AOAC SPIFAN Review Team (August 2016)

LC-MS/MS Analysis of Nicotinic acid

and Nicotinamide

Chemistry Method

Version 2

Page 2/15

Prepared by

S. Bhandari

Approved by

Issue Date

Implementation Date

________________________________________________________________________________

Uncontrolled When Printed

1. OVERVIEW

Purpose and background

Niacin (vitamin B3) is essential for the normal functioning of the human and animal body. Niacin is a

water soluble vitamin. Niacin (vitamin B3) is a water-soluble vitamin that is essential for converting

food to energy. It supports the function of the digestive system, skin and nerves, and improves

circulation. Niacin deficiency causes

which is characterized by a group of symptoms known

“the four Ds,” referring to

and death. Niacin plays a critical role in

the metabolism of fat, carbohydrates and amino acids. It functions in the body primarily in the form

of two coenzymes: NAD and NADPH. NAD is used in the breakdown of sugar and the oxidation of

fatty acids for energy production while NADPH is used in folate metabolism and the synthesis of

fatty acids, cholesterol, steroid hormones and important precursors of DNA. Outside of its functions

as a coenzyme, niacin is also involved in DNA repair and gene stability.

Principle

Free as well as bound nicotinic acid and nicotinamide are extracted using an ammonium

formate buffers employing protease and taka-diastase treatments. The sample extracts are

analyzed using liquid chromatography (LC) with tandem mass spectrometric detection.

Analyte Scope of the Method

The method is applicable for the analysis of total Nicotinic acid, Nicotinamide, Total vitamin

B3 in the sample can be calculated based on the results of the analysis.

Matrix Scope

Infant formula and adult nutrition formula and pet foods.

Analytical Range:

–

4375 mcg/100g (with no dilution of the extract).

2-10,000 mcg/100g (with 2.3 x dilution of extracts of samples containing niacin levels higher than 4000

mcg/100g).

LOD and LOQ of the method:

LOD and LOQ of the Method of total Vitamin B3 (niacin)analysis:

METHOD

FOR ERP USE ONLY

DO NOT DISTRIBUTE