中草药  2016, Vol. 47 Issue (10): 1665-1669
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引用本文doi: 10.7501/j.issn.0253-2670.2016.10.007
肖世基, 郭大乐, 徐德林, 张茂生, 陈放, 丁立生, 周燕 . 裸蒴化学成分研究[J]. 中草药, 2016, 47(10): 1665-1669.
XIAO Shi-ji, GUO Da-le, XU De-lin, ZHANG Mao-sheng, CHEN Fang, DING Li-sheng, ZHOU Yan . Chemical constituents of Gymnotheca chinensis[J]. Chinese Traditional and Herbal Drugs, 2016, 47(10): 1665-1669 .
裸蒴化学成分研究
肖世基1, 郭大乐2, 徐德林1, 张茂生1, 陈放3, 丁立生2, 周燕2     
1. 遵义医学院药学院, 贵州 遵义 563000 ;
2. 中国科学院成都生物研究所, 四川 成都 610041 ;
3. 四川大学生命科学学院, 四川 成都 610064
收稿日期: 2015-11-29
基金项目: 国家自然科学基金资助项目(31560102);贵州省教育厅自然科学研究项目(黔教合KY字[2015]362)
作者简介: 肖世基,男,博士,主要从事天然药物化学研究。E-mail:sjxiao@zmc.edu.cn
摘要: 目的 研究裸蒴Gymnotheca chinensis的化学成分。 方法 采用多种柱色谱及制备液相色谱分离纯化,经理化性质和波谱数据鉴定结构。 结果 从裸蒴的甲醇提取物中分离得到16个化合物,分别鉴定为2-(1-羟基-4-羰基环己-2,5-二烯基)乙腈(1)、N-苯甲酰-2-羟基酪胺(2)、N-苯甲酰酪胺(3)、N-苯甲酰基-2-苯基乙胺(4)、7,4'-二甲氧基山柰酚(5)、蜜桔黄素(6)、桔皮晶(7)、姜黄酮A(8)、6-羟基大柱香波龙-4,7-二烯-3,9-二酮(9)、5,6-环氧-3-羟基大柱香波龙-7-烯-9-酮(10)、9-羟基大柱香波龙-4-烯-3-酮(11)、9-羟基大柱香波龙-4,7-二烯-3-酮(12)、黑燕麦内酯(13)、6β-羟基-豆甾-4-烯-3-酮(14)、6β-羟基-豆甾-4,22-二烯-3-酮(15)、3β-羟基-豆甾-5-烯-7-酮(16)。 结论 化合物1和2为新的天然产物,其余化合物为首次从该特有属植物中分离得到。
关键词: 裸蒴     含氮化合物     多甲氧基黄酮     倍半萜     豆甾酮    
Chemical constituents of Gymnotheca chinensis
XIAO Shi-ji1, GUO Da-le2, XU De-lin1, ZHANG Mao-sheng1, CHEN Fang3, DING Li-sheng2, ZHOU Yan2     
1. School of Pharmacy, Zunyi Medical University, Zunyi 563000, China ;
2. Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, China ;
3. College of Life Science, Sichuan University, Chengdu 610064, China
Abstract: Objective To further study the chemical constituents of Gymnotheca chinensis. Methods The compounds were isolated and purified by silica gel column chromatography and preparative HPLC, and their structures were elucidated by spectral analysis. Results Sixteen compounds were isolated and identified as 2-(1-hydroxy-4-oxo-cyclohexa-2,5-dienyl)-acetonitrile (1), N-benzoyl-2-hydroxyltyramine (2), N-benzoyltyramine (3), N-benzoyl-2-phenylethylamine (4), 7,4'-dimethoxykaemferol (5), nobiletin (6), tangeretin (7), curculonone A (8), 6-hydroxy-4,7-megastigmadien-3,9-dione (9), 5,6-epoxy-3-hydroxy-7-megastigmen-9-one (10), 9-hydroxy-4-megastigmen-3-one (11), 9-hydroxy-4,7-megastigmadien-3-one (12), loliolide (13), 6β-hydroxy-stigmasta-4-en-3-one (14), 6β-hydroxy- stigmasta-4,22-dien-3-one (15), and 3β-hydroxy-stigmasta-5-ene-7-one (16). Conclusion Compounds 1 and 2 are new natural products, and the others are isolated from this genus for the first time.
Key words: Gymnotheca chinensis Decne.     nitrogen compound     polymethoxy flavonoid     sesquiterpene     stigmasterone    

