Applications
Separation of Testosterone and Epitestosterone by LC/MS/MS using Strata-X-A and Kinetex 1.7 µm C18
19761
Separation Mode: Reversed Phase
Reversed Phase
C18
HPLC
Androgens
Clinical

Applications

Separation of Testosterone and Epitestosterone by LC/MS/MS using Strata-X-A and Kinetex 1.7 µm C18
Analytes
1
2
3
Compound Name
Testosterone
Details
LC Conditions (App ID: 19761)
Column:
Brand Name:
Part No:
Elution Type:
Gradient
Mobile Phase:
A: 0.1% Formic Acid +1 mM Amm Formate in Water
B: 0.1% Formic Acid +1 mM Amm Formate in ACN
Gradient Profile:
Step No.Time(min)%A%B
109010
22.51090
33.51090
43.69010
Flow Rate:
0.4 mL/min
Temperature:
55°C
Detection:
Mass Spectrometer (MS) amu @ 22°C
Sample Preparation Details
Description:
Strata-X-A 33 µm Polymeric Strong Anion, 30 mg / 3 mL, Tubes, 50/Pk
Part Number:
Pre Treatment Note:
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
SPE Condition 1:
2 mL Methanol
SPE Condition 2:
2 mL Water
SPE Load:
Load 1.25 mL Sample diluted in Water
SPE Wash 1:
0.6 mL 50% Methanol in Water
SPE Dry:
Dry for 5 min under high vacuum
SPE Elution 1:
0.3 mL Methanol
SPE Elution 2:
0.3 mL Methanol
SPE Evaporation:
Evaporate to dryness
SPE Reconstitute:
250 µL 200 uL 5% aqueous formic acid

Order Items Used in This Application

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