[生物医药论文精品]大孔吸附树脂对贯叶金丝桃中苯并二蒽酮类成分的富集

大孔吸附树脂对贯叶金丝桃中苯并二蒽酮类成分的富集摘要目的考察大孔吸附树脂对贯叶金丝桃中苯并二蒽酮类成分的富集方法,并确定工艺路线和吸附参数。方法本文以贯叶金丝桃中苯并二蒽酮类成分为考察指标，采用动态和静态吸附试验相结合来考察大孔吸附树脂对贯叶金丝桃中苯并二蒽酮类成分的富集工艺。结果本文得到稳定可行的对贯叶金丝桃中苯并二蒽酮类成分富集的工艺路线和吸附参数。结论该方法可为贯叶金丝桃中苯并二蒽酮类成分的大孔树脂精制工艺的确定提供参考。关键词贯叶金丝桃；苯并二蒽酮；大孔吸附树脂；静态吸附；动态吸附所属专业中药与天然药物（中药与天然药物资源的开发利用和可持续发展）ENRICHMENTOFNAPHTHODIANTHRONESINHYPERICUMPERFORATUMLWITHMACROPOROUSRESINABSTRACTOBJECTIVETOSTUDYTHEENRICHMENTOFNAPHTHODIANTHRONESINHYPERICUMPERFORATUMLWITHMACROPOTOUSRESIN,ANDDEFINETHEREFINEMENTPROCESSANDADSORPTIONFACTORMETHODSTHEREFINEMENTPROCESSANDADSORPTIONFACTOROFTHESEPARATIONANDPURIFICATIONOFNAPHTHODIANTHRONESINHYPERICUMPERFORATUMLWITHMACROPOROUSRESINWERECOMPAREDBYSTATICANDDYNAMICADSORBINGEXPERIMENTSANDTHECONTENTOFNAPHTHODIANTHRONESWASDEVELOPEDASITSQUALITYSTANDARD,SCREENINGOUTTHEBESTREFINEMENTPROCESSANDADSORPTIONFACTORRESULTSTHEREFINEMENTPROCESSANDADSORPTIONFACTOROFTHEENRICHMENTOFNAPHTHODIANTHRONESINHYPERICUMPERFORATUMLWITHMACROPOROUSRESINARESTABLEANDFEASIBLECONCLUSIONTHEENRICHMENTMETHODCANOFFERINFORMATIONABOUTTHEDETERMINATIONOFREFINEMENTPROCESSOFA1A0A2A3A4A6A7A5A8A2A3A4A9A10A4A11A14A5A8LWITHMACROPOROUSRESINKEYWORDSHYPERICUMPERFORATUMLNAPHTHODIANTHRONESMACROPOTOUSRESINSTATICADSORPTIONDYNAMICADSORPTION贯叶金丝桃为藤黄科植物贯叶金丝桃HYPERICUMPERFORATUML的干燥地上2部分。贯叶金丝桃的主要化学成分包括苯并二蒽酮类化合物,如金丝桃素、伪金丝桃素、原金丝桃素、原伪金丝桃素等；黄酮类化合物,如芦丁、金丝桃甙、槲皮素、异槲皮素、阿曼托黄素等；间苯三酚类化合物,包括贯叶金丝桃素、加贯叶金丝桃素等；还含有挥发油类、香豆素类化合物等A12A13A15A16A17。据文献报道,贯叶金丝桃中的抗抑郁活性成分主要有苯并二蒽酮类、黄酮类、间苯三酚类等。其中，苯并二蒽酮类被普遍认为是抗抑郁G1328用G17751G3921的成分，G3281G1881G3818对贯叶金丝桃提G2474物G3822以金丝桃素G16757的苯并二蒽酮类含G18339G993G1314G1122G19G17G22G8G1328为G17148G18339G6523制指标。本文采用大孔吸附树脂来考察贯叶金丝桃中苯并二蒽酮类的吸附G10317性，G1186G13792G17839G980G8505确定苯并二蒽酮类活性成分富集方法,得到G17751G20652G13443G5242的贯叶金丝桃有G6940部G1313。1仪器、试剂和材料HP6890G8680相G14406G16901G1214，HP7694G20042G12366G17839G7691G1214；G4719G8953UV2401PC；金丝桃素对G10043G2709G17153G14270G13666G3281SIGMAG1856G2508；试G2070G3355为分G7524G13443。大孔树脂G3355为G3281G1147；贯叶金丝桃采G17153G1122G19497G16211G11477G8733中G5078。2实验部分21苯并二蒽酮类的含G18339G8991方法211供试G2709和对G10043G2709的制G3803供试G2709G9354G9094G726G2474G7691G2709G1343401G，精G4506G12228定，G1427950MLG7849G14406G18339G10954中，加G11014G1826740ML，G17241G3780G3800G10714G708G2163G10587350W、G20069G1058735KHZG70940分G19059，G6930G1931，加G11014G18267G14279G2063G5242，G6683G2260，G17819G9400。