Steroids

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Matrix

Urine, serum

Analytes

Cortisone, Cortisol, 21-Deoxycortisol, Corticosterone, 11-Deoxycortisol, Fluoxymesterone, Trenbolone, Boldenone, Androstenedione, Nandrolone, Methandienone, 17 α-Hydroxyprogesterone, Testosterone, 16 β-Hydroxystanozolol, Epitestosterone, 5 β-Estran-3α-ol-17-one, 17 α-Methyltestosterone, Methenolone, 5 α-Estran-3α-ol-17-one, Norethandrolone, Progesterone, Stanozolol

SPE Cartridge

SiliaPrep C8/SAX nec
200 mg / 6 mL
SPM-R022830B-06G

Sample Pretreatment before Loading

Serum: add 4 mL of Phosphate buffer (100 mM, pH 7.0) to 1 mL of sample.
Vortex.

Urine: add 1 mL of Acetate buffer (100 mM, pH 5.0) and 50 μL of β-glucuronidase to 1 mL of sample.
Vortex, heat at 65°C for 1 or 2 hours.
Add 2 mL of Phosphate buffer (100 mM, pH 7.0).
Vortex.

Conditionning

3 mL of Methanol

Equilibration

3 mL of Phosphate buffer (100 mM, pH 7.0)

Loading

Pretreated sample

Washing

3 mL of Water
3 mL of Methanol / Water (30:70)
Dry the cartridge

Elution

2 x 1.5 mL of Methanol

Further Treatment

Evaporation under Nitrogen, reconstitution with Methanol / Water (50:50) and quantification by LC/MS/MS.