氨基糖苷類(lèi)抗生素分析課件

氨基糖苷類(lèi)抗生素分析藥學(xué)院藥物分析教研室【大綱要求】1、掌握氨基糖苷類(lèi)抗生素代表藥物的體內(nèi)過(guò)程2、熟悉代表藥物的體內(nèi)分析方法SMGM主要代表藥物◆◆◆治療范圍窄6～20μmol/l

毒副作用較強(qiáng)，血藥濃度高于30μmol/L耳毒性和腎毒性T1/2較長(zhǎng)，動(dòng)物食品殘留檢測(cè)主要藥動(dòng)學(xué)特點(diǎn)Processinvivo■■■T1/2與身體健康狀況密切相關(guān)Concentration血藥濃度較低、幾乎完全從腎臟排泄Tissuedistribution主要分布于細(xì)胞外液、可通過(guò)胎盤(pán)屏障，不易通過(guò)血腦屏障Characteristic■■■NitrogenatomAlkali-OHgroupwatersolublepolarityNon-chromaphoreforUVdetection旋光特性－含有多個(gè)氨基糖，一般為右旋結(jié)合的苷鍵可發(fā)生水解AnalyticalmethodsImmuneanalysisIAUVorfluorescenceanalysisChromatographyanalysisLC/MSanalysisPreparationofthebiologicalsampleDerivationAnalyticalconditions條件摸索◆◆◆ChemicalDerivationUVderivationreagentFluorescencederivationreagent鄰苯二醛OPA丹酰氯DNS-Cl茚三酮2,4,6-三硝基苯磺酸TNBS1-氟-2,4-二硝基苯FDNBOptimizationsuitablederivationconditions

ReactionsolventandpHvalueMethanolorethanol含醇量50％為好pH=8.5~10.5■■Concentrationofderivationreagent對(duì)OPA：0.1

μg/ml；其他則無(wú)影響■Reactiontimeandtemperature20、30、60℃→60、40、10min。OPA-β-MP在30和60℃時(shí)可產(chǎn)生干擾的熒光物質(zhì)，需用CHCl3等提取后在測(cè)定OPA-β-MPRP-HPLCChromatogramPre-columnderivationSisMDeterminationofGMusingpost-columnderivation

byRP-HPLCinplasmaAnalyticalconditions:Analyticalcolumn:μBondapakC18Mobilephase:CH3CN-H2OFlowrate:1.5ml/minDerivationreagent:OPAFlowrateofderivationreagent:0.8ml/minTemperature:50℃

Detection:fluorescenceOptimizationsuitableconditionsProteinprecipitationmethanol10000rpmSamplepreparationAccuracyandprecisionRecoveryLinearrangeLimitdetection

LOQLODEvaluateddata

DeterminationofGMinplasmaandurinebyRP-HPLCusingsolidcolumnderivationtechniqueAnalyticalconditionsColumn:LichrosorbRP-18(12.5×4mm,I.D.,5μm)Mobile-phase:Methanol-10%HAcFlowrate:1.0ml/minDetection:excitationwavelength340nm,emissionwavelength418nmSamplesolutionISaddedComplexsolutionSilicagelcolumnchromatographyEthanolelutionsolutionOPAelutionsolutionHPLCanalysisSPEISGM-C1GM-C2分析方法的特異性高DeterminationofKMinserumandurinebyHPLCusingpre-columnderivationmethodColumn:ZorbaxODS(150×4.6mm,I.D.,5μm)Mobile-phase:methanol-water=72:28Flowrate:1.0ml/minDetector:FDExcitationwavelength-335nmEmissionwavelength-425nm方法學(xué)考察結(jié)