First-Order Derivative Spectrophotometry for Simultaneous Determination of Vitamin C and Nicotinamide: Application in Quantitative Analysis of Cocrystals.

Abstract

Vitamin C (l-ascorbic acid, ASC) and the amide form of vitamin B3 nicotinamide (NIC) can form cocrystals through hydrogen bonding. Currently, there is a lack of fast and reliable alternatives for precisely quantifying cocrystal components and their purity. Spectrophotometric analysis for quantifying such vitamin preparations is challenging due to overlapping absorbance bands in a narrow wavelength range in the ultraviolet (UV) region. Moreover, ASC undergoes progressive degradation if not diluted in a proper medium, requiring stability during quantitative analysis. This study adopted a fast, simple, and reliable two-component spectrophotometric assay for simultaneously determining ASC and NIC based on the first-order derivative spectrophotometry (FODS) method using sodium oxalate as a stabilizer for vitamin C. The FODS method showed linearity between 2 and 24 μg·mL-1 and good precision. The standard addition method was employed to validate FODS, with high recovery percentages (96.5 to 102.4% for ASC and 95.3 to 101.9% for NIC). The FODS method was successfully applied to quantify ASC and NIC in bulk powder produced by the gas antisolvent method. The proposed method could estimate cocrystal purity through mass balance regarding the expected 1:1 stoichiometry, confirmed by PXRD and DSC. Cocrystal purity determined by the FODS method (58-100%) aligned well with results from LC-MS (62-100%), with an accuracy exceeding 97%. The FODS method is as sensitive and accurate as high-performance liquid chromatography for simultaneously determining vitamin concentrations deriving from cocrystals. However, it is less costly, more efficient, and a suitable alternative to classical solid-state methods for estimating cocrystal purity.