裸蒴Gymnotheca chinesis Decne. 为三白草科中我国特有属裸蒴属Gymnotheca Decne. 草本植物，又名水百步还魂草，主要产于湖北、湖南、广东、广西、云南、贵州及四川等省区，全草可以药用，有消食积和解毒排脓等功效[1]。《中国中药资源志要》记载裸蒴全草辛、温，清热利湿、消肿利尿、止带，用于肺虚久咳、劳伤咳嗽、小便淋痛、水肿、带下病，外用于跌打损伤[2]。初步化学成分研究表明其含有木脂素类[3-8]、酚酸类[9]和甾体类化合物[10]。为了进一步阐明裸蒴的化学成分，指导民间科学用药，本课题组对其全草的甲醇提取物进行了更加深入的分离纯化，从中分离得到16个化合物，通过波谱分析鉴定了它们的结构，包括4个含氮化合物：2-(1-羟基-4-羰基环己-2,5-二烯基)乙腈 [2-(1-hydroxy-4-oxo-cyclohexa-2,5-dienyl)-acetonitril，1]、N-苯甲酰-2-羟基酪胺(N-benzoyl-2-hydroxyltyramine，2)、N-苯甲酰酪胺(N-benzoyltyramine，3)、N-苯甲酰基-2-苯基乙胺(N-benzoyl-2-phenylethylamine，4)；3个多甲氧基黄酮类化合物：7,4′-二甲氧基山柰酚(7,4′- dimethoxykaemferol，5)、蜜桔黄素(nobiletin，6)，桔皮晶(tangeretin，7)；6个倍半萜类化合物：姜黄酮A(curculonone A，8)、6-羟基大柱香波龙-4,7-二烯-3,9-二酮(6-hydroxy-4,7-megastigmadien-3,9- dione，9)、5,6-环氧-3-羟基大柱香波龙-7-烯-9-酮(5,6-epoxy-3-hydroxy-7-megastigmen-9-one，10)、9-羟基大柱香波龙-4-烯-3-酮(9-hydroxy-4-megastigmen-3-one，11)、9-羟基大柱香波龙-4,7-二烯-3-酮(9-hydroxy-4,7-megastigmadien-3-one，12)、黑燕麦内酯(loliolide，13)；3个豆甾酮类化合物：6β-羟基-豆甾-4-烯-3-酮(6β-hydroxy-stigmasta-4-en-3-one，14)、6β-羟基-豆甾-4,22-二烯-3-酮(6β-hydroxy-stigmasta-4,22-dien-3-one，15)、3β-羟基-豆甾-5-烯-7-酮(3β-hydroxy-stigmasta-5-ene-7-one，16)。化合物1和2为新的天然产物，其余化合物为首次从裸蒴属植物中分离得到。

1 仪器及材料

Finnigan LCQDECA型质谱仪(美国Thermo公司)；Bruker MicrO TOF-QII型高分辨质谱仪(德国Bruker公司)；Bruker AV-400型核磁共振仪，TMS为内标(德国Bruker公司)；LC3000型高效液相色谱仪(中国北京创新通恒公司)；Daisogel C18 HPLC制备柱(250 mm×30 mm，10 μm)；Kromasil C18 HPLC半制备柱(250 mm×10 mm，5 μm)；薄层色谱硅胶GF 254和柱色谱硅胶(200～300目，中国青岛海洋化工)；MCI树脂(日本Mitsubishi公司)；Sephadex LH-20凝胶(德国Merck公司)。

裸蒴采自重庆市南川金佛山，样品标本(20130125)由重庆市药物种植研究所易思荣研究员鉴定为三白草科裸蒴属植物裸蒴Gymnotheca chinesis Decne. 的干燥全草。

