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ID:26830090
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页数:7页
时间:2018-11-29
《rp-hplc柱前在线衍生法测定羊胎盘枸杞胶囊中16种氨基酸的含量》由会员上传分享,免费在线阅读,更多相关内容在教育资源-天天文库。
1、RP-HPLC柱前在线衍生法测定羊胎盘枸杞胶囊中16种氨基酸的含量RP-HPLC柱前在线衍生法测定羊胎盘枸杞胶囊中16种氨基酸的含量游景水(汤臣倍健股份有限公司,珠海519040)[摘要]目的:建立一种采用反相高效液相色谱法测定羊胎盘枸杞胶囊中16种氨基酸的含量。方法:样品中的蛋白质在6mol/L盐酸溶液中水解为氨基酸,其中一级氨基酸用邻苯二甲醛(OPA)、二级氨基酸用9-芴甲基氯甲酸酯(FMOC)进行柱前在线衍生;用AgilentHypersilAA-ODS柱(200mm×2.1mm,5μm)分离;用流动
2、相(0.16%醋酸钠)进行梯度洗脱;经荧光检测器(FLD)检测(激发波长:340nm,发射波长:450nm;15min时,激发波长:266nm,发射波长:305nm)。结果:16种氨基酸在5pmol/ul~250pmol/ul浓度范围内呈现良好的线性关系(r:0.99959~1.00000)(n=5),平均加标回收率(n=9)为91.6%~104.1%,相对标准偏差(RSD)为0.9%~3.7%,最低检测限为0.4pmol~1.2pmol。结论:该方法灵敏度高,具有良好的精密度与准确度,可以有效地控制产品质
3、量。[关键词]羊胎盘枸杞胶囊;柱前在线衍生;氨基酸;反相高效液相色谱法;邻苯二甲醛(OPA);9-芴甲基氯甲酸酯(FMOC);Onlinepre-columnderivazationdeterminationofsixteenkindsofaminoacidsinthecapsuleofsheepplacentaandmedlarbyRP-HPLCYOUJing-shui(By-HealthCo.Ltd,Zhuhai519040,China)[Abstract]Objective:Toestablisham
4、ethodfordeterminationofsixteenkindsofaminoacidsinthecapsuleofsheepplacentaandmedlarbyRP-HPLC。Methods:Proteinsinthesampleswerehydrolyzedintoaminoacidsin6mol/Lhydrochloricacidsolution,o-phthalaldehyde(OPA)and9-Fluorenylmethylchloroformate(FMOC)wereusedasonli
5、nepre-columnderivativereagentsfortheprimaryaminoacidsandsecondaryaminoacidsrespectively,andthechromatographicexperimentswereperformedonAgilentHypersilAA-ODScolumn(200mm×2.1mm,5μm).The0.16%sodiumacetatesolutionwasusedasmobilephase.Signalsweredetectedbyfluor
6、escencedetection(FLD)(excitationwavelength:第7页共7页RP-HPLC柱前在线衍生法测定羊胎盘枸杞胶囊中16种氨基酸的含量340nm,emissionwavelength:450nm),exciationwavelengthchangedto266nmandemissionwavelengthchangedto305nmatthe15thminute.Results:TheconcentrationofSixteenkindsofAminoAcidsdemonstr
7、atedgoodlinearityintherangeof5pmol/ul~250pmol/ul,(r:0.99959~1.00000),(n=5).Therecoverieswere91.6%~104.1%(n=9).TheRSDwere0.9%~3.7%.Thelimitsofdetectionwere0.4pmol~1.2pmol.Conclusion:Thismethodhasadvantagesofhighsensitivityandprecisionandaccuracy.It'ssuitabl
8、eforthedeterminationofsixteenkindsofaminoacidsinthecapsuleofsheepplacentaandmedlar.[Keywords]:Capsuleofsheepplacentaandmedlar;Onlinepre-columnderivazation;Aminoacid;RP-HPLC;O-phthaldialdehyde(OPA);9-Fluorenyl
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