Tobacco

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Matrix

Oral fluid

Analytes

Cotinine, Anabasine, Nicotine

Well Plate

SiliaPrepX SCX
30 mg / 2 mL
96W-P0005-AA

Sample Pretreatment before Loading

Collect oral fluid according to instructions on the device, place sample in adequate buffer solution and centrifugate. Add 1 mL of 1 % Formic Acid to 0.5 mL of supernatant.

Conditionning

1 mL of Methanol

Equilibration

1 mL of Water

Loading

Pretreated sample

Washing

1 mL of Water
1 mL of Acetone / Water (50:50)
Dry wells

Elution

2 x 500 μL of Ethyl acetate / Isopropanol / Ammonium Hydroxide (70:20:10)

Further Treatment

Evaporation under Nitrogen, reconstitution with Ammonium Bicarbonate (20 mM, pH 8.2) / Methanol (90:10) and quantification by LC/MS/MS.