2 提取分离

取干燥裸蒴全草1.45 kg，粉碎后用甲醇室温提取，减压浓缩提取液得甲醇提取物152 g，将甲醇提取物分散于热水，依次用石油醚、醋酸乙酯和正丁醇萃取。得到的石油醚萃取物(8 g)和醋酸乙酯萃取取物(19 g)合并，过MCI柱(90%甲醇-水洗脱)，脱去叶绿素后得到混合物15 g。该混合物经中压硅胶柱色谱分离，石油醚-丙酮(50：1、20：1、10：1、5：1、1：1)梯度洗脱，得到12个组分Fr. 1～12。Fr. 6经ODS柱(60%～100%甲醇-水)分为7个部分：Fr. 6.1～6.7。Fr. 6.2经半制备HPLC分离(45%乙腈-水，体积流量4.5 mL/min)得到化合物2(8 mg)。Fr. 6.7经中压硅胶柱反复分离得到化合物5(15 mg)和16(22 mg)。Fr. 7经制备HPLC(甲醇-水)分为3个部分Fr. 7.1～7.3。Fr. 7.1经半制备HPLC分离(66%甲醇-水，体积流量3 mL/min)得到化合物8(30 mg，tR＝15.9 min)。Fr. 7.3经半制备HPLC(95%乙腈-水，体积流量4.5 mL/min)反复分离得到化合物14(8 mg)和15(14 mg)。Fr. 8经Sephadex LH-20(氯仿-甲醇1：1)分为2个部分：Fr. 8A～8B。Fr. 8A经制备HPLC分离(70%甲醇-水)为4个亚组分：Fr. 8AA～8AD；Fr. 8AB进一步经半制备HPLC分离(40%甲醇-水，体积流量3 mL/min)得到化合物3(6 mg，tR＝35.9 min)和4(11 mg，tR＝38.7 min)。Fr. 9经Sephadex LH-20(氯仿-甲醇1：1)分为2个部分：Fr. 9A～9B。Fr. 9A经制备HPLC分离(甲醇-水梯度)为4个亚组分：Fr. 9AA～9AD；Fr. 9AA进一步经半制备HPLC分离得到化合物1(5 mg)、5(8 mg)、7(4 mg)、9(5 mg)、10(13 mg)、11(15 mg)、12(3 mg)、13(7 mg)。Fr. 11经制备HPLC(甲醇-水)分为4个部分：Fr. 11A～11D。Fr. 11A经半制备HPLC分离得到化合物6(7 mg)。

3 结构鉴定

化合物1：白色粉末，ESI-MS m/z: 148 [M－H]−, 172 [M＋Na]+；1H-NMR (400 MHz, CDCl3) δ: 6.91 (2H, d, J = 10.0 Hz, H-3, 5), 6.26 (2H, d, J = 10.0 Hz, H-2, 6), 3.73 (1H, s, OH), 2.77 (2H, s, H-7)；13C-NMR (100 MHz, CDCl3) δ: 184.9 (C-1), 149.6 (C-3, 5), 129.4 (C-2, 6), 116.8 (C-8), 67.5 (C-7)。波谱数据和文献对照一致[11]，故将化合物1鉴定为4-羟基-4-乙腈基-环己-2,4-二烯-1-酮，其为新的天然产物。

化合物2：白色粉末，C15H15NO3，ESI-MS m/z: 256 [M－H]−, 280 [M＋Na]+；HR-ESI-MS m/z: 280.094 6 [M＋Na]+；1H-NMR (400 MHz, DMSO-d6) δ: 9.28 (1H, s, 6-OH), 8.44 (1H, t, J = 5.4 Hz, CONH), 7.83 (2H, d, J = 7.2 Hz, H-3′, 7′), 7.51 (1H, t, J = 7.2 Hz, H-5′), 7.44 (2H, t, J = 7.2 Hz, H-4′, 6′), 7.16 (2H, d, J = 8.4 Hz, H-4, 8), 6.72 (2H, d, J = 8.4 Hz, H-5, 7), 5.33 (1H, d, J = 2.9 Hz, 2-OH), 4.67 (1H, brs, H-2), 3.43 (1H, m, H-1), 3.29 (1H, m, H-1)；13C-NMR (100 MHz, DMSO-d6) δ: 166.3 (C-1′), 156.4 (C-6), 134.6 (C-2′), 134.1 (C-3), 131.0 (C-5′), 128.2 (C-4′, 6′), 127.2 (C-4, 8), 127.1 (C-3′, 7′), 114.8 (C-5, 7), 70.9 (C-2), 47.7 (C-1)。通过HMBC谱及参考文献鉴定化合物2为N-苯甲酰-2-羟基酪胺[12]，其为新的天然产物。

