G005876

LC/MS/MS Analysis of Vitamin D Metabolites on Ascentis^® Express F5 (Monitored Ions)

application for HPLC

Properties

• technique(s): HPLC: suitable
• test parameters: column: Ascentis Express F5, 15 cm x 2.1 mm I.D., 2.7 μm particles (53571-U); mobile phase: [A] 5 mM ammonium formate water; [B] methanol; (25:75, A:B); flow rate: 0.4 mL/min; pressure: 5178 psi (357 bar); column temp.: 40 °C; detector: ESI(+), 100-1000 m/z; injection: 2 μL, each compound 300 ng/mL in 40:60 (water:methanol)
• suitability: application for HPLC
• application(s): clinical; vitamins, nutraceuticals, and natural products

Description

Analysis Note

The metabolic role and health implications of the various vitamin D isoforms are of current clinical interest. Therefore, it is important to have an analytical method that will resolve all of the known isoforms with necessary sensitivity and specificity.This application shows that a combination of Ascentis Express F5 with MS detection provided rapid resolution of seven such compounds. 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

1α,25-Dihydroxyvitamin D3 solution, 5 μg/mL in ethanol, ampule of 1 mL, certified reference material, Cerilliant^®
1α,25-Dihydroxyvitamin D2 solution, 5 μg/mL in ethanol, ampule of 1 mL, certified reference material, Cerilliant^®
25-Hydroxyvitamin D3 solution, 100 μg/mL in ethanol, ampule of 1 mL, certified reference material, Cerilliant^®
3-epi-25-Hydroxyvitamin D₃, 98% (CP)
3-epi-25-Hydroxyvitamin D₂ solution, 100 μg/mL in ethanol, 98% (CP)
1α-Hydroxyvitamin D₃, ≥97.0% (HPLC)