川鄂金絲桃的化學成分研究

鄧憬童，郝 吉，馬遠任，周童曦，黃慧琪，王 強，楊新洲**，龐克堅

(1. 中南民族大學 藥學院，湖北 武漢 430074；2. 和田維吾爾藥業(yè)股份有限公司，新疆 和田 848200)

金絲桃屬（Hypericum）歸屬于藤黃科植物，世界約有400余種，中國有55種8亞種，分布較廣泛，各地均有，特別是貴州、陜西、甘肅等地區(qū). 金絲桃屬植物由于其化學成分結(jié)構(gòu)新穎和藥理活性豐富，近年來備受關(guān)注[1]. 目前，關(guān)于金絲桃屬植物的研究發(fā)現(xiàn)其含有多種化學成分，已經(jīng)發(fā)現(xiàn)的有黃酮類、甾體類、萜類，尤其是近年來從金絲桃屬植物中發(fā)現(xiàn)的代表性間苯三酚衍生物類化合物，如李祖強等[2]在尖萼金絲桃中發(fā)現(xiàn)了間苯三酚及黃酮類化合物. 間苯三酚結(jié)構(gòu)新穎且具有有較好的生物活性，如Tian等[3]在連柱金絲桃（H.cohaerens）中分離得到一個具有二環(huán)[5.3.1]十一烷新穎骨架的間苯三酚hypercohin A和特殊三元碳環(huán)結(jié)構(gòu)的新穎骨架化合物hypercohin K以及一系列橋環(huán)間苯三酚. 活性研究表明，hypercohin A和hypercohin K都表現(xiàn)出一定的乙酰膽堿酯酶抑制活性和細胞毒活性.

川鄂金絲桃（HypericumwilsoniiN. Robson）作為藤黃科金絲桃屬植物，主要產(chǎn)地為四川東部及南部、湖北西部，生長于海拔1 000～1 750 m的山坡灌叢、林下或草地上，其果實可入藥，具有藥用價值，用于治療目赤[4]. 鑒于從金絲桃屬其他植物中分離得到有價值的化合物，而目前關(guān)于川鄂金絲桃的化學成分未見報道，本課題組認為川鄂金絲桃有較大的研究價值和前景. 綜合利用各種分離提取技術(shù)，對采自重慶市的川鄂金絲桃中的化學成分進行提取分離純化，從川鄂金絲桃的石油醚提取部位分離得到了15種化合物，且化合物1～15首次從該植物中分離得到，包括黃酮類、甾體類、三萜類、飽和脂肪酸酯類、氧雜蒽酮類、多環(huán)多異戊烯基取代間苯三酚衍生物等.

1 實驗材料與方法

1.1 儀器和材料、試劑Bruker DRX-600 MHz核磁共振儀（德國Bruker公司），F(xiàn)innigan MAT-95型質(zhì)譜儀，Q-TOF Micro LC-MS-MS質(zhì)譜儀，Waters制備型高效液相色譜儀（美國Waters公司），Waters 2998 DAD檢測器（美國Waters公司），Waters 2707自動進樣器（美國Waters公司），COSMOSIL C18(250 mm×φ10 mm，5 μm)半制備柱（日本COSMOSIL公司），COSMOSIL 5PFP (250 mm×φ10 mm，5 μm)半制備柱（日本COSMOSIL公司），HP20 大孔樹脂（日本三菱公司），GF254200-300，0.038 5～0.054 mm薄層硅膠板（煙臺江友硅膠開發(fā)有限公司），Sephadex LH-20葡聚糖凝膠（美國Amersham公司），色譜級甲醇及色譜級乙腈（美國TEDIA試劑公司）. 其余試劑均為市售分析純.

1.2 樣品來源川鄂金絲桃于2018年8月采集于重慶市，經(jīng)中南民族大學萬定榮教授鑒定為藤黃科金絲桃屬川鄂金絲桃(Hypericum wilsoniiN. Robson).現(xiàn)存放于湖北省武漢市中南民族大學藥學院植物標本庫(No.SC0758).