化合物3：白色粉末，ESI-MS m/z: 240 [M－H]−, 242 [M＋H]+, 264 [M＋Na]+；1H-NMR (400 MHz, CD3COCD3) δ: 8.13 (1H, s, 6-OH), 7.86 (2H, d, J = 7.4 Hz, H-3′, 7′), 7.74 (1H, brs, CONH), 7.50 (1H, t, J = 7.4 Hz, H-5′), 7.43 (2H, t, J = 7.4 Hz, H-4′, 6′), 7.08 (2H, d, J = 8.4 Hz, H-4, 8), 6.76 (2H, d, J = 8.4 Hz, H-5, 7), 3.57 (2H, dd, J = 14.5, 6.2 Hz, H-1), 2.83 (2H, m, H-2)；13C-NMR (100 MHz, CD3COCD3) δ: 167.2 (C-1′), 156.7 (C-6), 136.2 (C-3), 131.7 (C-5′), 131.2 (C-2′), 130.5 (C-4, 8), 129.1 (C-4′, 6′), 127.9 (C-3′, 7′), 116.1 (C-5, 7), 42.4 (C-1), 35.6 (C-2)。化合物的波谱数据和文献对照一致[13]，故鉴定化合物3为N-苯甲酰酪胺。

化合物4：白色粉末，ESI-MS m/z: 224 [M－H]−, 226 [M＋H]+, 248 [M＋Na]+；1H-NMR (400 MHz, CD3COCD3) δ: 7.86 (2H, d, J = 7.4 Hz, H-3′, 7′), 7.50 (2H, t, J = 7.4 Hz, H-4′, 6′), 7.43 (1H, t, J = 7.4 Hz, H-5′), 7.35～7.13 (5H, m, H-4～8), 3.63 (2H, dd, J = 14.2, 6.5 Hz, H-1), 2.93 (2H, t, J = 7.4 Hz, H-2)；13C-NMR (100 MHz, CD3COCD3) δ: 167.2 (C-1′), 140.6 (C-2′), 136.1 (C-3), 131.8 (C-5′), 129.6 (C-5, 7), 129.2 (C-4, 8), 129.1 (C-4′, 6′), 127.9 (C-3′, 7′), 126.9 (C-6), 42.1 (C-1), 36.45 (C-2)。波谱数据和文献对照一致[14]，故鉴定化合物4为N-苯甲酰基-2-苯基乙胺。

化合物5：黄色针状晶体(石油醚-丙酮)，mp 178～180 ℃。EI-MS m/z: 314 [M] +；1H-NMR (400 MHz, CDCl3) δ: 8.19 (2H, d, J = 8.9 Hz, H-2′, 6′), 7.05 (2H, d, J = 8.9 Hz, H-3′, 5′), 6.50 (1H, d, J = 1.8 Hz, H-8), 6.39 (1H, d, J = 1.8 Hz, H-6), 3.91, 3.92 (2×3H, s, 2×OCH3)；13C-NMR (100 MHz, CDCl3) δ: 175.2 (C-4), 165.7 (C-7), 161.1 (C-4′), 160.8 (C-5), 156.8 (C-9), 145.7 (C-2), 135.7 (C-3), 129.4 (C-2′, 6′), 123.2 (C-1′), 114.1 (C-3′, 5′), 103.9 (C-10), 97.9 (C-6), 92.2 (C-8), 55.8, 55.4 (2×OCH3)。波谱数据和文献对照一致[15]，故鉴定化合物5为7,4′-二甲氧基山柰酚。

