比格犬血浆中德谷胰岛素浓度检测方法的建立及应用  

Determination of insulin degludec concentration in Beagle dog plasma

作　　者：张家莹 李黎 李静 刘亮 李玉 郭林峰 李晓平 董立厚 王春梅 郭银汉 ZHANG Jia-ying;LI Li;LI Jing;LIU Liang;LI Yu;GUO Lin-feng;LI Xiao-ping;DONG Li-hou;WANG Chun-mei;GUO Yin-han(School of Life Sciences,Beijing University of Chinese Medicine,Beijing 102488;Institute of Lifeomics,Academy of Military Medical Science,Military Academy of Sciences,Beijing 102206;United-Power Pharma Tech Co.Ltd,Beijing 102206;HEC Pharm Co.,Ltd,Dongguan Guangdong 523871)

机构地区：[1]北京中医药大学生命科学学院,北京102488 [2]军事科学院军事医学研究院生命组学研究所,北京102206 [3]军科正源(北京)药物研究有限责任公司,北京102206 [4]广东东阳光药物研发有限公司,广东东莞523871

出　　处：《中南药学》2020年第6期947-952,共6页Central South Pharmacy

基　　金：重大新药创制科技重大专项(No.2020ZX09201026)。

摘　　要：目的建立测定比格犬血浆中德谷胰岛素浓度的液质联用(LC-MS/MS)方法,并用于其药代动力学研究。方法血浆样品经蛋白沉淀提取,流动相为乙腈-水(均含0.1%甲酸),液相分离采用Peptide ES-C18(50 mm×2.1 mm,2.7μm)色谱柱以梯度洗脱进行,通过配备电喷雾源的串联质谱,以多反应监测模式进行正离子检测,并考察了检测方法的专属性、线性、定量下限、准确度、精密度、提取回收率、稳定性及基质效应等。结果德谷胰岛素在2.00~500 ng·mL^-1与峰面积线性关系良好(r^2>0.990),定量下限为2.00 ng·mL^-1;方法的批内及批间的准确度和精密度均良好,提取回收率为66.9%~67.7%;基质效应为64.5%~79.0%,含0.2%全血的溶血基质对德谷胰岛素检测浓度无影响;样品在血浆中的稳定性良好。结论本方法简单、快速、灵敏及稳定性好,可用于测定血浆中德谷胰岛素的浓度。Objective To establish a liquid chromatography-tandem mass spectrometry(LC-MS/MS)method to determine insulin degludec concentration in the plasma of Beagle dogs,and application in the pharmacokinetic study.Methods The plasma samples were extracted by protein precipitation.The mobile phase was acetonitrilewater(containing 0.1%formic acid).The LC separation was conducted on a Peptide ES-C18(50 mm×2.1 mm,2.7μm)column with gradient elution.Positive ion was detected by electrospray ionization in multiple reaction monitoring mode.The specificity,linearity,lower limit of quantitation,accuracy,precision,extraction recovery,stability and matrix effects were validated.Results The insulin degludec showed a good linearity at 2.00~500 ng·mL^-1(r^2>0.990),with the lower limit of quantification at 2.00 ng·mL^-1.Both the intrabatch and inter-batch precisions were good.The extraction recoveries were 66.9%~67.7%.The matrix effects were 64.5%~79.0%The hemolytic matrix containing 0.2%whole blood had no effect on the determination of insulin degludec concentration.The samples were stable in the plasma.Conclusion The method is simple,rapid,sensitive,and stable,which can be used to determine the concentrations of insulin degludec in Beagle dog plasma.

关 键 词：德谷胰岛素 生物样品分析 LC-MS/MS 药代动力学 

分 类 号：R96[医药卫生—药理学]