HPLC-Q TOF MS法同时测定4种GLP-1类多肽及4种小分子减肥类添加物  

Simultaneous analysis of 4 GLP-1 peptide additives and 4 small molecule illegal anti-obesity drugs by HPLC-Q TOF MS

作　　者：凌明 舒展 金芩 王琤帅 LING Ming;SHU Zhan;JIN Qin;WANG Cheng-Shuai(Jinhua Institute for Food and Drug Control,Jinhua 321000,China)

机构地区：[1]金华市食品药品检验检测研究院,金华321000

出　　处：《药物分析杂志》2025年第2期280-289,共10页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:建立高效液相色谱-四极杆飞行时间质谱(HPLC-Q TOF MS)同时测定4种减肥类小分子及4种胰高血糖素样肽-1(GLP-1)多肽类非法添加物的分析方法。方法:样品以50%乙腈为提取溶剂进行超声提取,离心后取上清液,采用Agilent EC-C_(18)色谱柱(150 mm×3.0 mm,2.7μm)进行分离,以乙腈(0.1%甲酸)和水(0.1%甲酸)为流动相,梯度洗脱,流速为0.3 mL·min^(-1),柱温40℃。采用正离子全扫描及目标离子二级碎片扫描方式,碎裂电压150 V。其中一级质谱扫描范围为m/z 100~3 200,二级质谱扫描范围为m/z 50~3 200,扫描速度均为每秒1张质谱图。根据对照品的色谱保留时间、一级质谱和二级质谱信息建立数据谱库,通过数据库比对进行结构的确认。结果:多肽类物质定性筛查的检测限为0.5μg·mL^(-1),其他小分子物质为0.05μg·mL^(-1)。在全扫描模式下,8个化合物线性相关系数均> 0.995,回收率范围为79.4%~115.8%,RSD范围为0.21%~9.7%。采用本方法对20批减肥类样品进行检测,其中1批样品中检出西布曲明,4批样品中检出司美格鲁肽。结论:本方法扩大了国家食品药品监督管理局补充检验方法和检验项目批件2012005的筛查范围,能同时兼顾小分子物质和大分子多肽类物质(相对分子质量<5 000)检测的需要,具有高效、准确的特点。Objective:To establish a high-performance liquid chromatography quadrupole time-of-flight mass spectrometry(HPLC-Q TOF MS)method for the simultaneous analysis of illegal 4 anti-obesity small molecule drugs and 4 glucagon-like peptide-1(GLP-1)peptide additives.Methods:The samples were extracted by ultrasound using 50%acetonitrile as the extraction solvent.After centrifugation,the supernatant was taken and separated using an Agilent EC-c18 chromatographic column(150 mm×3.0 mm,2.7μm).Acetonitrile(0.1%formic acid)and water(0.1%formic acid)were used as mobile phases,with gradient elution at a flow rate of 0.3 mL·min^(-1) and column temperature of 40 C.Adopting positive ion full scanning and target ion secondary fragment scanning methods,with a fragmentation voltage of 150 V.The scanning range of the primary mass spectrometry was m/z 100-3200,and the scanning range of the secondary mass spectrometry was m/z 50-3200,with a scanning speed of 1 mass spectrum per second.Establish a data spectral library based on the chromatographic retention time,primary mass spectrometry,and secondary mass spectrometry information of the reference standard,and confirmed the structure through database comparison.Results:The screening detection limit for peptides was 0.5μg·mL^(-1),while small molecular drugs was 0.05μg·mL^(-1).The recoveries were in the range of 79.4%to 115.8%,with the relative standard deviations of 0.21%to 9.7%.Using this method,20 batches of anti-obesity drugs were tested,in which semaglutide was identified in 4 samples and sibutratmine was identified in 1 batche.Conclusion:Compared with the complementary method No.2012005 by the China Food and Drug Administration,the method established in this study can simultaneously analyze small and large molecules(the relative molecular mass<5000),featuring high efficiency and accuracy.

关 键 词：高效液相色谱-四极杆飞行时间质谱法 减肥类非法添加药物 胰高血糖素样肽-1受体激动剂(GLP-1 RA) 多肽类物质 裂解规律 

分 类 号：R917[医药卫生—药物分析学]