精G4506G18339G2474G13505G9400G90942ML，G1363410MLG7849G14406G18339G10954中，加G11014G18267G14279G2063G5242，G6683G2260。对G10043G2709G9354G9094G726精G4506G12228G2474G1128G8699化二G11979G1955G2399干燥G21G23G75的金丝桃素对G10043G2709，加G11014G18267G16G2549G2890G11G28G29G20G12G18209制成G8611G20G80G79含金丝桃素G21G19G的G9354G9094，G13634G7849G14406G18339G10954中，G1328为对G10043G2709G1660G3803G9094。212G8991定方法分G2047G2474供试G2709G9354G9094和对G10043G2709G9354G9094，以试G2070G1328G12366G11345，G10043分G1821G1821G5242法G708中G3281G146593G18682005G5192G10268G980部附G5417VBG709，G3324588NMG8886G19283G3800G8991定吸G6922G5242，G16757G12651，G2375得。213线性G14551G3272的考察分G2047精G4506G18339G2474对G10043G2709G1660G3803G909410ML、20ML、40ML、60ML、80MLG112210MLG7849G14406G18339G10954中，加G11014G18267G2549G289091G12244G18334G14279G2063G5242，G6683G2260，G10043分G1821G1821G5242法中G3281G14659G18682005G5192G10268G980部附G5417VB，G3324588NMG8886G19283G3800G8991定吸G6922G5242，以吸G6922G5242为G13449坐标，浓G5242为横坐标，绘制标准曲线。结果得到回归方程G726C029392353A，R09988。结果表明G3324198198G/MLG14551G3272G1881苯并二蒽酮类浓G5242与吸G6922G5242有良G3921的线性关系。22大孔吸附树脂有机残留物检查G10043有机G9354G2070残留G18339G8991定法G708中G3281G14659G18682005G5192G10268二部附G5417VIIIP第二法G709G8991定。221G14406G16901条件用键合/交联聚乙二G18267毛细管G14406G16901柱G70830M025MM025MHPINNOWAXAGILENTG1856G2508G709；柱温5G19，G17839G7691口及检G8991器温G5242分G2047为260及300；载G8680为氮G8680，流速G72620ML/MIN；G20042G12366G2474G7691G10954加热条件G726120加热15MIN。222供试G2709和对G10043G2709的制G3803供试G2709G9354G9094的制G3803G726G2474G7691G2709G1343402G，精G4506G12228定，G1363420MLG20042G12366G2474G7691G10954中，精G4506加入06十二烷基硫酸钠G9354G90945ML，G6683G2260，G2375得。对G10043G2709G9354G9094的制G3803G726精G4506G12228G2474正己烷，苯，G11014苯，对二G11014苯，邻二G11014苯，苯乙烯和G20,G21G16二乙基苯适G18339，加06十二烷基硫酸钠G9354G9094G18209制成G86111ML分G2047含15G，10G，12G，12G，13G，16G和13G的G9354G9094，G6683G2260，精G4506G18339G24745ML，G1363420MLG20042G12366G2474G7691G10954中，G2375得。223G8991定法分G2047G2474对G10043G2709G9354G9094和供试G2709G9354G9094，G3324120加热15MIN，依法G8991定，按G3818标法以峰面积G16757G12651，G2375得。23贯叶金丝桃提G2474G9094的制G3803G2474贯叶金丝桃G14659材，加6G19G8乙G18267回流提G2474二次，G8611次G20G175G75，合并提G2474G9094，G9400G17819，G9400G9094G1955G2399浓缩G142791MLG9354G9094G13434相当G11221GG14659材，离心，G3803用。24大孔树脂的预G3800G107144以95乙G18267湿法装柱,以95乙G18267洗脱，G993时检G8991流出的乙G18267，当流出的乙G18267与水混合后G993呈G11345G14406浑浊时G2375可，再以蒸馏水洗G14279无G18267味。