6-MAM by LC/MS/MS using Strata-X-Drug B and a Kinetex C18 2.6µm, 50x2.1mm

Sample Preparation Details

Part No:8B-S128-UCH
Solid Phase Extraction (SPE) procedure:Note: The solvent volumes shown below are for a 60 mg bed mass. The solvent volumes will need to be adjusted for a smaller or larger bed mass.
Load:Pre-treatment: To 2mL of urine add 1mL of B-glucuronidase solution (5000F units/mL in 100mM acetate buffer pH 5). Mix/vortex. Hydrolyze for 3 hours at 60ºC. Allow sample to cool and add 1mL of 100mM phosphate buffer (pH 6). Centrifuge for 10 minutes and discard pellet. Load pre-treated sample directly - no conditioning required
Dry:10 min at 10 inches of Hg
Final Prep and Analysis: Evaporate eluant to dryness and reconstitute in 1mL of 10% methanol in formic acid (0.1%) Inject: 5 µL on HPLC / Mass Spectrometer (MS) @ amu (ambient)


Name Spiked
Log P pKa % Rec %RSC (n=0)
1 6-MAM-d3 100 0.01
2 6-MAM 100.5 0.63

Application Detail (App ID: 19599)

Part No:00B-4462-AN
Dimensions:50 x 2.1 mm ID
Elution Type:Gradient
Elution A:0.1% Formic acid
Elution B:Acetonitrile
Gradient Profile:
Step No. Time (min) Pct A Pct B
1 0 90 10
2 1 30 70
3 3 30 70
4 3 30 70

Flow Rate:400 µL/min
Col. Temp: ambient
Detection: Mass Spectrometer (MS) @ amu (22°C)
Note:SecurityGuard™ ULTRA Guard Cartridge System extends column lifetime.
- SecurityGuard ULTRA Cartridges, UHPLC C18 for 2.1mm ID Columns, 3/Pk Part No.: AJ0-8782
- Holder Part No.: AJ0-9000

Analyte Details

Analyte Detail not available