抗生素类药物分析

TheAnalysisofSteroidHormoneDrugs甾体激素类药物分析Chapter10Themaincontents：甾体激素类药物的基本结构特点与分类Thestructure,characteristicsandclassification；甾体激素药物的鉴别试验Theidentificationtests；甾体激素药物的特殊杂质检查Thespecificimpuritytests；甾体激素药物的含量测定Assays.Fundamentalrequirement:掌握各类甾体激素药物的基本结构特点、鉴别实验及含量测定方法；Masteringthegeneralstructurecharacteristics、identificationtestsandassaysofsteroidhormones;熟悉HPLC在鉴别试验及含量测定中的应用；FamiliarizingwiththeapplicationofHPLCinidentificationtestsandassays;掌握IR在甾体激素类药物鉴别中的应用。MasteringtheapplicationofIRinidentificationtests.StructureIdentificationAssaySteroidhormonesSpecificimpurityBriefsummaryTorequirementTocontent基本结构与分类CC结构特点基本骨架:甾体激素类药物，一些为天然物，一些为人工合成品。但无论是天然的还是人工合成的本类药物，均含有环五烷骈多氢菲母核。多数药物都是半合成的。第一节GeneralstructuresCCStructurecharacteristicsSteroidskeleton:Steroidcompoundshaveacyclo-pentanoperhydrophenanthreneskeleton.Someofthesedrugsarenaturalproducts,andsomearesyntheticmaterials.Mostofthesedrugsareobtainedbysemi-syntheticmethods.Section1SortStructureTypicaldrugsAdrenocorticoidhormonesHydrocortisoneAcetate(醋酸氢化可的松）;DexamethasoneAcetate（醋酸地塞米松）;AndrogensMethyltestosterone（甲睾酮）;TestosteronePropinate（丙酸睾酮）;OOHCOCH2OHHOOHGalairanabolichormonesNandrolonePhenylpropionate(苯丙酸诺龙）;ProgestogensProgesterone;MegestrolAcetate（醋酸甲地孕酮）;Norethisterone（炔诺酮）;EstrogensEstradiol;Ethinylestradiol（炔雌醇）;Diethylstibestrol(己烯雌酚）.OOCOCH2CH2-C6H5OOCH3OHOH鉴别实验第二节Δ4-3-酮基—紫外分光光度法C17-α-醇酮基—呈色反应(与氧化剂四氮唑盐呈色)—沉淀反应酮基—呈色反应(与羰基试剂呈色)—制备衍生物测定其熔点—与强酸的呈色反应官能团—红外分光光度法甲基酮—呈色反应(与亚硝基铁氰化钠、间二硝基酚、芳香醛呈色)IdentificationtestsSection2Δ4-3-keto—Ultravioletspectrophotometryα-hydroxyacetone—Colorreaction(withtetrazolinesalt)—PrecipitationreactionKetonegroup—Colorreaction(withcarbonylreagents)—Meltingpointdeterminationofderivatives—ReactionwithstrongacidsFunctionalgroup—InfraredspectrophotometryMethylketonegroup—Colorreaction(withsodiumnitroferricyanide,m-dinitrophenol,aromaticaldehyde)鉴别实验Section2有机氟—呈色反应(经氧瓶燃烧后生成无机氟化物，再与茜素氟蓝及硝酸亚铈呈色）有机氯—沉淀反应(经有机破坏生成无机氯化物，在硝酸酸性下与硝酸银作用，生成白色沉淀)酚羟基—呈色反应(与重氮苯磺酸反应生成红色偶氮染料)—制备衍生物测定其熔点酯基—水解产物的反应炔基—沉淀反应(与硝酸银反应生成白色沉淀)IdentificationtestsSection2Organicfluorine—Colorreaction(withalizarincomplexoneandCerousnitrateafterdestroyedbyoxygenflaskcombustion）Organicchlorine—Precipitationreaction(withnitricacidandsilvernitrateafterdestroyedbyoxygenflaskcombustion)Phenolichydroxyl—Colorreaction(diazobenzenesulfonicacid)—MeltingpointdeterminationofderivativesEstergroup—HydrolysisproductreactionAlkynylgroup—Precipitationreaction(withsilvernitrate)NamesColorFluorescenceAddingwa-terdilutionCortisoneacetateYellowororange-ColordisappearsandclearsolutionformedPrednisoneOrange-Yellowtoblue-greenReactionwithsulfuricacidUltravioletspectrophotometryConjugatedbondingsystems:1.