Region

Applications

Separation of Testosterone and Epitestosterone by LC/MS/MS using Strata-X-A and Kinetex 1.7 µm C18

Sample Preparation Details

Condition 1 :2 mL Methanol
Condition 2 :2 mL Water
Load :Load 1.25 mL Sample diluted in Water
Wash 1 :0.6 mL 50% Methanol in Water
Dry :Dry for 5 min under high vacuum
Elution 1 :0.3 mL Methanol
Elution 2 :0.3 mL Methanol
Evaporate: :Evaporate to dryness
Reconstitute :250 µL 200 uL 5% aqueous formic acid
Notes :Following evaporation of elution solvent @ 50-55 C under gentle nitrogen stream; Add 50 uL 25% hydroxylamine solution and heat at 60-65 C for 5-10 min, then add 200 uL 5% aqueous formic acid and vortex the tubes Transfer the solution to autosampler vials and inject 25 uL on column

Application Detail (App ID: 19761)

Part No:00A-4475-AN
Dimensions:30 x 2.1 mm ID
Elution Type:Gradient
Elution A:0.1% Formic Acid +1 mM Amm Formate in Water
Elution B:0.1% Formic Acid +1 mM Amm Formate in ACN
Gradient Profile:
Step No. Time (min) Pct A Pct B
1 0 90 10
2 2.5 10 90
3 3.5 10 90
4 3.6 90 10

Flow Rate:0.4 mL/min
Col. Temp: 55 °C
Detection: Mass Spectrometer (MS) @ amu (22°C)
Detector Info: AB Sciex API 5000™

Analyte Details

Analyte Detail not available

1
Testosterone
search

2
Testosterone-d3
search

3
Epitestosterone
search