药学学报, 2021, 56(9): 2389-2393
引用本文:
韩盈, 魏国兰, 米楠, 郭林峰, 胡春云, 李爽, 毕吕存. UPLC-MS/MS法快速、灵敏、直接测定人血浆中的门冬胰岛素[J]. 药学学报, 2021, 56(9): 2389-2393.
HAN Ying, WEI Guo-lan, MI Nan, GUO Lin-feng, HU Chun-yun, LI Shuang, BI L�. Development of a fast and sensitive method for direct analysis of insulin aspart in human plasma[J]. Acta Pharmaceutica Sinica, 2021, 56(9): 2389-2393.

UPLC-MS/MS法快速、灵敏、直接测定人血浆中的门冬胰岛素
韩盈1, 魏国兰1, 米楠1, 郭林峰2, 胡春云2, 李爽3, 毕吕存1*
1. 徕博科医药研发 (上海) 有限公司, 上海 201203;
2. 东莞市东阳光生物药研发有限公司, 广东 东莞 523000;
3. 宜昌东阳光长江药业股份有限公司, 湖北 宜昌 443000
摘要:
本实验建立了一种超高效液相色谱-串联质谱法(UPLC-MS/MS)快速、灵敏、直接测定人血浆中的门冬胰岛素。血浆样品经过固相萃取(SPE)处理后,采用电喷雾电离离子源(ESI),多反应离子监测(MRM),正离子扫描进行测定。以牛胰岛素为内标,采用Waters ACQUITY UPLC CSH C18色谱柱(50 mm×2.1 mm,1.7 μm)进行色谱分离,以乙酸水溶液(A)-乙酸乙腈溶液(B)作为流动相,流速0.600 mL·min-1进行梯度洗脱。结果表明,门冬胰岛素在0.200~10.0 ng·mL-1内线性关系良好,精密度小于14.5%,方法的回收率在36.7%~41.7%。本方法灵敏度高,精密度重现性好,成功支持了采用葡萄糖钳夹技术在健康男性受试者中比较门冬胰岛素注射液与诺和锐®的药代动力学研究。试验获得四川大学华西医院临床试验伦理分委员会批准[批准号:2017年临床试验(西药)审(148)号]。
关键词:    门冬胰岛素      UPLC-MS/MS      固相萃取      生物等效性     
Development of a fast and sensitive method for direct analysis of insulin aspart in human plasma
HAN Ying1, WEI Guo-lan1, MI Nan1, GUO Lin-feng2, HU Chun-yun2, LI Shuang3, BI L�1*
1. Labcorp Pharmaceutical Research and Development(Shanghai) Co., Ltd., Shanghai 201203, China;
2. HEC Pharma Co., Ltd., Dongguan 523000, China;
3. HEC Pharma Co., Ltd., Yichang 443000, China
Abstract:
A fast and sensitive UPLC-MS/MS method was established for the direct quantification of insulin aspart in human plasma. The plasma samples were extracted by solid phase extraction (SPE), an ESI ion source was used and operated in the positive ion mode with multiple reaction monitoring (MRM). Bovine insulin was chosen as internal standard and the chromatographic separation of insulin aspart was performed on Waters ACQUITY UPLC CSH C18 column (50 mm×2.1 mm, 1.7 μm). A mixture of acetic acid aqueous solution and acetonitrile with acetic acid at a flow rate of 0.6 mL·min-1 in gradient elution mode was employed as mobile phase. We found that the method was validated over the range of 0.200-10.0 ng·mL-1 for insulin aspart and showed excellent linearity. The intra-and inter-assay accuracy and precision were below 14.5% and the recovery was 36.7%-41.7% over the three concentration levels evaluated. The UPLC-MS/MS method was selective, accurate, sensitive and robust, and the method was successfully applied in supporting the pharmacokinetic research of two insulin aspart injections (Test Product and NovoRapid®) in heathy male subjects. This clinical trial was approved according to the Ethics Committee of West China Hospital, Sichuan University (2017 Clinical Trial (Western Medicine) Approval (148)).
Key words:    insulin aspart    UPLC-MS/MS    solid phase extraction    bioequivalence   
收稿日期: 2021-06-30
DOI: 10.16438/j.0513-4870.2021-0972
通讯作者: 毕吕存,Tel:86-21-61925816,E-mail:Luke.Bi@labcorp.com
Email: Luke.Bi@labcorp.com
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