高效液相色谱法测定血浆中甲磺酸加替沙星的含量

Determination of gatifloxacin in human plasma by HPLC

目的 建立测定血浆中甲磺酸加替沙星浓度的高效液相色谱法。方法 待测血浆中加入内标环丙沙星后 ,以甲醇直接沉淀蛋白 ,进行高效液相荧光检测 ,色谱柱为ZorbaxSB C18,流动相为 0 .1mol·L-1磷酸盐缓冲液 甲醇 (6 8∶32 ) ,0 .1mol·L-1磷酸盐缓冲液中含有 0 .0 0 5mol·L-1离子对试剂氢氧化四丁基铵 (IPR A) ,pH值为 2 .35。流速 1.0mL·min-1,激发波长 2 95nm ,发射波长 4 95nm。结果 本测定方法的线形范围为 0 .0 6 2 5～ 1.0 0 0 0 μg·mL-1,r =0 .9990 ,日内、日间RSD均小于10 % ,回收率为 93.1%～ 94 .0 % ,血浆中甲磺酸加替沙星的最低检测限为 0 .0 313μg·mL-1。结论 本法精密、准确 ,适用于临床上血药浓度监测 ,人体药动学和生物利用度研究。

OBJECTIVE: A simple, accurate and reproducible HPLC procedure was developed for the determination of gatifloxacin in plasma. METHODS: Ciprofloxacin was used as the internal standard. A single step protein preciptitation procedure with methanol was employed for pretreatment. Zorbax SB-C18 was used as the analytical column. The mobile phase contained 0.1 mol&middotL-1 phosphate buffer solution: methanol(68::32, v/v), contained 0. 005 mol&middotL-1 tetrabutylummoinium hydroxide, with pH = 2.35. The flow rate was 1.0 mL&middotmin-1, with ex = 295 nm and em = 495 nm. RESULTS: The mean recovery of gatifloxacin was 93.1% ∼ 94.0%, and the relative standard deviation(RSD) of within day and between day were all less than 10%. The detection limit was 0.031 3 μg&middot mL-1. The assay developed was found to be linear for gatifloxacin(0.062 5∼1.000 0 μg&middotmL-1) with correlation coefficients was 0. 999 0. CONCLUSION: The method was found to be simple and quick with the high sensitivity.