实际操G1328中，乙G18267洗G14279柱体积的4倍时与水混合G993浑浊，洗G142796倍时，无树脂味。25影响大孔树脂静态吸附的因素251树脂型号的筛选选G24746种经预G3800G10714的湿态树脂5GG708D101、DA101、DA201、DB301、AB8、LSA20G709G13634100ML具塞锥形G10954中，G2474贯叶金丝桃提G2474G9094加入G10954中，G332425G6683床中振荡24H，吸附达到平衡后，G9400去树脂，定G18339移G2474清G9094，G8991定含G18339，G16757G12651吸附G10587。结果，吸附G10587依次为7G21G17G20G22，7G22G17G198，6G19G17G20G21，6G20G1778，65G175G20，8G19G17G225。本文根据吸附G10587结合树脂成本综合考虑，最终选择LSA20型大孔吸附树脂。252吸附时间的考察G247430G湿态LSA20型大孔吸附树脂，G13634G1122100ML具塞锥形G10954中，加入贯叶金丝桃提G2474G9094，G332425G6683床中振荡24H，G2474G7691时间依次为1，2，4，6，8，10，24H。G8991定吸附G10587，考察吸附时间对静态吸附的影响。结果为，结合吸附G10587和时间成本综合考虑，最终选择吸附时间为G21G75。26影响大孔树脂动态吸附的因素261洗脱G9354G2070的筛选精G4506吸G2474贯叶金丝桃提G2474G9094100MLG8611ML含生G146591G上柱,依次用蒸馏水、20乙G18267、50乙G18267、70乙G18267、95乙G18267梯G5242洗脱,G8611G10954G6922集G980个柱体积的洗脱G9094,依次按1，2，G22编号，分G2047浓缩G14279干,G8991定G8611G1233洗脱G9094中苯并二蒽酮G18339。以G6922集G18339G10954编号X为横坐标，苯并二蒽酮G18339Y为G13449坐标,绘制洗脱曲线,G16277G32821。5A181A19A20A21A22A23A24A25A26A27A28A29A30A31A32A33A34A35A36A37A38A39A40A41A42A21A22A23A24A26A27A28A29A30A31A33A34A35A36A37A38A39A4015A43A44A45A46A47A48610A49A5020A51A52A46A47A481115A49A5050A51A52A46A47A481622A49A5070A51A52A46A47A482326A49A5095A51A52A46A47结果G7186G12046，50、70、95乙G18267洗脱G9094中苯并二蒽酮G18339G2356G5647G18339的94，G16840明苯并二蒽酮主要集中G332450、70、95乙G18267洗脱G9094中。G13792其中50乙G18267洗脱G9094和70乙G18267洗脱G9094中苯并二蒽酮G18339G2356G5647G18339的84，95乙G18267洗脱G9094中苯并二蒽酮G18339G17751G4581。70乙G18267的G7509性与50乙G18267G6521G17829，G1000G4579G112250乙G18267，相对G13792G16340可洗脱G991的水G9354性G7446G17148G1075G17751G4581，可基本G9097G1143450乙G18267洗脱G9094可洗脱的苯并二蒽酮G18339。因G8504，综合考虑G1955G4581操G1328工G5219,G14422G13434生G1147成本等因素，G2033G8505确定洗脱条件为G1820用蒸馏水、20乙G18267洗去水G9354性G7446G17148，再以70乙G18267洗脱，G6922集70乙G18267洗脱部分。262洗脱G2070用G18339的确定精G4506吸G2474贯叶金丝桃提G2474G9094100MLG8611ML含生G146591G上柱,。G1820用蒸馏水、20乙G18267洗脱，再以70乙G18267洗脱，G8611G10954G6922集G980个柱体积的洗脱G9094,依次按1，2，G22编号，分G2047浓缩G14279干,G8991定G8611G1233洗脱G9094中苯并二蒽酮G18339。以G6922集G18339G10954编号X为横坐标，苯并二蒽酮G18339Y为G13449坐标,绘制洗脱曲线,G16277G32822。