FTIR determination of the degree of molar substitution for hydroxypropyl chitosan

Abstract

We developed and validated a novel Fourier transform infrared (FTIR) method to determine the degree of molar substitution (MS) for hydroxypropyl chitosan (HPCS) using nuclear magnetic resonance (1H NMR) as a reference, and investigated the factors influencing the MS assay. Through extensive screening of integration methods for candidate bands in the FTIR spectrum of HPCS using 20 HPCS samples with degrees of acetylation (DA) ranging from 0.003 to 0.139, we found that when using band area at 2970 cm−1 as a probe integral, the MS values obtained via the 1H NMR method exhibited linear correlations (R2 > 0.98) with at least 16 integral ratios derived from their FTIR spectra. The optimal reference bands with high reliability are located at 3440 cm−1 and 1415 cm−1, with R2 exceeding 0.99 and a MS range of 0.17–1.92. The band at 2875 cm−1 is less affected by the trace moisture present in HPCS samples than the others. The results of the method validation demonstrated a mean recovery of 98.9 ± 2.8 % and an RSD below 10 %, suggesting a simple, robust, and highly accurate and precise method. This method could be extendable for the determination of the MS of insoluble HPCS derivatives and other hydroxypropylated polysaccharides.