[生物医药论文精品]spe-hplc法测定藿香正气水中厚朴酚与和厚朴酚的含量

SPEHPLC法测定藿香正气水中厚朴酚与和厚朴酚的含量A1A0A2A3A4A5SPEHPLCA6A7A8A9A10A11A12A13A14A15A16A17A18A19A15A16A17A20A21A22A23A24A25A6A26A27A28A29A30SPEA31A32A33A34A35A36A37A38A26A39A40A32A41C18A42250MM46MMIDA435MA44A43A45A46A47A48A49A50A51A52A53A4270A5425A545A54025A44A55A56A57A58A43A56A59A5510MLMIN1A43A60A61A62A63A55294NMA43A32A64A55A65A64A26SPEHPLCA66A61A67A68A69A70A71A72A734080GML1R09998A43A74A68A69A70A71A72A732040GML1R09998A75A76A77A78A79A80A81A82A83A84A85A86A87A88A26A68A69A70A89A90A91A92A93A94A55998A43RSDA5508A43A74A68A69A70A89A90A91A92A93A94A55996A43RSDA5505A26A95A96A66A97A59A98A99A100A98A101A102A98A103A104A84A88A43A105A106A107A108A109A110A111A112A113A114A48A89A115A116A26A117A118A119A118A119A41SPEHPLCA111A112A113A114A48A68A69A70A74A68A69A70DETERMINATIONOFMAGNOLOLANDHONOKIOLINHUOXIANGZHENGQIWATERBYSPEHPLCABSTRACTASPEHPLCMETHODFORDETERMININGTHECONTENTSOFMAGNOLOLANDHONOKIOLINHUOXIANGZHENGQIWATERWASESTABLISHEDSAMPLESWEREPURIFIEDBYSILICAGELSPECOLUMNTHEHPLCCOLUMNUSEDINTHEEXPERIMENTWASC18A120250MM46MMID,5MA121,ANDTHEMOBILEPHASEWASMETHANOLWATERACETONITRILEACETICACID70255025THEFLOWRATEOFTHEMOBILEPHASEWAS10MLMIN1ANDDETECTIONWAVELENGTHWAS294NMTHEEXPERIMENTWASACCOMPLISHEDATROOMTEMPERATUREITHADGOODLINEARRELATIONSHIPBETWEENTHEPEAKAREASOFMAGNOLOLANDHONOKIOLANDTHEIRCONCENTRATIONSAT40800GML1R09998,20400GML1R09998,RESPECTIVELYTHEAVERAGERECOVERYOFHONOKIOLWAS996WITHRSDBEING05,ANDTHATOFMAGNOLOLWAS998WITHRSDBEING08THEMETHODWASRAPID,SIMPLE,ACCURATE,WITHGOODREPRODUCIBILITY,ANDITCANBEUSEDFORQUALITYCONTROLOFHUOXIANGZHENGQIWATERKEYWORDSSPEHPLCHUOXIANGZHENGQIWATERMAGNOLOLHONOKIOL藿香正气水系由甘草、陈皮、厚朴等多味中药制成的传统中成药，功能解表化湿、理气和中，常用于外感风寒、内伤湿滞或夏伤暑湿所致的感冒1，具有抗菌、抗病毒、抗过敏、影响胃肠活动、肌肉松弛和中枢抑制等作用2。中国药典2005年版一部1中以厚朴酚与和厚朴酚的含量作为藿香正气水的质量控制指标，以氯仿为溶剂，采用液液萃取技术对样品进行预处理，然后以高效液相色谱法HPLC进行含量测定。但液液萃取预处理技术费时、易乳化、需要样品量大,并G1000需要G17751多有毒或有G8757G7591的有G7438溶剂，易G8757G7591G10627G3671。G13792采用G3278相萃取技术G708G54G51G40G709G2499以G18003G1825G990G17860G13582G9869。G3278相萃取具有高的G3250G6922G10587和G4512G19610G1505G6980，G17836具有有G7438溶剂用量G4581，G7092相G2010G12175G6817作G2499G18003G1825G1147G10995乳化，G2010G7524G10301G8981G2010易于G6922G19610，能处理G4579G1319G12227G16809样，G6817作G12628G2345G5567G17907等G1260G986935。G7003G104986,7采用高效液相色谱法测定G1114藿香正气水中厚朴酚与和厚朴酚的含量，采用G11464G6521G12244G18334法对样品进行G2081处理。