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ObjectiveTo reveal the effects of different microbial agents on quality of Lycii Fructus by comparing the differences in the contents of multiple types of chemical components in Lycii Fructus after the application of different microbial agents. MethodTaking Ningqi No. 7 as experimental material， four microbial agents， namely Peiyuan combined with Xinterui（TP group）， Trichoderma harzianum combined with Bacillus subtilis（BW group）， Genwuyou（MT group） and Junyiduo（JYD group）， were applied， and no microbial agents was used as the blank group（CK group）. Then the contents of total phenolics， total flavonoids， saccharides， amino acids， nucleosides and bases， betaine and other components in Lycii Fructus were determined by ultraviolet spectrophotometry（UV）， high performance liquid chromatography（HPLC） and ultra-high performance liquid chromatography-mass spectrometry（UHPLC-MS/MS）， and the methods such as multiple comparisons， principal component analysis（PCA） and partial least squares-discriminant analysis（PLS-DA） were used to analyze the effect of different microbial agents on the quality of Lycii Fructus. ResultMicrobial agents had different effects on chemical components of Lycii Fructus. The content of total phenolics was the highest in the TP group， and it varied significantly from the CK group（P<0.05）. The total flavonoid content was the highest in the BW group， followed by the TP group. Both polysaccharide and alduronic acid contents were the highest in the JYD group. Betaine content in the TP and BW groups were significantly higher than that in the CK group（P<0.05）. For the determined 23 kinds of amino acids， most of them were the lowest in the JYD group， and the highest in the MT group， while the nucleoside bases were higher in the MT and BW groups. It indicated that Lycii Fructus from different treatment groups could be distinguished clearly based on the determined 45 chemical components. The result of PLS-DA showed that the major differential components in each group were polysaccharides， glucose， fructose， betaine， alduronic acid， asparagine， sucrose， threonine， total flavonoids， alanine and total phenolics. The results of PCA composite scores based on the main differential components showed that composite scores of chemical components in each group were BW group>TP group>MT group>CK group>JYD group. ConclusionThe application of microbial agents of BW， TP and MT can promote the quality improvement of Lycii Fructus， and the application of JYD can promote the accumulation of polysaccharides and alduronic acid to a certain extent， but the overall effect on the quality of Lycii Fructus is not clear. This study lays the foundation for the green and healthy development of Lycii Fructus industry.
RESUMO
OBJECTIVE: To establish a QTRAP-UPLC-MS/MS method for simultaneous determination of 10 kinds of nucleosides and nucleobases in Scrophulariae Radix processed by different methods, and investigate the influences of different processing methods on contents of nucleosides and nucleobases. METHODS: The contents of 10 kinds of nucleosides and nucleobases in Scrophulariae Radix processed with different methods were simultaneously determined by QTRAP-UPLC-MS/MS, and TOPSIS analysis was used for comprehensive evaluation of nucleosides and nucleobases. RESULTS: The contents of guanosine, uridine, adenosine and uracil were relatively high in Scrophulariae Radix. There were differences in contents of nucleosides and nucleobases of Scrophulariae Radix processed with different methods, and the samples processed by method of "sweating" had better quality. CONCLUSION: This study provides a scientific basis for investigation on the influence of processing methods on contents of nucleosides and nucleobases in Scrophulariae Radix and screening of suitable processing method for Scrophulariae Radix.
RESUMO
OBJECTIVE: To establish a QTRAP UPLC-MS/MS method for simultaneous determination of thirteen nucleosides and nucleobases in Pseudostellariae Radix, and analyze their dynamic changes at different periods. METHODS: The separation was carried out on a Waters Atlantis T3 column (2.1 mm × 150 mm, 3 μm)eluted by a mobile phase of 5 mmol · mL-1 of ammonium acetate (B)-methanol(A) at a flow rate of 0.4 mL · min-1. The elution program is as follows. 0 -4.5 min, 3% -4% A; 4.5-8 min, 4% -18% A; 8 -10 min, 18%A; 10 -10.1 min, 18% -3% A; 10.1 -13 min, 3%A. The target compounds were analyzed by the positive ion multiple reaction monitoring (MRM) mode. RESULTS: The calibration curves of the 13 nucleosides and nucleobases showed good linearity (r >0.991 0) in the ranges of the tested concentrations, and the average recoveries were between 96.46% - 105.07%. The contents of cytidine, uridine, inosine, guanosine, and thymidine in Pseudostellariae Radix harvested at different periods showed some differences, and the contents were higher in mid-June. CONCLUSION: The method is simple, sensitive, accurate and credible, and provides the basis for exploring the quality forming mechanism of Pseudostellariae Radix medicinal materials and the suitable harvest time.