technique(s)
HPLC: suitable
test parameters
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 °C
detector: MS, ESI(+), TIC, m/z 100-1000
injection: 5 μL
sample: 1000 ng/mL in 40:60 (water:methanol)
suitability
application for HPLC
application(s)
clinical
Analysis Note
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.
Legal Information
Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany
Analyte
Description
Testosterone solution, 1.0 mg/mL in acetonitrile, ampule of 1 mL, certified reference material, Cerilliant®
17α-Methyltestosterone solution, 1.0 mg/mL in 1,2-dimethoxyethane, ampule of 1 mL, certified reference material, Cerilliant®
5α-Dihydrotestosterone (DHT) solution, 1.0 mg/mL in methanol, ampule of 1 mL, certified reference material, Cerilliant®
6β-Hydroxytestosterone solution, 100 μg/mL in methanol, ampule of 1 mL, certified reference material, Cerilliant®
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