劉晶晶,陳 幸,魏志奇,李 彬

1四川省中醫(yī)藥科學(xué)院中藥研究所,成都 610041;2成都中醫(yī)藥大學(xué)藥學(xué)院,成都 611730

川木通的化學(xué)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(yī)藥科學(xué)院中藥研究所,成都 610041;2成都中醫(yī)藥大學(xué)藥學(xué)院,成都 611730

研究川木通的化學(xué)成分,根據(jù)結(jié)果對中國藥典川木通的薄層色譜鑒別內(nèi)容提出商榷意見。采用硅膠柱、SephadexLH-20分離及重結(jié)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(shù)據(jù)、理化性質(zhì)和對照參考文獻(xiàn),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(fā)現(xiàn)采集的 11批川木通藥材中均含有豆甾醇,而未發(fā)現(xiàn)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(nèi)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(jīng)下乳之功效。主治水腫、小便不利與經(jīng)閉乳少等癥,是常用中藥材[1]。中國藥典規(guī)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(jīng)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(jié)構(gòu)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(fā)現(xiàn)齊墩果酸。因此我們認(rèn)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(zhì)譜儀;Bruker Daltonics Bio TOF Q spectrometer高分辨質(zhì)譜儀;Nist 98標(biāo)準(zhǔn)質(zhì)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(zhì)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(fù)萃取,回收溶劑得浸膏 112.5 g。該浸膏經(jīng)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(jīng) TLC檢測,合并得 5個洗脫部分。A部分 (7.8 g)經(jīng) RP-18色譜柱層析分離,甲醇洗脫,經(jīng)過甲醇反復(fù)重結(jié)晶得到三個無色針狀結(jié)晶 2、3和 4。B部分 (2.579 g)經(jīng) RP-18色譜柱層析分離,甲醇∶水 (75∶25)為流動相洗脫,再經(jīng)過 Sephadex柱色譜層析純化,甲醇洗脫,得化合物1。

3 結(jié)構(gòu)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(shù)據(jù)通過Nist 98標(biāo)準(zhǔn)質(zhì)譜圖庫檢索,并結(jié)合有關(guān)文獻(xiàn)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(jié)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(shù)據(jù)與文獻(xiàn)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(shù)據(jù)與文獻(xiàn)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(shù)據(jù)與文獻(xiàn)值一致[5,6],鑒定化合物 4為β-谷甾醇。

4 TLC鑒別

根據(jù)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(huán)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(jié)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(jīng)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(guī)定以齊墩果酸為對照品薄層鑒別川木通,試驗結(jié)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(rèn)為藥典以齊墩果酸為對照品薄層鑒別川木通,其內(nèi)容有待商榷。

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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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