柱前衍生HPLC-FLD法测定人血浆中羧甲司坦

HPLC-FLD precolumn derivation determination of carbocisteine in human plasma

目的:建立血浆样品中羧甲司坦的柱前衍生 HPLC—FLD 测定法。方法:色谱柱:Agilent C_(18),(5μm,4.6 mm×150mm);柱温:30℃;流动相:0→25 min,A～B(89:11),A:40 mmol·L^(-1)磷酸二氢钠水溶液(1 mol·L^(-1)氢氧化钠调 pH 为7.8),B:甲醇-乙睛-水(45:45:10);25→32 min A 为100%;32→40 min,A—B(89:11);流速:0.8 mL·min^(-1);检测波长:激发波长338 nm,发射波长450 nm;进样量:2.0 μL,柱温30℃。结果:最低检测限为0.1018μg·mL^(-1);线性范围为0.1018～13.00μg·mL^(-1)(r=0.9999);低、中、高3种浓度日内(n=6)、日间(n=8)RSD 分别为4.0%,2.0%,0.7%和4.2%,2.6%,0.7%;3种浓度的绝对回收率分别为92.4%,97.2%,99.4%,RSD 分别为2.4%,1.3%,0.59%;相对回收率分别为96.96%,100.45%,100.35%,RSD 分别为2.91%,2.21%,0.50%。结论:本法准确、灵敏、易于操作,可用于羧甲司坦的体内分析及临床药学研究。

Objective：To establish a precolumn derivation HPLC -FLD method to determine carbocisteine in hu-man plasma. Method ： With the derived method use of OPA, the content of carbocisteine in human method is determined by external standard sample. Applied Agilent ODS - C18 column（5 μm,4.6 mm×150mm） ;The mobile phase was A - B（ 89：11 ） in 0→25 min, A = 40 mmol·L^-1 sodium dihydrogen phosphate WS（ adjusted to pH 7.8 with 1 mol·L^-1 sodium hydroxide） , B = methanol - acetonitrile - water（45： 45： 10） ;then the mobile phase was 100%A in 25→32min,in 32→40 min was A -B（89：11 ） ;the rate was 0.8 mL·min^-1 ;The λex= 338 nm, λem= 450 nm and column temperature was controlled about 30℃. Results：The regression line was linear in the concentration range of 0.1018 - 13.00μg·mL^-1（ r=0.9999） ; the limit of detection was 0.1018 μg·mL^-1; Intra-day and inter-day RSD for assaying the plasma sample containing three concentration of carbo-eisteine were 4.0% ,2.0% ,0.7% and 4.2% ,2.6% ,0.7% （n = 8 ） ;The absolute recovery of three concentration were 92.4% ,97.2% ,99.4% , RSD 2.4% , 1.3% ,0. 59% ; relative recovery were 96.96% , 100.45% , 100.35 % , RSD 2.91% ,2.21% , 0.50%. Conclusion ： The method is sensitive, accurate and simple. It can be used to determine the concentration of carbocisteine in plasma, and can be applied to the clinical pharmaceutical research.