Quantitative determination of natamycin by microbiological assay and HPLC

ANYL 171

Jianmei Luo, luojianmei@tust.edu.cn1, Zhihua Jin, luojianmei@tust.edu.cn2, peilin Cen2, and Min Wang1. (1) Department of Chemical and Biochemical Engineering, Tianjin Key Lab of Industrial Microbiology,Tianjin University of Science and Technology, Tianjin Key Lab of Industrial Microbiology, College of Biotechnology, Tianjin University of Science and Technology, Tianjin, 300, Tianjin, 300222, China, (2) Department of Chemical and Biochemical Engineering, Zhejiang university, Hangzhou, 310027, China
The agar diffusion bioassay technique for rapidly screening a large number of high-yielding strains from individual mutants was developed in natamycin fermentation. This method used Rhodotorula rubra AS 2.166 as a test organism and analytical conditions were as follows, sample volume: 70 µL, the test organism spore suspension concentration: 108 CFU/mL, the volume ratio between test organism spore suspension and upper medium: 1:10, cultivation time: 12h, temperature: 28°æ. Results showed that the relationship between the logarithm of natamycin content and the square of inhibitory zone diameter was linear in the concentration range of 0.2-1.2 g°¤L-1. The intra- and inter-day coefficient of variation (CV) were 0.42% (n=6) and 0.50% (n=6), respectively. The mean recoveries were 99.67%, 100.24% and 101.81% when the concentrations of spiked sample were 293, 586 and 1172 mg°¤L-1. Based on statistic analysis by ANOVA test, no significant difference was found between bioassay and HPLC methods with P<0.05. The microbiological assay is proved to be suitable for quickly determining the concentration of natamycin in fermentation broth and can be widely applied in selecting high-production isolates from enormous mutants. Compared with HPLC method, this approach is more simple, economical and less labor-intensive.

 

General Papers
7:00 PM-9:00 PM, Sunday, 10 September 2006 Moscone Center -- Hall D, Poster

Division of Analytical Chemistry

The 232nd ACS National Meeting, San Francisco, CA, September 10-14, 2006