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Nicotine using SPE with Gemini NX-C18, LC-MSMS (Fig.3a)

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Sample Preparation Details

Part Number :8B-S029-UBJ

Description :Strata-X-C 33 µm Polymeric Strong Cation, 60 mg / 3 mL, Tubes, 50/Pk

Condition 1 :2 mL Methanol at 1 mL/min

Condition 2 :2 mL Ammonium acetate buffer at 1 mL/min

Load :Load 2.00 mL Sample diluted in 0.5 diluted urine sample

Wash 1 :2 mL Ammonium acetate buffer at 1 mL/min

Wash 2 :2 mL 30% Methanol at 1 mL/min

Dry :> 10 Hg for 5 min to remove residual water

Elution 1 :2 mL 1.5% Ammonium hydroxide methanol solution at 1 mL/min

Elution 2 :2 mL 1.5% Ammonium hydroxide methanol solution at 1 mL/min

Evaporate: :Evaporate to dryness

Reconstitute :500 µL 500uL ACN/20mM Ammonium bicarbonate (10:90)

Notes :Reconstitution Solvent: 500uL Acetonitrile/20mM Ammonium bicarbonate (10:90)

Application Detail (App ID: 22036)

Column: Gemini® 3 µm NX-C18 110 Å, LC Column 50 x 2 mm, Ea

Part No:00B-4453-B0

Dimensions:50 x 2 mm ID

Elution Type:Gradient

Elution A:20mM Ammonium Bicarbonate

Elution B:100% Acetonitrile

Gradient Profile:

Step No.	Time (min)	Pct A	Pct B
1	0	90	10
2	3	25	75
3	3.1	90	10
4	5	90	10

Flow Rate:500 µL/min

Col. Temp: 25 °C

Detection: Tandem Mass Spec (MS-MS) @ (22°C)

Detector Info: SCIEX API 4000™

Analyte Details

Analyte Detail not available

Nornicotine

3-OH-Cotinine

Anabasine

Cotinine

Nicotine