二维液相色谱法测定人血清中丙戊酸浓度的不确定度  

Uncertainty evaluation in the determination of valproic acid in human serum by 2-dimensional high performance liquid chromatography

作　　者：钟彩妮[1] 曾环思[2] 蒋家兵 王裕锋 张璇[2] ZHONG Cai-ni;ZENG Huan-si;JIANG Jia-bing;WANG Yu-feng;ZHANG Xuan(Department of Pharmacy,Shenzhen Kangning Hospital,Shenzhen Guangdong 518020;Institute of Pharmacy,Shenzhen Kangning Hospital,Shenzhen Guangdong 518020)

机构地区：[1]深圳市康宁医院药学部,广东深圳518020 [2]深圳市康宁医院研究所,广东深圳518020

出　　处：《中南药学》2020年第7期1181-1186,共6页Central South Pharmacy

基　　金：深圳市卫生计生系统科研项目(No.SZFZ2018039);深圳市科技计划项目(No.JCYJ20150402093137766)。

摘　　要：目的 采用全自动二维液相色谱(2D-LC)测定人血清中丙戊酸浓度的不确定度。方法采用2D-LC法检测丙戊酸人血清药物浓度,第一维色谱柱为Aston SC2(25 mm×3.5 mm,5 μm),流动相为甲醇-乙腈-15.0 mmol·L^(-1)磷酸铵盐水溶液=2∶2∶3(v∶v∶v,pH=5.35);流速1.0 mL·min^(-1),第二维色谱柱为Aston SCB(125 mm×4.6 mm,5 μm),捕获柱为Aston SH(10 mm×3.5 mm,5 μm),流动相为乙腈-10.0 mmol·L^(-1)磷酸铵盐水溶液=44∶56(v∶v,pH=3.00),流速1.2 mL·min^(-1),柱温40℃,紫外检测波长215 nm。依据不确定度来源分类原则,建立不确定度模型,分析测定方法中重复性、样品称量、溶液配制、仪器允差、提取回收、标准曲线拟合等不确定度源拟合类型,选定散布函数,合成不确定度和扩展不确定度。结果 2D-LC法测定丙戊酸低、中、高血清药物浓度的拓展不确定度分别为U_L=1.8657 μg·mL^(-1),U_M=6.8542 μg·mL^(-1),U_H=13.9458 μg·mL^(-1)(P=95%),结果可分别表示为(7.52±1.87)、(123.85±6.85)、(249.33±13.95)μg·mL^(-1)。结论本法不确定度小,主要来源于标准曲线拟合,低浓度对不确定度贡献最大,溶液的配制和提取回收率过程对不确定度有部分影响。Objective To evaluate the uncertainty in the determination of valproic acid in human serum by 2-dimensional high performance liquid chromatography(2D-LC).Methods The concentration of valproate acid in human serum was detected by 2D-LC.The first dimension of chromatographic column was Aston SC2(25 mm×3.5 mm,5μm),and the mobile phase consisted of methanol∶acetonitrile∶15.0 mmol·L-1 ammonium phosphate solution=2∶2∶3(v∶v∶v,pH=5.35)with flow rate at 1.0 mL·min-1.The second dimension of chromatographic column was Aston SCB(125 mm×4.6 mm,5μm),and the captured column was Aston SH(10 mm×3.5 mm,5μm).The mobile phase of the second dimension consisted of acetonitrile∶10.0 mmol·L-1 ammonium phosphate solution=44∶56(v∶v,pH=3.0)with flow rate at 1.2 mL·min-1.The column temperature was maintained at 40℃,and the UV absorption wavelength was 215 nm.According to the principle of uncertainty source classification,the uncertainty model was established to analyze the repeatability,weighing,solution preparation,instrumental error,extraction recovery and standard curve fitting.Possible factors related to the uncertainty were included in the model.The measurement uncertainty and extended uncertainty were analyzed by the scatter function.Results The expanded uncertainty of valproic acid at low,middle and high concentrations were 1.8657,6.8542 and 13.9458μg·mL-1(P=95%),respectively.The measured results were expressed as(7.52±1.87),(123.85±6.85)and(249.33±13.95)μg·mL-1.Conclusion The uncertainty of this method is small and mainly comes from calibration curve fitting and especially from the low concentration(with the largest contribution).The preparation of solution and the rate of sample extraction have some impact on the uncertainty.

关 键 词：二维液相色谱法 丙戊酸 血清药物浓度 不确定度评定 

分 类 号：R965[医药卫生—药理学]