化合物6：浅黄色针状晶体(石油醚-丙酮)，mp 150～152 ℃。ESI-MS m/z: 403 [M＋H]+, 425 [M＋Na]+；1H-NMR (400 MHz, CD3COCD3) δ: 7.67 (1H, dd, J = 8.5, 2.1 Hz, H-6′), 7.59 (1H, d, J = 2.1 Hz, H-2′), 7.14 (1H, d, J = 8.5 Hz, H-5′), 6.62 (1H, s, H-3), 4.08, 4.04, 3.96, 3.92, 3.89, 3.85 (6×3H, s, 6×OCH3)；13C-NMR (100 MHz, CD3COCD3) δ: 176.6 (C-4), 161.5 (C-2), 153.2 (C-4′), 152.3 (C-7), 150.6 (C-3′), 149.1 (C-5), 148.5 (C-9), 145.0 (C-6), 139.1 (C-8), 124.7 (C-1′), 120.2 (C-6′), 115.8 (C-10), 112.5 (C-5′), 109.8 (C-2′), 107.2 (C-3), 62.3, 62.2, 61.9, 61.9, 56.2, 56.2 (6×OCH3)。波谱数据和文献对照基本一致[16]，故鉴定化合物6为蜜桔黄素。

化合物7：浅黄色针状晶体(石油醚-丙酮)，mp 135～137 ℃。ESI-MS m/z: 373 [M＋H]+；1H-NMR (400 MHz, CD3COCD3) δ: 8.04 (2H, d, J = 8.8 Hz, H-2′, 6′), 7.16 (2H, d, J = 8.8 Hz, H-3′, 5′), 6.58 (1H, s, H-3), 4.09, 4.05, 3.93, 3.91, 3.88 (5×3H, s, 5×OCH3)；13C-NMR (100 MHz, CD3COCD3) δ: 176.7 (C-4), 163.3 (C-4′), 161.7 (C-2), 152.3 (C-7), 149.1 (C-5), 148.5 (C-9), 144.4 (C-6), 137.5 (C-8), 128.6 (C-2′, 6′), 124.6 (C-1′), 115.4 (C-3′, 5′), 106.9 (C-10), 106.9 (C-3), 62.3, 62.3, 61.9, 61.9, 55.9 (5×OCH3)。波谱数据和文献对照基本一致[17]，故鉴定化合物7为桔皮晶。

化合物8：无色油状物，ESI-MS: m/z 259 [M＋Na]+；1H-NMR (400 MHz, CD3COCD3) δ: 5.75 (1H, s, H-3), 5.64 (1H, d, J = 15.5 Hz, H-10), 5.57 (1H, dt, J = 15.5, 6.4 Hz, H-9), 3.46 (1H, s, 11-OH), 1.92 (3H, s, 15-CH3), 1.23 (6H, s, 12, 13-CH3), 0.75 (3H, d, J = 6.8 Hz, 14-CH3)；13C-NMR (100 MHz, CD3COCD3) δ: 200.2 (C-2), 161.8 (C-4), 141.6 (C-10), 127.3 (C-3), 125.2 (C-9), 70.0 (C-11), 49.4 (C-1), 37.8 (C-8), 31.5 (C-7), 31.3 (C-5), 30.4 (C-12), 30.4 (C-13), 23.9 (C-15), 22.9 (C-6), 15.8 (C-14)。波谱数据和文献对照一致[18]，故鉴定化合物8为姜黄酮A。

化合物9：淡黄色油状物，1H-NMR (400 MHz, CD3COCD3) δ: 7.04 (1H, d, J = 15.9 Hz, H-7), 6.42 (1H, d, J = 15.9 Hz, H-8), 5.86 (1H, s, H-4), 2.60 (1H, d, J = 16.9 Hz, H-2), 2.28 (3H, s, 10-CH3), 2.22 (1H, d, J = 16.9 Hz, H-2), 1.88 (3H, d, J = 1.2 Hz, 13-CH3), 1.08 (3H, s, 12-CH3), 1.03 (3H, s, 11-CH3)；13C-NMR (100 MHz, CD3COCD3) δ: 197.1 (C-3), 196.2 (C-9), 160.8 (C-5), 146.2 (C-7), 130.7 (C-8), 127.0 (C-4), 78.8 (C-6), 49.3 (C-2), 41.2 (C-1), 26.6 (C-10), 23.8 (C-12), 22.6 (C-11), 18.0 (C-13)。波谱数据和文献对照一致[19]，故鉴定化合物9为6-羟基大柱香波龙-4,7-二烯- 3,9-二酮。