1.3 提取與分離取川鄂金絲桃干燥全草21 kg，粉碎，用80%的乙醇室溫浸提(4×20 L, 3 d/次)，合并多次提取液，減壓濃縮至無醇味即得浸膏1 947 g. 將浸膏加5倍體積熱水分散后，依次用石油醚、乙酸乙酯、正丁醇萃取得到石油醚部位(HWPE，452 g)、乙酸乙酯部位674 g、正丁醇部位712 g.取石油醚粗提物(430 g)用大孔樹脂柱層析進行粗分，以水-乙醇梯度洗脫(體積分數(shù)20%→30%→40%→50%→60%→65%→70%→75%→80%→82%→85%→90%→95%)，薄層色譜板檢測合并得到9個組分Fr. A～Fr. I，其中組分Fr. H中出現(xiàn)結(jié)晶，過濾得到單體化合物3(68.9 mg)；組分Fr. B用凝膠柱層析粗分，以甲醇(加入0.1%甲酸)洗脫，合并得12個組分(Fr.Ba～Fr.Bl)，F(xiàn)r.Bf組分結(jié)晶析出化合物7(4.6 mg)，經(jīng)高效液相色譜(乙腈-水，體積比30∶70→39∶61，22 min，流動相含有0.1%的甲酸)分離得到化合物8(5.2 mg)；Fr.Bg經(jīng)高效液相色譜(乙腈-水，體積比36∶64→49∶51，8 min; 49∶51→55∶45,14 min，流動相含有0.1%的甲酸)分離得到化合物1(55.7 mg)、2(7.0 mg). Fr. G(115 g)經(jīng)過正相硅膠柱層析初步分離，以石油醚-乙酸乙酯梯度洗脫(1∶0→0∶1)，經(jīng)TLC檢測合并分為Fr.Ga～Fr.Gj共10個組分，其中Fr.Gg中有結(jié)晶析出，過濾得化合物6(57.4 mg)，F(xiàn)r.Gi中有結(jié)晶析出，過濾得化合物4(1.887 1 g). Fr.Ge經(jīng)正相硅膠柱層析（石油醚-二氯甲烷體積比1∶1）等度洗脫，得8個組分(Fr.Ge1～Fr.Ge8). Fr.Ge3經(jīng)高效液相色譜(乙腈-水，體積比84∶16→95∶5, 40 min; 95∶5→100∶0,10 min；流動相含有0.1%的甲酸)進行粗分，得到11個組分，F(xiàn)r.Ge3～11經(jīng)高效液相色譜細分得到化合物10(4.2 mg)、12(8.9 mg)、13(2.6 mg). Fr.Ge7經(jīng)高效液相色譜(乙腈-水，體積比90∶10，等度洗脫，流動相含有0.1%的甲酸)粗分為9個組分，其中Fr.Ge7-2為化合物5(77.9 mg)，F(xiàn)r.Ge7-4為化合物11(133.7 mg)，F(xiàn)r.Ge7-6為化合物9(75.8 mg).Fr.Ge7-1經(jīng)高效液相色譜分離得到化合物15(5.1 mg)，F(xiàn)r.Ge7-3經(jīng)高效液相色譜分離得到化合物14(2.2 mg).

2 結(jié)果與討論

2.1 實驗結(jié)果綜合應(yīng)用大孔樹脂柱層析、正相硅膠柱層析、葡聚糖凝膠色譜、高效液相色譜等多種分離方法從川鄂金絲桃石油醚部位（HW-PE）中分離得到15個化合物. 分別為folecitin(1)、表兒茶素(2)、β-谷甾醇(3)、β-胡蘿卜素(4)、白樺脂酮酸(5), 單棕櫚酸甘油(6)、1,7-二甲氧基-3,6-二羥基氧雜蒽酮(7)、3,4-二羥基-2-甲氧基氧雜蒽酮(8)、27-epifurohyperforin isomer 1(9)、uralione H(10)、attenuatumione E(11)、sampsonione K(12)、attenuatumione C(13)、hypersampsonone C(14)和otogirinin B(15).按結(jié)構(gòu)分為黃酮類、甾體類、三萜類、飽和脂肪酸酯類、氧雜蒽酮類、多環(huán)多異戊烯基取代間苯三酚衍生物. 見圖1.

2.2 化合物的波譜數(shù)據(jù)化合物1：C21H20O11，黃色粉末；ESI-MS(m/z)：449[M+H]+.1H NMR (600 MHz,CD3OD)δH: 7.34 (1H, d,J=1.9 Hz, H-2′), 7.31 (1H,dd,J=8.3, 1.9 Hz, H-6′), 6.91 (1H, d,J=8.3 Hz, H-5′), 6.37 (1H, d,J=2.0 Hz, H-8), 6.20 (1H, d,J=2.0 Hz, H-6), 5.35 (1H, br s, H-1″), 4.22 (1H, m, H-2″),3.75 (1H, dd,J=3.2, 9.4 Hz, H-4″), 3.42 (1H, m, H-5″), 3.33 (1H, m, H-3″),0.94 (3H, d,J=6.1 Hz, H-6″).13C NMR (150 MHz, CD3OD)δC: 179.6 (C-4),165.9 (C-7), 163.2 (C-5), 159.3 (C-9), 158.5 (C-2),149.8 (C-4′), 146.4 (C-3′), 136.2 (C-3), 122.9 (C-1′), 122.9 (C-6′), 116.9 (C-2′), 116.4 (C-5′), 105.9(C-10), 103.5 (C-1″), 99.8 (C-6), 94.7 (C-8), 73.2(C-3″), 72.1 (C-5″), 72.0 (C-4″), 71.9 (C-2″), 17.7(C-6″). 其波譜數(shù)據(jù)與文獻報道的化合物一致[5]，故鑒定化合物為folecitin.

