【摘要】 目的建立简便快速测定唑尼沙胺血药浓度的高效液相色谱法。方法采用甲醇和醋酸铅沉淀蛋白法处理唑尼沙胺血清样本,以苯代三聚氰胺为内标,ACE UltraCore 2.5 SuperC 18（100.00 mm×3.00 mm）为色谱柱分离化合物。流动相A相为超纯水,B相为纯甲醇,梯度洗脱（0.01～5.00 min,18%～50%甲醇;5.00～5.50 min,50%～18%甲醇;5.50～7.50 min,18%甲醇）,流速为0.6 mL·min^-1,进样量20μL,柱温30℃,检测波长240 nm。并依据《中国药典》2020年版第四部通则中"生物样品定量分析方法验证指导原则"对所建方法的选择性、标准曲线、精密度与准确度、残留及稳定性等进行验证。结果本研究建立的方法测定唑尼沙胺血药浓度线性范围为3.92～125.55 mg·L^-1,R²>0.999;符合要求。经处理样品24 h内分析稳定,未处理样品4℃保存,1周内分析稳定。结论本研究建立的高效液相色谱法简便快速、准确度高、精密度好,适用于测定唑尼沙胺血药浓度。更多还原

【Abstract】 Objective To establish a simple and rapid high performance liquid chromatography（HPLC） method for determination of concentration of zonisamide in blood.Methods The serum samples of zonisamide were treated by protein precipitation with methanol and lead acetate,and the benzoguanamine was used as internal standard. Chromatographic separation was performed on a ACE UltraCore 2. 5 SuperC18（100. 00 mm×3. 00 mm） column. The mobile phase was ultrapure water（A） and pure methanol（B）. The flow rate of gradient elution（0. 01-5. 00 min,18%-50% methanol;5. 00-5. 50 min,50%-18% methanol;5. 50-7. 50 min,18% methanol） was 0. 6 mL·min^-1,the injection volume was 20 μL,the column temperature was 30 ℃,and the detection wavelength was 240 nm. The selectivity,standard curve,precision and accuracy,residue and stability of the method established in this study were verified according to " guidelines for the quantitative analysis of biological samples" in the general rules of Part IV of the Chinese pharmacopoeia（2020 edition）.Results The linear range of the method established in this study for detection of the plasma concentration of zonisamide was 3. 92-125. 55 mg·L^-1,R²>0. 999. The relative standard deviation of intra assay and inter assay precision of different concentrations of zonisamide were reach the standard. The results of the determination of zonisamide in pretreated serum samples were stable after placing at room temperature for 24 hours. The results of the determination of zonisamide in untreated serum samples were stable after storing at 4 ℃ in refrigerator for one week.Conclusion A simple and rapid HPLC method with high accuracy and precision is established for monitoring the blood concentration of zonisamide.更多还原