高效液相色谱法检测生物样本中苯肽胺酸的含量  

Determination of N-Phenyl-phthalamic Acid in Biological Samples by HPLC

作　　者：韦思华[1] 代玲玲[1] 周峰[1] 李龙[1] 

机构地区：[1]华中科技大学卫生毒理系,武汉430000

出　　处：《农药》2016年第1期33-35,38,共4页Agrochemicals

摘　　要：[目的]建立高效液相色谱法定量分析大鼠血样和组织样本中苯肽胺酸的含量,为该药进一步研究提供参考依据。[方法]用高效液相色谱-紫外检测法对含有苯肽胺酸的空白样本进行检测并优化色谱条件使药物色谱峰与杂峰实现有效的分离。色谱柱为XB-NH_2柱,检测波长254 nm,柱温40℃,进样量10μL,流速1 mL/min,等度洗脱。血样和组织样本分别使用不同的流动相进行洗脱,血样洗脱的流动相中乙腈和水体积比为65∶35(水相含甲酸0.4%),动物组织样本洗脱的流动相中乙腈和水比例为80∶20(水相含甲酸0.2%)。按照设定的色谱条件进行精密度、重现性和稳定性的研究,再取空白血样和动物组织样本进行药物回收率的研究。[结果]日间及日内精密度RSD≤2%,重现性和稳定性RSD≤5%,所有样本回收率范围85.2%~97.5%。[结论]实验建立的高效液相色谱法检测生物样本中的苯肽胺酸稳定性和重现性均符合实验要求,可作为相关实验的参考方法。预试验结果显示,增加流动相的pH值和降低水相的pH值均能使保留时间延长。此外,研究结果显示pH值会对药物色谱峰的峰型有影响。[Aims] The HPLC method for determination of N-phenylphthalamic acid in murine serum and tissue samples was established to provide reference for further study. [Methods] The blank biological samples which contained N-phenylphthalamic acid were detected by HPLC-UV, meanwhile the chromatographic condition was optimized to realize the separation of the drug chromatographic peak and impurity peak. The chromatographic conditions were： chromatographic column was XB - NH2 column, wavelength was 254 nm, column temperature was 40 ~C, sample size was 10 I^L and flow rate was 1 mL/min. Serum and tissue samples were eluted with different mobile phase. The mobile phase for serum samples was 65% of acetonitrile and 35% of water. The water phase contained 0.4% of formic acid. The mobile phase for tissue samples was 80% of acetonitrile and 20% of water, the water phase contained 0.2% of formic acid. According to the chromatographic condition, the accuracy, reproducibility and recovery were tested. [Results] The extra-day and intra-day accuracy variable coefficient was below 2%, and the reproducibility was below 5%. The recovery rates was between 85.2 and 97.5%. [Conclusions] The method has high stability and reproducibility. It could conform to demand of the study and reference for the analogous experiment. The results of the preliminary experiment suggested that with increasing the volume of the organic phase, the N-phenylphthalamic acid would retain more long in the chromatographic column. In addition, pH value of water phase can influence the shape of chromatographic peak of the drug.

关 键 词：高效液相色谱法 苯肽胺酸 生物样本 检测 

分 类 号：S482.8[农业科学—农药学] O657.72[农业科学—植物保护]