超高效液相色谱-串联四极杆-轨道阱高分辨质谱法测定甘精胰岛素单链前体残留量  

Determination of Insulin Precursor in Recombinant Insulin Glargine by UPLC-Q-Orbitrap-MS

作　　者：胡馨月 丁晓丽[1] 秦希月 孙悦[1] 李懿 李晶[1] 梁成罡[1] HU Xin-yue;DING Xiao-li;QIN Xi-yue;SUN Yue;LI Yi;LI Jing;LIANG Cheng-gang(Division of Hormone,National Institutes for Food and Drug Control,NHC Key Laboratory of Research on Quality and Standardization of Biotech Products,NMPA Key Laboratory for Quality Research and Evaluation of Chemical Drugs,Beijing 102629,China;Jilin University,Changchun 130015,China)

机构地区：[1]中国食品药品检定研究院激素室,国家卫生健康委员会生物技术产品检定方法及其标准化重点实验室,国家药品监督管理局化学药品质量研究与评价重点实验室,北京102629 [2]吉林大学,长春130015

出　　处：《中国药学杂志》2023年第7期611-618,共8页Chinese Pharmaceutical Journal

基　　金：中国食品药品检定研究院中青年发展研究基金课题资助(2022A3)。

摘　　要：目的 胰岛素单链前体是胰岛素生产工艺中的相关杂质,关系到该类产品的安全性和有效性,有必要建立准确度好、灵敏度高的检测胰岛素单链前体残留量的液质方法。本研究以甘精胰岛素单链前体为例,为胰岛素及其类似物单链前体液质联用方法开发提供参考与借鉴。方法 色谱柱ACQUITY UPLC peptide CSH C18(2.1 mm×150 mm, 130?,1.7μm);流动相A为体积分数0.1%甲酸-水溶液,流动相B为0.1%甲酸-乙腈溶液,梯度洗脱,流速0.3 mL·min^(-1),柱温60℃,检测波长214 nm。Orbitrap Exploris 480液质联用仪,ESI+电喷雾电离源,数据检测模式为平行反应监测(PRM),定量离子对1 155.6/1 151.5。结果 甘精胰岛素单链前体在1~100 ng·mL^(-1)内峰面积与浓度有良好的线性关系(r=0.999 2),方法平均回收率为97%(n=9)。结论 经方法学验证,本法能准确测定甘精胰岛素中单链前体胰岛素前体的残留量,方法精密度、重复性、准确度和耐用性良好。OBJECTIVE To establish an LC-MS/MS method with good accuracy and sensitivity for determination of insulin single chain precursor in insulin glargine,and provide reference for the development of determination method of single chain precursors of insulin and its analogues.METHODS ACQUITY UPLC peptide CSH C18 coulumn(2.1 mm×150 mm,130Å,1.7μm)was adopted.The mobile phase A was 0.1%formic acid/water and the mobile phase B was 0.1%formic acid/acetonitrile,gradient elution program was conducted at the flow rate of 0.3 mL·min^(-1),with column temperature of 60℃and detection wavelength of 214 nm.Orbitrap Exploris 480 was used,and parallel reaction monitoring was carried out by ESI+scanning and quantification ion pair 1155.6/1151.5.RESULTS Good linearity between peak area and concentration of insulin glargine single chain precursor was achieved in the range of 1-100 ng·mL^(-1)(r=0.9992).The average recovery of single chain precursor was 97%(n=9).CONCLUSION This method is suitable for the determination of insulin single chain precursor in recombinant insulin glargine,with good accuracy,repeatability,precision,and durability.

关 键 词：胰岛素单链前体 液质联用 工艺相关杂质 残留量 高分辨质谱 平行反应监测 

分 类 号：R917[医药卫生—药物分析学]