Study On Commodity Specification And Grade Standard Of Phyllanthi Fructus Based On "Three Step Classification"

Phyllanthi Fructus is a dry and mature fruit of Euphorbiaceae plant Phyllanthus emblica L.It is a characteristic medicinal material of Chinese and Tibetan medicine.It has the effects of clearing heat and cooling blood,eliminating food and strengthening stomach,generating body fluid and relieving cough.Because of its strong pharmacological activity,rich nutritional value and unique flavor,it has been listed in the list of"medicine and food homology"by the Ministry of health,and is the key variety of Rural Revitalization in Panxi area of Sichuan Province.With the development of Phyllanthi Fructus health products more and more,the lack of domestic Phyllanthi Fructus commodity specification and grade standards,the market circulation of sub file bargaining,high quality and good price lack of scientific basis.Appearance character is an important index for grading commodity specifications,and the processing of Phyllanthi Fructus from different producing areas has great influence on appearance characters such as color,texture and size.Therefore,it is of great significance to carry out the research on the commodity specification and grade standards and processing methods of Phyllanthi Fructus,and formulate the commodity specification and grade standards of Phyllanthi Fructus,so as to standardize and improve the commodity quality of Phyllanthi Fructus and help the healthy development of Phyllanthi Fructus industry.Based on the"three-step grading"method,the commodity specification grade standard of Phyllanthi Fructus was established,and on this basis,the innovation of processing method of Phyllanthi Fructus was discussed.The main research contents and results are as follows:（1）Evolution and market investigation of Materia MedicaAfter consulting and studying the ancient Chinese Materia Medica,modern published monographs and relevant standards,it is found that Phyllanthi Fructus is originated from Euphorbiaceae plant Phyllanthi Fructus,which has a complex historical and ethnic application background.The main producing areas of Phyllanthi Fructus are tropical and subtropical areas such as Guangdong,Guangxi,Hunan,Jiangxi and India.Based on the field investigation and research of Hehuachi medicinal materials market and Yunnan production area,the results show that In the market,they are sold by“Tonghuo”or“Routong”（pulp of Phyllanthi Fructus）,and the price hovers around 4-30 yuan/kg for many years.The quality standard mainly refers to the Chinese Pharmacopoeia,and there is a lack of commodity standard and related research.（2）Determination of mycotoxins in moldy Phyllanthi FructusIn order to find out whether the moldy Phyllanthi Fructus can produce highly toxic mycotoxins,we established a method for the determination of 22 mycotoxins in Phyllanthi Fructus based on ultra performance liquid chromatography triple quadrupole mass spectrometry methods.The samples of suspected moldy Phyllanthi Fructus were screened and determined.The results showed that the commercial Phyllanthi Fructus basically did not contain aflatoxin and ochratoxin,and the level of exogenous pollutants was very low,which could not be controlled.（3）Commodity specification division of Phyllanthi FructusThe core of Phyllanthus emblica accounts for nearly half of the total.Whether the core is removed or not becomes the key issue of the commodity specification of Phyllanthi Fructus.Literature,medicinal material standards and Market Research on the appearance of Phyllanthi Fructus were conducted based on existing databases;ultra-high performance liquid chromatography-quadrupole-electrostatic field orbital trap high resolution mass spectrometry（UPLC-Q-Orbitrap HRMS）technology was used to identify the constituents of fruit core and pulp.The antibacterial effects of Phyllanthi Fructus fruit core and pulp on Escherichia coli and Staphylococcus aureus were investigated by filter paper method.The antimicrobial activity of the core and pulp of Phyllanthi Fructus on Escherichia coli and Staphylococcus aureus were investigated by filter paper method and their antioxidant activity was compared by DPPH free radical scavenging test.The results showed that a total of 47 compounds were identified in core and pulp of Phyllanthi Fructus,mainly including tannins,flavonoids,phenolic acids,fatty acids,amino acids,organic acids,sugars and glycosides,most of which were mainly concentrated in the pulp,and the fatty acids in the nucleus accounted for a higher proportion.The contents of gallic acid,corilagin,chebulagic acid,ellagic acid and other phenolic compounds in the pulp of 20 batches of Phyllanthi Fructus were much higher than those in the core（2-50 times）.The results of antibacterial test showed that the core of different concentrations had no antimicrobial effect.The results of DPPH radical scavenging test showed that the antioxidant activity of core(the half maximal inhibitory concentration(IC₅₀):199.632μg·m L^-1)was much less than that of pulp(IC₅₀:12.688μg·m L^-1)（4）Commodity classification of Phyllanthi FructusThe diameter,thickness,weight of each grain,L^*,a^*,b^*values of pericarp color were taken as the appearance indexes for quantitative measurement.The contents of gallic acid,corilagin,chebulagic acid and ellagic acid were determined by HPLC.Descriptive statistical analysis,variance and principal component analysis were used to determine the importance of the main signs,and to explore the correlation between the appearance index and the internal components.Further formulate the commodity specification and grade standard of Phyllanthi Fructus.The Hep G2 cell experiment of liver cancer was used to verify the scientific nature of the commodity grade classification standard.The results of correlation analysis showed that L^*,a^*,b^*values of epidermis color were significantly negatively correlated with corilagin,chebulagic acid and ellagic acid（|r|>0,P<0.01）,but not with gallic acid（|r|<0.1）.Considering the coefficient of variation,the results of principal component analysis and the content of gallic acid as the quality control index of Phyllanthi Fructus in the Chinese Pharmacopoeia,it was determined that the content of gallic acid was one of the grading indexes in addition to the skin color L^*,a^*,b^*values.when the pericarp color L^*<44,a^*<7,b^*<10,and the content of gallic acid>1.6%,it can be judged as the first-class product;when the above conditions can not be met at the same time,it can be determined as the second-class product.The Hep G2 cell test of liver cancer showed that IC₅₀ of the first-class product was lower than that of the second-class product.Using the pericarp color L^*,a^*,b^*values to develop a color comparison card,which provides a visual tool for on-site evaluation of Phyllanthi Fructus commodity grade.