ACIDIC HYDROLYSIS OF PLASMALOGENS FOLLOWED BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY
ACIDIC HYDROLYSIS OF PLASMALOGENS FOLLOWED BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY
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DOI：
10.1007/bf02536090
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发表时间：
1993-06-01
期刊：
影响因子：
1.9
通讯作者：
HORROCKS, LA
中科院分区：
文献类型：
--
作者：
MURPHY, EJ;STEPHENS, R;HORROCKS, LA
本文报道了一种简单、定量测定小样品中血浆蛋白原含量的方法。该方法利用烷基、烯基和酰基对酸催化水解的不同敏感性，将血浆蛋白原亚类与其他两个非稳定亚类分开。血浆乙醇胺和血浆胆碱分别在酸处理4min和1min后完全水解。酸催化的水解不会改变磷脂脂肪酸的组成，使该方法适用于血浆蛋白原亚类的脂肪酸组成分析。采用高效液相色谱分离，磷脂定量采用脂磷法或直接峰面积法定量。使用这种方法，少量(10nmol)乙醇胺甘油磷脂和胆碱甘油磷脂经酸催化水解，然后从更稳定的烷基和二酰甘油磷脂组分中分离从血浆原中获得的溶质化合物。我们对商业胆碱和乙醇胺甘油磷脂制剂中血浆原的测定值与文献值一致。对于与培养的神经细胞样本相当的小甘油磷脂样本，证明了该方法的有效性。
A simple, quantitative method for determining the plasmalogen content of small samples is reported here. The method uses the different susceptibility to acid-catalyzed hydrolysis of the alkyl, alkenyl and acyl linkages to separate the plasmalogen subclass from the other two non-labile subclasses. Hydrolysis of plasmenylethanolamine and plasmenylcholine was complete after 4 and 1 min of acid treatment, respectively. The acid-catalyzed hydrolysis did not alter the phospholipid fatty acid composition, making this method useful for fatty acid compositional analysis of the plasmalogen subclass. High-performance liquid chromatography was used for separations, and phospholipids were quantitated by assay of lipid phosphorus or by direct quantitation of peak area. Using this method, small amounts (10 nmol) of ethanolamine glycerophospholipid and choline glycerophospholipid are subjected to acid-catalyzed hydrolysis and subsequent separation of the resulting lysocompounds obtained from plasmalogens from the more acid-stable alkylacyl and diacyl glycerophospholipid fractions. Our values for plasmalogens from commercial preparations of choline and ethanolamine glycerophospholipids agree with literature values. The usefulness of the method is demonstrated for small glycerophospholipid samples that are equivalent to samples from cultured neural cells.