Determination of Fat- Soluble Vitamins A, D3 and E in Infant Formula and Milk Powder using High Performance Liquid Chromatography with Photodiode Array Detection: Jordan Market as a Case Study

Document Type: Research Article


1 Department of Chemsitry, Mu'tah University, Jordan

2 Department of Medicinal Chemistry and Pharmacognosy, Jordan University of Science and Technology, Irbid 22110, Jordan

3 Department of Chemistry, Mut’ah University, Al-Karak 61710, Jordan

4 College of Pharmacy, Qatar University


A new, simple, rapid, and sensitive reversed-phase high performance liquid chromatography-photodiode array (HPLC-PDA) method was developed and validated for simultaneous analysis of fat-soluble vitamins A, D3, and E. The method required a simple sample preparation step of saponification with aqueous KOH and extraction with n-hexane. The method was validated in terms of linearity, accuracy, precision, stability, detection limits, and recovery. The method has the advantage of simultaneous determination of vitamins A, D3 and E in a short run time of 10 min. The method was applied successfully for determination of vitamins A, D3, and E in some infant formula and milk powder from Jordanian market. Based on this study, vitamin A content in all brandes was within 90.0-410% of the labeled value. Vitamin D3 content in the studied brands was within 100.0-850% , while vitamin E content in all brands was less than 48%. The results showed large variation and discrepancy of most vitamins’ contents, which were not in good agreement with the manufacturer label value, though they were below toxic levels.


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