衍生化-气相色谱-质谱法检测阿加曲班中氯甲酸异丁酯残留  

Determination of Isobutyl Chloroformate Residue in Agatroban by Derivatization-Gas Chromatography-Mass Spectrometry

作　　者：钱冲[1,2] 马博凯 牛闯 刘珊珊 黄雯雯 勾新磊 王尉 张梅[2] 曹学丽[1] QIAN Chong;MA Bo-Kai;NIU Chuang;LIU Shan-Shan;HUANG Wen-Wen;GOU Xin-Lei;WANG Wei;ZHANG Mei;CAO Xue-Li(Beijing Advanced Innovation Center for Food Nutrition and Human Health,School of Light Industry,Beijing Technology and Business University,Beijing 100048,China;Beijing Key Laboratory of Organic Materials Testing Technology&Quality Evaluation,Institute of Analysis and Testing,Beijing Academy of Science and Technology(Beijing Center for Physical&Chemical Analysis),Beijing 100094,China;Beijing City University,Beijing 100094,China)

机构地区：[1]北京工商大学轻工科学与工程学院,北京食品营养与人类健康高精尖创新中心,北京100048 [2]北京市科学技术研究院分析测试研究所(北京市理化分析测试中心),有机材料检测技术与质量评价北京市重点实验室,北京100094 [3]北京城市学院,北京100094

出　　处：《分析化学》2024年第1期113-120,共8页Chinese Journal of Analytical Chemistry

基　　金：北京市科学技术研究院创新工程项目(No.23CA005-03)资助。

摘　　要：基于衍生化反应,采用气相色谱-质谱联用方法(Gas chromatography-mass spectrometry,GC-MS)检测了阿加曲班原料药中残留的氯甲酸异丁酯(Isobutyl chloroformate,IBCF)。分别对提取和衍生试剂、衍生反应时间、定性和定量离子进行筛选和优化,推测了衍生反应机理和特征碎片离子的碎裂机理。阿加曲班样品经甲醇溶解及提取,其中残留的IBCF与甲醇发生衍生反应,生成甲基异丁基碳酸酯(Methyl isobutyl carbonate,MIBCB),室温静置衍生24 h后,IBCF完全转化为MIBCB,通过检测MIBCB间接实现了IBCF的准确测定。IBCF在25~500 ng/mL浓度范围内线性关系良好(R^(2)=0.9999),检出限(LOD,S/N=3)为0.75μg/g,定量限(LOQ,S/N=10)为2.50μg/g。低(2.50μg/g)、中(25.00μg/g)和高(50.00μg/g)3个浓度水平的加标回收率为95.2%~97.8%,相对标准偏差(Relative standard deviation,RSD)为2.0%~3.1%(n=6)。本方法的精密度良好,不同日期、不同实验人员测定加标样品溶液(25.00μg/g)结果的平均值为24.28μg/g,RSD为2.1%(n=12)。本方法的耐用性良好,测定条件的微小变动对结果影响较小,在原条件、初始柱温±5℃、升温速率±2℃/min和柱流量±0.1 mL/min的条件下分别对加标样品溶液(25.00μg/g)进行测定,检测结果为24.16μg/g,RSD为2.2%(n=7)。将本方法用于两家药业公司生产的8批次阿加曲班样品的检测,均未检出IBCF。本方法能同时实现阿加曲班的溶解以及IBCF的提取和衍生,操作简便、灵敏度高、基质效应低、结果准确可靠,为阿加曲班中IBCF残留的测定提供了有效方法。A derivatizaton method combined with gas chromatography-mass spectrometry(GC-MS)was established for detection of isobutyl chloroformate(IBCF)residue in active pharmaceutical ingredient of agatroban.The extraction and derivatization reagents,derivatization time,qualitative and quantitative ions were selected and optimized,respectively.The possible mechanism of derivatization and characteristic fragment ions fragmentation were speculated.The agatroban samples were dissolved and extracted by methanol,and the residual IBCF was derived with methanol to generate methyl isobutyl carbonate(MIBCB).After 24 h static derivatization at room temperature,IBCF was completely transformed into MIBCB,which could be used to indirectly detect IBCF accurately.The results showed that the linearity of this method was good in the range of 25–500 ng/mL(R2=0.9999).The limit of detection(LOD,S/N=3)was 0.75μg/g,and the limit of quantification(LOQ,S/N=10)was 2.50μg/g.Good recoveries(95.2%–97.8%)and relative standard deviations(RSDs)less than 3.1%(n=6)were obtained from agatroban samples at three spiked levels of IBCF(2.50,25.00,50.00μg/g),which showed good accuracy of this method.Good precision of detection results was obtained by different laboratory technicians at different times,the mean value of spiked sample solution(25.00μg/g)was 24.28μg/g,and the RSD was 2.1%(n=12).The durability was good,minor changes of detection conditions had little effect on the results.Under the original condition and conditions with initial column temperature±5℃,heating rate±2℃/min,column flow rate±0.1 mL/min,the IBCF content of spiked sample solution(25.00μg/g)was detected,the mean value of detection results was 24.16μg/g,and the RSD was 2.2%(n=7).Eight batches of agatroban samples from two manufacturers were detected using the established method,and the results showed that no IBCF residue was detected in any of these samples.The agatroban samples could be dissolved by methanol,and then the IBCF residue could be simultaneously extracted and d

关 键 词：阿加曲班 氯甲酸异丁酯 衍生反应 气相色谱-质谱 甲基异丁基碳酸酯 

分 类 号：O657.63[理学—分析化学] TQ460.72[理学—化学]