Analysis of Lutein in Spinach using HPLC

This demo video shows how to analyze lutein in spinach samples using Shimadzu HPLC. For sample preparation, spinach samples were cut and blended. After that, pyrogallol-ethanol and KOH were added to the samples and heated at 70℃ for 30 min. After cooling down, NaCl and n-hexane-ethyl acetate were added for extraction. The extract was dried by rotary evaporator and reconstituted with BHT-ethanol. Filter the solution with a PTFE membrane filter and collect the filtrate in an amber vial. The ready sample can be injected for HPLC analysis.
Check the status of the instrument and check the mobile phase and the connections of the tubing before HPLC running. Running the lutein standards and real samples in the same batch by HPLC. Process all the data obtained using Shimadzu LabSolutions software. The results show that in average, 5 - 8 mg of lutein were found in 100 g spinach samples.