化合物10：淡黄色油状物，1H-NMR (400 MHz, CD3COCD3) δ: 7.13 (1H, d, J = 15.8 Hz, H-7), 6.13 (1H, d, J = 15.8 Hz, H-8), 3.76 (1H, m, H-3), 2.32 (1H, m, H-4), 2.26 (3H, s, 10-CH3), 1.66 (1H, dd, J = 12.8, 8.7 Hz, H-2), 1.56 (1H, ddd, J = 12.9, 3.3, 1.6 Hz, H-4), 1.26 (1H, dd, J = 12.8, 10.5 Hz, H-2), 1.19 (3H, s, CH3), 1.15 (3H, s, CH3), 0.93 (3H, s, CH3)；13C-NMR (100 MHz, CD3COCD3) δ: 62.6 (C-3), 196.4 (C-9), 66.9 (C-5), 142.9 (C-7), 132.7 (C-8), 46.7 (C-4), 69.1 (C-6), 40.6 (C-2), 34.7 (C-1), 26.5 (C-10), 24.4 (C-12), 28.7 (C-11), 19.2 (C-13)。波谱数据和文献对照一致[20]，故鉴定化合物10为5,6-环氧-3-羟基大柱香波龙-7-烯-9-酮。

化合物11：无色油状物，1H-NMR (400 MHz, CDCl3) δ: 5.82 (1H, s, H-4), 3.75 (1H, m, H-9), 1.99 (3H, s, CH3), 1.20 (3H, d, J = 6.2 Hz, CH3), 1.06 (3H, s, CH3), 1.01 (3H, s, CH3)；13C-NMR (100 MHz, CDCl3) δ: 199.5 (C-3), 68.1 (C-9), 165.5 (C-5), 26.2 (C-7), 38.7 (C-8), 125.1 (C-4), 51.0 (C-6), 47.1 (C-2), 36.3 (C-1), 24.7 (C-10), 28.8 (C-12), 27.2 (C-11), 23.8 (C-13)。波谱数据和文献对照一致[20]，故鉴定化合物11为9-羟基大柱香波龙-4-烯-3-酮。

化合物12：无色油状物，1H-NMR (400 MHz, CDCl3) δ: 5.89 (1H, s, H-3), 5.67 (1H, dd, J = 15.3, 5.9 Hz, H-7), 5.55 (1H, dd, J = 15.4, 9.0 Hz, H-8), 4.34 (1H, p, J = 6.2 Hz, H-9), 2.52 (1H, d, J = 9.0 Hz, H-6), 2.33 (1H, d, J = 16.7 Hz, H-2), 2.08 (1H, d, J = 16.7 Hz, H-2), 1.89 (3H, s, 13-CH3), 1.28 (3H, d, J = 6.4 Hz, 10-CH3), 1.03 (3H, s, 11-CH3), 0.97 (3H, s, 12-CH3)；13C-NMR (100 MHz, CDCl3) δ: 199.2 (C-3), 68.3 (C-9), 161.8 (C-5), 126.6 (C-7), 138.4 (C-8), 125.9 (C-4), 55.4 (C-6), 47.4 (C-2), 36.1 (C-1), 23.5 (C-10), 27.9 (C-12), 27.1 (C-11), 23.5 (C-13)。波谱数据和文献对照一致[20]，故鉴定化合物12为9-羟基大柱香波龙-4,7-二烯-3-酮。

化合物13：白色粉末，1H-NMR (400 MHz, CD3COCD3) δ: 5.68 (1H, s, H-7), 4.29 (1H, m, H-3), 2.39 (1H, dt, J = 13.4, 2.5 Hz, H-4), 1.98 (1H, dt, J = 13.4, 2.6 Hz, H-2), 1.71 (1H, dd, J = 14.3, 3.6 Hz, H-4), 1.74 (3H, s, 11-CH3), 1.54 (1H, dd, J = 14.3, 3.6 Hz, H-2), 1.47 (3H, s, 9-CH3), 1.26 (3H, s, 10-CH3)；13C-NMR (100 MHz, CD3COCD3) δ: 36.6 (C-1), 47.8 (C-2), 66.8 (C-3), 46.4 (C-4), 87.0 (C-5), 183.4 (C-6), 113.3 (C-7), 171.6 (C-8), 31.0 (C-9), 26.8 (C-10), 27.5 (C-11)。波谱数据和文献对照一致[21]，故鉴定化合物13为黑燕麦内酯。

