Fat-Soluble Vitamins (A, D, E, K)

Fat-soluble vitamins A, D, E, and K are essential micronutrients absorbed with dietary fats and stored in fatty tissues. Their accurate determination is critical for nutritional labeling, fortification verification, and supplement quality control. Ovalab offers comprehensive fat-soluble vitamin testing.

Key Facts About Fat-Soluble Vitamins (A, D, E, K)

  • EU NRVs (Annex XIII, Regulation 1169/2011): Vitamin A 800 µg, Vitamin D 5 µg, Vitamin E 12 mg, Vitamin K 75 µg.
  • Method: Simultaneous HPLC-DAD (diode array detection) determines all four vitamins in a single chromatographic run.
  • Sample preparation: Saponification, liquid-liquid extraction, and purification before HPLC analysis.
  • Vitamin forms: A as retinol/retinyl esters; D as D2 and D3; E as α-tocopherol; K as phylloquinone (K1) and menaquinones (K2).
  • Claims: “Source of” requires ≥ 15% NRV per 100 g; “high in” requires ≥ 30% NRV (Regulation 1924/2006).

Testing Methods at Ovalab

Ovalab determines fat-soluble vitamins using HPLC-DAD and LC-MS/MS for simultaneous quantification. Sample preparation involves saponification and solvent extraction optimized per matrix. Under ISO/IEC 17025:2018 (ČIA, Certificate 537/2025) and GMP (SÚKL 74726/2023).

Frequently Asked Questions

Can all four fat-soluble vitamins be tested simultaneously?

Yes. Ovalab uses HPLC-DAD methods to determine vitamins A, D, E, and K in a single run after saponification and extraction — efficient and cost-effective.

Per Annex XIII, Regulation 1169/2011: Vitamin A 800 µg, Vitamin D 5 µg, Vitamin E 12 mg, Vitamin K 75 µg.

Saponification (alkaline hydrolysis with KOH) breaks the fat matrix and releases vitamins from ester forms, essential for extraction from dairy, oils, and fortified foods.

Fortified dairy/plant milks, margarines, infant formulas, cooking oils, supplements, fish oils, egg products, and liver products.

For vitamin D at low concentrations, Ovalab uses LC-MS/MS for superior sensitivity vs. HPLC-DAD — critical for non-fortified foods with naturally low vitamin D.