Vitamin B3 & Metabolites

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Matrix

Plasma, serum

Analytes

Nicotinic Acid, Nicotinuric Acid, Niacinamide

SPE Cartridge

SiliaPrep SCX nec
50 mg / 1 mL
SPE-R60430B-01B

Sample Pretreatment before Loading

Add 150 μL of 2 % aqueous Acetic Acid to 50 μL of serum, mix.

Conditionning

1 mL of Methanol

Equilibration

1 mL of 2 % Acetic Acid

Loading

Pretreated sample

Washing

2 x 1 mL of Water / Methanol / Acetic Acid (68:30:2)
2 x 1 mL of 2 % Acetic Acid in Methanol

Elution

2 x 400 μL of 5 % Ammonium Hydroxide in Methanol

Further Treatment

Evaporation under Nitrogen, reconstitution with 0.1 % Formic Acid and quantification by LC/MS/MS.