化 合 物2：C15H14O6,白 色 粉 末；ESI-MS(m/z)：289[M-H]-.1H NMR (600 MHz, CD3OD)δH: 6.97(1H, d,J=1.6 Hz, 3-OH), 10.48 (1H, s, 6-OH), 7.38(1H, s, H-8), 6.80 (1H, s, H-5), 6.38 (1H, br s, H-4),6.33 (1H, br s, H-2), 3.84(3H, s, 7-OCH3), 3.81(3H, s, 1-OCH3).13C NMR (150 MHz, CD3OD)δC: 172.6(C-9), 162.9 (C-3), 161.7 (C-1), 159.0 (C-4a), 152.9 (C-10a), 150.2 (C-6), 145.6 (C-7), 114.4 (C-8a), 105.8 (C-8), 104.9 (C-9a), 102.3 (C-5), 95.5 (C-2), 94.9 (C-4),55.9 (1-OCH3), 55.8 (7-OCH3). 其波譜數(shù)據(jù)與文獻報道的化合物一致[6]，故鑒定化合物為表兒茶素.

圖1 化合物1～15結(jié)構(gòu)Fig. 1 Chemical structures of compounds 1～15

化合物3：C29H50O，白色粉末；EI-MS(m/z)：397[M+H]+.1H NMR (600 MHz, acetone-d6)δH: 5.31(1H, br d,J=5.1 Hz, H-6), 3.39 (1H, m, H-3), 2.21(2H, m, H-4), 2.05 (1H, m, H-12a), 1.97 (1H, m, H-7a), 1.88 (1H, m, H-1a), 1.84 (1H, m, H-2a),1.76 (1H,m, H-16a), 1.69 (1H, m, H-26),1.44～1.63 (6H, m, H-2b, H-7b, H-8, H-11, H-15a), 1.20～1.44 (5H, m, H-16b, H-20, H-22a, H-23a, H-25a), 1.03～1.20 (6H, m,H-1b, H-12b, H-15b, H-17, H-23b, H-25b), 1.01 (3H,s, H-19), 0.91～0.99 (5H, m, H-14, 22b, 21),0.79～0.88 (11H, m, H-9, 24, 24′, 28, 27), 0.72 (3H,s, H-18).13C NMR (150 MHz, acetone-d6)δC: 142.4(C-5), 121.6 (C-6), 71.7 (C-3), 57.7 (C-14), 56.9 (C-17), 51.2 (C-9), 46.7 (C-24), 43.4 (C-13), 43.1 (C-4), 40.7 (C-12), 38.3 (C-1), 37.3 (C-10), 37.0 (C-20), 34.7 (C-22), 32.8 (C-7), 32.7 (C-8), 32.6 (C-2),29.9 (C-26), 29.0 (C-16), 26.7 (C-25), 25.0 (C-15),23.8 (C-23), 21.8 (C-11), 20.2 (C-19), 19.9 (C-28),19.4 (C-27), 19.3 (C-21), 12.3 (C-24′), 12.3 (C-18).其波譜數(shù)據(jù)與文獻報道的化合物一致[7]，故鑒定化合物為β-谷甾醇.

化合物4：C35H60O6，黃色油狀；ESI-MS(m/z)：575[M-H]-.1H NMR (600 MHz, DMSO-d6)δH: 5.32(1H, t,J=4.9 Hz, H-6), 4.21 (1H, d,J=7.8 Hz, H-1′),0.95 (3H, s,H-19), 0.90 (3H, d,J=6.4 Hz, H-21), 0.82(3H, d,J=6.8 Hz, H-27), 0.81 (3H, d,J=6.4 Hz, H-26),0.79 (3H, d,J=6.8 Hz, H-29), 0.65 (3H, s, H-18).13C NMR (150 MHz, DMSO-d6)δC: 140.5 (C-5), 121.3(C-6), 100.8 (C-1′), 76.9 (C-5′), 76.8 (C-3′), 76.8 (C-3), 73.5 (C-2′), 70.1 (C-4′), 61.1 (C-6′), 56.2 (C-14),55.5 (C-17), 49.6 (C-9), 45.2 (C-24), 41.9 (C-13),39.1 (C-12), 38.3 (C-4), 36.9 (C-1), 36.3 (C-10),35.5 (C-20), 33.4 (C-22), 31.5 (C-9), 31.5 (C-8),29.3 (C-2), 28.7 (C-25), 27.9 (C-16), 25.4 (C-23),23.9 (C-15), 22.6 (C-28),20.6 (C-11), 19.8 (C-26),19.2 (C-27), 19.0 (C-19), 18.7 (C-21),11.8 (C-29),11.7 (C-18). 其波譜數(shù)據(jù)與文獻報道一致[8]，故鑒定化合物為β-胡蘿卜苷.

