采用氨基柱的HPLC分离水稻线粒体膜磷脂组分

Separation of Phospholipids of Rice Mitochondrial Membrane by HPLC with Capcell Pak-SG NH_2 Column

比较了L iChrosorb C18(250×4.6 mm,5μm)和C apce ll Pak-SG NH2(250×4.6 mm,5μm)氨基柱分析红莲型细胞质雄性不育水稻(HL-CM S)线粒体膜磷脂组成的方法.结果表明:用C apce ll Pak-SG NH2氨基柱,流动相用乙腈-甲醇-85%磷酸(体积比100∶10∶1),二极管阵列(DAD)检测器,检测波长205 nm,在15 m in内使膜磷脂的4种主要成分达到基线分离.在此色谱条件下,样品测试精密度较高,重现性好.用面积归一化法对HL-CM S水稻不育系粤泰A(YTA)和保持系粤泰B(YTB)的线粒体膜磷脂组成成分含量进行了定量分析,结果表明两者膜磷脂组分相似,但其含量配比存在差异.

Two different columns, LiChrosorb C18（250× 4.6 mm, 5 μm）positive-phase column and Capcell Pak-SG NH2 column, were compared in HPLC analysis of rice mitochondrial membrane phospholipids. It was shown that the four essential and important phospholipids, PE, PI, PS and PC of mitochondrial membrane were separated better on Capcell Pak-SG NH2 column than on LiChrosorb C18. The optimized HPLC conditions with Capcell Pak-SG NH2 column were the mobile phase, acetonitrile-methanol -85% phosphoric acid （100 ： 10 ： 1, ratio of volume）, and DAD detector at detection wave length of 205 nm. Under this optimized chromatographic conditions, the four essential and important phospholipids, PE, PI, PS and PC of mitochondrial membrane could be separated in 15 minutes with the standard deviations of elution time from 0.21 % to 0.90 %. The quantitative analysis of the four important mitochondrial membrane phospholipids,PE, PI, PS and PC isolated from two rice lines, HL-CMS sterility line YTA and its＇ maintainer YTB, had been also achieved with area normalization method. The results indicated that the percentage of PE, PI, PS and PC in mitochondrial membrane in YTA was obviously different that in YTB although their components were similar with each other.