Qualitative and Quantitative Analyses of Phospholipids by LC-MS for Lipidomics
Qualitative and Quantitative Analyses of Phospholipids by LC-MS for Lipidomics
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DOI：
10.1007/978-1-60761-322-0_15
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发表时间：
2009-01-01
期刊：
影响因子：
--
通讯作者：
Taguchi, Ryo
中科院分区：
文献类型：
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作者：
Nakanishi, Hiroki;Ogiso, Hideo;Taguchi, Ryo
在本章中，我们将介绍脂质组学中的三种不同方法，以及如何有效地分析或计算从主要分子种类到次要分子种类的磷脂的量。1）通过数据依赖性LC-ESIMS/MS结合“Lipid Search”对磷脂的单个分子种类进行精确鉴定和分析。我们一直使用这种方法作为磷脂的全球分析。我们通常对新的生物样品至少应用一次这种方法。我们构建了一个自动化的搜索引擎，“脂质搜索”，磷脂的识别和分析。在应用该分析后，可以获得单个磷脂分子种类的每个洗脱峰的指定保留时间。因此，通过使用前体离子扫描或中性损失扫描的测量，可以有效地获得可重复的鉴定结果。2）LC-ESIMS鉴定酸性磷脂如磷脂酸和磷脂酰丝氨酸的有效分析方法。这是一种如何获得酸性脂质如磷脂酸和磷脂酰丝氨酸的尖锐色谱峰的方法，这些峰通常被检测为宽洗脱峰。通过这种改进，在次要酸性磷脂中有效地获得了非常少量的分子种类。3）在聚焦磷脂中的分子种类的鉴定和分析。第三部分是前体离子扫描或中性损失扫描等聚焦方法与高效液相色谱分离的组合分析。如前所述，通过反相LC-ESIMS，sn-1和sn-2上的脂肪酸的不同组合大多可以作为单独的峰被检测到。前体离子扫描或中性丢失扫描的检测限比不经LC分离的方法提高了十倍以上，这是因为离子抑制降低了。我们将提及这种方法在磷脂酰乙醇胺-缩醛磷脂集中分析中的应用。
In this chapter we are going to mention about three different approaches in lipidomics and how to effectively profile or calculate the amounts of phospholipids from major molecular species up to minor one.1) Precise identification and profiling of individual molecular species of phospholipids by data-dependent LC-ESIMS/MS combination with "Lipid Search". We have been using this method as a global analysis of phospholipid. We usually applied this method at least once for new biological samples. We constructed an automated search engine, "Lipid Search", for identification and profiling of phospholipids . Once after applying this analysis, a specified retention time can be obtained for each elution peak of individual phospholipid molecular species. Thus, reproducible identification results can be effectively obtained by survey by using precursor ion scanning or neutral loss scanning.2) An effective analytical method for LC-ESIMS for the identification of acidic phospholipids such as phosphotidic acid and phosphatidylserine. This is an approach of how to obtain sharp chromatographic peaks for acidic lipids such as phosphatidic acid and phosphatidylserine that are normally detected as broad elution peaks. With this improvement very small amount of molecular species in minor acidic phospholipids were effectively obtained.3) Identification and profiling of molecular species in focused phospholipids. Third one is a combination analysis of focused methods such a precursor ion scanning or neutral loss scanning and high efficient LC seperation. As reported previously, different combination of fatty acids on sn-1 and sn-2 can be mostly detected as separate peaks by reversed phase LC-ESIMS. Detection limit of precursor ion scanning or neutral loss scanning is more than ten times higher than that of the method without LC seperation, because of decreased ion suppression. We will mention about application of this methods for focused analysis on phosphatidylethanolamine-plasmalogens.