Abstract
An improved method for enzymatic hydrolysis, extraction, and high performance liquid chromatography (HPLC) separation of major carotenoids and retinol of mature human milk is described. After pretreatment with bile salts and protease, carotenoids and retinol are released by lipase treatment followed with brief chemical saponification. The HPLC method provides enhanced peak resolution and improved elution profiles. The procedure is nondestructive to provitamin-A carotenoids while effectively hydrolyzing retinyl esters. Using this method, we were able to significantly increase the recovery of both β-carotene and retinol from human milk over previously published procedures. The method is sensitive to picomolar quantities of carotenoids and retinol. Retinol, lutein/zeaxanthin, β-cryptoxanthin, lycopene, α-carotene, and β-carotene are effectively recovered and quantitated from a single 1-mL sample of milk.
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