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.