LC/MS Analysis of 25-Hydroxyvitamin D2 and 25-Hydroxyvitamin D3 on Ascentis® Express F5

LC/MS Analysis of 25-Hydroxyvitamin D2 and 25-Hydroxyvitamin D3 on Ascentis® Express F5

Conditions

column Ascentis Express F5, 10 cm x 2.1 mm I.D., 2.7 µm particles (53569-U)
column temp. 40 °C
mobile phase 5 mM ammonium formate in methanol:water (75:25)
flow rate 0.4 mL/min
sample 20 µg/mL in methanol
injection 1 µL
detector UV, 265 nm (MS, ESI(+), m/z 100-1000)

Description

Analysis Note The 25-OH forms of the secosteroids vitamin D2 and D3 are indicators of the vitamin D status of an individual. However, because they have different biological activity and clinical interest, they should be distinguishable. The Ascentis Express F5 phase was able to resolve the two compounds under highly LC-MS/MS compatible conditions. The highest grade LC-MS solvents were used to supply low background interference and low particulate contaminants for robust, trouble-free operation. Cerilliant CRMs provided reliable identification and quantification.
Categories Analytical Chromatography, Clinical, vitamins, Vitamins and Natural products
Featured Industry Clinical
Vitamins, Nutraceuticals, and Natural Products
Legal Information Ascentis is a registered trademark of Sigma-Aldrich Co. LLC
suitability application for HPLC

Materials

     
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