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The method of non-aqueous conductivity titration (NACT) of organic weak acids was applied to quickly and accurately determine the phenolic-hydroxyl and carboxyl-groups contents in humic acid. By varying the pH of the humic-acid sample, the concentration of the titrant, and the nitrogen-gas flow rate, the optimal titration conditions were determined to be a sample pH of 4, titrant concentration of 0.05 mol/L, and nitrogen-gas flow rate of 80 mL/min. Applying the detection method to p-hydroxybenzoic acid showed that its phenolic-hydroxyl content was 758.82±111.76 cmol/kg and carboxyl content was 744.44±51.11 cmol/kg. The theoretical phenolic-hydroxyl and carboxyl-groups contents of the p-hydroxybenzoic acid were 723.96 cmol/kg respectively, indicating that the method can accurately quantify the carboxyl and phenolic-hydroxyl groups in the sample. The NACT was used to measure the phenolic-hydroxyl and carboxyl-groups contents in humic acid quickly and accurately. In addition, 29 humic acid samples from 8 provinces of China covering the main humic-acid producing areas were collected and analyzed for acidic-groups content using the reported method.

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Prodigiosin is a red pigment produced by Serratia marcescens with anticancer, antimalarial, and antibacterial effects. In this study, we extracted and identified a red pigment from a culture of S. marcescens strain ZPG19 and investigated its effect on the growth performance and intestinal microbiota of Kunming mice. High-performance liquid chromatography/mass spectrometry revealed that the pigment had a mass-to-charge ratio (m/z) of 324.2160, and thus it was identified as prodigiosin. To investigate the effect of prodigiosin on the intestinal microbiota, mice (n = 5) were administered 150 μg/kg/d prodigiosin (crude extract, 95% purity) via the drinking water for 18 days. Administration of prodigiosin did not cause toxicity in mice. High-throughput sequencing analysis revealed that prodigiosin altered the cecum microbiota abundance and diversity; the relative abundance of Desulfovibrio significantly decreased, whereas Lactobacillus reuteri significantly increased. This finding indicates that oral administration of prodigiosin has a beneficial effect on the intestinal microbiota of mice. As prodigiosin is non-toxic to mouse internal organs and improves the mouse intestinal microbiota, we suggest that it is a promising candidate drug to treat intestinal inflammation.