Applications
LC-MS-MS Analysis of Testosterone on Gemini C18, 5u, 50x2.0 Column
18463
Separation Mode: Reversed Phase
Reversed Phase
C18
HPLC
Androgens
Clinical
Clinical Research
Therapeutic / Clinical

Applications

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

Analytes

1
Compound Name
Testosterone
CID
6013
Molecular Formula
C19H28O2
Molecular Weight
288
No. Hydrogen Bond Acceptors
2
No. Hydrogen Bond Donors
1

Conditions

LC Conditions (App ID: 18463)
Column:Gemini 5 µm C18 110 Å, LC Column 50 x 2 mm, Ea
Brand Name:Gemini
Part No:00B-4435-B0
Elution Type:Isocratic
Mobile Phase:A: 0.1% Formic acid in Water
B: 0.1% Formic acid in Acetonitrile
Gradient Profile:
Step No.Time(min)%A%B
113070
Flow Rate:0.5 mL/min
Temperature:22°C
Detection:Mass Spectrometer (MS) m/z @ 22°C

Sample Preparation Details
Description:Strata-X 33 µm Polymeric Reversed Phase, 30 mg / well, 96-Well Plates, 2/Pk
Part Number:8E-S100-TGB
Pre Treatment Note: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.
SPE Condition 1:400 µL Methanol at 1 mL/min
SPE Condition 2:400 µL Water at 1 mL/min
SPE Load:Load 1500.00 µL Sample diluted in Water
SPE Wash 1:800 µL Water at 1 mL/min
SPE Wash 2:800 µL Methanol:Water (50:50) at 1 mL/min
SPE Dry:1-2 mins at 10" of Hg
SPE Elution 1:200 µL Methanol
SPE Elution 2:200 µL Methanol
SPE Evaporation:Evaporate to dryness
SPE Reconstitute:500 µL Mobile Phase

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