川木通的化學成分及鑒別研究

劉晶晶,陳 幸,魏志奇,李 彬

1四川省中醫藥科學院中藥研究所,成都 610041;2成都中醫藥大學藥學院,成都 611730

川木通的化學成分及鑒別研究

劉晶晶1,2,陳 幸1＊,魏志奇1,李 彬1

1四川省中醫藥科學院中藥研究所,成都 610041;2成都中醫藥大學藥學院,成都 611730

研究川木通的化學成分,根據結果對中國藥典川木通的薄層色譜鑒別內容提出商榷意見。采用硅膠柱、SephadexLH-20分離及重結晶等方法,從川木通 Clem atis ar m andiiFranch.的乙醇提取物中,分離純化得到四種化合物。通過波譜數據、理化性質和對照參考文獻,分別鑒定為 9,12-octadecadienoic acid(Z,Z)-,2,3-dihydroxypropyl ester(1)、麥角甾醇 (2)、豆甾醇 (3)和β-谷甾醇 (4),其中 1和 2為首次分離得到。用 TLC和HPLC圖譜比對發現采集的 11批川木通藥材中均含有豆甾醇,而未發現齊墩果酸。中國藥典以齊墩果酸為對照品薄層色譜法定性鑒別川木通的內容有待商榷。

川木通;麥角甾醇;豆甾醇;β-谷甾醇;鑒別

《中國藥典》2005版收載川木通為毛茛科植物小木通 Clem atis ar m andiiFranch.或繡球藤 Clem atis m ontanaBuch.-Ham.的干燥藤莖,具有清熱利尿、通經下乳之功效。主治水腫、小便不利與經閉乳少等癥,是常用中藥材[1]。中國藥典規定以齊墩果酸為對照品薄層鑒別川木通。在我們以往的研究中,按藥典方法試驗,薄層色譜顯示小木通和繡球藤均不含齊墩果酸[2]。本次經過成分分離、鑒定,從小木通的莖中分離得到 4個化合物,其結構分別鑒定為9,12-octadecadienoic acid(Z,Z)-2,3-dihydroxypropyl ester(1)、麥角甾醇 (2)、豆甾醇 (3)和β-谷甾醇(4),其中 1和 2為首次從小木通中分離得到。通過 HPLC和 TLC圖譜比對,9批川木通和 2批繡球藤均含有豆甾醇而未發現齊墩果酸。因此我們認為中國藥典以齊墩果酸作為對照品薄層鑒別川木通有待商榷。

1 儀器與材料

Auto Spec Ult ima-Tof質譜儀;Bruker Daltonics Bio TOF Q spectrometer高分辨質譜儀;Nist 98標準質譜圖庫;Varian INOVA-400型核磁共振儀;I R-8300型紅外光譜儀;UV-2401PC型 (島津)分光光度計。小木通 Clem atis ar m andiiFranch.采自四川省洪雅縣、峨眉山魚洞、峨眉四峨山,繡球藤 Clem atis m ontanaBuch.-Ham.采自四川省康定縣,由四川省中藥研究所生藥室方清茂副研究員鑒定。

2 提取與分離

小木通木質藤莖 50 kg粉碎過 10目篩,用 95%乙醇回流提取兩次,每次 2 h,濾過,減壓濃縮,濃縮液用石油醚反復萃取,回收溶劑得浸膏 112.5 g。該浸膏經硅膠柱色譜層析分離,石油醚:乙酸乙酯 (25∶1-1∶1)梯度洗脫。流出組分經 TLC檢測,合并得 5個洗脫部分。A部分 (7.8 g)經 RP-18色譜柱層析分離,甲醇洗脫,經過甲醇反復重結晶得到三個無色針狀結晶 2、3和 4。B部分 (2.579 g)經 RP-18色譜柱層析分離,甲醇∶水 (75∶25)為流動相洗脫,再經過 Sephadex柱色譜層析純化,甲醇洗脫,得化合物1。

