Vitamin D2 and D3 Separation by New Highly Hydrophobic UHPLC/HPLC Phase

Separations of structurally similar compounds such as vitamin D₂ and vitamin D₃ are very challenging.

Vitamin D₂ (ergocalciferol) and D₃ (cholecalciferol) can be found in different foods including fatty fishes, meat, egg, and some mushrooms. Both compounds are (indirectly) involved in a number of biological functions in the body, including bone metabolism and enhancement of intestinal absorption of calcium, iron, magnesium, phosphate, and zinc. A regular intake of vitamin D therefore is essential.

Figure 1: Structures of vitamin D2 and vitamin D3.

Standard C18 columns are not able to fully separate the two vitamins. A very hydrophobic phase with a higher carbon coverage and therefore a greater density of C18 chains is required. The highly hydrophobic phase YMC-Triart C18 ExRS (carbon load 25%!) is able to separate these two.

Figure 2: Easy method transfer HPLC <-> UHPLC as a result of full scalability.

The isocratic high performance liquid chromatography (HPLC) method for vitamin D₂ and D₃ separation, using a 5 μm YMC-Triart C18 ExRS column, can easily be transferred to a ultrahigh-performance liquid chromatography (UHPLC) method using a 1.9 μm column, thereby reducing the analysis time by 50%. Furthermore, the resolution can be increased, resulting in a full baseline separation.
 

YMC-Triart C18 ExRS (5 µm, 8 nm), 150 × 3.0 mm

YMC-Triart C18 ExRS (1.9 µm, 8 nm),
        75 × 2.1 mm

YMC Europe GmbH
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E-mail: info@ymc.de
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