G005858

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

application for HPLC

Properties

• technique(s): HPLC: suitable
• test parameters: 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
• suitability: application for HPLC
• application(s): clinical; vitamins, nutraceuticals, and natural products

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

Analyte

25-Hydroxyvitamin D3 solution, 5 μg/mL in ethanol, ampule of 1 mL, certified reference material, Cerilliant^®
25-Hydroxyvitamin D2 solution, 50 μg/mL in ethanol, ampule of 1 mL, certified reference material, Cerilliant^®