ABSTRACT
OBJECTIVE：To establis h HPLC fingerprint of Chinese gall leaven ，simultaneous determination of 5 components and optimization of distillers ’grains. METHODS ：HPLC method was adopted. The determination was carried out on Waters Symmetry Shield TM RP18 column with mobile phase consisted of acetonitrile- 0.1％ trifluoroacetic acid aqueous （gradient elution ）at the flow rate of 0.8 mL/min. The column temperature was 30 ℃，and detection wavelength was set at 280 nm. The sample size was 5 µL. Using gallic acid as reference ，HPLC fingerprints of 14 batches of sample were drawn. The similarity evaluation was performed by using Similarity Evaluation System of Chromatogram Fingerprint of TCM （2012 edition）to determine common peaks；SPSS 20.0 software was used for cluster analysis. RESULTS ：There were 11 common peaks in 14 batches of samples ，and the similarity range of 14 batches of samples was 0.424-0.998，A1-A5 was less than 0.850；5 components were identified in B 1-B9 sample，i.e. gallic acid ，（－）-epigallocatechin，methyl gallate ，2，4，6-tri-O-galloyl-α-D-glucose and 2，4，6-tri-O-galloyl- β-D-glucose. The results of cluster analysis showed that 14 batches of samples could be grouped into 3 categories，i.e. A1-A5 into one category，B1 and B 6 into one category ，B2-B5 and B 7-B9 into one category. The linear ranges of above 5 components were 29.96-599.2 μg/mL（r＝0.999 6），0.832-416 μg/mL（r＝0.999 6），0.102-51 μg/mL（r＝0.999 8），0.286 4-143.2 μg/mL（r＝0.999 8）， 0.286 4- 143.2 μg/mL（r＝0.999 8），respectively. The limits of quantitation were 0.060，0.104，0.017，0.029，0.057 μg/mL；the limits of detection were 0.018，0.031，0.005，0.009，0.017 μg/mL，respectively. RSDs of precision ，stability，reproducibility and durability tests were all lower than 3％. The recoveries were 97.16％-101.88％（RSD＝1.60％，n＝6），96.98％-99.24％（RSD＝ 0.85％，n＝6），97.7％-101.64％（RSD＝1.54％，n＝6），97.77％-103.08％（RSD＝1.82％，n＝6），98.16％-101.88％（RSD＝1.24％， n＝6），respectively. CONCLUSIONS：The established HPLC fingerprint and cluster analysis can be used to evaluate the quality of Chinese gall leaven. The established method is simple to Δ 基金项目：国家重点研发计划项目（No.2018YFC1707200）；公 operate and can be used to determine the contents of 5 益性行业科研专项项目（No.201507004-03）；河南中医药大学研究生 components simultaneously ；rice husk distillers ’grains were 科研创新基金项目（No.YJS2018B14） ＊硕士研究生 。研究方向 ：中药饮片及新药研究 。E-mail： not suitable for fermented Chinese gall leaven . 401327039@qq.com
ABSTRACT
Objective To study and establish the HPLC fingerprint standard for the quality analysis and compare effects on the chemical composition of gallnut by ferment of Chinese gall leaven. Methods The fingerprint of Chinese gall leaven was built by Waters Symmmetry ShieldTM RP18 (250 mm × 4.6 mm, 5 μm) C18 column, and acetonitrile-0.1% trifluoroacetic acid aqueous in gradient as mobile phase, the flow rate was 0.8 mL/min, and the detecting wavelength was set at 280 nm. The chemical fingerprint similarity of 10 batches of Chinese gall leaven was calculated with the Chromap Chromafinger 2005 beta 0.1 standard substance comparison and HPLC-MS were adopted to identify the common peaks. Results The fingerprint chromatography for the 10 batches of Chinese gall leaven included 10 common peaks, with a good separation at each peak. The relative retention time for common peaks of each batch was less than 1.0%, and the similarities among 10 samples were greater than 0.90. Gallic acid (peak 1), (-)-epigallocatechin (EGC, peak 2), methyl gallate (peak 3), ethyl gallate (peak 5), epigallocatechin gallate (EGCG, peak 6), 2,4,6-tri-O-galloyl-β-D-glucose (peak 7), and 2,4,6-tri-O-galloyl-α-D-glucose (peak 9) were identified. The gallnut fermented made the content of gallic acid and 2,4,6-tri-O-galloyl-α-D-glucose increased and the contents of methyl gallate, ethyl gallate, and (-)-epigallocatechin gallate decreased. It was found that (-)-epigallocatechin and 2,4,6-tri-O-galloyl-β-D-glucose had formed in the process for the first time. Conclusion The processing mechanism of Chinese gall leaven is related to gallnut fermentation process change and create new chemical composition and fingerprint can be used to monitor the quality of fermentation processing of Chinese gall leaven.
