高效液相色谱法测定血清硝基安定浓度的方法研究

Determination of nitrazepam in human-serum with high performance liquid chromatography

目的建立适于临床普遍应用的血清硝基安定浓度的测定方法。方法应用反相高效液相色谱法，以茶碱为内标，采用窄径柱LUNAC18（2）（3μm，150mm×2mm）分离，流动相为水和甲醇（25：75），二极管阵列紫外检测器选择各组分最大吸收波长检测。结果在4～256μg／L范围内，硝基安定和内标的色谱峰高比与浓度呈线性关系（r＝0．9986）；平均回收率为102．8％，变异系数为4．7％。最低检测浓度为4μg／L。对18例癫痫患者进行治疗药物监测，取得良好疗效。结论该方法灵敏、准确、用血量少且简便快速，适于科研和临床应用。

Objective To establish a reversed phase-high performance liquid chromatography(RP-HPLC)method for the analysis of nitrazepam in human serum, so as to be widely suitable for the clinical application and research. Methods A RP-HPLC method was used with theophylline as the internal standard. With a narrow-pore column of LUNA C18(2) (3 μp, 150mm×2 mm)and with a mobile phase of water-methanol (25:75), the constituents were detected at its maximum adsorbance-wavelenth with the diodearray detector. Results The ratio of chromatographic peak heights of nitrazepam and internal standard correlated linearly with the concentration of nitrazepam ranging from to 256 μg/L, r=0.9986. The mean recovery we 102.8%, and the coefficient of variability was 4.7%. The minimum concentration of detection was 4 μg/L. The method is applied in the therapeutic drug monitoring of 18 patients with epilepsy and satisfactory effect was obtained. Conclusions The method is sensitive, accurate, simple, fast, and suitable for clinical application and research.