LC/MS/MS Analysis of Testosterone and Metabolites on Ascentisยฎ Express C18
Materials
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5ฮฑ-Dihydrotestosterone (DHT) solution
1.0 mg/mL in methanol, ampule of 1 mL, certified reference material, Cerilliantยฎ6ฮฒ-Hydroxytestosterone solution
100 ฮผg/mL in methanol, ampule of 1 mL, certified reference material, Cerilliantยฎ17ฮฑ-Methyltestosterone solution
1.0 mg/mL in 1,2-dimethoxyethane, ampule of 1 mL, certified reference material, CerilliantยฎTestosterone solution
1.0 mg/mL in acetonitrile, ampule of 1 mL, certified reference material, CerilliantยฎCONDITIONS
column
Ascentis Express C18, 5 cm x 2.1 mm I.D., 2.7 μm particles (53822-U)
mobile phase
[A] 5 mM ammonium formate water pH 4.0 formic acid; [B] 0.1% formic acid acetonitrile
gradient
10 to 70% B in 5 min; held at 70% B for 3 min
flow rate
0.6 mL/min
pressure
3698 psi (255 bar)
column temp.
35 ยฐC
detector
MS, ESI(+), TIC, m/z 100-1000
injection
5 μL
sample
1000 ng/mL in 40:60 (water:methanol)
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๋ถ์ ๋ฉ๋ชจ
An Ascentis Expres F5 column provided rapid, baseline resolution of testosterone and metabolites. The highest quality 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.
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Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany