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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 a LC / MS method for the determination of carnosine in the lens of the rabbit. Methods: Rabbit eye lens was weighed and homogenized. Acetonitrile was used for extraction. The extract was analyzed by HPLC-MS / MS. The mobile phase was a 0.2% formic acid solution (formic acid, acetonitrile and water 0.2: 9: 100 by volume) -acetonitrile (65:35) using a Waters Xterra MSC18 (3.5 μm, 3.0 mm × 150 mm) column. The ESI positive ion mode was used for detection. The ion collection mode was multi-reaction monitoring mode (MRM, m / z 227.0 → 110.0). Results: The linear range of detection of carnosine in lens homogenates was 0.0407 ~ 9.9μg.mL-1. The recovery of carnosine in the lens was close to 100% (n = 5). The recovery rate was 80% ~ 120% Intra-RSD and inter-lot RSD were less than 15%. Conclusion: This method is specific, accurate and reproducible, and can be used to determine the concentration of carnosine in the lens quickly and accurately.