LC/MS/MS Analysis of Vitamin D Metabolites on Ascentis® Express F5 (Monitored Ions)
Materialien
Analytische Säule
Komponente der mobilen Phase
Standard
25-Hydroxyvitamin D2 -Lösung
50 μg/mL in ethanol, ampule of 1 mL, certified reference material, Cerilliant®25-Hydroxyvitamin D3 -Lösung
100 μg/mL in ethanol, ampule of 1 mL, certified reference material, Cerilliant®1α,25-Dihydroxyvitamin D3 -Lösung
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®CONDITIONS
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)
Beschreibung
Hinweis zur Analyse
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.
Rechtliche Hinweise
Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany