三甲基硅烷衍生化GC-MS测定兔血浆中的游离脂肪酸

doi:10.5246/jcps.2020.06.039

中国药学(英文版) ›› 2020, Vol. 29 ›› Issue (6): 411-421.DOI: 10.5246/jcps.2020.06.039

三甲基硅烷衍生化GC-MS测定兔血浆中的游离脂肪酸

杜嘉琳¹, 杨凉九¹, 卫克昭², 龚灿^1*, 高建平^2*, 许旭^1*

1. 上海应用技术大学化学与环境工程学院, 上海 201418
2. 上海中医药大学中药学院, 上海 201203

收稿日期:2020-01-16 修回日期:2020-04-10 出版日期:2020-06-30 发布日期:2020-04-15
通讯作者: Tel.: +86-21-60877215, +86-21-60873372, E-mail: gongcancan@yeah.net (Gong), xuxu3426@sina.com (Xu)
基金资助:
National Natural Science Foundation of China (Grant No. 31671928) and Natural Science Foundation of Shanghai (Grant No. 15ZR1440800).

Determination of free fatty acids in rabbit plasma by GC-MS after trimethylsilylation derivatization

Jialin Du¹, Liangjiu Yang¹, Kezhao Wei², Can Gong^1*, Jianping Gao^2*, Xu Xu^1*

1. School of Chemical and Environmental Engineering, Shanghai Institute of Technology, Shanghai 201418, China
2. School of Pharmacy, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China

Received:2020-01-16 Revised:2020-04-10 Online:2020-06-30 Published:2020-04-15
Contact: Tel.: +86-21-60877215, +86-21-60873372, E-mail: gongcancan@yeah.net (Gong), xuxu3426@sina.com (Xu)
Supported by:
National Natural Science Foundation of China (Grant No. 31671928) and Natural Science Foundation of Shanghai (Grant No. 15ZR1440800).

摘要/Abstract

摘要：

建立了分析兔血浆中游离脂肪酸(Free fatty acid, FFA)的三甲基硅烷衍生化GC-MS方法。兔血浆样品经N,O-双(三甲基甲硅烷基)三氟乙酰胺(BSTFA)-三甲基氯硅烷(TMCS)衍生化后, 用GC-MS测定了兔血浆中的游离脂肪酸(棕榈酸, 硬脂酸, 油酸, 亚油酸和花生四烯酸)。考察了从脂质萃取到三甲基硅烷化反应在内各项实验条件的影响, 并选定实验条件。经方法学验证, FFA峰面积及其浓度之间的线性相关系数(r²)均大于0.999, 回收率在82%–111%之间。不同浓度血浆样品中的FFA日内变化RSD均小于6%。16种兔血浆样品的FFA分析结果表明, 该方法可以较好用于体内血浆样品的测定, 与传统的FFA衍生化方法相比, 所建立的三甲基硅烷衍生化方法简单、特异性好, 且完全不受已酯化脂肪酸(例如甘油三酯)的干扰, 可用于临床实验室或药理研究中的FFA谱分析。

关键词: 游离脂肪酸, 三甲基硅烷, 衍生化, GC-MS, 兔血浆

Abstract:

In the present study, free fatty acids (FFAs, including palmitic acid, stearic acid, oleic acid, linoleic acid and arachidonicacid) in rabbit plasma were determined by gas chromatography-mass spectrometry (GC-MS) after trimethylsilylation derivatizationusing N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) – trimethylchlorosilane (TMCS) as derivatization reagent. The experimental conditions, including extraction and silylation reaction, were investigated. The method was experimentally validated. The linearity between fatty acids’ peak areas and their concentrations was obtained with the corelative coefficient (r²) all more than 0.999, and the recoveries were between 82% and 111%. The intra-day variations of FFAs’ in plasma samples at different concentrations were all less than 6%. FFA analysis results of 16 rabbit plasma samples showed that the method could be well applied in the determination of plasma samples in vivo. In contrast to the traditional method of FFA derivatization, the established trimethylsilylationmethod presented simplicity, high specificity, and completely free from the interference of the esterified fatty acid, such as triacylglyceride. The method could be applied for analyzing FFA profiles in the clinical laboratory or pharmacological research.

Key words: Free fatty acid, Trimethylsilylation, Derivatization, Gas chromatography-mass spectrometry, Rabbit plasma

中图分类号:

R917

Supporting:

杜嘉琳, 杨凉九, 卫克昭, 龚灿, 高建平, 许旭. 三甲基硅烷衍生化GC-MS测定兔血浆中的游离脂肪酸[J]. 中国药学(英文版), 2020, 29(6): 411-421.

Jialin Du, Liangjiu Yang, Kezhao Wei, Can Gong, Jianping Gao, Xu Xu. Determination of free fatty acids in rabbit plasma by GC-MS after trimethylsilylation derivatization[J]. Journal of Chinese Pharmaceutical Sciences, 2020, 29(6): 411-421.

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三甲基硅烷衍生化GC-MS测定兔血浆中的游离脂肪酸

Determination of free fatty acids in rabbit plasma by GC-MS after trimethylsilylation derivatization

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