[生物医药论文精品]雷公藤多苷片高效液相指纹图谱的研究

1雷公藤多苷片高效液相指纹图谱的研究A1A0A1A0A2A3A4A5A6A7A8A9A10A11A12A13A14A15A16A17A18A19A20A21A22A23A19A20A24A25A26A27A28A29A30A31A32A33A34A35A36A13A37A38A39A40A41A42A37A43A44A39A24A45A46A19A20A34A47A39A48A49A50A51A52A53A5A6A7A8A9A10A54A55A56A16A57A58A59A60A61A62A63A27A64A65A5A6A7A8A9A10A48A66A67A23A68A69A70A71A72A73A74A75A76A77A11A67A78A56A16A5A6A7A8A9A10A48A54A55A27A79A80A81A26A49A48A11A67A34A21A82A75A34A11A83A84A48A85A86A51A87A88A87A88A89A60A90A91A92A93A94A48A5A6A7A8A9A10A27A95A26A66A67A23A68A96A72A97A27A96A72A98A74DIOMONISLC18A99A100A98A27A101A102A103A104A74218NMA27A98A105A7435A106A27A107A108A109A110A111A74A112A113A68A2710MLMIN1A48A112A114A73A115A116A117A118A51A119A120A119A120A121A122A5A6A7A8A9A10A48A69A70A71A72A51A123A69A70A71A72A48A124A97A11A125A126A48A127A128A34A37A129A130A116A21A131A132A34A51A60A90A133A134A48A66A67A23A68A69A70A71A72A48A135A136A137A138A139A140A141A27A142A143A60A90A93A94A14A144A145A48A5A6A7A8A9A10A48A135A136A146A99A138A139A68A90A27A147A148A55A11A149A60A90A51A150A132A151A152A153A154A137A74A5A6A7A8A9A10A48A155A11A137A27A156A157A64A65A151A5A6A7A8A9A10A48HPLCA69A70A71A72A27A74A158A55A123A54A55A159A160A142A161A151A162A163A51A119A164A119A164A165A26A5A6A7A8A9A10HPLCA69A70A71A72A166A107A125A126A78A167A168A5A6A7A8A9A10A48A169A170A54A55A27A171A172A124A97A173A174A27A175A114A27A11A67A27A176A177A27A11A178A179A5A6A7A8A9A10A48A54A55A56A16A51A180A181A182A180A181A183A66A67A23A68A96A72A97A27A69A70A71A72A27A5A6A7A8A9A10A27A5A6A7A169A184A185雷公藤多苷片是卫矛科（CELASTRACEAE）雷公藤属雷公藤TRIPTERYGIUMWILFORDIIHOOKF的干燥去皮根经提取、纯化后得到的有效组分。雷公藤多苷片与其它雷公藤制剂相比具有用量小,不良反应少等优点1，雷公藤多苷片广泛应用于治疗类风湿性关节炎、肾病综合征、白塞氏三联征、麻风、自身免疫性肝炎及皮肤病等疾病，疗效显著，其生理活性是由二萜内酯、生物碱、三萜等多种成分协同产生2。雷公藤多苷片是目前临床上使用较多的非甾体类免疫抑制剂。目前国内雷公藤多苷片至今还没有很完善的质量控制标准,仅对雷公藤内酯G18267G6122雷公藤内酯G11014G17839G15904控制3,4,5，G1209至不同G2390G4490的产G2709G4396G3324不同G12255G5242的G5058G5334，G1038G1114准G11842G3332控制雷公藤多苷片G1025有效成分G13688的G2559量，G5326G12447雷公藤多苷片的G6363G13453G3282G169016，对其临床用G14659的有效性G2656G4445G1852性有很G3835的G5859G1053。1仪器与材料GILLSONG3G20652效G9094相G14406G16901G1214G13007G13491，G2265G6336G726306G3423G8905，119G13055G3818/G2499G16277G1821G7828G8991G3132，715G14406G16901G3800理G13007G13491，RHEODYNE9125G3423G17839G7691G19412，G18209G120920LG18331G7691G10627，805G8991G2399G1214，811CG2172G5589G9163合G3132，GILSON818G7621G9213G12677。EYELAG7071G17728G14988G2469G1214N1001G3423G3（G7097G7424G1008G1152理化G3132G7812G7678G5347G1262G12050）。B2500SMTHG3423G17241G3780G8886G9177G8939G1214（上G9035G5529G14033G1461G17241G3780有G19492公G2508）。AL204G11017G4388G3837G5191（G7769G10317G2214G16G6188G2045多G1214G3132有G19492公G2508）。