| [Objective] Objective : Optimizing the processing technology of S.glabra,standardizing the production of traditional Chinese medicine decoction pieces,providing relevant process parameters for the production of S.glabra decoction pieces,improving the quality evaluation system of S.glabra,and improving the clinical application effect of traditional Chinese medicine decoction pieces;Looking for the best fresh processing method,the pilot study verified the feasibility of the processing technology of S.glabra and the implementation of the fresh processing technology,providing certain reference for the integration of S.glabra processing and production area processing;The concept of pharmacodynamic equivalence was introduced to explore the feasibility of S.glabra fresh processing.It provides new ideas for the development and utilization of S.glabra and provides certain theoretical basis for its quality evaluation.Method:1.On the basis of single factor experiment,the “comprehensive score” of neochlorogenic acid,chlorogenic acid,4-dicaffeoylquinic acid,caffeic acid,Isofraxidin,Astilbe chinensis glycoside and rosmarinic acid content was used as evaluation index by AHP,and drying temperature and drying time were used as influencing factors.The purification process of S.glabra leaves was determined by single factor investigation.Taking soaking time,moistening time,cutting length and drying temperature as influencing factors,the optimal cutting process of S.glabra stems was optimized by central composite design-effect flour method.Multi-index analytic hierarchy process(AHP)is used to analyze and verify the optimal process.2.Establish ultra-high phase liquid chromatography(UPLC)fingerprint of S.glabra,compare 6 effective components of 15 batches of S.glabra at the same wavelength,SPSS18.0 analysis software was used for quality cluster analysis and principal component analysis of S.glabra.To provide scientific basis for quality evaluation and control of S.glabra.3.On the basis of traditional processed methods,UPLC optimized the best fresh processed technology of S.glabra.Three batches of fresh S.glabra were tested in small scale,and the contents of 7 effective components were compared according to different processing methods.According to the actual situation of Chinese herbal medicine production enterprises,a batch of Chinese herbal medicine production workshop certified by GMP of Jiangxi Guhan Refined Chinese Herbal Medicine Company was selected for pilot production to verify the pilot production process of fresh processing of S.glabra pieces.4.Headspace gas chromatography-mass spectrometry(HS-GC-MS)was used to analyze the volatile oil components of traditional herbal pieces and fresh processed herbal pieces of S.glabra.5.The anti-inflammatory and analgesic effects of the decoction pieces processed by the two methods were compared with mouse pain induced by acetic acid and mouse auricle inflammation induced by xylene respectively.Using S.glabra anti-inflammatory experiment,the equivalence of anti-inflammatory efficacy of two processing decoction pieces was studied by fitting the regression equation of equal effect dose(or equal dose effect).Results:1.The optimized processing method of S.glabra is as follows: the stem and leaf of S.glabra are separated,the stem is cleaned with water once,soaked in 12 times of water for 1 h,moistened and softened for 3 h,cut into 15 mm segments after taking out,and baked at 50℃ for 2 h;Cleaning leaves with water once,drying at 50℃ for 4 h;If the stem and leaf are used together as medicine,Mixing the processed S.glabra stems and leaves at a ratio of 2: 1.2.UPLC fingerprints of 15 batches of S.glabra samples were established and studied by similarity evaluation,cluster analysis and principal component analysis.The fingerprint similarity of S.glabra decoction pieces is between 0.782 ~ 0.995,and 13 common peaks are determined.Cluster analysis results are basically consistent with principal component analysis results.3.The optimized fresh processing technology of S.glabra is as follows: leaves and stems of S.glabra are separated,leaves are cleaned and dried at 50℃ for 6 h;After stem cleaning,cut into 15 mm long sections and dry at 50℃ for 9 h.The OD value of fresh processed decoction pieces is higher than that of traditional processed decoction pieces in laboratory trial process.The quality of S.glabra pieces produced in pilot-scale production has reached the requirements under the 2015 edition of "Chinese Pharmacopoeia".The average comprehensive score of fresh processed pieces is 8.3593,and that of traditional processed pieces is 5.8284.4.There are 41 volatile oil components in the stems of S.glabra processed by the two methods,and 0.25 ml of the stems of S.glabra processed fresh are collected,accounting for 90.24% of the total volatile oil components;Collecting 0.2 ml of traditional processed S.glabra stems,accounting for 84.64% of the total volatile oil components;Collecting 0.9ml of fresh processing S.glabra leaves,and the fresh S.glabra leaves accounted for 93.40% of the total volatile oil components.collecting 0.7ml of traditional processed S.glabra Leaves,accounting for 92.15% of the total volatile oil components.5.Compared with the model control group,fresh processed S.glabra decoction pieces can significantly reduce the number of writhing reactions caused by glacial acetic acid in mice,inhibit xylene-induced auricle inflammatory reactions in mice,and relieve auricle swelling degree;The anti-inflammatory and analgesic effects of fresh processed S.glabra pieces are more obvious than those of traditional processed pieces.The fitting equation of dose-effect curve is Y(%)= 4.753 X(g/kg)+29.515(R2 = 0.586,R=0.766,P <0.01).The fitting equation of traditional processed decoction pieces is Y(%)=13.052 In X(g/kg)+ 26.920(R2=0.546,R=0.739,P <0.01).The anti-inflammatory effect equivalence results show that in the range of 34.862%~55.596% anti-inflammatory effect,DEE the fresh processed slices(g/kg)= 0.615× DEE the traditional processed slices(g/kg)+ 0.453(R2 = 0.959,R = 0.979,p <0.01);Within the dose range of 1.125 g/kg~9.0 g/kg mice,EED the fresh processed slices(%)= 1.390× EED the processed slices(%)?10.544(R2 = 0.927,R = 0.963,P <0.01).Conclusion:The optimized processing technology is simple,stable and feasible.The obtained parameters can provide scientific basis for research and production of S.glabra,and AHP can provide reference for multi-index evaluation of traditional Chinese medicine.The UPLC fingerprint of S.glabra decoction pieces combined with cluster analysis can effectively distinguish different batches of S.glabra and provide reference basis for identification and quality control of S.glabra decoction pieces.Response Surface Methodology was used to optimize the processing technology of S.glabra and the processing technology of fresh S.glabra were simple and stable,and the pilot production results were stable and feasible.HS-GC-MS combined technology can be effectively used to determine the volatile oil components of S.glabra.The fresh processed decoction pieces and the traditional processed decoction pieces have obvious anti-inflammatory and analgesic effects,and the fresh processed decoction pieces have better effect and have the feasibility of fresh processing.Studies on the equivalence of anti-inflammatory and pharmacodynamic effects in mice show that the equivalent dose(equivalent dose effect)of fresh processed slices and traditional processed slices is not equivalent,while the equivalent dose of fresh processed slices is smaller than traditional processed slices,while the equivalent dose effect of fresh processed slices is larger than traditional processed slices.It can provide certain reference for the integration of S.glabra processing and production area processing,and has certain popularization and application value. |
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