HPLC-MS/MS法检测兔眼晶状体内肌肽的浓度

HPLC-MS/MS determination of carnosine in rabbit lens

目的:建立液质联用法测定兔眼晶状体中肌肽的浓度。方法:取兔眼晶状体精密称重后匀浆,加入乙腈提取,提取液采用HPLC-MS/MS进行分析。采用Waters Xterra MSC18(3.5μm,3.0mm×150mm)色谱柱,流动相为0.2%甲酸溶液(甲酸、乙腈及水的体积比为0.2:9:100)-乙腈(65:35)。采用ESI正离子模式进行检测;离子采集方式为多反应监测模式(MRM,m/z227.0→110.0)。结果:晶状体匀浆液中肌肽检测方法的线性范围为0.0407～9.9μg.mL-1,晶状体中肌肽检测的提取回收率接近100%(n=5),方法回收率为80%～120%,批内RSD及批间RSD均小于15%。结论:本方法专属性强,准确度高、重现性好,可快速准确地测定晶状体中肌肽的浓度。

Objective：To establish an HPLC-MS/MS method to determine the concentration of carnosine in rabbit lens.Methods：The collected lens were weighed exactly,homogenized with deionized water and mixed with acetonitrile.The supernatant of the mixture were analyzed by HPLC-MS/MS.Carnosine were eluted by the mobile phase which consisted of 65% formic acid-acetonitrile-water（0.2：9：100,v/v/v,） and 35% acetonitrile with a Waters Xterra MS C18 column（3.5 μm,3.0 mm×150 mm）.Ion mode of ESI＋ was used for carnosine determination and quantitation were performed in multiple reaction monitoring mode（MRM,m/z 227.0→110.0）.Results：Working solutions over the concentration range 0.0407-9.9 μg·mL-1 worked well for the determination of carnosine in lens homogenate.The average extraction recovery was close to 100%（n=5） and the method recovery was 80%-120%;The RSDs of intra-day and inter-day were less than 15%.Conclusion：The method is specific,sensitive and repeatable for determination of carnonsine in lens.