雷公藤多苷片G36G794G43G1861G27G1022G6221G2507（上G9035G3809G7098G3809G2338制G14659G2390G2656G8755G14499G8900G5042G13666G17902制G14659G2390）G727雷公藤内酯G18267对G10043G2709,G2559量G28G28G17G24G22G8,雷公藤内酯G11014，G2559量G28G28G17G207G8，均购自上G9035科卫医疗技术研究所。G1025性氧化铝，上G9035五四化学试剂G2390生产G727微孔滤膜，孔径0G174G24M，上G9035半岛实业有G19492公G2508净化G3132材G2390生产。乙酸乙酯,分析纯乙腈（G14406G16901纯G3，G13666国FISHER公G2508）G727自制纯水G727其他溶剂均G1038分析纯。G322方法与结果21供试品溶液的制备G18331用均匀取G7691法从雷公藤多苷片取出G20G24片，研细成粉，精密称取400MG，置于2G24ML三角烧瓶G1025，加入20ML乙酸乙酯，G17241G3780提取G220MIN，过滤，滤G9094过G20G24G3GG1025性氧化铝G7621，用G220ML乙酸乙酯G8939脱，收集G1852部过G7621后的G8939脱G9094，回收乙酸乙酯，得残G4396物，加G20G17G24ML乙腈溶解，0G174G24M微孔滤膜过滤，备用。22雷公藤内酯醇对照品溶液的制备精密称取干燥恒重的雷公藤内酯G18267对G10043G27090G17000G27G20G，用乙腈溶解并定容G3324G200ML容量瓶G1025，制成浓G5242G10380G170G27G20MGMLA186A187对G10043G2709溶G9094，备用。23色谱条件G14406G16901G7621DIKMA公G2508DIAMONISLG3C18G32G240G3MM4G176MM,G24MG727流G2172相G3水（G36）G16乙腈（B），梯G5242G8939脱G7260G79470MIN，G36G726G36G24G8G7940G8，BG726G22G24G8G794G2000G8G727流速G20G170MLMINA186A187G3，G7828G8991G8886长G32G20G27NMG727G7621G9213G22G24G727G17839G7691体积20L。G324指纹图谱方法学建立与考察2G174G17G20G7828G8991G8886长的比较G726G3G7424实验选择G1114210，215，218，219，225NM五G1022G8886长G17839G15904G8991定，结果G3324G8886长218NM下出现的G14406G16901峰数目适G1025，内参物雷公藤内酯G18267的G14406G16901峰完整，且基线G5191稳，故选择218NM作G1038雷公藤多苷片HPLCG6363G13453G3282G16901的G7828G8991G8886长。G32G174G172不同流G2172相及G8939脱方法的比较G726G18331用乙腈G16水、乙腈G160G17G20G8磷酸等不同流G2172相G17839G15904等G5242G6122梯G5242G8939脱，结果G2469现G2559有磷酸的流G2172相并不G14033改变峰G3423。G18331用梯G5242G8939脱,G3G2499兼得二者的优点。G16277表G20。A188A189A190A191A192A193A194A195A194A195A196A197MINA198A199A20011A2011515A201403535A2011006565A201020A2014040A20110060A201030A2016040A20110060A201040A2013030A2016060A201702020A2011001008080A2010050A2017040A20110060A201060A2017035A20110065A2010结果G2469现G3324方法6的条件下，雷公藤内酯G18267的保留时间G2656峰G19766积较稳定，与相G18063其他G14406G16901峰分G12175较G3921，且其它G2520G14406G16901峰G1055间的分G12175效果G1075较G3921，且基线G5191稳，G2520峰峰G5430G1075较G3921，故G7380G13468选择6作G1038G7424实验G20652效G9094相G14406G16901法的流G2172相。G32G174G17G22不同提取方法的比较G726由于雷公藤多苷成分G3809G7446，G3252G8504G18331用不同溶剂、G8939脱G7621、G89393脱溶剂G17839G15904分G12175，G16277表2，G1209G1427G14731得良G3921的G14406G16901G3282。G3A188A202A203A204A205A206A207A208A209A210A211A194A212A194A212A210A211A213A214A192A193A215A192A193A213A214A189A216A217A218A219A220A221A222A216A217A202A216A217A218A219A220A221A222A223A224A225A216A217A226A227A216A217A228A216A217A226A227A223A224A229A216A217A226A227A223A224A230A216A217A231A189A232A233A234A235A216A217A226A227A223A224A230A216A217A231A189A232A189A234A236A216A217A218A219A220A221A222A223A224A230A216A217A231A189A232A233A234A237A216A217A218A219A220