3 結構鑒定

化合物 1 C21H38O4,無色油狀物。UV(E tOH) λmax(logε):210.6 nm。IR(KBr)νmaxcm-1:3258, 2854,1715,1650。EI-MSm/z:354[M]+,336,262, 81,67(100)。1H NMR(CDCl3,400 MHz)δ:5.39 (2H,J=11.2,6.8 Hz),5.33(2H,J=10.8,6.4 Hz),4.16(2H,m),2.34(2H,t,J-7.6),0.89(3H,t, CH3);13C NMR(CDCl3,100 MHz)δ:174.3,130.2, 130.0,128.1,127.9,70.2,65.1,63.4,34.1,31.5, 29.6,29.3,29.1,29.1,27.2,25.6,24.9,22.5, 14.0。以上數據通過Nist 98標準質譜圖庫檢索,并結合有關文獻人工圖譜解析,鑒定化合物 1為 9,12-Octadecadienoic acid(Z.Z)-,2,3-dihydroxypropyl ester。

化合物 2 C28H44O,白色粉末狀結晶。EI-MS m/z:396[M]+(45),378[M-H2O]+(4),363(27), 337(12),271(26),253(16),157(18),143(22), 105(37),95(52),81(67),69(91),55(100)。13C NMR(CDCl3,100 MHz)δ:141.3(C-8),139.8(C-5),135.6(C-22),132.0(C-23),119.6(C-6),116.3 (C-7),70.5(C-3),55.8(C-17),54.6(C-14),46.3 (C-9),42.8(C-13),42.3(C-24),40.8(C-4),40.4 (C-20),39.1(C-21),38.4(C-1),37.0(C-10),33.1 (C-25),32.0(C-2),28.2(C-16),23.1(C-15),21.1 (C-11),21.1(C-12),19.9(C-28),19.6(C-27), 17.6(C-26),16.2(C-19),12.0(C-18)。以上數據與文獻值一致[3],鑒定化合物 2為麥角甾醇 (ergosterol)。

化合物 3 C29H48O,無色針晶。mp.142～144℃(CH3OH)。EI-MSm/z:412[M]+(100),397 (9),394(28),379(10),327(3),300(40),273 (19),271(45),255(73),231(10),213(23)。13C NMR(CDCl3,100 MHz)δ:140.8(C-5),138.3(C-22),129.3(C-23),121.7(C-6),71.8(C-3),56.9 (C-17),56.0(C-14),51.2(C-24),50.2(C-9),42.3 (C-4),42.2(C-13),40.4(C-20),39.7(C-12),37.3 (C-1),36.5(C-10),31.9(C-25),31.9(C-7),31.9 (C-8),31.6(C-2),28.9(C-16),25.4(C-28),24.4 (C-15),21.2(C-27),21.1(C-11),21.0(C-21), 19.4(C-19),19.0(C-26),12.2(C-29),12.0(C-18)。以上數據與文獻值一致[4],鑒定化合物 3為豆甾醇 (stigmasterol)。

圖 1 小木通、甾醇混合對照品、齊墩果酸對照品、白木通、繡球藤薄層色譜圖Fig.1 TLC chromatogram of ergosterol,stigmasterol,β-sitosterol,the Caules ofC.ar m andiiandC.M ontana

化合物 4 C29H50O,無色針晶。mp.138～140℃(CH3OH)。EI-MSm/z:414[M]+(71),396 (51),381(21),329(20),273(17),255(23),231 (13),213(26),159(20),81(69),69(57),55 (100)。13C NMR(CDCl3,100 MHz)δ:140.8(C-5), 121.7(C-6),71.8(C-3),56.8(C-14),56.1(C-17), 50.2(C-9),45.9(C-4),42.3(C-13),39.8(C-12), 37.3(C-24),36.5(C-1),36.1(C-10),34.0(C-22), 31.9(C-8),31.9(C-20),31.6(C-7),29.2(C-2), 29.2(C-16),28.2(C-11),26.2(C-15),24.3(C-23),23.1(C-27),21.1(C-28),19.8(C-19),19.4 (C-21),19.1(C-25),18.8(C-29),12.0(26),11.9 (18)。以上數據與文獻值一致[5,6],鑒定化合物 4為β-谷甾醇。

