Preparation Process And Quality Evaluation Of Sanqi Decoction Slices Based On Fresh Steaming

Panax Notoginseng（Burk.）F.H.Chen is a plant of the Panax genus in the Acanthopanax family,widely distributed in Yunnan,Guangxi,and other places.Its dry roots and rhizomes are often used as the main medicinal parts.As a traditional and precious Chinese medicinal material in China,Panax notoginseng is said to be"raw and cooked to nourish".After being processed,its hemostatic and stasis dispersing functions are weakened and its blood enriching functions are enhanced.Raw Panax notoginseng powder and its extract,Panax notoginseng saponins,have been widely used in the prevention and treatment of cardiovascular and cerebrovascular diseases,but the blood tonic effect of Steamed Panax Notoginseng has rarely been developed.Currently,there are only a few products such as Steamed Panax Notoginseng powder（including drugs and pieces）,Steamed Panax Notoginseng pills,and the processing technology is to use dried Panax notoginseng to beat powder and then evaporate.The2020 edition of the Chinese Pharmacopoeia contains a simple specification for Panax Notoginseng decoction pieces,with only Panax Notoginseng powder and no standard for steamed Panax notoginseng.Red Notoginseng（RPN）is a processed product of fresh roots and rhizomes of Panax Notoginseng after being steamed.Processing while fresh has great potential for ensuring the quality of medicinal materials,reducing production costs,and promoting industrial development.Therefore,this thesis takes the fresh processing of Panax notoginseng from its producing area as the starting point,develops the preparation process for the prepared pieces of Panax notoginseng,establishes the quality standards for the prepared pieces of Panax notoginseng,and conducts a comparative study on the blood enriching effect and metabonomic mechanism of the prepared pieces of Panax notoginseng and traditional steamed Panax notoginseng,providing theoretical support for the fresh processing of Panax notoginseng from its producing area,the development and clinical application of new types of Panax notoginseng.The main elements of this thesis study are as follows.:1.Study on the preparation process of Red NotoginsengAfter cleaning and slicing fresh Panax notoginseng,the slice thickness of Panax notoginseng was determined based on the total content of the main saponin components（R₁,Rg₁,Re,Rb₁,Rd）in raw Panax notoginseng.Using fresh Panax notoginseng thick tablets as raw materials,the total content of the main saponin components（R₁,Rg₁,Rb₁）in raw Panax notoginseng and the content of the blood tonic related saponin components（Rk₃,Rh₄,and 20-（R）-Rg₃）in Steamed Panax Notoginseng were selected as evaluation indicators to determine the steaming temperature,steaming time,and drying time and temperature.Based on the single factor experimental results,Based on the Box-Behnken response surface method,the optimal processing conditions for preparing Red Notoginseng were obtained as follows:steaming temperature 130℃,steaming time 4h,drying temperature 60℃,drying time 48h.Under this process condition,the total saponin content obtained was8.326%,and the RSD was 0.087%.After verification,this method is stable and feasible,and can be used for the standardized production of Red Notoginseng slices,providing a basis for the fresh processing and development of Red Notoginseng slices in the origin of notoginseng.2.Study on the quality standard of Red NotoginsengUsing the prepared slices of Red Notoginseng produced by optimizing the process as raw materials,referring to the relevant provisions of the Chinese Pharmacopoeia（2020）and the Steamed Panax Notoginseng powder standard of Yunnan Provincial Drug Administration,the microscopic and thin layer chromatographic identification,content,moisture,alcohol soluble extracts,and other indicators of the prepared slices of Red Notoginseng were tested,and it was determined that the total content of ginsenosides Rk₃,Rh₄,and 20-R-Rg₃in the prepared slices of Red Notoginseng should not be less than 5.4%.The content of alcohol soluble extracts in the prepared slices of Red Notoginseng shall not be lower than 29.0%and other control indicators,and based on this,a preliminary quality standard（draft）for the prepared slices of Red Notoginseng shall be established to provide a scientific basis for the quality evaluation of the prepared slices of Red Notoginseng.3.Pharmacodynamic Study on the Treatment of Blood Deficiency by Red Notoginseng and Traditional Steamed Panax Notoginseng PiecesA mouse model of blood deficiency was established using acetylphenylhydrazine（APH）and cyclophosphamide（CTX）.The blank group,the blood deficiency model group,the positive（FEJ）group,the Red Notoginseng decoction piece（1.2 g/kg,0.6g/kg,0.3 g/kg）treatment group,and the traditional Steamed Panax Notoginseng decoction piece（1.2 g/kg,0.6 g/kg,0.3 g/kg）treatment group were established,respectively.The physical signs of the mice were observed,and the body mass of（1）mice was measured;（2）Peripheral blood picture;（3）Organ index;（4）IL-6 and IL-1 in serumβ、TNF-α、Bcl-2,IL-2,TPO,EPO,G-CSF,M-CSF,GM-CSF,and other related factors;（5）Pathological section of spleen;（6）Expression of CD4+and CD8+in spleen tissue;（7）Expression of JAK/STAT pathway related m RNA in spleen tissue;（8）The expression of JAK/STAT pathway related proteins and other evaluation indicators were tested to investigate the difference in blood tonic effects between Red Notoginseng and traditional Steamed Panax Notoginseng slices.Research has found that both Red Notoginseng and traditional Steamed Panax Notoginseng can treat blood deficiency caused by APH-CTX,and the blood tonic effect of Red Notoginseng decoction pieces is better than that of traditional Steamed Panax Notoginseng decoction pieces.Among them,the high-dose group of Red Notoginseng decoction pieces can significantly improve the changes in physical signs caused by blood deficiency in mice,reduce body mass,reverse splenomegaly,improve the decrease in blood routine indicators（WBC,RBC,Hb,PLT）caused by modeling,and upregulate the positive expression of CD4⁺and CD8⁺in mice spleen;At the same time,it can activate the JAK/STAT signaling pathway and significantly increase the serum hematopoietic related factors（IL-2,TPO,EPO,G-CSF,M-CSF,GM-CSF,etc.）and immune cytokines（IL-6,IL-1β,TNF-α,etc.）.While improving the hematopoietic capacity of mice and enhancing their immune capacity,the two have a synergistic effect,thereby exerting their blood tonic effect.4.A metabonomic study of Red Notoginseng and traditional Steamed Panax Notoginseng slices in the treatment of blood deficiencyUsing the plasma of a mouse model of blood deficiency caused by acetylphenylhydrazide-cyclophosphamide as the experimental material,and using UPLC-Q-TOF-MS/MS as the platform,metabonomic analysis methods were used to detect the plasma metabolites of each group of mice.To screen metabolites through PCA,PLS-DA,OPLS-DA,and other analytical methods,identify differential metabolites and related metabolic pathways,and explore the potential metabolic pathways and mechanisms of Red Notoginseng and traditional Steamed Panax Notoginseng decoction pieces in enriching blood.22 differential metabolites such as L-valine,L-leucine,and tryptophan were identified in plasma.The results of KEGG enrichment analysis showed that these endogenous metabolites can regulate metabolic pathways such as cholesterol metabolism,linoleic acid metabolism,phenylalanine,tyrosine,and tryptophan biosynthesis,protein digestion and absorption,and vitamin digestion and absorption.Among them,the prepared pieces of Red Notoginseng mainly regulate cholesterol metabolism and linoleic acid metabolism,while the prepared pieces of Steamed Panax Notoginseng mainly regulate cholesterol metabolism,playing a therapeutic role in mice with blood deficiency caused by APH-CTX.