化合物5：C30H46O3，白色粉末；ESI-MS(m/z)：453[M-H]-.1H NMR (600 MHz, CDCl3)δH: 4.73 (1H,s, H-29β), 4.61 (1H, s, H-29α), 3.00 (1H, m,H-19),2.48 (1H, m, H-2β), 2.40 (1H, m, H-2α), 2.28 (1H, m,H-16β), 2.21 (1H, m, H-13), 1.99 (2H, m, H-15β, 22β),1.89 (1H, m, H-1β), 1.71 (1H, m, H-12β), 1.68 (3H, s,H-30), 1.62 (1H, m, H-18), 1.54 (1H, m, H-21β), 1.52(2H, m, H-6), 1.47 (1H, m, H-22α), 1.44 (4H, m, H-7,11β, 16α), 1.42 (1H, m, H-15α), 1.38 (2H, m, H-1α, 9),1.35 (1H, m, H-5), 1.34 (1H, m, H-11α), 1.22 (1H, m,H-21α), 1.06 (3H, s, H-23), 1.05 (1H, m, H-12α), 1.00(3H, s, H-24), 0.98 (3H, s, H-27), 0.96 (3H, s, H-26),0.92 (3H, s, H-25).13C NMR (150 MHz, CDCl3)δC:218.6 (C-3), 182.4 (C-28), 150.5 (C-20), 109.9 (C-29),56.5 (C-17), 55.0 (C-5), 49.9 (C-9), 49.3 (C-18), 47.5(C-4), 47.0 (C-19), 42.6 (C-14), 40.7 (C-8), 39.7 (C-1),38.6 (C-13), 37.2 (C-22), 37.0 (C-10), 34.3 (C-2), 33.7(C-7), 32.2 (C-16), 30.6 (C-15), 29.8 (C-21), 26.7 (C-23), 25.6 (C-12), 21.5 (C-11), 21.1 (C-24), 19.7 (C-6),19.5 (C-30),16.1 (C-25), 15.9 (C-26), 14.7 (C-27). 其波譜數(shù)據(jù)與文獻報道一致[9]，故鑒定化合物為白樺脂酮酸.

化合物6：C19H38O4，白色粉末；ESI-MS(m/z)：331[M+H]+.1H NMR (600 MHz, CDCl3)δH: 4.21 (1H,dd,J=11.7, 4.5 Hz, H-1α), 4.15 (1H, dd,J=11.7, 6.2 Hz,H-1β), 3.93 (1H, m, H-2), 3.70 (1H, dd,J=11.5, 4.0 Hz,H-3α), 3.60 (1H, dd,J=11.5, 5.8 Hz, H-3β), 2.35 (2H,t,J=7.6 Hz, H-2′), 0.88 (3H, t,J=7.0 Hz, H-16′).13C NMR (150 MHz, CDCl3)δC: 174.6 (C-1′), 70.4 (C-2), 65.3 (C-1), 63.4 (C-3), 34.3 (C-2′), 32.1 (C-3′),29.9 (C-5′), 29.8 (C-6′), 29.8 (C-7′), 29.8 (C-8′),29.8 (C-9′), 29.6 (C-10′), 29.5 (C-11′), 29.4 (C-12′),29.3 (C-13′), 25.1 (C-14′),22.8(C-15′), 14.3 (C-16′).其波譜數(shù)據(jù)與文獻報道一致[10]，故鑒定化合物為單棕櫚酸甘油.

化合物7：C15H12O6，黃色粉末；ESI-MS(m/z)：287[M-H]-.1H NMR (600 MHz, DMSO-d6)δH: 10.74(1H, s, 3-OH), 10.48 (1H, s, 6-OH), 7.38 (1H, s, H-8),6.80 (1H, s, H-5), 6.38 (1H, br s, H-4), 6.33 (1H, br s,H-2), 3.84(3H, s, 7-OCH3), 3.81 (3H, s,1-OCH3).13C NMR (150 MHz, DMSO-d6)δC: 172.6 (C-9), 162.9 (C-3), 161.7 (C-1), 159.0 (C-4a), 152.9 (C-10a), 150.2 (C-6), 145.6 (C-7), 114.4 (C-8a), 105.8 (C-8), 104.9(C-9a), 102.3 (C-5), 95.5 (C-2), 94.9 (C-4), 55.9 (1-OCH3), 55.8 (7-OCH3). 其波譜數(shù)據(jù)與文獻報道一致[11]，故鑒定化合物為1,7-二甲氧基-3,6-二羥基氧雜蒽酮.

化合物8：C14H10O5，黃色粉末；ESI-MS(m/z)：257[M-H]-.1H NMR (600 MHz, acetone-d6)δH: 8.24(1H, dd,J=8.0, 1.6 Hz, H-8), 7.79 (1H, dt,J=8.6, 1.8 Hz, H-6), 7.59 (1H, dd,J=8.6, 1.0 Hz, H-5), 7.43 (1H,dt,J=8.0, 1.0 Hz, H-7), 7.25 (1H, s, H-1), 3.96 (3H, s,2-OCH3).13C NMR (150 MHz, acetone-d6)δC: 176.0(C-9), 156.8 (C-5a), 146.6 (C-2), 143.0 (C-3), 141.7 (C-4), 135.0 (C-6), 134.1 (C-4a), 126.9 (C-8), 124.5 (C-7), 122.2(C-8a), 118.8 (C-5), 114.7 (C-1a), 97.0 (C-1),56.5 (2-OCH3). 其波譜數(shù)據(jù)與文獻報道一致[12]，故鑒定化合物為3,4-二羥基-2-甲氧基氧雜蒽酮.

