Plant Lipids - Methods and Protocols
Plant Lipids - Methods and Protocols
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植物脂质 - 方法和方案
DOI：
10.1007/978-1-0716-1362-7_11
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Broughton R
中科院分区：
文献类型：
--
作者：
Broughton R
总脂提取物皂化后，用气相色谱(GC)法测定植物组织中总甾醇的含量和组成。然而，在成油组织中，相当大比例的甾醇被酯化成脂肪酸，而GC方法无法提供关于完整甾酯(Ses)的比例和分子种类组成的信息。在这里，我们描述了电喷雾电离串联质谱仪(ESI-MS/MS)和多反应监测(MRM)方法，该方法与GC分析相结合，可以准确地测定种子中游离和酯化的甾醇含量和组成。用正己烷提取种子油后，游离的甾醇用十一酰氯衍生，三酰甘油(TAG)中的总酯再用液相色谱纯化，然后以氨加合物的形式进样和电离，在有内标的情况下，通过裂解鉴定和定量分子种类。
Total sterol content and composition in plant tissues can be easily determined by gas chromatography (GC) after saponification of the total lipid extract. However, in oleogenic tissues a significant proportion of the sterol is esterified to fatty acids, with GC methodologies unable to provide information about the proportion and the molecular species composition of intact steryl esters (SEs). Here we describe an electrospray ionization-tandem mass spectrometry (ESI-MS/MS) and Multiple Reaction Monitoring (MRM) method which, in parallel with GC analysis, allows for the accurate determination of both free and esterified sterol content and composition in seeds. After extraction of seed oil with hexane, free sterols are derivatized with undecanoyl chloride, total steryl esters are then purified from triacylglycerol (TAG) by liquid chromatography, infused and ionized as ammonium adducts, with molecular species identified and quantified by fragmentation in the presence of internal standards.