Research On The Correlation Between The Chemical Components And Invigorating Spleen And Promoting Diuresis Therapeutic Effects Of Poria And Its Quality Control

Research background and purpose:Poria,a traditional Chinese medicine,is known for its mild nature,harmonizing the spleen and stomach,promoting diuresis and dampness removal,and invigorating the spleen and kidneys.It has remarkable therapeutic effects,extensive applications,and a large market demand.The current quality standards of Poria in the Chinese Pharmacopoeia are relatively outdated,lacking the determination of indicator components and some unreasonable limit standards,which cannot effectively control its quality.Improving the quality standards of Poria and its quality control methods is of great significance for guiding medicinal material production and ensuring therapeutic effectiveness.Studies have shown that the main active ingredients in Poria are polysaccharides(water-soluble)and triterpenoids,which may be the key components for Poria to exhibit traditional functions such as invigorating spleen and promoting diuresis.They can be used for the quality control of Poria.Studies found that the amino acids and trace elements in Poria may be closely related to the content of active ingredients and quality of the herbs.To establish a comprehensive and accurate quality control method for Poria,the author studied the components that contribute to its traditional functions.Through a variety of subjective and objective analysis methods,a method for comprehensively evaluating the quality of Poria based on active ingredient was established,and the differences in components and quality of Poria from different producing areas were compared.The relationships between various components in Poria were studied,and the rationality of current quality standards for Poria was discussed.These achievements provide a basis for improving the quality standards and optimizing the quality control methods for Poria to ensure the therapeutic effects of Poria.Materials and Methods:1.Efficacy StudyA rat model of spleen deficiency and dampness obstruction was established using the "purgative and qi-breaking + irregular diet" method,in which the purgative and qi-breaking medicine was Xiao Cheng Qi Decoction(Magnolia officinalis: Aurantii immaturus: raw Rheum officinale = 3:3:4).We used total protein(TP),albumin(ALB),D-xylose absorption capacity,water metabolism capacity,and organ indices as measurement indicators to study the diuretic and spleen-invigorating effects of various active components in Poria.We established a method for comprehensive efficacy evaluation and compared the efficacy differences of principal active components in Poria to preliminarily explore the optimal effective doses of each component.2.Component StudyWe conducted a study on 13 batches of Poria samples from different main production areas and determined the content of moisture,total ash,extract,poria polysaccharides(PAC),total Poria triterpenes,eight triterpenic acids,trace elements,and amino acids.We established quantitative analysis methods for multiple components in Poria and established an HPLC fingerprint of Poria triterpene components.We applied correlation analysis,principal component analysis(PCA),cluster analysis,and gray relational degree analysis(GRA)to compare the component differences among different batches of Poria samples and to explore the relationships among the components.3.Comprehensive Evaluation of Poria QualityBy combining subjective and objective weighting methods,we used the Analytic Hierarchy Process(AHP),CRITIC method,and entropy method to establish a comprehensive evaluation method for Poria quality,determined the weight coefficients of each component of Poria,and used the weighted sum method to calculate the scores to compare the quality differences of Poria medicinal materials from different production areas and batches.4.Improvement of Poria Quality StandardsCombining the quality standards of Poria in multiple countries and regions,we discussed the existing problems in the Poria quality standards in the current Chinese Pharmacopoeia,proposed relevant suggestions,and promoted the improvement of Poria quality standards.Result:1.Significant differences in related indices were observed between the blank group and the model group of rats after modeling,indicating the successful establishment of a rat model of spleen deficiency and dampness obstruction.Water-soluble polysaccharide extracts and total triterpene extracts from different doses of Poria showed improvement effects on the water metabolism rate,serum D-xylose content,spleen and thymus indices,total protein(TP),and albumin(ALB)content in rats with spleen deficiency and dampness obstruction.The comprehensive scoring results indicated that poria polysaccharides(PAC)extract exhibited stronger spleen-invigorating and diuretic effects than Poria triterpene extract.2.Accurate,efficient,and high extraction rate quantitative analysis methods for poria polysaccharides(PAC)and triterpene active components were established,as well as an HPLC analysis method for simultaneously determining multiple triterpenic acids in Poria.The precision,stability,and repeatability of these methods met the relevant requirements.An HPLC fingerprint of Poria triterpene components was established,with methodological validation results and similarity judgment results meeting the technical requirements.3.The component research results of Poria showed that the highest chemical component content in Poria is alkaline-soluble polysaccharides,up to 80%～90%,followed by water-soluble polysaccharides,accounting for approximately 8% of the total mass.Poria triterpenes only account for 0.4%～0.7%,with the highest content being pachymic acid,ranging between 0.06% and 0.13%.The amino acid content in Poria is around 0.8 g/100 g,with aspartic acid,glutamic acid,and serine being the highest in content.The trace element content in Poria ranges from 192.68 to 885.04 mg/kg,with K,Mg,and Ca having the highest content,with differences up to several tens of times.Statistical analysis results showed that there were significant correlations between multiple components in Poria,and there were no obvious regional differences in the chemical components of Poria medicinal materials.4.The consistency of both subjective and objective weighting methods used met the requirements and was suitable for comprehensive evaluation of Poria quality.The objective weighting method had the highest weight index for extract and alkaline-soluble polysaccharides,which was related to the large differences in the measured values of the related indicators of each batch of Poria.The CRITIC method and entropy method in the objective evaluation method were able to better distinguish between different batches of samples.The final weight results of each indicator were:moisture 8.019%,total ash 7.194%,extract 16.948%,water-soluble polysaccharides20.433%,alkaline-soluble polysaccharides 10.955%,total triterpenes 19.284%,and 8triterpenic acids 17.168%.The weight coefficients of active components,water-soluble polysaccharides,and total triterpenes were the highest.The comprehensive scoring results showed that the quality of the collected Poria samples was in the following order from high to low: S7 > S12 > S8 > S3 > S2 > S10 > S1 > S11 > S5 > S4 > S6 > S9 >S13.5.According to current standards,the total ash content of commercially available Poria is far lower than the specified value,and occasionally,the water content and extract contents do not meet the standards.Referring to domestic and foreign standards,the author believes that the total ash limit set at ≤1% is more reasonable,and the limit on pachymic acid content(≥0.04%)still needs to be increased.The limits on moisture(≤18%)and extract content(≥2.5%)can remain unchanged,and restrictions on the amount of Poria polysaccharides and triterpenoids should be added,but specific values still need further research.Conclusion:This study successfully established a rat model of spleen deficiency and damp obstruction.The pharmacological research results showed that the spleen-strengthening and diuretic effects of poria polysaccharides(PAC)are superior to those of Poria triterpenes.The established component analysis and fingerprinting methods meet the relevant technical requirements.The measurement results show that there are significant differences in the content of various components in Poria,and multi-index analysis results show significant correlations between multiple components in Poria.There are no significant regional differences in the chemical composition of Poria,making it difficult to distinguish Poria samples from different production areas through chemical composition.This study has established a method for comprehensively evaluating the quality of Poria through multiple indicators,among which water-soluble polysaccharides,total triterpenes,8 triterpenic acids,and extracts have a significant impact on the overall quality assessment score.The scoring results show that the quality of commercially available Poria varies greatly,with the top three being Yunnan,Hubei,and Sichuan Poria.The researchers suggest revising the quality standards for Poria,referring to the Taiwan Pharmacopoeia.The standards should include Poria acid as an indicator,and increase its specified value(originally 0.04%).The total ash content should be reduced to 1%,and regulations for Poria polysaccharides and triterpenes should be added.