A53A54A55A56A57A58A59A60A61A62A63A64A65A64A66A64A67A64A68A64A69A64A70A64A71A64A72A64A63A73A64A63A63A64A63A65A64A63A66A64A63A67A64A63A68A64A63A69A64A63A70A64A63A71A64A63A72A64A65A73A64A65A63A64A65A65A64A65A66A64A65A67A64A65A68A64A65A69A64A74A75A76A77A78A79A80A81A82A83A846A85A86A87A88A89A90A85A90A86A90A87A90A88A90A89A90A91A86A91A92A91A87A91A93A91A88A91A94A91A89A91A95A91A90A85A91A90A90A91A90A86A91A90A92A91A90A87A91A90A93A91A90A88A91A90A94A91A90A89A91A90A95A91A96A91A97A98A99A100A101A102A103A104A105A106A1072A108A10970A110A111A112A113A114A115A116A117A118A119A120A121A122A123A112A113A124A125A126A127A128A110A111A113A114A115A116A117A118A120A121A122A123A124A125A127A12815A129A130A131A132A133610A12920A134A135A131A132A1331119A136A13770A138A139A140A141A142G32822结果G7186G12046，当以水洗脱4倍柱体积、20乙G18267洗脱3倍柱体积时，洗脱曲线G5062G17829横G17736G1000G6164G6922集的洗脱G9094G1972G1058无G14406，因G13792确定G19512去G7446G17148的水用G18339为4倍柱体积，20乙G18267用G18339为3倍柱体积。采用70乙G18267洗脱，洗脱G142797倍柱体积时曲线G5062G6521G17829横G17736，G1000G20817倍柱体积洗脱G9094中苯并二蒽酮G18339G13434G235670乙G18267G5647洗脱G18339的96以上，G6937确定洗脱G9354G4198为7倍柱体积的70乙G18267。263影响大孔树脂吸附性G14033因素的G11752G123622631大孔吸附树脂的吸附性G14033的考察大孔吸附树脂的吸附性G14033与G14659G9094浓缩G8616、上G7691流速、柱G5464G20652G8616等因素有关。本文采用L934正交试验筛选最G1351的上柱工艺条件。正交G16786G16757的因素水平表G16277表1。A1431L9A14434A145A146A147A148A149A150A151A152A153A154A147A148A152A153A155A156A157A158A159A160A161A162A163AA164A165A166A167A168A163BA169A170A171A172C11051A173201BV/H21A17311A173102BV/H31A17321A17353BV/H72632试验方法及结果本正交试验以70乙G18267洗脱G9094中苯并二蒽酮G18339G708以100G贯叶金丝桃G14659材G16757G709为考察指标，结果G16277表2。A1742A175A176A177A178A179A180A181A182A183A175A176A178A179A180A181A184A185A186A187A188A189A190A191AA192A193A194A195A196A191BA197A198A199A200CA201A202DA203A204A205A206A207A208A209A210102G1111148621222447313333324212364952231498623124297313248283213422933213251X12651617133713092X15761367142113583X12291086131214031X4225394464362X5254564744533X40362437468R115177037032上G17860数据表明G726G3324G16786G13634水平G14551G3272G1881，以苯并二蒽酮G18339为指标,G11013G1122G7509G5058R的大G4579G7186G12046影响因素的主次，G6164以其影响G20046G5219为BAC；综合分G7524，B为主要因素，A次G1055。G17902G17819G11464G16278分G7524，最G1260G13464合G5224为B1A2C2，G2375G2334G5464G726柱G20652G8616为120；G14659G9094浓缩G8616为11；G14659G9094上G7691流速为2BV/H。2633方G5058分G75248G11013G1122G14659G9094上G7691流速CG2027与G12366G11345DG2027的RG1552相G17829，G16840明G14659G9094上G7691流速对结果的影响G17751G4579。