第十六章 药品质量控制中的现代分析方法与技术

药品质量控制中的

现代分析方法与技术Modernanalyticalmethods&techniquesinqualitycontrolofdrugs第十六章现代分析方法与技术，为药学的发展提供了适时而有效的手段与动力。 色谱及其联用技术：

药学研究－－分子水平。手性分析：毛细管电泳及手性色谱技术－－药物研究与质量控制提供了保障。现代光谱技术：药物结构鉴定，微量杂质检定。第一节概况Capillaryelectrophoresis,CEModernchromatogr&itsapplication药物现代色谱法及其应用UPLCUltraPerformanceLC®(UPLC®)technologystartswithunique1.7µmsmall-particlechemistries.Chromatographersnolongerneedtochoosebetweenspeedandresolution—withUPLCyougetboth.MassspectroscopyMS

NuclearmagneticresonancespectrometryNMR

X-raydiffractionmethod

NearinfraredspectrometryNIRS现代光谱法及其应用Modernspectroscopy&itsapplicationinpharmaceuticalanalysisGC-FTIRGC-MSUPLC-MSHPLC-NMRHyphenatedTechniquesinChromatography现代联用技术及其应用HPLC-MSCE-MS++

样品与溶剂脱离及电离EIESIAPCILC/MS接口

离子源质量分析器检测离子HPLC数据系统质谱离子识别

QuadrapoleTimeofFlightFourierTransform…+++++++离子检测++-++++++-+++第二节液质联用技术与应用2.1离子化方式2.2离子分离与测定模式Full-ScanMassSpectrometryAdvantageProvidesMWInformationFull-ScanMSofBuspironeBuspirone

(丁螺环酮)C21H31N5O2MW=385150200250300350400450500m/z255075100RelativeAbundance386408(M+H)+(M+Na)+SingleIonMonitoring(SIM)AdvantagesTargetedAnalyteMonitoringHighDutyCycleSimpleDisadvantagesCansufferfrominterferencesNotassensitiveorselectiveasSRM(seebelow)Fixedm/zPassAllPassAllProductIonScanning:ATandemMSMethodAdvantageProvidesStructuralInformationDisadvantageLowdutycycleFixedm/zPassAllScanningProductIonSpectrumQ3Q2Q1ProductIonSpectrumofBuspirone100150200250300350400m/z255075100RelativeAbundance122386222150265180(M+H)+PrecursorIonScanningAdvantageIDcompoundsproducingspecificfragmention(e.g.,PO3−forphosphopeptides)DisadvantageLowdutycycleFixedm/zPassAllScanningPrecursorIonSpectrumQ3Q2Q1PrecursorIonScanModeforBuspironeMetabolitesPrecursorIonScan:Q3settom/z122100200300400500m/zRelativeAbundance402386910111213141516Time(min)255075100RelativeAbundance11.6213.8413.1614.4012.1310.4515.45NeutralLossScanningAdvantageScreenforcompoundsproducingspecificneutralloss(e.g.,lossof176forglucuronideconjugates)DisadvantageLowdutycycleScanningPassAllScanningNeutralLossSpectrumLinkedQ3Q2Q1NeutralLossScanofBuspironeMetabolitesNeutralLossScan:Q1/Q3differencesetto121Da100200300400500m/zRelativeAbundance402386910111213141516Time(min)255075100RelativeAbundance13.9211.6913.2115.5010.58SelectedReactionMonitoring(SRM)AdvantagesTargetedAnalyteMonitoringHighDutyCycle“Simultaneous”MonitoringofMultipleTransitionsDisadvantageNo“advanced”structuralinformationFixedm/zPassAllFixedm/zQ1Q2Q3MS/MSSelectivityinComplexMatrices息斯敏——阿斯咪唑(astemizole)Chlroamphenicol(氯霉素，CAP)残留测定黄杨生物碱成分鉴定苯甲酸利扎曲普坦人体药代动力学研究2.3药物分析中的典型应用C11H12Cl2N2O5

FMW=323.13 【类别】酰胺醇类抗生素 【适应症】本品是治疗伤寒、副伤寒的首选药物，外用可治疗沙眼。因脑脊液浓度高，故常用于治疗细菌性脑膜炎和脑脓肿。此外，尚可外用治疗痤疮、酒糟鼻、脂溢性皮炎等。被农业养殖滥用！肉食品中严格检查。2.3.1Chlroamphenicol(氯霉素，CAP)残留测定HPLCanalysiswasperformedontheFinniganSurveyorHPLCmodulewithMSPumpandAutosamplerColumn:

ThermoHypersilGoldC18(100×2.1mm,5µ)MobilePhase:A:Water;B:AcetonitrileColumnTemperature:

40

oCGradientProgram:0.25mL/minInjection:20uLwithloopTime(min)%A%B080202.520803.020803.180205.08020OperationConditionsforCAPIonsource:ESIIonpolarity:NegativeSprayvoltage:4000VSheathgaspressure:45Auxiliarygaspressure:15Iontransfercapillarytemperature:300oCSourceCID:8VScanType:SRM,3transitionsof[M-H]-(m/z:321)(321

152,321

194and321

257)Q1peakwidth0.7DainSRMor0.2DainH-SRMQ3peakwidth0.7DaCollisionPressure:Arat1.3mTorrQ1peakwidthandH-SRMexperimentEnablingtheH-SRMexperiment

HighlySelectiveSelectedReactionMonitoring(H-SRM)Reduces“isobaric”chemicalnoiseIncreasesconfidenceofanalysis&improvedLOQQ1peakwidth0.7DainSRMor0.2DainH-SRMQ3peakwidth0.7DaCAPSRMResult:CAPStandard

