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
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篇名: 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
TITLE:
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20 μl。结果:VitA血药浓度在0.012 4~3.210 μg/ml范围内线性关系良好(r=0.997 2,n=5),定量下限为0.012 4 μg/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VitA浓度的测定。
ABSTRACT: 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 determined 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 recoveries 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). CONCLUSIONS: 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.
期刊: 2016年第27卷第35期
作者: 徐凤梅,于美丽,苏瑞,陆伟
AUTHORS: XU Fengmei,YU Meili,SU Rui,LU Wei
关键字: 高效液相色谱法;维生素A;血药浓度;肝硬化
KEYWORDS: HPLC; Vitamin A; Blood concentration; Hepatocirrhosis
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