A221A222A223A224A230A216A217A231A189A232A189A234A233A216A217A226A227A230A218A219A220A221A222A238A239A215A231A189A232A189A234A216A217A189A240A241A200A198A224A218A219A220A221A222A198A242A198A243A189A189A244A245A200A246A247A248A249A250A251A246A218A219A220A221A222A198A242A198A243A189A202A198A242A198A243A218A219A220A221A222A198A242A198A243G3结果G2469现，使用乙酸乙酯作G1038提取溶剂G2656G8939脱剂，G1025性氧化铝作G1038G8939脱G7621G1038G7380G1351的G7691G2709提取方法。G32G174G174稳定性试验G726G3取雷公藤多苷片G7691G2709G36的G1391试G2709溶G9094，分G2047G33240、2、4、G27、G202HG17839G7691G8991定。G1209雷公藤内酯G18267G1038内参比峰，G16757G12651G2520G1861有峰的G14406G16901相对保留时间G2656相对峰G19766积。结果显G12046，G20G27G1022G1861有峰的G14406G16901相对保留时间的G53G54D均小于0G17G246G8、相对峰G19766积的G53G54D均小于G24G1767G8。G32G174G17G24精密G5242试验G726G3取雷公藤多苷片G7691G2709B的G1391试G2709溶G9094，G17842G13505G17839G7691G24G8437，G16757G12651G2520G1861有峰的G14406G16901相对保留时间G2656相对峰G19766积。结果G20G27G1022G1861有峰的G14406G16901相对保留时间的G53G54D均小于0G17G242G8、相对峰G19766积的G53G54D均小于4G1766G8。G32G174G176重现性试验G726G3取雷公藤多苷片G7691G2709G36，G5191G15904取G24G1233，G6365G1391试G2709溶G9094制备方法提取，得G1391试G2709溶G9094，分G2047G17839G7691G7828G8991，G16757G12651G2520G1861有峰的G14406G16901相对保留时间G2656相对峰G19766积。结果显G12046，G20G27G1022G1861有峰的G14406G16901相对保留时间的G53G54D均小于06、同时G1209雷公藤内酯G18267峰G19766积G1038G16278G4531G6363标，其峰G19766积的RSDG1038346G32G174G177G1861有G6363G13453峰的G11842定G726G3G3324G14406G16901G6363G13453G3282G16901研究G1025，不同G7691G2709G1025相对保留时间相同的峰，称G1038G14406G16901G6363G13453G3282G16901G1861有峰。G6365上G17860方法制备G11142G1022G2390G4490G27G6221雷公藤多苷片的G1391试G2709，G8892入G14406G16901G1214，G16772G5417G14406G16901G3282，其G1868G3423的G43G51LG38的G14406G16901G6363G13453G3282G16901G16277G3282G20。G34T/MINAA252A253A254A255A1A0A2A3A4A5SA12A6A7A4A5A152A252A253A254A8A9A1018A252A253A254A8A9A11A691A203A204A205A206A207A208A13A14A209A16A17A69A18G3G3结果显G12046，G1861出现G14406G16901峰G17829G240G1022，其G102522G1022G1038G2520G7691G2709所有。G13783G15397到溶剂对保留时间小于G24MIN的G14406G16901峰的G5445G2721，并G1209G2345峰G19766积G3835于G6122等于G20G8G1038选择标准，G11842定G1114G20G27G1022峰G1038G1861有G14406G16901G6363G13453峰，其G1861有G6363G13453峰的G5647G19766积G2356G5647峰G19766积的G280G3G8G1209上。248雷公藤多苷片G7691G2709相对保留时间的分析G726选择2G2507峰（雷公藤内酯G18267）G1038参G10043物，G8504峰G1038所有G7691G2709G1025G1861有，分G2047G8726出G2520G10317征峰的G16855整保留时间G1055比G162G1552，结果G16277表G22。