化合物9：C35H52O5，無 色 油 狀；[α]D20+16.5(c1.0, CHCl3)；ESI-MS(m/z)：551[M-H]-.1H NMR(600 MHz, CDCl3)δH: 5.03 (1H, m, H-17), 4.98 (1H,m, H-22), 4.94 (1H, m, H-32), 4.80 (1H, dd,J=10.0,8.4 Hz, H-27), 2.98 (1H, dd,J=13.6, 10.0 Hz, H-26α),2.94 (1H, dd,J=13.6, 8.4 Hz, H-26β), 2.51 (1H, m, H-31α), 2.45 (1H, m, H-31β), 2.12 (1H, m, H-21α), 2.09(1H, m, H-11), 2.04 (1H, m, H-16α), 1.91 (1H, dd,J=13.6, 4.3 Hz, H-5α), 1.87 (1H, m, H-15α), 1.85 (2H,m, 16β), 1.77 (1H, m, H-21β), 1.70 (3H, s, H-24), 1.69(6H, s, H-34, 35), 1.64 (1H, m, H-4), 1.63 (3H, s, H-19), 1.58 (3H, s, H-20), 1.56 (3H, s, H-25), 1.52 (1H,m, H-15β), 1.42 (1H, m, H-5β), 1.29 (3H, s, H-30),1.17 (3H, s, H-29), 1.13 (3H, d,J=6.5 Hz, H-12), 1.05(3H, d,J=6.5 Hz, H-13), 1.00 (3H, s, H-14).13C NMR(150 MHz, CDCl3)δC: 209.7 (C-10), 206.6 (C-1),187.2 (C-9), 176.4 (C-7), 135.0 (C-33), 133.6 (C-23),131.3 (C-18), 124.7 (C-17), 122.5 (C-22), 120.2 (C-32), 119.3 (C-8), 93.3 (C-27), 83.9 (C-2), 71.8 (C-28),54.6 (C-6), 48.5 (C-3), 42.7 (C-4), 42.5 (C-11), 38.6(C-5), 36.6 (C-15), 29.4 (C-31), 27.5 (C-21), 27.0 (C-26), 26.6 (C-30), 26.1 (C-24), 26.0 (C-34), 25.8 (C-19), 24.9 (C-16), 23.2 (C-29), 21.7 (C-13), 20.7 (C-12), 18.3 (C-25), 18.2 (C-35), 17.8 (C-20), 14.1 (C-14). 其波譜數(shù)據(jù)與文獻報道一致[13]，故鑒定化合物為27-epifurohyperforin isomer 1.

化 合 物10：C38H50O5，無色油狀；[α]D20-92.2(c3.5, CHCl3)；ESI-MS(m/z): 587[M+H]+.1H NMR(600 MHz, CDCl3)δH: 7.59 (2H, d,J=7.4 Hz, H-12,16), 7.45 (1H, t,J=7.4 Hz, H-14), 7.28 (2H, t,J=7.4 Hz,H-13, 15), 5.15 (1H, t,J=7.2 Hz, H-35), 4.95 (1H, t,J=7.0 Hz, H-25), 4.86 (1H, t,J=6.6 Hz, H-30), 3.72(1H, d,J=9.1 Hz, H-20), 3.18 (1H, dd,J=14.5, 6.6 Hz,H-29α), 3.04 (1H, dd,J=14.5, 6.6 Hz, H-29β), 2.54(2H, d,J=7.2 Hz, H-34), 2.38 (1H, dt,J=13.5, 2.0 Hz,H-18α), 2.10 (2H, m, H-4, 24α), 2.00 (1H, dd,J=13.5,3.9 Hz, H-18β), 1.91 (1H, dd,J=13.8, 4.7 Hz, H-5α),1.86 (2H, m, H-19), 1.68 (6H, s, H-27, 37), 1.67 (1H,m, H-24β), 1.66 (3H, s, H-38), 1.64 (1H, m, H-5β),1.64 (3H, s, H-33), 1.63 (3H, s, H-32), 1.56 (3H, s, H-28), 1.27 (3H, s, H-17), 0.96 (3H, s, H-22), 0.93 (3H,s, H-23).13C NMR (150 MHz, CDCl3)δC: 206.6 (C-1), 196.7 (C-10), 194.4 (C-7), 167.3 (C-9), 137.2 (C-11), 134.8 (C-36),133.7 (C-31), 133.3 (C-26), 132.7 (C-14), 128.6 (C-12), 128.6 (C-16), 128.2 (C-13),128.2 (C-15), 127.4 (C-8), 122.1 (C-25), 120.9 (C-30), 119.7 (C-35), 87.4 (C-20), 72.3 (C-21), 72.0 (C-2), 64.9 (C-6),46.4 (C-3), 40.1 (C-5), 36.8 (C-4), 33.7 (C-18), 29.3(C-34), 27.2 (C-24), 26.2 (C-37), 26.0 (C-27), 25.7 (C-32), 25.7 (C-22), 25.0 (C-23), 24.2 (C-19), 22.8 (C-29), 18.3 (C-38), 18.1 (C-33), 18.0 (C-28), 17.4 (C-17). 其波譜數(shù)據(jù)與文獻報道一致[14]，故鑒定化合物為uralione H.