Q1peakwidth=0.7DaTIC321->152321->194321->257CAPPeakAreaCounts=2.4E4CAPSRMResult:KidneyBlankTIC321->152321->194321->257CAPSRMResult:KidneySpiked(0.5ng/g)TIC321->152321->194321->257CAPNotaccurateforconfirmationCAPdetectedCAPH-SRMResult:CAPStandard

Q1peakwidth=0.2DaTIC321->152321->194321->257PeakAreaCounts=7.3E3CAPH-SRMResult:KidneyBlankTIC321->152321->194321->257NoCAPdetectedCAPH-SRMResult:KidneySpiked(0.5ng/g)TIC321->152321->194321->257CAP2.3.2黄杨宁生物碱HPLC-MS联用鉴定黄杨科植物小叶黄杨BuxusmicrophllaSieb.et.Zucc.var.sinicaRehd.etWils中含有具有较强心血管疾病治疗活性的孕甾烷生物碱，主要含环维黄杨星D、环黄杨碱D和环常绿黄杨碱C等生物碱成分。

黄杨生物碱HPLC-ELSD色谱图色谱条件色谱柱：LichrospherSiO2(250mm×4.6mm,5µm)流动相：四氢呋喃-甲醇-乙腈-氨水(32:50:13:3)流速：1mL·min-1柱温：30℃ELSD参数：漂移管温度70℃雾化气体（N2）流速：1.5L·min-1环维黄杨星D和有关生物碱含量测定结果次数环维黄杨星D含量%峰1生物碱含量%峰2生物碱含量%峰4生物碱含量%峰5生物碱含量%有关生物碱总含量%186.853.578.090.790.9213.37287.083.848.540.800.9814.15385.833.468.820.810.9514.03485.703.548.760.870.9314.10585.053.528.640.740.8513.75684.843.749.030.790.9714.53Mean85.893.618.650.800.9313.99RSD%1.073.653.384.624.632.8环维黄杨星D及其有关生物碱的鉴别

质谱条件电喷雾离子化正离子检测喷口电压5000V雾化气压35psi辅助气压力5psi毛细管温度350℃碰撞气氩气压力1.5mTorr色谱条件色谱柱：LichrospherSiO2(250mm×4.6mm,5µm)流动相：四氢呋喃-甲醇-乙腈-氨水(32:50:13:3)流速：1mL·min-1柱温：30℃黄杨宁LC-MS/MS全扫描色谱图

黄杨宁LC-MS/MS全扫描色谱放大图

123456789峰1母离子质荷比峰1二级质谱图[M+H]+=370可能为峰1的黄杨宁有关生物碱CyclobuxomicreineKCyclobuxosuffrineKBuxenoneMCyclobuxoviridineB峰2母离子质荷比峰2二级质谱图[M+H]+=431可能为峰2的黄杨宁有关生物碱CyclomicrophyllineBBuxazidineBCyclobuxoxazineCCyclomicrophyllineC峰3母离子质荷比峰3二级质谱图[M+H]+=415可能为峰3的黄杨宁有关生物碱CycloprotobuxineACyclokreanineBCyclovirobuxeineB16-deoxybuxidienineC峰4母离子质荷比峰4二级质谱图环常绿黄杨碱C[M+H]+=417峰5母离子质荷比峰5二级质谱图[M+H]+=401383.34可能为峰5的黄杨宁有关生物碱CycloprotobuxineCBuxaminolEBuxocyclamineACyclobuxineB峰6母离子质荷比峰6二级质谱图环黄杨碱D[M+H]+=387峰7母离子质荷比峰7二级质谱图环维黄杨星D[M+H]+=403峰8母离子质荷比[M+H]+=375357.15峰8二级质谱图峰9二级质谱图

371.17m/z=375m/z=357m/z=344m/z=326m/z=309

357.15峰9母离子质荷比[M+H]+=389峰9二级质谱图

371.17m/z=389m/z=371m/z=358m/z=340m/z=309

371.17峰位号tR(min)[M+H]+(m/z)特征碎片离子(m/z)可能生物碱名称13.92370339，325，283，135，70，58CyclobuxomicreineK,CyclobuxosuffrineK,BuxenoneM,CyclobuxoviridineB24.03431413，382，323，86，70，58BuxazidineB,CyclomicrophyllineB,CyclobuxoxazineC,CyclomicrophyllineC35.35415384，84，58CycloprotobuxineA,CyclokreanineB,CyclovirobuxeineB,16-deoxybuxidienineC45.92417399，386，368，84，58CylcyclovirobuxineC56.11401370，352，326，171，58CycloprotobuxineC,BuxaminolE,BuxocyclamineA,CyclobuxineB67.03387369，356，338，171，58CyclobuxineD77.63403385，372，354，70，58CyclovirobuxineD88.28375357，344，326，309，58未有相关文献报道99.77389371，358，340，173，70，58可能为CyclobuxineD双键加氢还原产物HPLC-ELSD法黄杨宁有关生物碱归属表苯甲酸利扎曲普坦(RizatriptanBenzoate)MW:391.47

分子式：C15H19O5·C7H6O25-HT受体拮抗剂2.3.3苯甲酸利扎曲普坦人体药代动力学研究药理作用刺激大脑血管壁的后接点5-HT1B受体收缩血管，降低颅内血管通透性；刺激三叉神经前突触5-HT1D受体，调节神经递质的释放，抑制硬膜的神经原性炎症反应和血浆外渗；阻止血管肽的释放，使血管口径正常化，通过收缩颅内血管并抑制神经炎症；刺激脑干5-HT1B或5-HT1D受体，