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Extraction of Steroids Using Strata-X on Kinetex 2.6µm C18 50 x 2.1mm ID by LCMS

Sample Preparation Details

Part No:8B-S100-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:0.5 mL Human plasma (spiked with analyte at 50ng/mL) was diluted with 1 mL of 0.1M Acetic acid
Dry:2 mins at 10" of Hg pressure
Final Prep and Analysis: Blown down samples (after evaporation) were reconstituted in 100 uL of initial mobile phase (A/B=80/20). Inject: 10 µL on HPLC / Mass Spectrometer (MS) @ amu (ambient)

Analytes:

Name Spiked
Conc.
Log P pKa % Rec %RSC (n=0)
1 Triamcinolone 0.25 
2 Prednisolone 0.25 
3 Cortisone 0.25 
4 Betamethasone 0.25 
5 Corticosterone 0.25 
6 Triamcinolone acetonide 0.25 
7 11-alpha-Hydroxyprogesterone 0.25 
8 Cortisone acetate 0.25 
9 Testosterone 0.25 
10 11-Ketoprogesterone 0.25 
11 Betamethasone 17-valerate 0.25 

Application Detail (App ID: 18848)

Part No:00B-4462-AN
Dimensions:50 x 2.1 mm ID
Elution Type:Gradient
Elution A:0.1%FA in H2O
Elution B:0.1%FA in Acetonitrile
Gradient Profile:
Step No. Time (min) Pct A Pct B
1 0 80 20
2 2.5 20 80
3 2.51 5 95
4 2.8 5 95
5 2.81 80 20
6 4.6 80 20

Flow Rate:450 µL/min
Col. Temp: 25 °C
Detection: Mass Spectrometer (MS) @ amu (22°C)
Detector Info: ESI conditions
Positive Ionization
NEB: 10.00
CUR: 8.00
IS: 4500.00
TEM: 425.00
CAD: 7.00
EP 10.00
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

1
Triamcinolone
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2
Prednisolone
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3
Cortisone
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4
Betamethasone
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5
Corticosterone
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6
Triamcinolone acetonide
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7
11-alpha-Hydroxyprogesterone
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8
Cortisone acetate
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9
Testosterone
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10
11-Ketoprogesterone
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11
Betamethasone 17-valerate
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