Unfolding of Poly-L-glutamic Acid by Microbubbling of Supercritical Carbon Dioxide
The conformational changes in α-helical poly-L-glutamic acid caused by microbubbling supercritical CO2 were investigated with circular dichroism spectra. After microbubbling using a micropore filter at 35 and 30 MPa for 30 min, α-helix content decreased to 37%, while without the filter it was 68%. The α-helix structure was significantly decomposed by a high density of CO2. No important changes were observed in heating, autoclaving, or pH-lowering.
Inactivation of Enzymes in Namazake Using Micro-bubble Supercritical Carbon Dioxide
Glucoamylase, acid protease, and acid carboxypeptidase in namazake were inactivated with microbubble super critical carbon dioxide (SC CO2) at 25 MPa, 35°C for 30 min. After the treatment, their activities decreased from 330, 20.1, and 115 U/ml sake to 51.8, 1.8, and 0 U/ml sake, respectively. During the storage at 20°C for 60 days, no increase in amino acid value was observed. On the other hand, glucose concentration in the sake treated with SC CO2 increased as well as namazake. The formation of lactic acid, acetic acid, and ethanol were depressed, and acidity was constant during storage. On the contrary, the decrease of pyruvic and malic acid after 20 days of storage could be depressed. Analysis of volatile compounds suggests that the treatment with SC CO2 did not cause the formation of volatile compounds in contrast to heat-treatment. As a result of sensory evaluation, the sake treated with SC CO2 at 20 MPa, 35°C for 30 min was given sensory scores near to those of namazake.
HPLC determination of chlorhexidine gluconate and p-chloroaniline in topical ointment.
A novel fast isocratic reversed-phase HPLC method for simultaneous determination of chlorhexidine and its degradation product p-chloroaniline was developed. Zorbax SB Phenyl column (75 mm x 4.6 mm, 3.5 microm) was used for the separation. Mobile phase composed of acetonitrile and buffer solution of 0.08 M sodium phosphate monobasic containing 5 ml of triethylamine (0.5%) and adjust with 85% phosphoric acid to pH 3.0 in ratio 35:65 (v/v) pumped isocratically at flow rate 0.6 ml min(-1) was used. UV detection was performed at 239 nm, the total analysis time was about 10 min. The method is suitable for practical routine analysis of topical ointment in the quality control laboratory.