反相高效液相色谱法同时测定4种苯二氮类药物

Simultaneous Determination of Four Kinds of Benzodiazepines by Rversephase High Performance Liquid Chromatography

提出了一种反相高效液相色谱法同时测定4种苯二氮卓艹类药物的分析方法.考察了4种苯二氮卓艹类药物的保留值与流动相组成、流速、pH及柱温等色谱条件的关系,优化了色谱条件.确定了以ODSHypersil为色谱柱,甲醇-水(51∶49;V/V,用氨水调pH7.8)为流动相,流速为0.8mL/min,柱温为45℃,检测波长为220nm的最佳色谱条件.甲苯作为内标物,以内标法峰面积定量.硝西泮,地西泮,三唑仑和艾司唑仑线性范围分别是0.1mg/L～320mg/L,0.2mg/L～320mg/L,0.1mg/L～320mg/L和0.08mg/L～320mg/L;检测限分别为0.1mg/L,0.2mg/L,0.1mg/L和0.08mg/L.该方法用于尿液中的苯二氮卓艹类药物的测定.其回收率为97.0%～108.7%,标准偏差为1.7%～3.0%(n=7).该方法简单、快速、精确、灵敏、重复性好.

An analytical method for the simultaneous separation and determination of 4 kinds of Benzodiazepines by RP-HPLC with Multiple Wavelength detector was developed. The effects of mobile phase components, pH value, flow speed, column temperature and wavelength on retention parameters of 4 kinds of Benzodiazepines were reviewed. Chromatographic conditions were optimized. A ODS Hypersil column (5μm, 100 × 4.6 mm i. d.) was used with methanol-water (51∶49, V/V) as mobile phase. The optimized condition was as follows the pH value was 7.8, the flow rate was 0.8 ml/min at the column temperature 45 ℃ with the wavelength 220 nm. Methylbenzene was used as the internal stand. The detection limits were 0.1mg/L, 0.08mg/L, 0.1mg/Land 0.2mg/L for Nitrazepam, Estazolam, Triazolam and Diazepam, respectively. The linear rang were 0.1mg/L～320 mg/L, 0.1mg/L～320mg/L, 0.08mg/L～320 mg/L and 0.2mg/L～320 mg/L for Nitrazepam, Estazolam, Triazolam and Diazepam, respectively. The method has been applied to the determination of 4 kinds of Benzodiazepines in human urine. The recoveries were 97.0 %～108.7 %, RSD 1.7 %～3.0 % (n=7). The advantages of this method are simple, fast, accurate, sensitive and reproducible.