G8504外，G17836有对藿香正气水中G11345G14475、G8237皮G14539、甘草G14539、水G2524G8699化G2081G14005G13044、G14469G7910毒G13044、甘草G18252、G8237G21656G18242或G5334G8237G21656G18242、G8443G2081G14005G13044等的G11752G12362811。G7424G7003G3324药典G7053法的G3534G11796G990，G5326G12447G1114采用SPE样品预处理技术G13443化样品，G1889G2045用G43G51G47G38法对厚朴酚与和厚朴酚进行定量G2010G7524，并进行G1114系统的G7053法G4410G13783G4531，G16792G1227SPE技术用于G16825药品预处理的G4466用G5627。G31实验部分11仪器与试药111仪器G3高效液相色谱G1214G708CXTH3000，G2283G1152G2031G7044G17902G5670G12197技有G19492G1856G2508G709、G17241G3780G8886G9177G8939G3132G708DS2060，G3837G8953G5078G1008G5259G12197技有G19492G1856G2508G709、G11017G4388G3837G5191G708AG204，G10802G3775METTLERTOLEDOG709、SPEG7621G708500MG/G6915，G14270G3647装G709。G3112试药厚朴酚与和厚朴酚对照品G708G990海云G990G10995G10301有G19492G1856G2508G709、藿香正气水G70810ML，G5078售G709；乙腈G708色谱G13443G709、甲醇G708G1260级G13443和G2010G7524G13443G709，氯仿、G5334丙醇、正己烷和羧甲G3534纤维G13044钠等均为G2010G7524G13443，硅胶G708200300目，青岛海洋化工厂G709，重蒸水。G312色谱条件色谱G7621C18G70825046MMID，5MG709；G8981动相甲醇水乙腈乙G18252G70870255025G709；G8981G179071MLMIN1；检测G8886长294NM；室温；进样量20L。G313溶液的配制131对照品溶液的配制G3精密称取厚朴酚对照品20MG、和厚朴酚对照品10MG，配制成含厚朴酚80GML1，和厚朴酚40GML1的甲醇溶液，做为对照品储备液。精密量取G990G17860对照品储备液25ML置10ML量瓶中，甲醇定容，摇匀，即得对照品溶液。G3132样品溶液的配制1321药典法G3根据药典法处理样品1精密量取样品5ML，加盐G182522滴，用三氯甲烷振摇提取三次，每次10ML，G2524并三氯甲烷提取液，蒸干，残渣用甲醇溶解并转移到10ML量瓶中，加甲醇至刻度，摇匀。精密量取5ML，置10ML量瓶中，加甲醇至刻度，摇匀，经045M微孔滤膜滤过，取续滤液即得。G31322SPE法G31SPE柱的制备称取硅胶05G，加入075ML01的羧甲G3534纤维G13044钠G708CMCNAG709溶液，G3324G11752钵中G11752匀，装入G7621中，60活化1H，放入干燥G3132中备用。G32固相萃取精密量取样品溶液01ML，加入预先活化的SPEG4579G7621，用3ML正己烷G8939脱，G6922G19610G8939脱液，加入1MLG5334丙醇，用甲醇定容至5ML量瓶中，摇匀。经045M微孔滤膜过滤，取续滤液即得。G31323直接稀释法精密量取样品01ML，置5ML量瓶中，加甲醇适量G17241G378030MIN，放冷至室温，甲醇定容，摇匀即得。G3G2010别取对照品溶液及样品溶液，按G990G17860色谱条件测定，记录色谱图，见图1。G32结果与讨论21系统适用性实验和厚朴酚与厚朴酚的保留时间各约为93MIN和140MIN，理论塔板G6980按厚朴酚计算为8764，拖尾因G4388104，G2010G12175度206，样品溶液G3324待测组G2010峰处G7092干扰，另外由色谱图G2499见，经SPE处理的样品中其它组G2010峰已明显减G4581。G322线性关系考察G3从对照品储备液中依次精密量取05、1、2、25、5ML溶液，用甲醇定容至10ML量瓶中，连同对照品储备液共得到6个不同G8999度的对照品溶液，按G990G17860色谱条件测定，连续进样3次，记录峰G19766G12227，G8726G5191均G1552。G2010别以厚朴酚与和厚朴酚的G8999度G708CG709为G8190G3364标，峰G19766G12227G708AG709为G13449G3364标G3250G5414，得标G1946G7366G13459G7053G12255G3218822522514269035973941934014079221577071421283542495663246810121416182022MIN28363931933214083221291470714212835424956246810121416182022MIN3928926914004707142128246810121416182022MINA1221A123A124A125A126A127A125A128A129A130HPLCA131A122