Potential of lactic acid bacteria from Pico cheese for starter culture development.

Autochthonous lactic acid bacteria may provide a means of promoting the quality and safety of traditional fermented food products, in particular, artisanal cheeses. Pico cheese is an artisanal, dairy specialty of the Azores in risk of disappearing. Efforts to maintain its quality to the requirements of the modern markets are, thus, necessary. Lactic acid bacteria were isolated from artisanal Pico cheese, identified by sequencing of the 16S rRNA gene, and their potential as starter cultures was evaluated by studying their acidification ability, enzymatic activities (caseinolysis, lipolysis and API-ZYM profile), diacetyl and expolysaccharide production, autolysis, antimicrobial activity against Listeria monocytogenes ATCC 7466, Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 29523, Pseudomonas aeruginosa ATCC 27853 and Clostridium perfringens ATCC 8357, sensory evaluation of odour formation in milk, syneresis and firmness of the curd. Several of the studied lactic acid bacteria isolates showed interesting properties for practical application as starters in artisanal cheese production. The isolates with the highest number of positive traits and, therefore, the most promising for starter development were Lactococcus lactis ssp. lactis L1C21M1, Lactobacillus paracasei L1B1E3, Leuconostoc pseudomesenteroides L1C1E6, Lactobacillus casei L1A1E5 and L1C1E8.

Evaluation and comparison of two improved techniques for the on-line detection of antioxidants in liquid chromatography eluates.

Two methods for the on-line detection in HPLC eluates of analytes possessing radical scavenging activity were improved and compared. The instrumental set-up of the method that is based on on-line inhibition of luminol chemiluminescence (CL) by antioxidants was improved using better quality syringe pumps, employing a diode array detector, and introducing a mixing/neutralisation coil and a pulse damper. Sensitivity of the HPLC-CL detection increased by a factor of 4. Post-column neutralisation of eluates improved compatibility of this detection method with acidified HPLC eluents. The second method, which is based on the post-column quenching of 2,2'-diphenyl-1-picrylhydrazyl radical (DPPH*), was improved by readjusting composition and flow-rate of the reagent, mounting an additional pulse damper and detecting unreacted DPPH* with a detector equipped with a tungsten lamp. Purging of the DPPH* solution with He gas prior to analysis was introduced. This led to 30-fold better detection limits. The improved methods were compared with respect to limits of detection, the radical scavenging mechanism involved, compatibility with common HPLC solvents and pH range, and some technical aspects. The techniques described have high potential for the rapid identification of radical scavengers in complex samples like plant extracts.

On-line detection of antioxidative activity in high-performance liquid chromatography eluates by chemiluminescence.

Luminol chemiluminescence (CL) was employed for the on-line detection of radical scavengers in HPLC eluates. Optimization of CL reagents and instrumental setup resulted in a steady postcolumn luminol photochemical reaction in the presence of microperoxidase and hydrogen peroxide at pH 10. Quenching of the CL signal was utilized to detect radical scavenging activity of both natural and synthetic antioxidants at the nanogram level. The detection system can be used with isocratic or gradient elution. Several antioxidative compounds were detected in thyme and sage acetone extracts. Quantitative results can be obtained when antioxidants are analyzed at certain concentrations. The method is suitable for rapid screening of antioxidants in crude extracts.