G3A188A225A203A204A205A206A207A208A13A19A66A20A21A22A196A197A209A46A23A130A24A66A20A21A22A196A197SA5A214A25A216A217A218A26A27A28A189A240A201A235A229A236A189A223A240A201A235A235A236A228A240A201A235A235A225A225A240A201A225A237A202A189A201A240A240A240A240A189A201A240A240A240A240A240A201A240A240A225A189A201A228A228A225A228A223A189A201A228A235A236A229A189A201A228A235A240A228A240A201A237A237A228A189A201A229A189A233A233A223A189A201A229A235A225A225A189A201A229A228A235A236A189A201A228A233A229A189A201A235A202A233A228A223A189A201A235A229A236A236A189A201A235A228A237A237A189A201A189A225A235A189A201A237A228A236A235A223A189A201A237A233A189A233A189A201A237A236A237A240A189A201A229A225A236A189A201A233A202A189A225A223A189A201A233A228A236A240A189A201A233A225A236A228A189A201A240A240A237A202A201A240A189A235A237A223A202A201A240A236A202A233A202A201A240A229A235A237A189A201A237A235A233A202A201A189A240A228A236A223A202A201A189A235A236A233A202A201A189A228A233A228A202A201A240A237A189A240A202A201A202A240A236A233A223A202A201A202A237A240A229A202A201A202A229A236A229A202A201A225A225A189A189A202A201A228A240A202A189A223A202A201A228A233A189A235A202A201A228A235A228A236A202A201A233A189A189A202A202A201A228A229A228A189A223A202A201A229A228A236A240A202A201A229A189A237A229A202A201A233A233A189A225A202A201A229A228A225A237A223A202A201A235A225A240A233A202A201A229A233A236A237A202A201A229A237A189A228A202A201A235A235A202A236A223A202A201A236A189A202A235A202A201A235A237A202A235A189A201A235A237A189A229A202A201A237A240A225A235A223A202A201A237A235A240A202A202A201A237A225A229A240A189A201A233A228A189A235A225A201A240A189A240A189A223A225A201A240A236A235A225A225A201A240A228A236A189A202A201A225A189A189A236A225A201A240A236A229A235A223A225A201A189A228A228A229A225A201A189A189A240A240A202A201A229A233A189A237A229A201A202A189A229A225A223A229A201A202A233A235A228A229A201A202A235A240A229A202A201A229A237249G1861有G6363G13453峰的相对峰G19766积G726G3G1209G3282G16901G1025雷公藤内酯G18267G1038内参物，G4570G2520G14406G16901峰峰G19766积与同G980G3282G16901G1025内参物的峰G19766积比较，其比G1552G1038G2520G14406G16901峰的相对峰G19766积，G16277表4。G3A188A228A237A147A203A204A205A206A207A208A13A19A224A29A231A130A234A139A46A66A20A30A31A130A24A13A19A209A66A20A130A126A32A226A227A248A27A228A229A230A231A189A225A201A237A233A189A240A228A201A235A240A235A237A237A201A225A240A233A189A235A201A237A240A228A229A189A225A201A233A233A202A189A229A201A237A240A236A235A237A201A202A228A229A228A202A201A235A202A237A228A202A189A189A189A189A189A189A189A189A225A240A201A229A228A229A240A240A201A225A233A235A202A240A201A229A237A202A225A189A201A202A189A235A229A240A201A202A229A237A189A240A201A202A189A236A228A240A201A202A228A189A228A240A201A202A240A225A225A228A240A201A235A189A240A240A240A201A228A228A233A229A240A201A236A225A202A189A189A201A189A228A189A235A240A201A202A235A236A236A240A201A189A225A236A236A240A201A229A229A235A228A240A201A229A229A236A233A229A240A201A237A237A233A189A240A201A235A233A233A189A189A201A237A189A202