AA132A133SPEA134A135A127A125A130HPLCA131A122BA133SPEA134A135A127A125A130HPLCA131A122CA123A124A125A128A129HPLCA131A122DA136A124A137A138A139A140A134A135A127A125A130HPLCA131A1221A126A141A142A143A1442A141A142A143FIGURE1THEHPLCSPECTRAOFTHEREFERENCEANDTHESAMPLESOLUTIONSATHEHPLCSPECTRUMOFTHESAMPLESOLUTIONUNTREATEDBYSPEBTHEHPLCSPECTRUMOFTHESAMPLESOLUTIONTREATEDBYSPECTHEHPLCSPECTRUMOFTHEREFERENCESOLUTIONDTHEHPLCSPECTRUMOFTHESAMPLESOLUTIONTREATEDBYTHEPHARMACOPOEIAMETHOD1HONOKIOL2MAGNOLOL21272325253592901401870714212835246810121416182022MINC1122DBA2112212厚朴酚A54963C45503G708R09998，N6G709，和厚朴酚A104175C86963G708R09998，N6G709，表明厚朴酚、和厚朴酚的检测G8999度G2010别G33244080GML1、2040GML1的G14551G3272内与各G14270峰G19766G12227G13459G5627G1863系G14403G3921。厚朴酚的检测G19492G708S/N3G709和定量G19492G708S/N10G709G2010别G7171022GML1和074GML1，和厚朴酚的检测G19492和定量G19492G2010别G7171016GML1和053GML1。23日内精密度实验取对照品溶液G3324一G3837内重G3809进样6次，记录峰G19766G12227，G13479G7536厚朴酚峰G19766G12227的RSDG72907G705，和厚朴酚的RSDG72904G705，表明G16825G7053法G7097内精密度G14403G3921。24日间精密度实验取对照品溶液连续进样5D，记录峰G19766G12227，G13479G7536厚朴酚峰G19766G12227的RSD16，和厚朴酚峰G19766G12227的RSD07，表明G16825G7053法G7097间精密度G14403G3921。G325稳定性实验G3按SPE法处理样品溶液，连续三G3837内G2010时进样，以峰G19766G12227作为G12295定G5627G16792G1227指标，G13479G7536厚朴酚与和厚朴酚的峰G19766G12227G3534G7424不G2476，并G1000G7092G7446峰G1998G10628，表明样品溶液至G4581G332448H内G12295定。G326重现性实验取同一G6221G2507样品5G1233按SPE法G5191行处理，按G990G17860色谱条件测定，G13479G7536厚朴酚峰G19766G12227的RSD16，和厚朴酚峰G19766G12227的RSD06，表明G16825G7053法重G10628G5627G14403G3921。27回收率实验G3取同一G6221G2507样品按照G11464G6521G12244G18334法进行G6817作，同时按照SPE法G5191行G6817作。G5191行G1128G1233，G2010别记录峰G19766G12227，计算G3250G6922G10587，G13479G7536厚朴酚的G5191均G3250G6922G10587为998，RSDG708G709为08；和厚朴酚的G5191均G3250G6922G10587为G28G28G17G25G8，G53G54G39G708G70905。G4466G20576G13479G7536显G12046G5191均G3250G6922G10587G33249911011以内，表明G16825G2010G7524G7053法G1946G11842度G9397G5859。G329样品测定按SPE法和药典G7053法，G2010别制备厂G4490一和厂G4490G1120不同G6221G2507样品的G1391G16809品溶液，记录峰G19766G12227，计算和厚朴酚与厚朴酚的含量。G13479G7536表明，G7424G7003G7053法与药典G7053法的测定G13479G7536G3534G7424一致。G3G3表1样品含量测定G13479G7536G3TABLE3DETERMINATIONRESULTSOFTHESAMPLESMANUFACTURERBATCHNUMBERPHARMACOPOEIAMETHODG708MGML1G709THEMETHODG708MGML1G709HONOKIOLMAGNOLOLHONOKIOLMAGNOLOL107062304860395049204050707010451035304570351060502037105190381053507061504640362048503742200705080643047206510487210讨论G32101SPE柱填料的选择采用C18为G3647G7021，G2499G1363待G2010G7524G10301厚朴酚与和厚朴酚G17751G5390G3332保留G3324G2572G19480剂G990，G4570G7446质先G9119G8939G991G7481，G1889G8939脱待G2010G7524G10301。