A simple and robust HPLC method to analyze lutein in wheat

A method for the determination of lutein, the main component of carotenoid in wheat by high performance liquid chromatography (HPLC) was established. The samples were extracted with acetone and methanol (acetone : methanol 7 : 3, v / v, 0.1 % BHT w / v), and fully dissolved in methanol / acetonitrile / n-hexane (7 : 2 : 1, v / v / v) mixed solution. Poroshell 120 EC-C18 column (4.6 mm× 150 mm, 4μm) was used as the separation column, acetonitrile,methanol and n-hexane solution were used as mobile phase with gradient elution, flow rate was 1.0 mL·min(−1), column temperature was 25 ℃, injection volume was 50 μL, and detection wave length was 450 nm. The mass concentration of lutein had a good linear relationships with the chromatographic peak area in the range of 0.5-5.0 μg·mL(−1), the correlation coefficients were all not less than 0.999 8, and the detection limits was 12 ng·mL(−1). The recoveries rate of standard addition in sampling were 95.50%-97.33%, and the relative standard deviations of determination results were 1.20%-1.66%(n=5). The method is sensitive, precise, rapid, accurate and efficient. The advantages of this study can be summarized in the next bullet points: • Can be used for the determination of lutein in wheat. • The detection time is short. • Limit of detection (LOD) and limit of quantification (LOQ) for lutein was 12 ng·mL(−1) and 42 ng·mL(−1).