A Novel Approach for Quantitation of Nonderivatized Sialic Acid in Protein Therapeutics Using Hydrophilic Interaction Chromatographic Separation and Nano Quantity Analyte Detection

Aerosol-based detectors for liquid chromatography

“…Sialic acid quantitation is required in characterization of biopharmaceutical proteins but lacks a strong UV chromophore. Chemmalil et al [80] described a novel HPLC-NQAD method for quantitation of nonderivatized sialic acid -N-acetylneuraminic acid (NANA) and N-glycolylneuraninic acid (NGNA) in glycoproteins. Using HILIC, they found the method to have a linear dynamic range (312.5-10,000 ng injected), excellent precision (grand RSD of 0.95%), repeatability (0.96% RSD), and accuracy (96-107%).…”

Nanotechnology and sialic acid biology

“…This sialic acid quantitative sensitive method lacking requirement of active chromophore or fluorophore finds importance over conventional methods and HPLC/NQAD method offers advantage in reproductive results over the conventional HPLC/DMB method. While HPLC/DMB method involves derivatization of glycoproteins using 1, 2-diamino-4, 5-methylenedioxybenzene (DMB) and Dionex-based high-pH anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD), the HPLC/NQAD method is designed with elimination of the derivatization step and efficient PolyGLYCOPLEX amide column adding to the sensitivity to the detection method [42].…”