Testosterone from Male Human Plasma by LC/MS/MS using Strata-X-A and Kinetex 1.7 µm C18

Sample Preparation Details

Part No:8B-S123-TBJ
Solid Phase Extraction (SPE) procedure:Note: The solvent volumes shown below are for a 30 mg bed mass. The solvent volumes will need to be adjusted for a smaller or larger bed mass.
Load:Into individually labeled test tube combine 0.25 mL serum sample (or calibrator or QC sample),1 mL DI water and 0.1 mL working internal standard solution (2 ng/mL).
Proceed to SPE procedure
Dry:Dry for 5 min under high vacuum
Final Prep and Analysis: 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 Inject: 20 µL on HPLC / Mass Spectrometer (MS) @ amu (ambient)


Name Spiked
Log P pKa % Rec %RSC (n=0)
1 Testosterone
2 Testosterone-d3 0.5 

Application Detail (App ID: 19760)

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