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高效液相色谱法对精胺脂的微尺度测定及其在公猪精子中精胺脂含量测定中的应用

Micro-scale determination of seminolipid by high performance liquid chromatography and its application for determination of the seminolipid content in boar spermatozoa.

作者信息

Suzuki A, Handa S, Ishizuka I, Yamakawa T

出版信息

J Biochem. 1977 Jan;81(1):127-34. doi: 10.1093/oxfordjournals.jbchem.a131427.

Abstract

Seminolipid (1-O-alkyl-2-O-acyl-beta-3'sulfogalactosyl glycerol) was detected on a micro-scale by high performance liquid chromatography (HPLC) using a UV detector. The benzoylation method of McCluer and Evans ((1973) J. Lipid Res. 14, 611-617) was used to convert seminolipid to a derivative with UV absorption. Total lipid from germinal tissue was benzoylated and the product was separated by Florisil and Sephadex LH-20 column chromatographies. The seminolipid obtained in the effluent from the Sephadex LH-20 column was determined by HPLC. Eighty percent recovery of 35S-labelled seminolipid was obtained by this method. The method was used to determine the seminolipid content of boar spermatozoa: 980 nmoles of seminolipid was found per gram of packed cells.

摘要

使用紫外检测器,通过高效液相色谱法(HPLC)在微观尺度上检测了半乳糖神经酰胺(1-O-烷基-2-O-酰基-β-3'-磺基半乳糖甘油)。采用麦克卢尔和埃文斯的苯甲酰化方法((1973年)《脂质研究杂志》14卷,611 - 617页)将半乳糖神经酰胺转化为具有紫外吸收的衍生物。对生殖组织中的总脂质进行苯甲酰化处理,产物通过弗罗里硅土柱色谱和葡聚糖凝胶LH - 20柱色谱进行分离。通过HPLC测定从葡聚糖凝胶LH - 20柱流出液中获得的半乳糖神经酰胺。用该方法获得了35S标记的半乳糖神经酰胺80%的回收率。该方法用于测定公猪精子中的半乳糖神经酰胺含量:每克压实细胞中发现980纳摩尔的半乳糖神经酰胺。

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