Optimization of extraction process for hyaluronic acid from Rana chensinensis by response surface methodology

With Rana chensinensis David skin as the research object,the hyaluronic acid(HA) rate was set as a measure indicator. Under the condition of solid-liquid ratio 1∶40(g/m L),three extraction parameters including enzyme temperature,enzyme hydrolysis time(time of double enzyme was equal) and enzyme amount(dosage ratio of double enzyme was 1∶1) of double enzyme(alkaline proteinase and trypsin enzyme) hydrolysis method as the respective variables and hyaluronic acid rate as response value were explored by using Box-Benhnken central composite design and response surface analysis theory based on single factor experiment,so as to study the interaction of the respective variables and their influence on the HA extraction rate and set up the simulated quadratic polynomial regression equation of prediction model. The optimum extraction condition was enzyme amount 55000 U/g under the optimal temperature 50 ℃ for a duration of 14 h. Under these conditions,the average extraction yield of HA was up to 27.6839 mg/g,glucuronide content was 13.8420 mg/g,namely whose purity 42.2% in extraction. Besides,the response surface methods were also proved to be available to extract HA exactly.