Kutajarista is an Ayurvedic fermented herbal formulation prescribed for gastrointestinal disorders. This herbal formulation undergoes a gradual fermentative process and takes around 2 months for production. In this study, microbial composition at initial stages of fermentation of Kutajarista was assessed by culture independent 16S rRNA gene clone library approach. Physicochemical changes were also compared at these stages of fermentation. High performance liquid chromatography-mass spectrometry analysis showed that Gallic acid, Ellagic acid, and its derivatives were the major chemical constituents recovered in this process. At 0 day of fermentation, Lactobacillus sp., Acinetobacter sp., Alcaligenes sp., and Methylobacterium sp. were recovered, but were not detected at 8 day of fermentation. Initially, microbial diversity increased after 8 days of fermentation with 11 operational taxonomic units (OTUs), which further decreased to 3 OTUs at 30 day of fermentation. Aeromonas sp., Pseudomonas sp., and Klebsiella sp. dominated till 30 day of fermentation. Predominance of c-Proteobacteria and presence of gallolyl derivatives at the saturation stage of fermentation implies tannin degrading potential of these microbes. This is the first study to highlight the microbial role in an Ayurvedic herbal product fermentation.
Flavonoids (in particular unsubstituted B ring flavanones) in Eucalyptus foliage play an important role in mediating animal plant interactions, and there is a need for methods to analyse the diverse profiles found in leaves. A simple, high-performance liquid chromatographic (HPLC) method with in-line connected photodiode-array (PDA) detection was developed and validated to identify and quantify nine Bring unsubstituted and three Bring substituted flavonoids in ten Australian species of Eucalyptus. Of these, eight compounds were detected and quantified in the crude methanolic extracts of leaves of various Eucalyptus species (E. sieberi, E. rossii, E. fastigata, E. macrorhyncha, E. fraxinoides, E. agglomerata, E. consideniana, E. pauciflora, E. dives and E. obliqua) based on comparison with the retention times and λmax values of pure compounds. This rapid and sensitive HPLC/PDA method was coupled with electrospray ionization mass spectrometry (HPLC-ESI-MS) for qualitative analysis to corroborate the identification of compounds by HPLC/PDA analysis.
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