浊点萃取-高效液相色谱法同时测定药片与尿样中异烟肼、利福平和吡嗪酰胺

Simultaneous Determination of Isoniazid,Rifampicin and Pyrazinamide in Pharmaceuticals and Human Urine by Cloud Point Extraction High Performance Liquid Chromatography

建立了以聚乙二醇(PEG)6000为萃取剂的浊点萃取(CPE)-反相高效液相色谱同时测定异烟肼、利福平和吡嗪酰胺的方法,由于异烟肼不易被浊点萃取方法富集,使异烟肼与对氯苯甲醛在酸性条件下反应衍生为异烟肼腙,异烟肼腙在液相色谱上的信号与异烟肼浓度存在着线性关系。在优化的CPE条件下,采用梯度洗脱的方式,用紫外检测器在310 nm波长下,异烟肼、吡嗪酰胺和利福平的保留时间分别为14.5、34.4和38.1 min,定量范围分别是0.01～10 mg/L、0.08～25 mg/L和0.05～20 mg/L。线性相关系数r>0.99。3种药的平均回收率范围为91.50%～98.75%,精密度(RSD)为0.22%～1.54%。异烟肼、利福平和吡嗪酰胺的检出限分别为4×10-3、2×10-2和3×10-2mg/L。本法用于药片和尿样中3种药物的检测,结果满意。

Isoniazid（IN） can not be sufficiently extracted by the cloud point extraction（CPE） of polylethyle- neglyeol（PEG） 6000, and reacts with para-chlorobenzaldehyde to form a derivative（para-chlorobenzaldehyde- isonicotiny- hydrazone, CIH）. It was proven that there was linear relationship between the signal of CIH and the concentration of IN. A cloud point extraction system of polylethyleneglycol 6000（NH4）2SO4 coupled with high performance liquid chromatography was developed for the separation and the simultaneous determination of isoniazid（IN） , pyrazinamide（PZ） and rifampicin（RF）. Under the optimum conditions, the analytes were eluted with a gradient mode and detected with a UV-deteetor at 310 nm, the retention time of isoniazid, pyraz- inamide and rifampiein was 14.5, 34.4 and 38.1 rain, respectively, the linear calibration curves in the range of 0.01 -10 mg/L for isoniazid and 0.05 -20 mg/L for pyrazinamide and 0.08 -25 mg/L for rifampicin, respectively（r 〉 0.99）. Average recoveries of three drugs ranged from 91.5% to 98.6% with relative stand- ard deviations from0.2% to 1.5%. The limits of detection of IN, PZ and RF was 4 ×10-3, 2 ×10-2 and 3 ×10-2 mg/L, respectively. The CPE-HPLC-UV method had been applied to the determination of IN, PZ and RF in pharmaceuticals and in urine samples.