氨基糖苷类抗生素分析课件

氨基糖苷类抗生素分析药学院药物分析教研室【大纲要求】1、掌握氨基糖苷类抗生素代表药物的体内过程2、熟悉代表药物的体内分析方法SMGM主要代表药物◆◆◆治疗范围窄6～20μmol/l

毒副作用较强，血药浓度高于30μmol/L耳毒性和肾毒性T1/2较长，动物食品残留检测主要药动学特点Processinvivo■■■T1/2与身体健康状况密切相关Concentration血药浓度较低、几乎完全从肾脏排泄Tissuedistribution主要分布于细胞外液、可通过胎盘屏障，不易通过血脑屏障Characteristic■■■NitrogenatomAlkali-OHgroupwatersolublepolarityNon-chromaphoreforUVdetection旋光特性－含有多个氨基糖，一般为右旋结合的苷键可发生水解AnalyticalmethodsImmuneanalysisIAUVorfluorescenceanalysisChromatographyanalysisLC/MSanalysisPreparationofthebiologicalsampleDerivationAnalyticalconditions条件摸索◆◆◆ChemicalDerivationUVderivationreagentFluorescencederivationreagent邻苯二醛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％为好pH=8.5~10.5■■Concentrationofderivationreagent对OPA：0.1

μg/ml；其他则无影响■Reactiontimeandtemperature20、30、60℃→60、40、10min。OPA-β-MP在30和60℃时可产生干扰的荧光物质，需用CHCl3等提取后在测定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分析方法的特异性高DeterminationofKMinserumandurinebyHPLCusingpre-columnderivationmethodColumn:ZorbaxODS(150×4.6mm,I.D.,5μm)Mobile-phase:methanol-water=72:28Flowrate:1.0ml/minDetector:FDExcitationwavelength-335nmEmissionwavelength-425nm方法学考察结