Simultaneous determination of nicotinamide mononucleotide, niacinamide, and nicotinic acid in health supplements using liquid chromatography method

Abstract

This study aimed to develop and validate an HPLC-PDA method for simultaneous quantification of three compounds: nicotinamide mononucleotide (NMN), nicotinic acid, and niacinamide in dietary supplements. The sample preparation procedure was simple, fast, and used environmentally green extracted solvents. The HPLC conditions were as follows: C18 column (250 mm x 4.6 mm, 5 µm); mobile phase: 10 mM phosphate buffer pH = 3: methanol (90:10, v/v), UV detection at wavelength 261 nm. The method met the AOAC criteria for validation, including selectivity, linearity (R2> 0.999 for all analytes in the range of 0.1-50.0 µg/mL), sensitivity (LOD: 0.4-0.45 mg/kg for NMN, 0.1-0.61 mg/kg for nicotinic acid, and 0.1-0.6 mg/kg for niacinamide; LOQ: 1.3-1.5 mg/kg for NMN, 0.49-2.00 mg/kg for nicotinic acid, and 0.47-1.90 mg/kg for niacinamide), accuracy (recovery: 95.5-103.0 % for NMN, 92.5-104.1 % for nicotinic acid, and 95.3-103.8 % for niacinamide), precision (RSDr: 0.54-3.79 % for NMN, 0.39-4.82 % for nicotinic acid, and 0.81-5.14% for niacinamide; RSDR: 1.03-3.70 % for NMN, 1.75-4.10 % for nicotinic acid, and 1.83-4.48 % for niacinamide). The method was successfully applied to analyze 20 supplement products collected in Hanoi.