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A new method for simultaneous determination of puerarin, daidzin, daidzein and genistein in Radix puerariae by ultra-high performance liquid chromatography was established. The target analytes were extracted from Radix puerariae by 70% ethylene glycol with the assistance of ultrasonication, purified by the absorption of N-propyl ethylenediamine (PSA), and separated on a Supersil ODS column (4.6 mm × 250 mm × 2.5 μm). Gradient elution in 12 min was performed with the mobile phase 0.1% formic acid(A)–acetonitrile(B). The column temperature was 25 °C and the flow rate was 1 mL/min. The detection wavelength of the four target analytes was 250 nm. The limits of detection (LODs) of puerarin, daidzin, daidzein and genistein were 0.086 mg/L, 0.020 mg/L, 0.027 mg/L and 0.037 mg/L, respectively, and limits of quantitation (LOQs) were 0.29 mg/L, 0.065 mg/L, 0.090 mg/L and 0.12 mg/L, respectively. The recovery of the four substances ranged from 90.5% to 109.6%, and the relative standard deviation (n = 6) was less than 7.7%. With the established methods, puerarin, daidzin, daidzein and genistein in Radix puerariae from 11 origins were determined. The contents of the four compounds varied with the origin and variety. It provides basic data and technical means for quality control and regulation of Radix puerariae.

The in vitro digestion and fermentation behaviors of Lactobacillus helveticus LZ-R-5- and L. pentosus LZ-R-17-sourced exopolysaccharides (LHEPS and LPEPS) were investigated by stimulated batch-culture fermentation system. The results illustrated that LHEPS was resistant to simulated saliva and gastrointestinal (GSI) digestion, whereas LPEPS generated a few monosaccharides after digestion without significant influence on its main structure. Additionally, LHEPS and LPEPS could be consumed by the human gut microbiota and presented stronger bifidogenic effect comparing to α-glucan and β-glucan, as they promote the proliferation of Lactobacillus and Bifidobacterium in cultures and exhibited high values of selectivity index (13.88 and 11.78, respectively). Furthermore, LPEPS achieved higher contents of lactic acid and acetic acid (35.74 mM and 45.91 mM, respectively) than LHEPS (35.20 mM and 44.65 mM, respectively) during fermentation for 48 h, thus also resulting in a larger amount of total SCFAs (110.86 mM). These results have clearly indicated the potential prebiotic property of EPS fractions from L. helveticus LZ-R-5 and L. pentosus LZ-R-17, which could be further developed as new functional food prebiotics to beneficially improve human gut health.

A new rapid and accurate method was developed for simultaneous determination of pyridoxine and pyridoxal in ginkgo seeds, using ultra-performance liquid chromatography (UPLC) equipped with a fluorescence detector. Diluted hydrochloric acid solution was used as the extracting solvent. For the pretreatment of extracts, a zeolitic imidazolate framework material (ZIF-8) was prepared and characterized. An ODS-BP column (4.6 mm × 250 mm × 5 μm) was used for separation. The conditions of sample extraction, cleaning and separation were optimized. The linear correlation coefficient (R2) of the analyte was better than 0.9999, indicating good linearity. The limits of detection (LODs) of pyridoxal and pyridoxine were 0.0065 mg/kg and 0.0057 mg/kg, respectively, and limits of quantitation (LOQs) were 0.022 mg/kg and 0.019 mg/kg, respectively. The recovery of the two substances ranged from 86.2% to 110.4%, and the relative standard deviation (n = 6) was less than 7.5%. The method was applied to determine the contents of pyridoxine and pyridoxal in actual ginkgo seed samples with satisfactory results.

In this study, a high-throughput sequencing technique was used to analyze bacterial and fungal diversity of two traditional Tibetan kefir grains from Linzhi (K1) and Naqu (K2) regions. Comparative bioinformatic analyses indicated that Lactobacillus kefiranofaciens, L. kefiri and Kluyveromyces marxianus were the main dominant strains in K1 and K2. In order to research the relationship of the growth of kefir grains, the biofilm and the extracellular polysaccharides (EPS) produced by microorganisms, the proliferation rate of kefir grains, the yield and chemical structure of EPS and the optimal days for biofilm formation were determined. The results showed that the growth rate, the yield of EPS and the biofilm formation ability of K1 were higher than K2, and the optimal day of their biofilm formation was the same in 10th day. Additionally, the live cells, dead cells and EPS in biofilm formation of K1 and K2 were observed by fluorescence microscope to clarify the formation process of kefir grains. To determine the influence of microbial interactions on biofilm and the formation of kefir grains, the essential role of microbial quorum sensing needs further attention.