HPLC Analysis of Statins in Rat Plasma on Ascentis^® Express C18 after SPE using Supel^™-Select HLB

sample preparation

SPE (Solid Phase Extraction)

sample/matrix

rat plasma spiked with provastin and atorvastin at 5 and 100 ng/mL

SPE tube/cartridge

Supel-Select HLB SPE, 30 mg/1 mL (54181-U)

condition

0.5 mL methanol:acetonitrile (1:1, v/v); 0.5 mL DI water

sample addition

0.5 mL

washing

0.5 mL 5% methanol

elution

0.5 mL methanol:acetonitrile (1:1, v/v)

column

Ascentis Express C18, 5 cm x 2.1 mm, 2.7 μm particles (53822-U)

eluate post-treatment

evaporate to dryness with nitrogen at 37 °C, reconstitute in 0.5 mL DI water

mobile phase

0.1% acetic acid diluted in 60% methanol

flow rate

0.2 mL/min

column temp.

35 °C

detector

ABI 3200 QT; ESI(+), MRM (423.3/321.3, 423.3/101.0, 557.3/397.2 and 557.3/453.4)

injection

5 μL