Department of Pharmacology, University of Colorado Denver, Aurora, Colorado 80045, USA.
Anal Chem. 2009 Aug 15;81(16):6633-40. doi: 10.1021/ac900583a.
A set of four (D(0), D(4), D(6), and D(10)) deuterium enriched 4-(dimethylamino)benzoic acid (DMABA) N-hydroxysuccinimide (NHS) ester reagents was developed that react with the primary amine group of glycerophosphoethanolamine (PE) lipids to create derivatives where all subclasses of DMABA labeled PE are detected by a common precursor ion scan. The positive ion collision induced dissociation data from (D(0), D(4), D(6), and D(10))-DMABA labeled PE standards indicated that a precursor ion scan of m/z 191.1, 195.1, 197.1, and 201.1 could be used to selectively detect (D(0), D(4), D(6), and D(10))-DMABA modified PE, respectively, in a complex biological mixture. The PE lipids from a time course (0, 30, 60, and 300 min) of 2,2'-azobis-(2-amidinopropane) hydrochloride (AAPH) treatment of liposomes made of RAW 264.7 cell phospholipids were each labeled with the D(0)-, D(4)-, D(10)-, and D(6)-DMABA NHS ester reagents, respectively. The DMABA derivatives revealed loss of endogenous PE lipids and an increase in oxidized PE lipid throughout the time course of AAPH treatment. These DMABA NHS ester reagents provide a universal scan for diacyl, ether, and plasmalogen PE lipids that cannot be readily observed otherwise, enable differential labeling, and provide an internal standard for each PE lipid.
开发了一套四组（D(0)、D(4)、D(6)和 D(10)）氘代 4-（二甲氨基）苯甲酸（DMABA）N-羟基琥珀酰亚胺（NHS）酯试剂，它们与甘油磷酸乙醇胺（PE）脂质的伯胺基团反应，生成衍生物，其中所有 DMABA 标记的 PE 亚类都可以通过共同的前体离子扫描检测到。（D(0)、D(4)、D(6)和 D(10)）-DMABA 标记的 PE 标准品的正离子碰撞诱导解离数据表明，m/z 191.1、195.1、197.1 和 201.1 的前体离子扫描可分别用于选择性检测复杂生物混合物中的（D(0)、D(4)、D(6)和 D(10)）-DMABA 修饰的 PE。来自 RAW 264.7 细胞磷脂脂质体 2,2'-偶氮双（2-脒基丙烷）盐酸盐（AAPH）处理的时间过程（0、30、60 和 300 min）的 PE 脂质分别用 D(0)-、D(4)-、D(10)-和 D(6)-DMABA NHS 酯试剂标记。DMABA 衍生物揭示了内源性 PE 脂质的损失和氧化的 PE 脂质的增加，整个 AAPH 处理过程中都是如此。这些 DMABA NHS 酯试剂为不能轻易观察到的二酰基、醚和血浆衍生的 PE 脂质提供了通用扫描，实现了差异标记，并为每种 PE 脂质提供了内部标准。
