高效液相色谱法测定健康人尿液中苯乙基异硫氰酸酯乙酰半胱氨酸缀合物的浓度

Determination the concentration of the N-acetylcysteine conjugate of phenethyl isothiocyanate in healthy human urine by HPLC

目的建立高效液相色谱法测定健康人尿液中苯乙基异硫氰酸酯乙酰半胱氨酸缀合物的浓度。方法色谱柱为Sepax Amethyst-C18柱,流动相为0.2%磷酸水溶液-甲醇(35∶65),流量:1 mL.min-1,紫外检测波长250 nm;氯雷他定为内标。结果尿液中代谢物在0.8～64.0μg.mL-1内线性关系良好,回归方程为y=0.08x-1.28×10-3(n=8,γ=0.9982),提取回收率大于80%,批内和批间精密度RSD<10%。结论该方法可用于测定人尿液中苯乙基异硫氰酸酯乙酰半胱氨酸缀合物的浓度。

Objective To establish a HPLC method for the determination the concentration of N-acetylcysteine conjugate of phenethyl isothiocyanate（PEITC-NAC） in healthy human urine.Methods Separation was carried on a Sepax Amethyst-C18 and the mobile phase was consisted of 0.2% phosphoric acid-methanol（35∶65） with the flow rate of 1 mL·min-1.The detection wavelength was 250 nm.Results A good linearity was demonstrated between 0.8-64.0 μg·mL-1 by linear equation y=0.08x-1.28×10-3（n=8,γ=0.9982）.The within-batch and between-batch deviation was showed by RSD10%.Conclusion The method can be applied for the determination the concentration of PEITC-NAC in human urine.