化合物11 (11)：C35H52O5，無色油狀；[α]D20+95.02(c1.25, CHCl3)；ESI-MS(m/z): 551[M-H]-.1H NMR (600 MHz, CDCl3)δH: 4.97 (2H, m, H-20,32), 4.93 (1H, m, H-25), 4.76 (1H, t,J=10.6 Hz, H-11), 2.98 (2H, dd,J=10.6, 1.9 Hz, H-10), 2.47 (1H, t,J=6.5 Hz, H-16), 2.42 (1H, dd,J=11.6, 7.2 Hz, H-19α),2.05 (1H, dd,J=13.4, 4.9 Hz, H-24α), 2.00 (1H, dd,J=12.8, 6.5 Hz, H-31α), 1.92 (1H, dd,J=12.6, 6.5 Hz,H-31β), 1.82 (1H, dd,J=13.8, 4.3 Hz, H-6α), 1.70 (1H,m, H-24β), 1.68 (1H, m, H-7), 1.66 (3H, s, H-22), 1.65(3H, s, H-27), 1.65 (2H, m, H-30), 1.64 (6H, s, H-23,34), 1.56 (3H, s, H-35), 1.53 (3H, s, H-28), 1.37 (1H,m, H-6β), 1.37 (3H, s, H-13),1.26 (1H, m, H-19β),1.24 (3H, s, H-14), 1.14 (3H, d,J=6.5 Hz, H-17), 1.14(3H, d,J=6.5 Hz, H-18), 1.12 (3H, s, H-29).13C NMR(150 MHz, CDCl3)δC: 208.9 (C-15), 205.9 (C-9),190.8 (C-4), 171.7 (C-2), 134.4 (C-21), 133.6 (C-26),131.8 (C-33), 124.5 (C-32), 122.3 (C-25), 120.7 (C-3),119.6 (C-20), 94.0 (C-11), 74.1 (C-1), 71.2 (C-12),64.0 (C-5), 47.7 (C-8), 42.3 (C-7), 40.9 (C-16),39.6 (C-6), 38.2 (C-30), 29.4 (C-19), 28.2 (C-24), 26.8(C-13), 26.8 (C-10), 26.1 (C-27), 26.1 (C-22), 25.8 (C-34), 25.4 (C-14), 24.4 (C-31), 21.2 (C-17), 21.2 (C-18), 18.2 (C-23), 18.1 (C-35), 17.9 (C-28), 14.6 (C-29). 其波譜數(shù)據(jù)與文獻報道一致[15]，故鑒定化合物為attenuatumione E.

化合物12：C38H50O5，無色油狀；[α]D20+79.56(c1.00, CHCl3)；ESI-MS(m/z): 587[M+H]+.1H NMR(600 MHz, CDCl3)δH: 7.45 (2H, d,J=7.5 Hz, H-28,32), 7.38 (1H, t,J=7.5 Hz, H-30), 7.21 (2H, d,J=7.5 Hz, H-29, 31), 5.12 (1H, t,J=7.1 Hz, H-17), 5.03(1H, dd,J=8.1, 7.0 Hz, H-22), 4.89 (1H, t,J=7.1 Hz, H-36), 4.63 (1H, dd,J=10.5, 5.8 Hz, H-3), 3.08 (1H, dd,J=14.1, 7.0 Hz, H-21α), 2.99 (1H, dd,J=14.1, 8.1 Hz,H-21β), 2.72 (1H, dd,J=13.2, 10.5 Hz, H-2α), 2.30(1H, d,J=14.0 Hz, H-11β), 2.17 (4H, m, H-11α, 15β,35), 2.08 (2H, m, H-16), 1.80 (1H, dd,J=13.2, 5.8 Hz,H-2β), 1.71 (3H, s, H-19), 1.70 (3H, s, H-38), 1.64(3H, s, H-20), 1.63 (3H, s, H-25), 1.62 (3H, s, H-24),1.55 (3H, s, H-39), 1.52 (1H, m, H-15α), 1.50 (1H, m,H-10), 1.48 (3H, s, H-33), 1.41 (3H, s, H-34), 1.36(3H, s, H-14).13C NMR (150 MHz, CDCl3)δC: 205.7(C-12), 194.1 (C-26), 193.6 (C-7), 172.7 (C-5), 137.0(C-27), 133.0 (C-23),132.7 (C-18), 132.7 (C-37),132.1 (C-30), 128.2 (C-29), 128.2 (C-31), 128.1 (C-28),128.1 (C-32), 124.6 (C-36), 123.7 (C-17), 120.0 (C-22), 115.6 (C-6),89.9 (C-3), 77.1 (C-8), 73.0 (C-13),58.9 (C-1), 49.4 (C-9), 47.9 (C-10), 37.1 (C-11), 36.8(C-15), 30.8 (C-2), 29.1 (C-35), 27.1 (C-34), 26.0 (C-24), 25.9 (C-38), 25.9 (C-19), 23.4 (C-14), 22.4 (C-21), 22.4 (C-33), 22.0 (C-16), 17.9 (C-39), 17.9 (C-25), 17.9 (C-20). 其波譜數(shù)據(jù)與文獻報道一致[16]，故鑒定化合物為sampsonione K.

