谷胱甘肽欧洲药典5.1标准

GLUTATHIONEGlutathionumC10H17N3O6SMr307.3DEFINITIONL-?-Glutamyl-L-cysteinylglycine.Content:98.0percentto101.0percent(driedsubstance).谷胱甘肽C10H17N3O6SMr307.3定义描述L－r谷氨酰基－L－半胱氨酰基甘氨酸含量：按干燥品计算，98.0%-101.0%CHARACTERSAppearance:whiteoralmostwhite,crystallinepowderorcolourlesscrystals.Solubility:freelysolubleinwater,veryslightlysolubleinethanol(96percent)andinmethylenechloride.性状外观性状：白色或几乎白色结晶性粉末或无色的结晶。溶解度：易溶于水，微溶于96%乙醇及二氯甲烷。IDENTIFICATIONA.Itcomplieswiththetestforspecificopticalrotation(seeTests).B.Infraredabsorptionspectrophotometry(2.2.24).Comparison:glutathioneCRS.鉴别A比旋度符合特定的光学旋转测定（见测定项）。B红外吸收光谱检测（2.2.24）对比：谷胱甘肽CRS.TESTSSolutionS.Dissolve5.0gindistilledwaterRanddiluteto50mlwiththesamesolvent.Appearanceofsolution.SolutionSisclear(2.2.1)andcolourless(2.2.2,MethodII).Specificopticalrotation(2.2.7):?15.5to?17.5(driedsubstance).Dissolve1.0ginwaterRanddiluteto25.0mlwiththesamesolvent.测定溶液S：取本品5.0g，蒸馏水R溶解并稀释至50ml。溶液澄清度溶液S澄清(2.2.1)，无色(2.2.2,MethodII)比旋度（2.2.7)－15.5。～－17.5°（干品物质）取本品1.0g，蒸馏水R溶解并稀释至25.0ml。Relatedsubstances.Capillaryelectrophoresis(2.2.47).Preparethesolutionsimmediatelybeforeuse.Internalstandardsolution(a).Dissolve0.100gofphenylalanineRintheelectrolytesolutionanddiluteto50.0mlwiththesamesolution.Internalstandardsolution(b).Dilute10.0mlofinternalstandardsolution(a)to100.0mlwiththeelectrolytesolution.Testsolution(a).Dissolve0.200gofthesubstancetobeexaminedintheelectrolytesolutionanddiluteto10.0mlwiththesamesolution.Testsolution(b).Dissolve0.200gofthesubstancetobeexaminedininternalstandardsolution(b)anddiluteto10.0mlwiththesamesolution.Referencesolution(a).Dissolve20mgofthesubstancetobeexaminedininternalstandardsolution(a)anddiluteto10.0mlwiththesamesolution.Referencesolution(b).Dilute5.0mlofreferencesolution(a)to50.0mlwiththeelectrolytesolution.Referencesolution(c).Dissolve0.200gofthesubstancetobeexaminedin5mloftheelectrolytesolution.Add1.0mlofinternalstandardsolution(a),0.5mlofa2mg/mlsolutionofL-cysteineR(impurityB)intheelectrolytesolution,0.5mlofa2mg/mlsolutionofL-glutathione,oxidisedR(impurityC)intheelectrolytesolutionand0.5mlofa2mg/mlsolutionofL-?-glutamyl-L-cysteineR(impurityD)intheelectrolytesolution.Diluteto10.0mlwiththeelectrolytesolution.有关物质采用毛细管电泳法（2.2.47）溶液应临用前新鲜配制内标液（a）取苯丙氨酸0.100g，加电解质溶液溶解并稀释至50.0ml内标溶液（b）量取内标溶液（a）10.0ml，用电解质溶液稀释至100.0ml供试溶液（a）取待检物质0.200g，加电解质溶液溶解并稀释至10.0ml。