高效液相色谱法测定氨基葡萄糖拟肽的含量

Abstract

目的采用WATErS 2695型高效液相色谱仪建立定量分析氨基葡萄糖拟肽的方法。方法以HyPErSIlOdS2(4.6MMx250MM,5μM)为分离柱,流动相为5 MMOl/l PH=7.5的磷酸二氢钾-乙腈体系(70∶30,体积分数),检测波长为200nM,流速为1.0Ml/MIn。结果 4种物质的浓度分别在25~800μg/Ml(r=0.999 9,TAg-PHE),40~1 000μg/Ml(r=0.999 3,TAg-lEu),27~600μg/Ml(r=0.999 1,TAg-glu)及50~600μg/Ml(r=0.999 1,TAg-VAl)时呈良好的线性关系,对应的定量限(S/n=10)分别为:20.00,35.54,25.00和49.00μg/Ml。检测限(S/n=3)依次为:4.50,15.30,13.45和19.30μg/Ml。平均回收率在98.75%~101.56%之间,日内及日间精密度均小于1.37%。结论该方法简便,快速,专属性强,适用于氨基葡萄糖拟肽类物质的含量测定。

Objective To establish a method to quantitatively analyze glucosamine peptide mimetics with HPLC.Methods Samples were separated on a Hypersil ODS2column(5μm,4.6mm×250mm),the mobile phase consisted of 5mmoL potassium dihydrogen phosphate-acetonitrile(pH=7.5,V/V).The flow rate was 1.0mL/min.Results The standard curves of four samples were respectively linear over the range in 25~800.00μg/mL(r=0.999 9,TAG-Phe),40~1 000μg/mL(r=0.999 3,TAG-Leu),27~600.00μg/mL(r=0.999 1,TAG-Glu)and 50 ~ 600.00μg/mL(r=0.999 1,TAG-Val).The limits of quantitation were 20.00,35.54,25.00 and 49.00μg/mL in turn(S/N=10).And limits of detection were 4.50,15.30,13.45 and 19.30μg/mL(S/N=3).The average recovery rate of four samples was 98.75%~101.56%,day and inter-day precision were less than 1.37%.Conclusion The method was simple,rapid,specific,and suitable for the determination of glucosamine peptide mimetic.