LC/MS Analysis of Omeprazole and Metabolites in Plasma on Ascentis^® Express RP Amide after Sample Prep using HybridSPE^®-Phospholipid

sample preparation

SPE (Solid Phase Extraction)

sample/matrix

rat plasma spiked with omeprazole, phenacetin and metabolites at 2 μg/mL

SPE well plate

HybridSPE-Phospholipid, 96-well plate, 15 mg/0.8 mL (575656-U)

sample addition

100 μL plasma to each well, followed by 300 mL of 1% formic acid in acetonitrile

elution

apply vacuum

eluate post-treatment

eluate was collected, evaporated, and reconstituted in mobile phase.

column

Ascentis Express RP-Amide, 5 cm x 2.1 mm I.D., 2.7 μm particles (53911-U)

mobile phase

[A] water, 10 mM ammonium formate, pH 3.4; [B] methanol, 10 mM ammonium formate, pH 3.4; (65:35, A:B)

flow rate

0.3 mL/min

column temp.

35 °C

detector

ESI(+), m/z = 50-400

injection

1 μL