高效液相色谱法测定人血浆中羧甲司坦浓度

Determination of carbocisteine in human plasma by HPLC

目的:建立高效液相色谱法测定人血浆中羧甲司坦浓度.方法:采用邻苯二甲醛为柱前衍生化试剂,反相高效液相色谱法检测羧甲司坦衍生物:Hypersil BDS C18柱(4.6 mm×150 mm,5 μm);流动相:甲醇-0.05 mol·L-1醋酸钠缓冲液(pH 5.9)-四氢呋喃(15:84:1);流速:1. 0 mL·min-1;进样量:5μL,使用荧光检测器检测激发波长:338 nm,发射波长:450 nm.结果:标准曲线线性、范围1.0～40.0 mg·L-1(r=0.999 9,n=6),血浆中羧甲司坦最低检测限为0.5 mg·L-1,日内RSD为0.3%～1.3%,日间RSD为0.4%～4.5%,准确度为97.9%～102.7%,羧甲司坦回收率为99.5%～103.0%.结论:该方法分离效果好,方法学结果满意,适用于人血浆中羧甲司坦的检测.

OBJECTIVE To establish a method for determining concentration of carboeisteine in human plasma by high-performance liquid chromatography. METHODS Carbocisteine was converted into fluorescent derivative with o-phthalaldehyde and quantitatively determined by reversed-phase high-performance liquid chromatography. Hypersil BDS C18 column （4. 6 mm× 150mm, 5μm） was used. The mobile phase consisted of methanol-sodium acetate buffer （pH 5. 9）-tetrahydrofuran （15：84：1 ）. The flow-rate was 1.0 mL·min^-1, 5μL of sample solution was injected and detected by the fluorescence detector with a 338 nm excitation filter and a 45nm emission filter. RESULTS The linear range of carbocisteine was 1.0 - 40.0 mg·L^-1 （r= 0. 999 9,n = 6）. The limit of detection was 0. 5 mg＇L-1. The RSD of intra-day tests were 0. 3% - 1.3% and that of inter-day tests were 0.4% -4. 5%. The accuracy was 97.9 % - 102. 7 %. The recovery of carbocisteine was 99.5 % - 103.0 ^. CONCLUSION The results of methodology validation were satisfied. It is suitable for determination of carbocisteine in human plasma.