基于两步衍生化-HPLC-MS的体内游离脂肪酸定性分析研究  

Qualitative Study of Free Fatty Acids Based on Two-step Derivatization-HPLC-MS Method

作　　者：肖雯 任海慧 田媛[1] 张尊建[1] XIAO Wen;REN Haihui;TIAN Yuan;ZHANG Zunjian(China Pharmaceutical University,Key Laboratory of Drug Quality Control and Pharmacovigilance,Nanjing 210009,China)

机构地区：[1]中国药科大学,药物质量与安全预警教育部重点实验室,南京210009

出　　处：《中国现代应用药学》2024年第20期2789-2798,共10页Chinese Journal of Modern Applied Pharmacy

摘　　要：目的建立两步衍生化-高效液相色谱质谱联用方法,对生物样本中的游离脂肪酸进行定性分析,重点区分双键位置异构体。方法采用间氯过氧苯甲酸和N,N-二乙基乙二胺对6种具有不同饱和度的代表性游离脂肪酸进行两步衍生化,采用高效液相色谱串联三重四极杆质谱检测。结果经标准品验证,证明采用质谱裂解数据回溯游离脂肪酸结构的分析思路可行、可靠;从比格犬血浆中共鉴定出游离脂肪酸38种,其中单不饱和的13种,多不饱和的17种,饱和的8种;可区分双键位置异构体10种,并准确定位低丰度的二十二碳六烯酸双键位置。结论建立的两步衍生化-高效液相色谱质谱联用方法为体内游离脂肪酸的定性分析,特别是区分双键位置异构方面提供了新的思路,也为相关代谢组学、脂质组学研究提供了方法学支撑。OBJECTIVE To establish a two-step derivatization and high performance liquid chromatography-mass spectrometry method,and to qualitatively study the free fatty acids in biological samples,with a focus on distinguishing double bond isomers.METHODS Six representative free fatty acids with different saturation were subjected to two-step derivatization using m-chloroperoxybenzoic acid and N,N-diethylethylenediamine.High performance liquid chromatography-mass spectrometry was used for analysis and detection.RESULTS Verified by 6 free fatty acid standards,the analysis approach of mass spectrometry fragmentation data backtracking structure was feasible and reliable.A total of 38 kinds of free fatty acids were identified from Beagle plasma,including 13 monounsaturated,17 polyunsaturated and 8 saturated.It could distinguish 10 kinds of double bond isomers and accurately analyze the double bond positions of low abundance docosahexadilute acids.CONCLUSION The established two-step derivatization and high performance liquid chromatography mass spectrometry method provides a new idea for the qualitative analysis of free fatty acids in vivo,especially for the differentiation of double bond isomerism,and also provides methodological support for related metabolomics and lipidomics studies.

关 键 词：游离脂肪酸 衍生化 高效液相色谱-质谱法 

分 类 号：R917[医药卫生—药物分析学]