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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 application for HPLC

Materials

analytical column

Product No.
Description
Pricing

53569-U

Ascentis® Express F5, 2.7 μm HPLC Column

2.7 μm particle size, L × I.D. 10 cm × 2.1 mm

standard

Product No.
Description
Pricing

H-083

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

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.

Legal Information

Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany

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