LC/MS Analysis of 25-Hydroxyvitamin D2 and 25-Hydroxyvitamin D3 on Ascentis® Express F5
Materiales
analytical column
standard
25-Hydroxyvitamin D3 solution
100 μg/mL in ethanol, ampule of 1 mL, certified reference material, Cerilliant®CONDITIONS
column
Ascentis Express F5, 10 cm x 2.1 mm I.D., 2.7 μm particles (53569-U)
mobile phase
5 mM ammonium formate in methanol:water (75:25)
flow rate
0.4 mL/min
column temp.
40 °C
detector
UV, 265 nm (MS, ESI(+), m/z 100-1000)
injection
1 μL
sample
20 μg/mL in methanol
Descripción
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.
Legal Information
Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany