Comparison of Sensitivity and Specificity of A Commercial ELISA Test and HPLC Method for Detection of Human Serum Carotenoids

Objectives: The levels of differential sensitivity in the use of high-performance liquid chromatography (HPLC) or enzyme linked immunosorbent assay (ELISA) method in the detection of carotenoids in human tissues remains unknown.This study compared the specificity/sensitivity of the two methods in the determination of lutein and zeaxanthin in human serum. Methods: Thirty-two subjects made up of 17 males (18-63 years old) and 15 females (16-71 years old) were recruited for the study. The blood samples were collected aseptically into plain tubes, centrifuged, and their serum samples were collected for determination of lutein and zeaxanthin concentrations. Results: The concentration range of lutein and zeaxanthin in serum by HPLC method was 0.00-0.17ng/mL and 0.00-0.13ng/mL respectively, while the range concentration of lutein and zeaxanthin inserum by ELISA method was 32.98-103.82ng/mL and 213.53-1677.95ng/mL respectively. Therewas also, no relationship between age and concentration of zeaxanthin (R2HPLC =0.011, R2ELISA = -0.069) and lutein (R2HPLC =0.005, R2ELISA = 0.000) when determined by eitherHPLC or ELISA method. Conclusion: ELISA method demonstrateda more sensitive and specificestimation method than theHPLC methodin determination of lutein and zeaxanthinowing to its ability to detect the two carotenoid levelsin greater number of participants.