Analysts rely on laborious derivatization procedures and complementary analytical instrumentation to test products. The QC process is often time-consuming and expensive due to the lack of a single platform capable of analyzing necessary parameters. Physicochemical properties such as homogeneity, size and composition serve as important QC parameters for carbohydrates. It is essential to have proper QC of carbohydrate-based therapeutics to ensure their efficacy and safety.
Additionally, SEC-CAD chromatograms indicated the heterogeneous nature of the samples.Ĭarbohydrates have become increasingly important commercially not only as food or structural building blocks, but also as natural health products. HILIC-CAD analysis of commercially purchased hyaluronic acid products indicated that samples were composed of GlcNAc and GlcA. The optimized gradient method demonstrated good linearity (R 2 > 0.99), precision (RSD < 8%), LOD (< 85 ng/mL) and LOQ (< 280 ng/mL). We separated common uronic acids (GlcA, GalA, LIdoA and Neu5Ac), amino sugars (GlcN, GalN and GlcNAc) and select neutral sugars (LRha, LFuc, Man and Gal) using a gradient HILIC-CAD method. Additionally, since CAD is suitable for SEC experiments, we tested the homogeneity of hyaluronic acids using a SEC-CAD method. As an application of the HILIC-CAD method, we performed composition analysis of commercially purchased hyaluronic acid products.
In this study, we developed a gradient method for simultaneous analysis of acidic, amino and select neutral monosaccharides. Although, several neutral sugars have been separated and detected using HILIC-CAD, there has not been any report on acidic and amino sugar analysis using this method. HILIC-CAD has emerged as an effective platform for direct monosaccharide composition analysis of complex carbohydrates without derivatization. QC analysis of carbohydrates has been historically cumbersome due to lengthy and laborious derivatization techniques and the requirement of complimentary instrumentation.
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