Chitin assay to estimate the growth of Candida albicans in organs of infected mice

Abstract

Two methods have been used to estimate chitin. In both, the chitin was first converted into chitosan. The insoluble chitosan was then either depolymerized and deaminated with HNO2 and the product colorimetrically determined with 3-methylbenzo-2-thiazolone hydrazone and Fe3+ (method A), or hydrolysed in HCl and the glucosamine determined with p-dimethylaminobenzaldehyde (method B). Method B showed a better correlation between chitin concentration and absorbance, and its reliability was higher. This method was equally convenient to estimate the chitin content of germ tubes of Candida albicans grown in vitro, showing good correlation between mycelial growth and chitin content. Finally, method B was used to measure the growth of C. albicans in organs of animals infected experimentally.