HPLC-MS/MS法检测兔眼晶状体内肌肽的浓度___

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 MS C₁₈(3.5 μm,3.0 mm×150 mm)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/z 227.0→110.0)。 结果: 晶状体匀浆液中肌肽检测方法的线性范围为0.0407~9.9 μg·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.

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