供试溶液（b）取待检物质0.200g，加溶液(b)溶解并稀释至10.0ml对照溶液（a）取待检物质20g，加标准溶液（a）溶解并稀释至10.0ml对照溶液（b）取对照溶液（a）5.0ml，加电解质溶液稀释至50.0ml对照溶液(c)取待检物质0.200g，加电解质溶液5ml溶解，加入1.0ml对照溶液（a），0.5ml2mg/mlL-半胱氨酸R的电解质溶液（杂质B），0.5ml2mg/ml氧化型谷胱甘肽R电解质溶液（杂质C）及0.5ml2mg/mlL－r谷氨酰基－L－半胱氨酰基甘氨酸R的电解质溶液（杂质D），然后用电解质溶液稀释至10.0mlCapillary:-material:uncoatedfusedsilica,-size:lengthtothedetectorcell=0.5m;totallength=0.6m;?=75μm.Temperature:25°C.Electrolytesolution.Dissolve1.50gofanhydroussodiumdihydrogenphosphateRin230.0mlofwaterRandadjusttopH1.80withphosphoricacidR.Diluteto250.0mlwithwaterR.CheckthepHand,ifnecessary,adjustwithphosphoricacidRordilutesodiumhydroxidesolutionR.Detection:spectrophotometerat200nm.Preconditioningofanewcapillary:rinsethenewcapillarybeforethefirstinjectionwith0.1Mhydrochloricacidfor20minat138kPaandwithwaterRat138kPafor10min;forcompleteequilibration,conditionthecapillarywiththeelectrolytesolutionfor40minat350kPa,andsubsequentlyfor60minatavoltageof20kV;毛细管-材料：石英（未涂层）-尺寸：有效长度0.5m，总长：0.6m，内径75μm温度：25℃电解质溶液：取无水磷酸二氢钠R1.50g，加水R230.0ml，用磷酸R调节PH值为1.80。加水R稀释至150.0ml，检测PH值，如有必要，用磷酸R或氢氧化钠溶液R调节PH值。检测：采用分光光度计在200nm处检测新毛细管的预处理：在首次进样前在压力138Kpa下用0.1mol/L盐酸溶液前冲洗毛细管20min，138Kpa压力下用水R冲洗10分钟已达到全平衡状态，用电解质溶液在350kpa条件下冲洗40min，在电压20kv条件下保持60min。Preconditioningofthecapillary:rinsethecapillaryat138kPawiththeelectrolytesolutionfor40min;Between-runrinsing:rinsethecapillarywithwaterRat138kPafor1min,with0.1Msodiumhydroxidefor2min,withwaterRat138kPafor1min,with0.1Mhydrochloricacidfor3minandwiththeelectrolytesolutionat138kPafor10min.Injection:underpressure(3.45kPa)for5s;injecttheelectrolytesolution(blanksolution),referencesolutions(b)and(c)andtestsolutions(a)and(b).Migration:applyavoltageof20kV.Runtime:45min.预处理的毛细管：138kPa压力下，用电解质溶液冲洗毛细管40min；批间冲洗：138kPa压力下，依次用水R冲洗毛细管1min，0.1mol/L氢氧化钠溶液冲洗2min；水R冲洗1min，0.1mol/L盐酸溶液冲洗3min，最后用电解质溶液冲洗10min；进样：3.45kPa压力下维持5s；注入电解质溶液（空白溶液），对照溶液（b）、（c）和供试溶液a、b迁移：电压为20KV运行时间：45minRelativemigrationwithreferencetotheinternalstandard(about14min):impurityA=about0.77;impurityB=about1.04;impurityE=about1.2;impurityC=about1.26;impurityD=about1.3.相对迁移率：相对于内标物质（约14min）杂质A=约0.77杂质B=约1.04；杂质E=约1.2杂质C=约1.26；杂质D=约1.3Systemsuitability:-resolution:minimum1.5betweenthepeaksduetotheinternalstandardandimpurityBinthechromatogramobtainedwithreferencesolution(c);ifnecessaryincreasethepHwithdilutesodiumhydroxidesolutionR;-peak-to-valleyratio:minimum2.5,whereHp=heightabovethebaselineofthepeakduetoimpurityDandHv=heightabovethebaselineofthelowestpointofthecurveseparatingthispeakfromthepeakduetoglutathioneinthechromatogramobtainedwithreferencesolution(c);ifnecessarylowerthepHwithphosphoricacidR;-checkthatintheelectropherogramobtainedwithtestsolution(a)thereisnopeakwiththesamemigrationtimeast