LC-MS-MS Analysis of Testosterone on Gemini C18, 5u, 50x2.0 Column

Sample Preparation Details

Part No:8E-S100-TGB
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:500 uL of Human plasma was diluted with 1000 ul of water(1:2 dilution)
Dry:1-2 mins at 10" of Hg
Final Prep and Analysis: Liquid Chromatography/Mass spectrometryAnalysis was performed using an HP 1100 LC system from Agilent Technologies, Palo Alto, CA, equipped with a quaternary pump, in-line degasser, multi-wavelength detector and autosampler. HP Chemstation software was used to analyze the data. The HPLC column used was a Gemini 5 um, C18, 50 x 2.0 mm from Phenomenex, Torrance, CA.
Conditions: Flow rate 0.5 mL/min; Mobile phase: 0.1% formic acid in water (pH 2.7, mobile phase A), 0.1% formic acid in acetonitrile (mobile phase B); A/B 30/70; total run time 1 min; injection volume = 5 uL; Autosampler: G1329A ALS Pump: G1312A (binary); MS detector: API 3000 Lc/MS/MS with ESI (TurbolonSpray®)(www.Applied, positive ion mode in APCI; MRM, heater gas flow 7000 cc/min; heater temperature 425o C.
Inject: 5 µL on HPLC / Mass Spectrometer (MS) @ 289/97 m/z (ambient)


Name Spiked
Log P pKa % Rec %RSC (n=0)
1 Testosterone 70

Application Detail (App ID: 18463)

Part No:00B-4435-B0
Dimensions:50 x 2 mm ID
Elution Type:Isocratic
Elution A:0.1% Formic acid in Water
Elution B:0.1% Formic acid in Acetonitrile
Gradient Profile:
Step No. Time (min) Pct A Pct B
1 1 30 70

Flow Rate:0.5 mL/min
Col. Temp: ambient
Detection: Mass Spectrometer (MS) @ 289/97 m/z (22°C)
Note:SecurityGuard™ Guard Cartridge System extends column lifetime.
- SecurityGuard Cartridges, Gemini C18 4 x 2.0mm ID, 10/Pk Part No.: AJ0-7596
- Holder Part No.: KJ0-4282

Analyte Details

Analyte Detail not available