化合物13：C38H50O5，無色油狀；[α]D20+20.9(c0.15, CHCl3)；ESI-MS(m/z): 587[M+H]+.1H NMR(600 MHz, CDCl3)δH: 7.45 (2H, d,J=7.5 Hz, H-28,32), 7.38 (1H, t,J=7.5 Hz, H-30), 7.22 (2H, t,J=7.5 Hz, H-29, 31), 5.15 (1H, t,J=7.0 Hz, H-17), 5.04 (1H,dd,J=7.4 Hz, H-22), 4.88 (1H, t,J=6.9 Hz, H-36),4.67 (1H, dd,J=10.7, 5.7 Hz, H-3), 3.09 (1H, dd,J=14.2, 6.9 Hz, H-21α), 2.99 (1H, dd,J=14.2, 8.2 Hz,H-21β), 2.73 (1H, dd,J=13.0, 10.7 Hz, H-2α), 2.30(1H, d,J=14.1 Hz, H-11β), 2.22 (1H, m, H-11α), 2.18(2H, m, H-35), 2.14 (3H, m, H-15α, 16), 1.80 (1H, dd,J=13.0, 5.7 Hz, H-2β), 1.72 (3H, s, H-24), 1.70 (3H, s,H-38), 1.67 (1H, m, H-15β), 1.65 (3H, s, H-20), 1.64(3H, s, H-19), 1.63 (3H, s, H-25), 1.55 (3H, s, H-39),1.50 (1H, m, H-10), 1.48 (3H, s, H-33), 1.41 (3H, s, H-34), 1.18 (3H, s, H-14).13C NMR (150 MHz,CDCl3)δC: 205.7 (C-12), 194.0 (C-7), 193.6 (C-26),172.7 (C-5), 137.0 (C-27), 133.0 (C-23),132.8 (C-18),132.7 (C-37), 132.2 (C-30), 128.3 (C-28), 128.3 (C-32), 128.1 (C-29),128.1 (C-31), 124.6 (C-36), 123.7 (C-17), 120.0 (C-22), 115.7 (C-6),89.2 (C-3), 77.1 (C-8),73.1 (C-13), 58.9 (C-1), 49.4 (C-9), 47.9 (C-10), 39.6(C-15), 36.6 (C-11), 31.0 (C-2), 29.1 (C-35), 27.1 (C-34), 26.0 (C-38), 25.9 (C-24), 25.9 (C-19), 22.5 (C-16), 22.5 (C-33), 22.4 (C-21), 21.1 (C-14), 17.9 (C-25), 17.9 (C-20), 17.9 (C-39). 其波譜數(shù)據(jù)與文獻報道一致[15]，故鑒定化合物為attenuatumione C.

