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

Sample Preparation Details

Condition 1 :400 µL Methanol at 1 mL/min
Condition 2 :400 µL Water at 1 mL/min
Load :Load 1500.00 µL Sample diluted in Water
Wash 1 :800 µL Water at 1 mL/min
Wash 2 :800 µL Methanol:Water (50:50) at 1 mL/min
Dry :1-2 mins at 10" of Hg
Elution 1 :200 µL Methanol
Elution 2 :200 µL Methanol
Evaporate: :Evaporate to dryness
Reconstitute :500 µL Mobile Phase
Notes :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 Biosystems.com), positive ion mode in APCI; MRM, heater gas flow 7000 cc/min; heater temperature 425o C.

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

1
Testosterone
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