鸟氨酸、瓜氨酸的HPLC测定方法优化

Optimized Determination Method of Ornithine and Citrulline by HPLC

目的:建立一种简单快速测定鸟氨酸、瓜氨酸的柱前衍生化方法。方法:比较文献中不同方法的优劣,选取异硫氰酸苯酯(PITC)作为柱前衍生试剂,高效液相色谱(HPLC)法分离测定,以保留时间定性,峰面积定量。结果:PITC柱前衍生化HPLC梯度洗脱测定鸟氨酸、瓜氨酸,流动相A:0.1 mol/L乙酸钠∶乙腈(93∶7),流动相B:水∶乙腈(1∶4);流速:1.0 m L/min;检测波长:254 nm;柱温:40℃;洗脱条件:0 min^0(B%),3 min^34(B%),15 min^100(B%),16 min^0(B%),20 min^0(B%);鸟氨酸的保留时间为8.5 min,瓜氨酸的保留时间为6.0 min,样品分析时间为20 min。结论:PITC柱前衍生化HPLC方法测定鸟氨酸、瓜氨酸前处理简单,分析速度快于文献报道的方法。

To establish a PITC pre-column derivative and HPLC method which was thst and simple for the determination of ornithine and citrulline？ Through compare the different methods in literatures, at last we selected phenylisothiocyanate （PITC） as derivative reagent and HPLC as the detection, the retention time was used as a qualitative standard and peak area as a quantitative standard. HPLC showed that ornithine retention time was 8.5 min and citrulline was 6.0 min. Mobile phase A： 0.1mol/L CH3COONa ： CH3CN （93 ： 7）, Mobile phase B： H20 ： CH3CN（1 ： 4）; flow velocity： 1.0 mL/min; λ=245 nm; Column temperature： 40 ℃： Elution requirement： 0 min-0 （B%）, 3 min-34 （B%）, 15 min-100 （B%）, 16 min-0 （B%）, 20 min-0（B%）. There is 20 min for per sample. The method of this article to detect ornithine and citrulline was fast and simple, and faster than the method of the present reported.