化合物14：白色粉末，mp 206～208 ℃。ESI-MS m/z: 451 [M＋Na]+；1H-NMR (400 MHz, CDCl3) δ: 5.81 (1H, s, H-4), 4.35 (1H, m, H-6)；13C-NMR (100 MHz, CDCl3) δ: 37.1 (C-1), 34.3 (C-2), 200.4 (C-3), 126.3 (C-4), 168.5 (C-5), 73.3 (C-6), 38.5 (C-7), 29.7 (C-8), 53.6 (C-9), 38.0 (C-10), 21.1 (C-11), 39.6 (C-12), 42.5 (C-13), 55.9 (C-14), 24.2 (C-15), 28.2 (C-16), 56.1 (C-17), 12.0 (C-18), 19.5 (C-19), 36.1 (C-20), 18.8 (C-21), 33.6 (C-22), 25.4 (C-23), 45.1 (C-24), 29.2 (C-25), 20.2 (C-26), 19.0 (C-27), 22.7 (C-28), 12.0 (C-29)。波谱数据和文献对照一致[22-23]，故鉴定化合物14为6β-羟基-豆甾-4-烯-3-酮。

化合物15：白色粉末，mp 209～211 ℃。ESI-MS m/z: 449 [M＋Na]+；1H-NMR (400 MHz, CDCl3) δ: 5.81 (1H, s, H-4), 5.15 (1H, dd, J = 15.1, 8.6 Hz, H-22), 5.03 (1H, dd, J = 15.1, 8.5 Hz, H-23), 4.35 (1H, m, H-6)；13C-NMR (100 MHz, CDCl3) δ: 37.1 (C-1), 34.3 (C-2), 200.4 (C-3), 126.3 (C-4), 168.4 (C-5), 73.3 (C-6), 38.5 (C-7), 29.7 (C-8), 53.6 (C-9), 38.0 (C-10), 21.1 (C-11), 39.5 (C-12), 40.5 (C-13), 55.9 (C-14), 24.1 (C-15), 28.8 (C-16), 56.0 (C-17), 12.2 (C-18), 19.0 (C-19), 39.5 (C-20), 21.2 (C-21), 138.1 (C-22), 129.5 (C-23), 51.2 (C-24), 31.9 (C-25), 19.5 (C-26), 19.0 (C-27), 25.4 (C-28), 12.2 (C-29)。波谱数据和文献对照一致[22-23]，故鉴定化合物15为6β-羟基-豆甾-4,22-二烯-3-酮。

化合物16：白色粉末，mp 203～205 ℃。ESI-MS m/z: 451 [M＋Na]+；1H-NMR (400 MHz, CDCl3) δ: 6.19 (1H, brs, H-6), 4.35 (1H, ddd, J = 12.0, 5.4, 1.5 Hz, H-3)；13C-NMR (100 MHz, CDCl3) δ: 36.3 (C-1), 33.8 (C-2), 68.7 (C-3), 42.5 (C-4), 171.5 (C-5), 119.6 (C-6), 199.5 (C-7), 34.1 (C-8), 53.7 (C-9), 39.0 (C-10), 21.0 (C-11), 39.4 (C-12), 42.4 (C-13), 55.9 (C-14), 24.2 (C-15), 28.1 (C-16), 55.5 (C-17), 11.9 (C-18), 18.3 (C-19), 36.1 (C-20), 18.7 (C-21), 33.8 (C-22), 26.1 (C-23), 45.8 (C-24), 29.1 (C-25), 19.8 (C-26), 19.0 (C-27), 23.1 (C-28), 12.0 (C-29)。波谱数据和文献对照一致[24]，故鉴定化合物16为3β-羟基-豆甾-5-烯-7-酮。

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