為8-甲氧基槲皮素-3--β-吡喃葡萄糖苷。

化合物11：黃色無定形粉末，ESI-MS/: 773 [M＋H]+，分子式為C33H40O21；1H-NMR (500 MHz, DMSO-6): 12.67 (1H, s, 5-OH), 7.61 (1H, dd,= 8.5, 2.0 Hz, H-6?), 7.56 (1H, d,= 2.0 Hz, H-2?), 6.85 (1H, d,= 8.5 Hz, H-5?), 6.76 (1H, d,= 2.0 Hz, H-8), 6.43 (1H, d,= 2.0 Hz, H-6), 5.66 (1H, d,= 7.5 Hz, H-1?), 5.09 (2H, m, H-1???, 1??), 0.76 (1H, d,= 6.5 Hz, H-6???)；13C-NMR (125 MHz, DMSO-6): 177.5 (C-4), 162.8 (C-7), 160.9 (C-5), 156.9 (C-2), 156.0 (C-9), 148.6 (C-4?), 144.9 (C-3?), 133.2 (C-3), 121.8 (C-6?), 121.1 (C-1?), 116.3 (C-5?), 115.1 (C-2?), 105.7 (C-10), 100.5 (C-1???), 99.7 (C-1??), 99.4 (C-6), 98.3 (C-1?), 94.3 (C-8), 77.7 (C-3??), 77.4 (C-5??), 77.2 (C-5?), 77.2 (C-2?), 76.4 (C-3?), 73.1 (C-2??), 71.9 (C-4???), 70.7 (C-2???), 70.6 (C-3???), 70.3 (C-4??), 69.6 (C-5???), 68.3 (C-4?), 61.0 (C-6??), 60.6 (C-6?), 17.3 (C-6???)。以上數(shù)據(jù)與文獻(xiàn)報道基本一致[16]，故鑒定化合物11為槲皮素-3--α--吡喃鼠李糖基- (1→2)-β--吡喃半乳糖基-7--β--吡喃葡萄糖苷。

化合物12：黃色無定形粉末，ESI-MS/: 361 [M＋H]+，分子式為C18H6O8；1H-NMR (500 MHz, methanol-4): 7.22 (1H, d,= 2.0 Hz, H-2?), 7.18 (1H, d,= 2.0 Hz, H-6?), 6.98 (1H, s, H-3), 6.30 (1H, s, H-6), 3.89 (3H, s, 5?-OCH3), 3.96 (3H, s, 4?-OCH3), 3.77 (3H, s, 8-OCH3)；13C-NMR (125 MHz, methanol-4): 181.8 (C-4), 162.8 (C-2), 156.9 (C-7), 155.9 (C-5), 153.4 (C-5?), 150.6 (C-3?), 149.6 (C-9), 139.7 (C-4?), 127.6 (C-8), 125.9 (C-1?), 107.3 (C-3), 104.6 (C-6?), 103.6 (C-10), 102.3 (C-2?), 98.9 (C-6), 61.2 (8-OCH3), 60.1 (4?-OCH3), 56.1 (5?-OCH3)。以上數(shù)據(jù)與文獻(xiàn)報道基本一致[17]，故鑒定化合物12為5,7,3?-三羥基-8,4?,5?-三甲氧基黃酮。

化合物13：黃色無定形粉末，ESI-MS/: 477 [M＋H]+，分子式為C22H20O12；1H-NMR (500 MHz, DMSO-6): 7.23 (1H, dd,= 8.5, 2.0 Hz, H-6?), 7.11 (1H, d,= 2.0 Hz, H-2?), 6.76 (1H, d,= 8.5 Hz, H-5?), 6.83 (1H, s, H-3), 6.76 (1H, d,= 2.0 Hz, H-8), 6.66 (1H, d,= 2.0 Hz, H-6), 4.99 (1H, d,= 7.5 Hz, H-1?), 3.30～4.10 (4H, overlapped, H-2?～5?), 3.54 (3H, s, -OCH3)；13C-NMR (125 MHz, DMSO-6): 182.0 (C-4), 169.2 (C-6?), 164.2 (C-2), 162.5 (C-7), 161.2 (C-5), 157.0 (C-9), 151.3 (C-4?), 146.8 (C-3?), 122.9 (C-1?), 118.9 (C-6?), 113.1 (C-2?), 112.2 (C-5?), 105.6 (C-10), 103.9 (C-3), 99.4 (C-1?), 99.1 (C-6), 94.6 (C-8), 75.4 (C-5?), 75.2 (C-3?), 72.7 (C-2?), 71.3 (C-4?), 55.8 (-OCH3)。以上數(shù)據(jù)與文獻(xiàn)報道基本一致[18]，故鑒定化合物13為木犀草素-7--葡萄糖醛酸苷-6??-甲酯。

