| abstract |
The invention relates to a method of determination of hyaluronic acid in complex samples by cleavage with SpnHyl enzyme followed by the reaction with 3-methyl-2-benzothiazolinone hydrazone (MBTH) to form a coloured product analysed by spectrophotometry. In case of a high content of glycosaminoglycans, these can be separated by precipitation before the cleavage, e.g. with the use of cetyltrimethylammomumbromide (CTAB) in acetate buffer and salt medium. In case of a high content of reducing sugars, e.g. glucose, their influence can be eliminated, after removing the glycosaminoglycans and before the cleavage, by precipitating hyaluronic acid from the formed supernatant and then redissolving this sediment in acetate buffer and performing the cleavage step. |