因G8504本文采用正交G16786G16757G2173G6175IIV31G17731件，G17839G13792以CG2027和DG2027G1328为G16835G5058G20045G17839行方G5058分G7524，结果G16277表3。A2113A212A213A214A215A216A213A214A215A216A217A218A219A220A221A218A222A217A223A224A225A226FA227FA228A229A230A231005A232A233A234A235A24321334694A236B47122585694A236A237A2380364G11013方G5058分G7524结果可G11705，AG2027G708G14659G9094浓缩G8616G709、BG2027G708G2334G5464A239柱G20652G8616G709对洗脱结果有G7186G14891性G5058异，CG2027G708G14659G9094上G7691流速G709无G7186G14891性G5058异。综合G11464G16278分G7524和方G5058分G7524结果及生G1147实际，最终确定上柱工艺条件为G2334G5464G726柱G20652G8616为120；G14659G9094浓缩G8616为11；G14659G9094上G7691流速为3BV/H。27验G16789试验按G10043上G17860实验条件，G17839行验G16789试验，G8991定70乙G18267洗脱G9094的G9036G14179得G10587，G9036G14179中苯并二蒽酮类含G18339和大孔树脂有机残留G18339，结果G16277表4。A2404A241A242A243A241A244A245A243A241A244A245A246A247A248A249A250A251A252A253A254A255A55A0A1A249A44GA4570A57A58A2A3A4A5A62A6A7A8A9A252A44GA45A7A8A6A64A44A57A45A24A10A11A12A13A29A14A252A57A24A10A11A12A13A29A15A16A64A57A71A17A18A19A20A21A77A44A57A45110003833831465592A22A23A2500037A26A27A28A30A31210003333331525062A22A23A2500013A26A27A28A30A31310003783781485594A22A23A2500016A26A27A28A30A31上G17860数据可以G11487出，该条件稳定可行。大孔树脂残留G1075G12538合G457G14659G2709G8892G1888的G3281际G6228G7427要G8726G708G17148G18339部分G709G458G708ICH指G4560G4008G2604G1262G2620G9035G19067主G16805G709。93讨论G22G17G20苯并二蒽酮类的含G18339G8991定方法主要参考G13666G3281G14621G14659G18684,G7691G2709G9354G9094经G8886G19283G6207G6563，其最大吸G6922G1552与对G10043G2709G980G14280,G3355为588NM，G12366G11345G9354G9094经G6207G6563G3324588NMG8886G19283G3800无吸G6922。因G8504，选择G8991定G8886G19283为588NM。G22G17G21综合上G17860实验G11752G12362，大孔吸附树脂对贯叶金丝桃中苯并二蒽酮类成分的分离G13443化工艺归G13447如G991G726选用LSA20型大孔吸附树脂，上柱G14659G9094浓缩G8616为11，柱G20652G8616为120，G14659G9094上G7691流速为3BV/H，吸附时间为G21G75。G1820用4倍柱体积蒸馏水和3倍柱体积20乙G18267洗脱G5335去，再选用7倍柱体积70乙G18267洗脱，G6922集洗脱G9094，G1955G2399干燥，G2375得。本文G25622贯叶金丝桃提G2474G9094的制G3803G257G20045G991G6164用贯叶金丝桃G14659材含苯并二蒽酮类G13434为010，G6164制G3803的上柱G14659G9094含苯并二蒽酮类G13434为026。本文G17902G17819大孔吸附树脂富集后，G7691G2709中苯并二蒽酮类含G18339G13434达到148,为G14659材含G18339的148倍和上柱G14659G9094含G18339的57倍。G17902G17819验G16789试验G16840明该富集方法的工艺路线稳定可行，吸附参数准确可G19764。G22G17G22本文还G17839G980G8505考察G1114大孔树脂的再生。为考察树脂再生后是G2554G12538合要G8726,本文G17839行G1114树脂再生后有G6940部G1313的G1457留G10587试验。经G17819三次平行试验，苯并二蒽酮类的G1457留G1