化合物14：黃色無定形粉末，ESI-MS/: 463 [M＋H]+，分子式為C21H18O12；1H-NMR (500 MHz, DMSO-6): 7.43 (1H, d,= 2.0 Hz, H-2?), 7.41 (1H, dd,= 8.5, 2.0 Hz, H-6?), 6.88 (1H, d,= 2.0 Hz, H-5?), 6.77 (1H, d,= 2.0 Hz, H-8), 6.72 (1H, s, H-3), 6.42 (1H, d,= 2.0 Hz, H-6), 5.11 (1H, d,= 7.5 Hz, H-1?), 3.70 (1H, d,= 10.0 Hz, H-3?), 3.14 (1H, overlapped, H-2?), 3.30 (1H, overlapped, H-4?), 3.35 (1H, overlapped, H-5?)；13C-NMR (125 MHz, DMSO-6): 181.9 (C-4), 172.4 (C-6?), 164.5 (C-2), 162.9 (C-7), 161.0 (C-5), 157.0 (C-9), 150.2 (C-4?), 145.9 (C-3?), 121.2 (C-1?), 119.1 (C-6?), 116.1 (C-5?), 113.5 (C-2?), 105.3 (C-10), 103.7 (C-3), 99.6 (C-1?), 99.5 (C-6), 94.5 (C-8), 76.3 (C-3?), 74.2 (C-5?), 72.9 (C-2?), 71.9 (C-4?)。以上數(shù)據(jù)與文獻(xiàn)報道基本一致[19]，故鑒定化合物14為木犀草素-7--葡萄糖醛酸苷。

化合物15：黃色無定形粉末，ESI-MS/: 301 [M＋H]+，分子式為C16H12O6；1H-NMR (500 MHz, DMSO-6): 7.57 (1H, d,= 2.0 Hz, H-2?), 7.56 (1H, dd,= 8.5, 2.0 Hz, H-6?), 6.93 (1H, d,= 2.0 Hz, H-5?), 6.91 (1H, s, H-3), 6.51 (1H, d,= 2.0 Hz, H-8), 6.19 (1H, d,= 2.0 Hz, H-6), 3.89 (3H, s, -OCH3)；13C-NMR (125 MHz, DMSO-6): 181.8 (C-4), 164.2 (C-7), 163.7 (C-2), 161.4 (C-5), 157.3 (C-9), 150.7 (C-3?), 148.0 (C-4?), 121.5 (C-6?), 120.4 (C-1?), 115.8 (C-5?), 110.2 (C-2?), 103.7 (C-3), 103.2 (C-10), 98.8 (C-6), 94.1 (C-8), 56.0 (-OCH3)。以上數(shù)據(jù)與文獻(xiàn)報道基本一致[20]，故鑒定化合物15為金圣草黃素。

利益沖突 所有作者均聲明不存在利益沖突

[1] 顏世利, 巴杭, 阿吉艾克拜爾. 新疆醉馬草化學(xué)成分的研究 [J]. 天然產(chǎn)物研究與開發(fā), 2004, 16(5): 395-398.

[2] 張偉, 李冠, 李小飛. 醉馬草毒性成分的提取研究 [J]. 生物技術(shù), 2006, 16(6): 60-62.

[3] 張蕾, 金靖, 王嫚, 等. 醉馬草活性成分苦馬豆素抗腫瘤作用研究概況 [J]. 中國民族民間醫(yī)藥, 2017, 26(17): 25-27.

[4] Shi Y, Zhang X, Lou K. Isolation, characterization, and insecticidal activity of an endophyte of drunken horse grass,[J]., 2013, 13: 151.

[5] 張偉, 李冠, 田聰. 醉馬草提取物的抑菌特性研究 [J]. 食品科學(xué), 2007, 28(4): 92-94.

[6] 李雨蔚, 李榮嬌, 袁綠益, 等. 西藏秦艽花的化學(xué)成分研究 [J]. 中草藥, 2015, 46(14): 2052-2056.

[7] Isaza J H, Ito H, Yoshida T. A flavonol glycoside-lignan ester and accompanying acylated glucosides from[J]., 2001, 58(2): 321-327.

[8] Yang Y Y, Lin M M, Wang Y X,. Compounds from the pods of[J]., 2020, 56(2): 328-330.

[9] Sang S, Lapsley K, Jeong W S,. Antioxidative phenolic compounds isolated from almond skins (Batsch) [J]., 2002, 50(8): 2459-2463.