A189A189A201A228A233A240A237A189A201A228A202A235A233A240A201A236A233A229A229A240A201A233A228A235A189A240A201A235A229A236A233A235A189A201A240A229A235A189A189A201A189A229A240A236A202A201A237A228A233A189A225A201A189A225A237A236A189A201A233A229A236A229A189A201A236A237A233A189A189A201A229A237A233A228A189A201A228A229A236A237A236A240A201A236A225A189A233A240A201A236A237A233A202A189A201A229A233A240A237A189A201A233A235A202A189A240A201A233A202A235A229A240A201A235A229A225A236A240A201A236A228A233A229A240A201A236A237A225A236A237A229A201A240A228A189A236A229A201A235A237A236A235A189A240A201A237A240A235A228A189A189A201A202A233A233A233A235A201A235A235A189A202A228A201A189A233A240A228A229A201A229A228A228A236A235A201A189A189A225A229A233A228A201A228A202A240A225A228A201A229A225A202A240A189A240A201A225A228A236A236A233A201A225A235A189A237A228A201A228A233A233A228A225A201A202A202A236A228A225A201A229A236A240A189A229A201A228A229A235A236A189A240A228A201A237A228A236A202A229A201A189A225A233A228A189A202A201A228A237A240A237A189A240A201A236A228A189A189A228A201A189A235A189A189A202A201A225A225A225A202A228A201A240A228A236A228A235A201A235A228A235A225A189A189A189A201A189A233A202A237A189A201A228A237A189A236A202A201A229A225A228A235A225A201A233A225A235A229A189A201A237A229A237A189A201A240A233A189A189A201A228A189A240A235A189A201A189A236A225A233A189A202A202A201A240A202A228A237A202A201A235A225A235A237A236A201A189A202A225A189A228A201A235A235A189A202A236A201A233A228A235A236A229A201A202A228A229A202A235A201A235A235A240A235A225A201A235A235A235A189A189A225A235A201A202A235A236A189A229A201A236A229A228A229A189A236A201A237A225A225A233A189A202A201A228A189A225A236A228A201A189A237A240A202A202A201A228A229A235A229A228A201A189A229A228A236A237A201A240A202A229A229A189A228A189A240A201A202A228A237A225A189A240A201A233A236A229A189A202A189A201A237A233A228A229A189A229A201A237A235A233A202A202A189A201A225A225A240A225A189A235A201A225A237A240A202A189A237A201A237A202A189A225A189A235A201A237A202A240A233A189A229A202A201A237A228A236A189A202A201A229A236A228A229A236A201A229A237A225A237A229A201A233A235A229A233A225A201A225A233A233A233A202A201A189A235A228A229A225A201A240A229A189A228A201A240A228A229A237A189A235A236A201A189A240A236A233A235A201A233A225A189A189A189A236A201A229A233A240A189A189A189A201A235A233A202A229A236A201A240A237A236A235A225A201A236A229A235A235A201A202A225A225A228A189A240A201A189A202A236A189A189A236A202A201A237A233A228A240A225A201A202A189A228A189A236A201A240A235A228A236A229A201A202A236A225A202A225A201A240A233A233A189A202A201A189A235A225A229A225A201A202A240A233A229A229A201A189A237A240A228A189A237A240A201A189A233A240A237A240A201A189A237A225A237A240A201A228A225A228A240A240A201A202A229A235A202A240A201A225A225A233A229A240A201A225A235A237A225A240A201A228A233A202A233A240A201A225A233A202A2023讨论6雷公藤多苷片是从雷公藤根提取精制G13792成的G14038溶性成分G9163合物,目前国内不同G2390G4490生产的雷公藤多苷片G3252G13582G1059有效的质量控制方法，G3252G13792其化学成分及G2559量G4396G3324G11540G980定的G5058G5334。