HPLC法测定丙戊酸血药浓度两种衍生方法的比较研究  

Comparison of Two Pre-column Derivatization HPLC Methods for the Determination of Valproic Acid in Serum

作　　者：王麟奇 张倩影 吕立勋 纪硕 周珊珊 秦锟 王洪民[3] Wang Linqi;Zhang Qianying;Lv Lixun;Ji Shuo;Zhou Shanshan;Qin Kun;Wang Hongmin(School of Pharmacy,North China University of Science and Technology,Hebei Tangshan 063210,China;Department of Pharmacy,Affiliated Hospital of North China University of Science and Technology;Department of Pharmacy,Tangshan Maternal and Child Health Hospital)

机构地区：[1]华北理工大学药学院,河北唐山063210 [2]华北理工大学附属医院药学部 [3]唐山市妇幼保健院药剂科

出　　处：《中国药师》2022年第12期2268-2271,共4页China Pharmacist

基　　金：河北省卫生厅科技项目(编号:20191148)。

摘　　要：目的:比较两种衍生方法的最佳衍生条件测定人血中丙戊酸(VPA)浓度的差异及相关性,为临床VPA血药浓度监测方法选择提供参考依据。方法:使用高效液相色谱(HPLC)法测定VPA血药浓度。血样分别采用2-溴苯乙酮和2-溴-对硝基苯乙酮对VPA进行衍生,以环己烷羧酸为内标,色谱柱采用Aglient Capcell Pak C18(250 mm×4.6 mm, 5μm),保护柱采用Scienhome C18(600220保护柱组件),流动相为甲醇-水(80/20,V/V),流速:1.0 ml·min^(-1),进样量10μl,柱温35℃,保留时间15 min。通过扫描全波长最终确定,2-溴苯乙酮为衍生试剂时的检测波长为248 nm, 2-溴-对硝基苯乙酮时的检测波长为265 nm。采用统计学方法比较两种方法测定63例患者VPA血药浓度的相关性和差异。结果:两种衍生方法经方法学考察均符合要求,两种方法测得VPA血药浓度相关性较好(Y=0.971 3X+1.528 9,r=0.989 6),且差异无统计学意义(P>0.05)。结论:两种方法均可用于临床上VPA血药浓度监测,相比2-溴苯乙酮作为衍生试剂,2-溴-对硝基苯乙酮具有低成本、高效率的特点,更适合用于临床VPA的血药浓度监测。Objective: To investigate the differences and correlation between two pre-column derivatization HPLC methods for the determination of valproic acid(VPA) in serum, and provide reference for the clinical monitoring of VPA concentration. Methods: VPA in human serum was determined by high performance liquid chromatography(HPLC) with cyclohexanecarboxylic acid as internal standard, and the samples were derived respectively by 2-bromoacetophenone and 2-bromo-4’-nitroacetophenone. Chromatographic separation was achieved in approximately 15 min on an Aglient Capcell Pak C18column(250 mm×4.6 mm, 5 μm) coupled with Scienhome C18(600220 safe-keeping column group of parts) guard precolumn. The mobile phase consisted of methanol-water(80/20,V/V) at 1.0 ml·min^(-1). The column temperature was set at 35 ℃. The injection volume was 10 μl, and the detection wavelengths were 248 nm and 265 nm. The correlation and differences of the VPA serum concentration in 63 patients respectively detected by the two methods were evaluated by statistical methods.Results: The two derivative methods were investigated in order to validate the analytical methods. The serum concentrations of VPA obtained by the two methods showed good correlation coefficient(Y=0.971 3X+1.528 9, r=0.989 6). There were not statistically significant differences between the results determined by the two methods(P>0.05).Conclusion: Both methods can be used to monitor the concentration of VPA in clinical practice. However, due to the characteristics of low cost and high efficiency, 2-bromo-4’-nitroacetophenone as a derivatizing reagent is more suitable for the determination of VPA in serum than 2-bromoacetophenone.

关 键 词：丙戊酸 柱前衍生高效液相色谱法 2-溴苯乙酮 2-溴-对硝基苯乙酮 治疗药物监测 

分 类 号：R969.1[医药卫生—药理学]