HPLC法测定维生素A在健康受试者及肝硬化患者体内的血药浓度  

Determination of Serum Concentration of Vitamin A in Healthy Volunteers and Hepatocirrhosis Patients by HPLC

作　　者：徐凤梅[1] 于美丽[2] 苏瑞[3] 陆伟[3] 

机构地区：[1]天津市第三中心医院药剂科,天津300170 [2]天津市第三中心医院/天津市人工细胞重点实验室,天津300170 [3]天津市第二人民医院/天津市肝病研究所,天津300192

出　　处：《中国药房》2016年第35期4927-4930,共4页China Pharmacy

基　　金：天津市卫生行业重点攻关项目(No.13KG110;14KG109);天津市卫生局科技基金项目(No.2015KY02);王宝恩肝纤维化研究基金科研课题(No.CFHPC20151026)

摘　　要：目的：建立测定人血清中维生素A（VitA）浓度的方法，并将其应用于健康受试者及肝硬化患者。方法：血清样品经液-液萃取后，采用高效液相色谱法测定。以维生素A醋酸酯为内标，色谱柱为Kromasil C18，流动相为甲醇-水（98∶2，V/V），流速为1 ml/min，检测波长为325 nm，柱温为室温，进样量为20μl。结果：VitA血药浓度在0.0124～3.210μg/ml范围内线性关系良好（r＝0.9972，n＝5），定量下限为0.0124μg/ml，日内、日间RSD为1.66％～2.97％，加样回收率为98.18％～99.56％，提取回收率为89.59％～91.38％。采用该法测得24例健康受试者和24例肝硬化患者VitA的平均血药浓度分别为（0.71±0.08）、（0.28±0.06）μg/ml；不同年龄段健康受试者与肝硬化患者VitA平均血药浓度比较，差异均有统计学意义（P＜0.05）。结论：该方法简便、快速，且灵敏度高、准确性好，可用于健康受试者及肝硬化患者血清中Vit A浓度的测定。OBJECTIVE：To establish a method for the concentration determination of vitamin A（VitA）in human serum,and apply it in healthy volunteers and hepatocirrhosis patients. METHODS：After liquid-liquid extraction,serum sample was deter-mined by HPLC. Using VitA acetic ester as internal standard,the separation was performed on Kromasil C18 column with mobile phase consisted of methanol-water （98∶2,V/V） at the flow rate of 1 ml/min. The wavelength was set at 325 nm,and the column temperature was room temperature. The sample size was 20 μl. RESULTS：The linear range of VitA were 0.012 4-3.210 μg/ml（r=0.997 2,n=5）,and the limit of quantification was 0.012 4 μg/ml. Inter-day and intra-day RSD ranged 1.66%-2.97%;sample re-coveries were 98.18%-99.56%;extraction recoveries were 89.59%-91.38%. Average blood concentration of VitA were（0.71±0.08）μg/ml in 24 healthy volunteers and （0.28 ± 0.06）μg/ml in 24 hepatocirrhosis patients. There was statistical significance in average blood concentration of VitA between healthy volunteers and hepatocirrhosis patients in different age groups （P〈0.05）. CONCLU-SIONS：The method is simple,rapid,sensitive and accurate,and can be used for the concentration determination of VitA in serum of healthy volunteers and hepatocirrhosis patients.

关 键 词：高效液相色谱法 维生素A 血药浓度 肝硬化 

分 类 号：R969.4[医药卫生—药理学]