G3926G1321G3324提G20652G14659效G2656G19489G1314G8614G2115作用G1055间G6226到G980种G5191G15925,并G6466G8504制定G980G1022有效的质控标准,对于雷公藤多苷片G17837G980非甾体类免疫抑制剂的临床应用G2656G14659效G16792G1227G18129具有重G16213G5859G1053。G8504G8437试验G2033G8505G5326G12447G1114雷公藤多苷片的HPLCG6363G13453G3282G16901，由试验结果G2499G1209G11487出不同G7691G2709的G20652效G9094相G6363G13453G3282G16901的G1039G16213峰的G19766积基G7424G980G14280，提G12046不同G2390G4490及G6221G2507的雷公藤多苷片的G1039G16213成分基G7424相同，G1306G2559量有所不同。G2045用雷公藤多苷片HPLCG6363G13453G3282G16901G2499G1209较G3921G3332反G7156雷公藤多苷片的内G3324质量，G1038今后雷公藤多苷片的质量控制提G1391G1114G1393G6466。G14406G16901G6363G13453G3282G16901实验条件应G14033G9397G17287G6363G13453G3282G16901的G19668G16213，不G4464G12628G2345G3883用G2559量G8991定的实验条件，G1038G14731取G17287G1209G1207表雷公藤多苷片G10317征的G6363G13453G3282G16901。参G13783有关G7003G10498后，G20330G1820选择G1114等G5242G8939脱G1038流G2172相，G3926乙腈G16水G220G72670，乙腈G16水G22G24G7266G24，乙腈G16水40G72660G3，乙腈G16水60G72640,G3乙腈G160G17G20G8磷酸40G72660等等G5242G8939脱，G1306分G12175效果不理G5831，G2520峰重G2484G10587较G20652，内参物雷公藤内酯G18267的G14406G16901峰不G14033完G1852分G12175。G5415流G2172相G1025乙腈比G1375G3324G220G8G79460G8时，G19555G11540乙腈比G1375G1955少，G6363G13453G3282G16901G1025G2520G14406G16901峰保留时间G2588G5322长G17247G2195。G3雷公藤多苷片G1025G2559有较多的化学成分,G3G5415流G2172相G18331用等G5242G8939脱及G18331用不同比G1375的流G2172相时，G2029出峰时间G6301长G6122G14406G16901峰不G14033G17810到基线分G12175。G18331用梯G5242G8939脱,G3G2499兼得二者的优点，雷公藤内酯G18267的保留时间G2656峰G19766积较稳定，与相G18063其他G14406G16901峰分G12175较G3921，且其它G2520G14406G16901峰G1055间的分G12175效果G1075较G3921，且基线G5191稳，G2520峰峰G5430G1075较G3921。G18331用不同的提取溶剂时，G20330G1820G4593试G1114用G8707G1235G17839G15904分G12175提取，结果G2520G14406G16901峰不G14033G3827较G3921的分G12175，内参物雷公藤内酯G18267峰G19766积较小，且与其它G14406G16901峰重G2484，不G14033准G11842G17788G2047出雷公藤内酯G18267的G14406G16901峰。G5415G18331用G7092水乙G18267G6122G14988G20323水作G1038提取溶剂时，G14406G16901峰出峰的效果较G8707G1235G1351。G18331用用乙酸乙酯G7481取G1207G8707G1235、G7092水乙G18267G6122G14988G20323水所制得的G1391试G2709，G7446质较少，G2520G14406G16901峰基G7424G2499G1209完G1852分G12175，且雷公藤内酯G18267的相对峰G19766积较G3835，与相G18063其它G14406G16901峰分G12175较G3921。G3G18331用不同的G8939脱溶剂时，G5415使用G8707G1235作G1038提取溶剂，G11014G18267G6122G11014G18267G16G8707G1235作G1038G8939脱剂时，内参物雷公藤内酯G18267的峰G19766积较G3835，G1306其它G7446质峰G19766积变得G7368G3835，反G13792使得雷公藤内酯G18267相对峰G19766积G1955小。相比G1055下G5415乙酸乙酯作G1038G89