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Determination of Chondroitin Sulfate by Thiazine Dyes using Flow Injection Analysis with Spectrophotometric Detection

Abstract A new method for the flow‐trough spectrophotometric determination of chondroitin sulfate in presence of Azure B was developed. A study was done under both static and flow‐trough conditions. Statistic factorial design was used to find the significant experimental parameters for the flow‐trough configuration and their optimal values were found: pH=5.5, c Azure B=4×10−5 mol l−1, the flow rate 5.8 ml min−1, sample volume injected of 100 µl, reaction coil length of 20 cm. The calibration curves are linear in the concentration range of 0–5 µg ml−1 of chondroitin sulfate, the limit of detection of about 0.29 µg ml−1, the limit of determination of about 1.22 µg ml−1. The stoichiometry of an ion aggregate chondroitin sulfate:Azure B was determined in ratio of 1:3 per one disaccharide unit under the optimal conditions.