[10] N?rbaek R, Kondo T. Flavonol glycosides from flowers ofand[J]., 1999, 51(8): 1113-1119.

[11] Kazuma K, Noda N, Suzuki M. Malonylated flavonol glycosides from the petals of[J]., 2003, 62(2): 229-237.

[12] Ahn D, Lee E B, Ahn M S,. Flavonoids from the underground parts of[J]., 2013, 44(3): 253-256.

[13] Wu H, Dushenkov S, Ho C T,. Novel acetylated flavonoid glycosides from the leaves of[J]., 2009, 115(2): 592-595.

[14] Dongmo A B, Miyamoto T, Yoshikawa K,. Flavonoids fromand their cyclooxygenase (COX-1 and COX-2) inhibitory activities [J]., 2007, 73(11): 1202-1207.

[15] Wu T S, Furukawa H. Flavonol glycosides from[J]., 1983, 22(4): 1061-1062.

[16] Kaouadji M, Doucouré A, Mariotte A M,. Flavonol triglycosides fromperoliata [J]., 1990, 29(4): 1283-1286.

[17] 張曉峰, 胡伯林, 周炳南. 藏藥薰倒牛的活性物質(zhì)研究 [J]. 藥學(xué)學(xué)報, 1995, 30(3): 211-214.

[18] Terencio M C, Sanz M J, Fonseca M L,. Phenolic compounds fromL [J]., 1992, 47(1/2): 17-20.

[19] Ozgen U, Sevindik H, Kazaz C,. A new sulfated α-ionone glycoside fromMatthews [J]., 2010, 15(4): 2593-2599.

[20] 李玉潔, 唐浩軒, 李娟, 等. 紅樹植物瓶花木枝葉中化學(xué)成分研究[J]. 中草藥, 2019, 50(23): 5677-5682.

Flavonoids chemical constituents from

YUAN Xiang1, 2, LI Zhen-yuan1, MEI Li-juan2, ZHANG Pei-cheng1

1. State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China 2. Qinghai Provincial Key Laboratory of Tibetan Medicine Research, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810008, China

To investigate the chemical constitutes from.The compounds were isolated and purified by a combination of various chromatographic techniques including HP-20 macroporous resin, ODS MPLC, Sephadex LH-20, and semi-preparative HPLC. Their structures were identified by spectroscopic data and comparing the spectral data reported in literature.Fifteen compounds were isolated and identified from the ethanol extract ofand their structures were elucidated as isoorientin (1), hyperoside (2), quercetin-3--β--glucopyranoside (3′→O-3′′′)quercetin-3--β-- galactopyranoside (3), 3'--methylquercetin-3--α-L-rhamnopyranosyl (1→6)-β--glucopyranoside (4)、isorhamnetin-3--α-- rhamnopyranosyl (1→2)-β--glycopyranoside (5), quercetin-3--α--rhamnopyranosyl (1→2)-β--glucopyranoside (6), quercetin- 3--β--glucopyranoside-7--α--rhamnopyranoside (7), kaempferol 3--α-L-rhamnopyranosyl (1→2)-β--glucopyranoside (8), kaempferol 3--α--rhamnopyranosyl (1→4)-β--glucopyranoside (9), 8-methoxyquercetin 3--β-glucoside (10), quercetin-3--α--rhamnopyranosyl(1→2)-β--galactopyranoside-7--β--glucopyranoside (11), 5,7,3?-trihydroxy-8,4?,5?-trimethoxyflavone (12), luteolin-7--glucuronide-6''-methyl ester (13), luteolin-7--glucuronide (14) and chrysoeriol (15).All compounds are isolated from this plant for the first time.

(Hance) Keng.; flavonoids; isoorientin; hyperoside; luteolin-7--glucuronide; chrysoeriol

R284.1

A

0253 - 2670(2021)04 - 0937 - 06

10.7501/j.issn.0253-2670.2021.04.005

2020-09-15

“重大新藥創(chuàng)制”科技重大專項(xiàng)（2018ZX09711001-001）；國家自然科學(xué)基金項(xiàng)目（81773588）；博士后科學(xué)基金項(xiàng)目（2019M650579）；中國醫(yī)學(xué)科學(xué)院醫(yī)學(xué)與健康科技創(chuàng)新工程（2016-I2M-1-010）

苑 祥，主要從事天然產(chǎn)物化學(xué)成分研究。E-mail: yuanxiang@imm.ac.cn。

張培成，研究員，主要從事天然產(chǎn)物化學(xué)成分研究。Tel/Fax: (010)63165231 E-mail: pczhang@imm.ac.cn。

[責(zé)任編輯 王文倩]