Nakagawa Y, Kishida K, Kodani Y, Matsuyama T
Department of Applied Biological Chemistry, Faculty of Agriculture, Niigata University, Japan.
Microbiol Immunol. 1997;41(1):27-32. doi: 10.1111/j.1348-0421.1997.tb01169.x.
Through the adoption of a chiral stationary phase in high-performance liquid chromatography and a simple derivatization method for hydroxy fatty acids, it became easy to separate and identify the optical isomers of 2- and 3-hydroxy fatty acids composing several kinds of microbial lipids. The 2- and 3-hydroxy fatty acids were converted with dinitrophenyl isocyanate to their 3,5-dinitrophenyl urethane derivatives (DU-derivatives), which were analyzable by HPLC using a chiral column. By varying the composition of an eluent, separation of the DU-derivatives of hydroxy fatty acids differing in optical configuration, chain length and position of hydroxyl group was achieved. The general elution orders of these DU-derivatives were determined with authentic 2- and 3-hydroxy fatty acids. Small amounts (approximately 300 micrograms) of ornithine-containing lipids isolated from the Serratia marcescens strains were examined by this method to identify 3-hydroxy fatty acids of the lipids as D isomers.
通过在高效液相色谱中采用手性固定相以及对羟基脂肪酸采用简单的衍生化方法，就能够轻松分离和鉴定构成几种微生物脂质的2-羟基脂肪酸和3-羟基脂肪酸的光学异构体。2-羟基脂肪酸和3-羟基脂肪酸用异氰酸二硝基苯转化为它们的3,5-二硝基苯基脲衍生物（DU-衍生物），这些衍生物可用手性柱通过高效液相色谱进行分析。通过改变洗脱液的组成，实现了不同光学构型、链长和羟基位置的羟基脂肪酸的DU-衍生物的分离。用纯正的2-羟基脂肪酸和3-羟基脂肪酸确定了这些DU-衍生物的一般洗脱顺序。用该方法检测了从粘质沙雷氏菌菌株中分离出的少量（约300微克）含鸟氨酸脂质，以鉴定脂质中的3-羟基脂肪酸为D异构体。
