LC/MS/MS Analysis of Testosterone in Human Serum on Ascentis^® Express C18 after Extraction with BioSPME using C18 SPME LC Tips

sample/matrix

human plasma

SPME fiber

SPME LC Tips C18 (57234-U)

extraction

60 min with 1 mL sample

desorption process

30 min with 200 μL of methanol

column

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

mobile phase

[A] 0.1% formic acid in water ; [B] methanol

gradient

5% B to 70% B in 3 min, then to 90% B in 0.1 min, hold 90% B for 0.9 min

flow rate

0.5 mL/min

pressure

4351 psi (300 bar)

column temp.

40 °C

detector

MS/MS, ESI(+), MRM (See Figure for transitions)

injection

10 μL

sample

10 ng/mL in methanol