但G7171由于样品溶剂为40G79450的乙醇，G7424G17535G8939脱能G2159G17751大，G1363其不能G3324G2572G19480剂G990G5468G3921保留，与G7446质一G17227G15999G9119G8939，G17810不到预G7411的目的，G13792G6393干样品溶剂G2460G3698加G1114G6817作的G13333G10724。G6925用G7509G5627G2572G19480剂硅胶，G4570G7446质保留G3324G2572G19480剂G990，G13792G20330先G4570待G2010G7524G10301G8939脱G991G7481，G12628化G1114G6817作G8505G20600，取得G1114G9397G5859效G7536。G32102SPE柱的制备和活化G3324硅胶中加入适量G21667G2524剂01CMCNA溶液，G1363硅胶G21667G2524，制备G54G51G40G4579G7621G2499不G1363用G12591板，并G1000所G990样品G2010G5079均匀。SPEG7621G17885定G2572G19480剂用量为05G，60干燥1H活化。G32103淋洗剂的选择及用量G7378G17885用甲醇、乙腈、乙G18252乙G18243、G1120氯甲烷、G11719G8845G18286和正己烷等溶剂，或其中一定G8616G1375的G1016G12193溶剂G9163G2524进行G8939脱，G7380后G17885用正己烷作为G8939脱剂。G11464G6521用3ML正己烷G8939脱后，G8939脱液用甲醇G12244G18334时，G1126溶G5627不G7171G5468G3921，因G8504加入1MLG5334丙醇，G3698加溶剂间的G1126溶G5627。G32104上样量的选取SPEG4579G7621的G990样量一G14336不得G17241过G3647G7021高度的1/3，同时G13520G2524G13783G15397样品中待测成G2010的G8999度和检测G9801敏度，G7424G4466G20576中G990样量G17885取01ML。G33结论G3G3278相萃取法G1823G7393G1114一G14336液液萃取的G13582G9869，G2499G14422约时间和溶剂，G1940化效G7536G3921，对G6817作G13785G4445G1852，G1000G10627G3671G2463G3921，重G10628G5627G9397G5859，G7171一G12193G5567G17907高效的样品G2081处理G7053法。G3参考文献1STATECOMMITTEEOFPHARMACOPOEIAPHARMACOPOEIAOFPEOPLESREPUBLICOFCHINA,PART1BEIJINGCHEMICALINDUSTRYPRESSA145A146A147A148A149A150A151A152A153A154A155A156A157A158A146A148A149A159A160A161A162A163A164A165A166A167A168A169A170A171A1722005A1636612STATECOMMITTEEOFPHARMACOPOEIANOTICEOFMEDICINESCLINICALUSEVOLUMETCMBEIJINGPEOPLESMEDICALPUBLISHINGHOUSEA145A146A147A148A149A150A151A152A173A174A175A148A176A177A153A148A178A161A162A163A155A156A179A180A168A169A170A171A1722005A163263TIANJ,ZHAIB,YULPRINCIPLESOFSPEMODESSELECTIONJJOURNALOFDALIANINSTITUTEOFLIGHTINDUSTRYA145A181A182A183A166A167A165A184A165A185A171A1721996A17215A163414GAOW,WUZP,XUCXSTUDYONSPETECHNOLOGYJJIANGSUFOODANDFERMENTATIONA145A186A187A188A189A190A191A192A171A1722006A1723A163175WANGL,GUOLBSITUATIONOFTHEAPPLICATIONSPEFORMSJJOURNALOFHENANUNIVERSITYOFCHINESEMEDICINEA145A193A194A153A195A165A184A165A185A171A1722007A17224A163866LIULZ,GUOLDETERMINATIONOFMAGNOLOLANDHONOKIOLCONTENTSINHUOXIANGZHENGQISOLUTIONBYHPLCJ