4 TLC鑒別

根據中國藥典薄層色譜法(附錄V IB),分別吸取樣品和對照品溶液各 5μL,點于同一硅膠 G薄層板上,以環己烷-丙酮(4∶1)為展開劑,展開,取出,晾干,噴以 10%的硫酸乙醇溶液,在 105℃加熱至斑點清晰。結果顯示小木通和繡球藤均不含齊墩果酸,但在薄層色譜上顯示出一紫紅色共同斑點 (圖 1中A點),經與對照品斑點比對,該斑點為麥角甾醇、豆甾醇、β-谷甾醇的混合物。

5 討論

中國藥典規定以齊墩果酸為對照品薄層鑒別川木通,試驗結果表明不能檢出齊墩果酸。該薄層條件下檢出的主要斑點是麥角甾醇、豆甾醇、β-谷甾醇的混合物,因此我們認為藥典以齊墩果酸為對照品薄層鑒別川木通,其內容有待商榷。

1 Committee of Chinese Phar macopoeia.Chinese Pharmacopoeia(中華人民共和國藥典),2005,1:25.

2 LiB(李彬),Chen X(陳幸),LiWS(黎萬壽).Discussion on ChuanMutong’s TLC essay contents recorded in Chinese pharmacopoeia.J Chin M ed M at(中藥材),2005,28:1147-1148.

3 Wang F(王飛),Liu JK(劉吉開).The chemical constituents of basidiomyceteCalodon Suaveolens.Nat Prod Res Dev (天然產物研究與開發),2004,16:204-206.

4 Yan LH(閆利華),Xu LZ(徐麗珍),Zou Z M(鄒忠梅),et al.Chemical constituents from stems ofClem atis ar m andii (Ⅰ).Chin Tradit Herb D rugs(中草藥),2007,38:340-342.

5 Guo XM(郭學民),Hong YF(洪永福),Zhang L(張玲),et al.Studies on the chemical constituents of common floweringquince(Chaenom eles lagenaria).Chin Tradit Herb D rugs (中草藥),1997,28:584-585.

6 Tian J(田軍),Wu FE(吳鳳鍔),Qiu MH(丘明華),et al. Chemical constituents ofPterocephalus Hookeri.Nat Prod Res Dev(天然產物研究與開發),2000,12:31-38.

Chem ical Constituents and Identification of the Caules ofClem atis arm andiiFranch.andClem atis m ontanaBuch.-Ham.

L IU Jing-jing1,2,CHEN Xing1＊,WEI Zhi-qi1,L IBin11Sichuan Academ y of ChineseM edicine Sciences,Sichuan 610041,China;2School of Phar m aceutical Sciences,Chengdu University of Traditional ChineseM edicine,Chengdu 611730,China

Four compounds,9,12-octadecadienoic acid(Z,Z)-,2,3-dihydroxypropyl ester(1),ergosterol(2),stigmasterol(3),andβ-sitosterol(4),were isolated from the ethanol extracts ofClem atis ar m andiiFranch.,and identified on the basis of the spectroscopic and physicochemical datawith those reported previously.Compounds 1 and 2 were isolated from the stems ofC.ar mandiifor the first time.However,oleanolic acid,an indexed component in Chinese Pharmacopoeia for the identification ofClem atisspp.was not detected in our conditions.To determinewhetheroleanolic acid presented inC.am andiiandC.M ontana,we collected 11 batch samples include nine forC.amandiiand two forC.M ontana.There was no trace of oleanolic acid in all 11 samples based on HPLC and TLC analysis,indicating the need for reinvestigation the chemical constituents and the indexed components ofC.spp..

Clem atis ar m andiiFranch.;Clem atis montanaBuch.-Ham.;ergosterol;stigmasterol;β-sitosterol;identification

1001-6880(2010)06-0998-03

2009-01-20 接受日期:2009-07-01

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