化合物14：C38H50O7，無色油狀；[α]D20+9.89(c2.00, CHCl3)；ESI-MS(m/z): 617[M-H]-.1H NMR(600 MHz, CDCl3)δH: 7.58 (2H, d,J=7.4 Hz, H-12,16), 7.38 (1H, t,J=7.4 Hz, H-14), 7.26 (2H, t,J=7.4 Hz,H-13, 15), 5.48 (1H, dd,J=10.7, 4.8 Hz, H-23), 5.09(1H, d,J=2.8 Hz, H-17), 5.05 (1H, m, H-28), 4.35(1H, s, 4-OH), 4.01 (1H, d,J=2.8 Hz, H-18), 2.88 (1H,dd,J=14.8, 10.7 Hz, H-22β), 2.52 (1H, m, H-22α),2.50 (1H, m, H-33), 2.47 (1H, m, H-6β), 2.26 (1H, dd,J=14.2, 6.3 Hz, H-6α), 2.12 (2H, m, H-27), 2.10 (2H,m, H-25), 2.08 (2H, m, H-32), 2.03 (1H, m, H-7), 1.73(3H, s, H-26), 1.69 (3H, s, H-31), 1.61 (3H, s, H-30),1.48 (3H, s, H-37), 1.39 (3H, s, H-38), 1.35 (6H, s, H-35, 36), 1.33 (3H, s, H-21), 1.28 (3H, s, H-20).13C NMR (150 MHz, CDCl3)δC: 207.7 (C-9), 203.6 (C-2),194.6 (C-10), 140.8 (C-24), 136.8 (C-11), 132.1 (C-29), 132.0 (C-14), 129.7 (C-12), 129.7 (C-16), 127.9(C-13), 127.9 (C-15), 123.9 (C-28),120.1 (C-23), 106.8(C-4), 89.6 (C-18), 87.0 (C-34), 83.2 (C-17),82.4 (C-1), 76.8 (C-3), 70.0 (C-19), 57.1 (C-5), 52.3 (C-8),50.2 (C-33), 44.8 (C-7), 41.3 (C-6), 40.2 (C-25), 32.2(C-35), 29.0 (C-32), 28.7 (C-22), 27.4 (C-20), 27.0 (C-38), 26.5 (C-21), 26.4 (C-27),26.0 (C-31), 24.3 (C-36),23.1 (C-37), 17.9 (C-30), 16.4 (C-26). 其波譜數(shù)據(jù)與文獻報道一致[17]，故鑒定化合物為hypersampsonone C.

化合物15：C38H50O7，無色油狀；[α]D20+10.65(c1.67, CHCl3)；ESI-MS(m/z): 619[M+H]+.1H NMR(600 MHz, CDCl3)δH: 7.41 (1H, t,J=7.5 Hz, H-28),7.29 (2H, t,J=7.5 Hz, H-27, 29), 7.16 (2H, d,J=7.5 Hz,H-26, 30), 5.15 (1H, t,J=7.0 Hz, H-32), 5.06 (1H, t,J=6.9 Hz, H-36), 4.57 (1H, dd,J=11.6, 2.9 Hz, H-3),3.55 (1H, dd,J=14.9, 11.6 Hz, H-2α), 2.78 (1H, dd,J=10.8, 8.0 Hz, H-7), 2.62 (2H, d,J=7.0 Hz, H-31),2.58 (1H, dd,J=14.3, 6.3 Hz, H-15α), 2.42 (1H, ddd,J=15.5, 10.8, 4.7 Hz, H-8α), 2.10 (1H, m, H-9), 2.03(2H, m, H-35), 1.99 (2H, m, H-34), 1.88 (1H, d,J=14.3 Hz, H-15β), 1.69 (3H, s, H-39), 1.66 (3H, s, H-38), 1.58 (3H, s, H-40), 1.54 (1H, m, H-2β), 1.51 (1H,m, H-8β), 1.48 (3H, s, H-23), 1.36 (3H, s, H-22), 1.30(3H, s, H-20), 1.13 (3H, s, H-18), 1.08 (3H, s, H-21),1.07 (3H, s, H-19).13C NMR (150 MHz, CDCl3)δC:208.1 (C-12), 205.3 (C-13), 204.5 (C-16), 192.3 (C-24), 139.1 (C-33), 134.8 (C-25),132.6 (C-28), 131.7 (C-37), 128.9 (C-26), 128.9 (C-30), 128.2 (C-27), 128.2(C-29), 124.2 (C-36), 118.8 (C-32), 89.0 (C-3), 88.6(C-6), 81.9 (C-11),73.2 (C-17), 68.6 (C-14), 66.7 (C-1), 50.4 (C-10), 44.5 (C-9), 42.7 (C-7), 41.3 (C-15),40.2 (C-34), 31.5 (C-8), 31.2 (C-2), 29.7 (C-31), 28.2(C-20), 26.8 (C-35), 26.0 (C-19), 25.9 (C-38), 25.0 (C-18), 25.0 (C-23), 22.8 (C-22), 18.0 (C-21), 17.8 (C-40), 16.7 (C-39). 其波譜數(shù)據(jù)與文獻報道一致[18]，故鑒定化合物為otogirinin B.

3 結(jié)論

采用各種波譜技術(shù)，以及質(zhì)譜等對所得化合物進行了結(jié)構(gòu)鑒定，從藤黃科金絲桃屬植物川鄂金絲桃中分離并鑒定出15個化合物，包括黃酮類、甾體類、三萜類、飽和脂肪酸酯類、氧雜蒽酮類和多環(huán)多異戊烯基取代間苯三酚衍生物. 分離所得的化合物uralione H、attenuatumione C在已有報道中表明具有良好的生物活性：uralione H的細胞實驗表明該化合物對PC12細胞的損傷有誘導保護作用[10]；attenuatumione C對人體癌細胞系中的SMMC7721和U2OS表現(xiàn)出中等細胞毒性活性[13]. 研究結(jié)果為合理有效利用該植物提供了科學依據(jù).