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As national strategic resources,traditional Chinese medicine (TCM) resource is the material basis for the pharmaceutical industry and health services.The high-efficiency utilization of TCM resource is a major strategic issue that realize the resource conservation and environment friendly recycling economy,guarantee the sustainable development of medicine.But currently,on one hand,the wild TCM resource was seriously damaged and in a serious shortage of stock,the cultivation variety was degenerated,the tending of endangered medicinal materials and the development of alternative varieties were faced many problems.On the other hand,the limited resources cannot be effectively utilized,which results in the waste of resources.Therefore,two ecological restricted resources,poria cocos and Chinese gall,were studied for comprehensive utilization of resources to explore the comprehensive exploitation and utilization of TCM resource as well as the pattern of sustainable development.We suggested that TCM resource should be taken into account as a whole for comprehensive utilization.We should especially pay attention to basic research for the sustainable development of TCM resource,discovery of bioactive substance,excavation and transformation for functional components,the use of biotechnology,the product development,the formation and extension of industry chain.So as the comprehensive exploitation and utilization of TCM resources and sustainable development would be realized.
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Objective: To optimize the processing technology for Chinese gall leaven by orthogonal test. Methods: The L9(34) orthogonal test was used taking the main contents of gallic acid, ellagic acid, and in vitro antibacterial activity as indexes of evaluation. The affecting factors contains factor A, the species of zymocyte, such as Rhizopus koji, Angel yeast, and Aspergillus niger, factor B, the species of tea, such as green tea, black tea, and puer tea, factor C, the material ratios of quantity of raw material, yeast, and tea, such as 25:7.5:2.5, 25:6.25:1.1, and 25:2.5:1.9. Multi-index comprehensive weighted mark method was used in the data analysis. Results: The best processing technology was set at A1B2C1, that strain selection was R. koji, with green tea, and the material ratio was 25:7.5:2.5. Conclusion: The studies show that this processing technology is stable and feasible. The research provides the reference data for processing Chinese gall leaven.
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OBJECTIVE:To study the spermicidal and antibacterial effects of tannic acid from Chinese gall.METHODS:Human sperm samples of 20 subjects were taken for spermicidal tests in vitro in accordance with the standard method recommended by WTO.The minimum inhibitory concentration and minimum bactericidal concentration(MIC and MBC)of tannic acid from Chinese gall on bacterium coli and staphylococcus aureus were detected.RESULTS:The lowest effective spermicidal concentration of tannic acid at 20s was 20mg? mL-1.The MIC and MBC of tannic acid against bacterium coli(3 strains)were 0.195～ 0.390mg? mL-1 and 0.390～ 0.780 mg? mL-1 respectively,against staphylococcus aureus(2 strains)were 0.049～ 0.098mg? mL-1 and 0.195～ 0.390mg? mL-1 respectively.CONCLUSION:Tannic acid from Chinese gall showed potent coagulating effect on spermatines and remarkable antibacterial effect,and which is expected to be an effective and safe vaginal spermicidal and antibacterial agent,further study on which remains to be carried out.