[Determination of lutein and zeaxanthin in serum by high performance liquid chromatography].
To establish a simultaneous determination method for measuring lutein, zeaxanthin and β-carotene in serum by internal standard on C(30)-HPLC.Experimental data were as follows: stationary phase, Develosil carotenoid column C(30) (250 mm×4.6 mm, 5 μm); mobile phase A, acetonitrile:methanol (3:2, v/v); mobile phase B, MTBE; grads elution; flow rate, 1 mL/min; monitoring wavelength, 450 nm; injection volume, 20 μL; column temperature, 25 °C.Lutein, zeaxanthin and β-carotene were thoroughly separated with the average retention time of 9.9 min, 10.3 min and 21.2 min, respectively. The intra-day relative standard deviation (RSD) values were 3.22%, 3.81% and 1.60%. The linear ranges of serum concentrations of lutein and β-carotene were both 0.012 5-12.5 mg/L (r=0.999 5, r=0.999 7), and that of zeaxanthin was 0.005-5.0 mg/L (r=1). The mean serum concentrations of lutein, zeaxanthin and β-carotene for 58 healthy elder inhabitants (>50 years) were 0.410 μmol/L, 0.054 μmol/L and 0.128 μmol/L, respectively.This established method can be used for determination of lutein, zeaxanthin and β-carotene in serum.