药学学报, 2021, 56(9): 2383-2388
孙筱初, 林菲菲, 万咪咪, 佟悦, 常路, 袁梦, 冯莹莹, 滕国生, 刘佳. 液相色谱-串联质谱法测定大鼠血浆中赖脯胰岛素及其药动学研究[J]. 药学学报, 2021, 56(9): 2383-2388.
SUN Xiao-chu, LIN Fei-fei, WAN Mi-mi, TONG Yue, CHANG Lu, YUAN Meng, FENG Ying-ying, TENG Guo-sheng, LIU Jia. Determination of insulin lispro in rat plasma by LC-MS/MS and its application in a pharmacokinetics study[J]. Acta Pharmaceutica Sinica, 2021, 56(9): 2383-2388.

孙筱初1,2, 林菲菲2, 万咪咪2, 佟悦2, 常路2, 袁梦2, 冯莹莹2, 滕国生1*, 刘佳2*
1. 长春工业大学化学与生命科学学院, 吉林 长春 130012;
2. 中国科学院上海药物研究所, 上海 201203
赖脯胰岛素皮下注射吸收较快,与人胰岛素相比,其降血糖作用起效更快,血药浓度峰值更高,可更好地控制餐后高血糖。本文建立了液相色谱-串联质谱法(LC-MS/MS)测定大鼠血浆中的赖脯胰岛素,并评价其药动学特征。采用牛胰岛素作为内标,血浆样品经过固相萃取后,使用ACQUITY UPLC Peptide CSH C18柱(2.1 mm × 50 mm,1.7 μm)进行色谱分离。采用电喷雾离子源(ESI源),在正离子模式下以多反应监测模式检测,赖脯胰岛素和牛胰岛素(内标)定量的离子对分别为m/z 1 162.5→217.2和m/z 1 157.5→136.0。方法学验证结果表明,大鼠血浆中赖脯胰岛素的线性范围为0.1~100 ng·mL-1,日内、日间准确度和精密度均符合生物样品分析相关要求,回收率在63.1%~68.1%之间。与文献报道方法相比,色谱分离得到改善,回收率显著提高。本方法成功应用于大鼠的单剂量皮下注射赖脯胰岛素的药代动力学研究。本研究中实验动物的使用已获得中国科学院上海药物研究所实验动物管理与使用委员会的批准。
关键词:    赖脯胰岛素      LC-MS/MS      药代动力学      固相萃取     
Determination of insulin lispro in rat plasma by LC-MS/MS and its application in a pharmacokinetics study
SUN Xiao-chu1,2, LIN Fei-fei2, WAN Mi-mi2, TONG Yue2, CHANG Lu2, YUAN Meng2, FENG Ying-ying2, TENG Guo-sheng1*, LIU Jia2*
1. School of Chemistry and Life Sciences, Changchun University of Technology, Changchun 130012, China;
2. Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China
Compared with human insulin, insulin lispro shows a faster hypoglycemic effect and a higher peak plasma concentration, which can better control postprandial hyperglycemia. In this study, we used a solid phase extraction pretreatment method and liquid chromatography-tandem mass spectrometry (LC-MS/MS) to quantify insulin lispro in rat plasma. Bovine insulin was used as an internal standard. Plasma samples were separated on an ACQUITY UPLC Peptide CSH C18 column (2.1 mm × 50 mm, 1.7 μm) after solid phase extraction. Positive electrospray ionization was performed using multiple reaction monitoring (MRM) with transitions of m/z 1 162.5→217.2 for insulin lispro and m/z 1 157.5→136.0 for insulin bovine (internal standard). The method validation results showed that the linear range was 0.1 ng·mL-1-100 ng·mL-1; intra- and inter-day accuracy and precision met the acceptance criteria for biological sample analysis. The recovery of insulin lispro ranged from 63.1% to 68.1%. The method was applied in a pharmacokinetic study of insulin lispro following a single-dose subcutaneous administration to rats. Animal experiments were approved by the Experimental Animal Ethics Committee of Shanghai Institute of Materia Medica, Chinese Academy of Sciences.
Key words:    insulin lispro    LC-MS/MS    pharmacokinetics    solid phase extraction   
收稿日期: 2021-04-30
DOI: 10.16438/j.0513-4870.2021-0670
通讯作者: 滕国生,Tel:18946545897,E-mail:gshteng@163.com;刘佳,Tel:86-21-58559563,E-mail:jia.liu@simm.ac.cn
Email: gshteng@163.com;jia.liu@simm.ac.cn
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