α,β-unsaturatedketonegroup2.SteroidswitharomaticringATheUVspectraofhydrocotisoneandbetamethasonePrecipitationreactionReactionwithsilvernitratepreparedinammoniaCorticalhormonesSilvernitrateAmmoniaSilverprecipitate(black)ReactionwithsilvernitrateSteroidhormones(alkynyl)SilvernitrateSilversaltprecipitate(white)MeltingpointsofderivativesTheformulationofketoxime（酮肟）OCOCH3CHONNOHCH3+2HONH2+2H2O（Progesterone黄体酮）（Meltingpoint:235~240℃）MeltingpointofderivativesTheformulationofsemicarbazone（缩氨基脲）+H2O+HCl（NandrolonePhenylpropionoate苯丙酸诺龙）（Meltingpoint:180℃(dec.)）OOCOCH2CH2NOCOCH2CH2H2NCONH+NH2CONHNH2·HClMeltingpointsofderivativesTheformulationofesterOHOHCCH+COCl+KOHOHOCCHCO+KCl+H2O（Meltingpoint:200-202℃）Ethinylestradiol炔雌醇Infraredspectrophotometry–OHstretchingeverywhere~3600aCH2&CH3stretchingeverywhere2970~2850b=C–Hstretchingaromaticring3040~3010≡C–Hstretching~3320aliphaticketonestretchhexa-typering1720~1705cfive-membered1749~1742C20－1710~1706–OCOCH3everywhere1742~1735–C=C–C=Ohexa-typering1684~1620d1585~1620(Δ4–3–keto)Assignmentlocationwavenumber(cm-1)comments–C–OH(alcohol)everywhere1230~1000–C–OH(hydroxybenzene)1300~1200–OCOR1200~1000–C=C–Heverywhere900~650Commentsa:Ifhydrogenbondswiththehydroxygroups,thebandshiftsto3550~3330cm-1.b:Oftenappearsasashoulder.c:C=OstretchinginCCl4occursatalowwavenumber.d:AsaresultofhydrogenbondingwiththehydroxygroupstheνC=Obandofthe3-ketogroupsappearsatrelativelylowfrequency,andthustheνC=Obandcannotbeobserved.υC=C1615，1590，1505cm－1苯环的骨架振动；υ≡CH3300cm－1炔的特征峰；υ－OH3610cm－1游离酚羟基的伸缩振动；υ－OH3505cm－1C17-羟基的伸缩振动。Ethinylestradiol(炔雌醇)IRspectrumIRspectrumofDexamethasone(地塞米松)特殊杂质检查甾体激素药物多由其它甾体化合物或结构类似的其他甾体激素经结构改造而来，因而可能带来原料、中间体、异构体、降解产物以及试剂和溶剂等杂质。甾体激素类药物在纯度检查时，除一般杂质外，检查其特殊“其他甾体”的限度，是一个重要的项目。第三节TheSpecificImpuritytestsSteroidhormonedrugsareobtainedbymodifyingothersteroidcompoundsorotherstructurallyrelativesteroidhormones.Impurities,suchasrawmaterials、intermediatecompounds、isomers、degradationproducts、reagentsandsolventsetc,maybeintroducedintotheirfinalproducts.Exceptforgeneralimpurities,thelimitofothersteroidsisalsoanimportantiteminthepuritytests.Section3•地塞米松磷酸钠、氢化可的松磷酸钠——游离磷酸——测吸收度•地塞米松磷酸钠——甲醇和丙酮——气相色谱法•炔雌醇——雌酮——比色法•醋酸地塞米松、醋酸氟轻松——硒——氧甁法-比色法•甾体激素——其它甾体——薄层色谱法、高效液相色谱法举例•Dexamethasonesodiumphosphate、hydrocortisonesodiumphosphate——freephosphonicacid——determinationofA•Dexamethaso