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@#【Objective】 To study the effects of soil environment on the growth, yield, and quality of Sharen (Amomi Fructus) under different planting patterns. 【Methods】 Soil physical and chemical indices and enzyme activities in four periods including early flowering (March), full flowering (June), fruit ripening (September), and late fruit picking (December), were measured under three planting patterns including natural forest, greenhouse, and rubber forest in Xishuangbanna, China. The changes in soil indices during the growth periods of Sharen (Amomi Fructus) under different planting patterns were analyzed, and the differences in plant growth, yield, and quality under different planting patterns were explored. Pearson correlation analysis was used to analyze the relationship between soil indices and Sharen (Amomi Fructus) growth, yield, and quality. Principal component analysis was used to investigate the effects of soil environment under different planting patterns on Sharen (Amomi Fructus) growth, yield, and quality. 【Results】 The soil moisture, available potassium content, and urease activity of the three planting patterns of Sharen (Amomi Fructus) increased initially and decreased afterwards throughout the year; pH and organic matter content showed little change in the whole year. Exchangeable manganese content and acid phosphatase activity gradually increased throughout the year. Hydrolyzed nitrogen content, exchangeable calcium content, available zinc content, protease activity, and sucrase activity decreased initially and increased afterwards throughout the year. Exchangeable magnesium content, available iron content, and catalase activity decreased annually. Total nitrogen content, total phosphorus content, and available phosphorus content fluctuated throughout the year. The total potassium content under natural forest and greenhouse planting decreased throughout the year, while the total potassium content under rubber forest showed an upward trend all year round. The organic matter content, total nitrogen content, total potassium content, available potassium content, available zinc content, urease activity, acid phosphatase activity, and catalase activity under greenhouse were significantly lower than those under natural and rubber forests (P < 0.05). Correlation analysis showed that plant growth, yield, and quality of Sharen (Amomi Fructus) were significantly correlated with soil organic matter, total nitrogen, hydrolyzed nitrogen, total phosphorus, available phosphorus, total potassium, available potassium, exchangeable manganese, exchangeable magnesium, exchangeable calcium, available zinc, urease, acid phosphatase, and invertase (P < 0.05). The results of the principal component analysis indicated that the soil environment of Sharen (Amomi Fructus) under natural forest was the best, followed by rubber forest and greenhouse. The order of its advantages and disadvantages is consistent with the growth index of Sharen (Amomi Fructus), but contrary to the yield of Sharen (Amomi Fructus), indicating that the soil environment directly affects the growth index and nutritional components of plants. 【Conclusion】 Different planting patterns of Sharen (Amomi Fructus) have different soil nutrient content, and the change rules in the growths period are not similar, with some differences. Soil indices have impacts on plant growth, yield, and quality of Sharen (Amomi Fructus). Soil ecological environment is positively correlated with the growth characteristics of Sharen (Amomi Fructus) plants, but has no direct correlation with yield and quality.
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ObjectiveTo investigate the relative content changes of differential metabolites and reducing sugars during the processing process of Rehmanniae Radix Praeparata （RRP） processed with Amomi Fructus （AF） and Citri Reticulatae Pericarpium （CRP）， and to lay the foundation for revealing the processing principle of this characteristic variety. MethodThe samples of the 0-54 h processing process of RRP processed with AF and CRP were taken as the research object， and their secondary metabolites were detected by ultra performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry （UPLC-Q-TOF-MS）. The 0.1% formic acid aqueous solution （A）-acetonitrile （B） was used as the mobile phase for gradient elution （0-1 min， 1%-3%B； 1-10 min， 3%-9%B； 10-15 min， 9%-12%B； 15-22 min， 12%-18%B； 22-31 min， 18%-24%B； 31-35 min， 24%-100%B； 35-36 min， 100%-5%B； 36-40 min， 5%-1%B； 40-45 min， 1%B）， column temperature was 40 ℃， injection volume was 3 μL， flow rate was 0.3 mL·min-1. Electrospray ionization （ESI） was used to scan and collect MS data in the negative ion mode， the scanning range was m/z 50-1 250. Data analysis was carried out using PeakView 1.2 software， and the chemical composition of RRP processed with AF and CRP was identified by combining the literature information and chemical composition databases. The MS data were normalized by MarkerView 1.2， and then the multivariate statistical analysis was applied to screen the differential metabolites， and the changes of the relative contents of the differential metabolites with different processing times was analyzed， finally， correlation analysis was performed between the differential metabolites， the change of the reducing sugar content was combined to determine the most suitable processing time of RRP processed with AF and CRP. ResultA total of 121 compounds were identified from RRP processed with AF and CRP at different processing times， and 12 differential metabolites were screened out by multivariate statistical analysis， including catalpol， hesperidin， isoacteoside， acteoside， narirutin， echinacoside， isomartynoside， decaffeoylacteoside， 6-O-E-feruloylajugol， dihydroxy-7-O-neohesperidin， jionoside D， and rehmapicroside. With the prolongation of processing time， the relative contents of these 12 differential metabolites and reducing sugars changed slightly at 52-54 h. ConclusionUPLC-Q-TOF-MS can comprehensively and accurately identify the chemical constituents of RRP processed with AF and CRP at different processing times， and the suitable processing time of 52-54 h is determined according to the content changes of different metabolites and reducing sugars， which provides a basis for revealing the scientific connotation of the processing principle of this variety.
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ObjectiveTo investigate the quality of Amomi Fructus in the market， and to compare the difference between the seed mass and shell， so as to provide a basis for standardizing the usage of Amomi Fructus. MethodThe properties， thin layer identification， moisture， the content of bornyl acetate were determined by the methods in the 2020 edition of Chinese Pharmacopoeia， and the ash and extract content were determined according to the collection method of the 2020 edition of Chinese Pharmacopoeia. ResultAmong the 17 batches of samples， except the content of bornyl acetate in 2 batches of Amomum longiligulare， 2 batches of A. longiligulare and A. villosum mixture was lower than the standard， the quality of other samples all met the standard of the 2020 edition of Chinese Pharmacopoeia， but there were two specifications with shell and without shell. The husk rate， volatile oil， extract and bornyl acetate contents of the seed mass and shell were tested. It was found that the content of volatile oil in three kinds of Amomi Fructus seed mass was 1.8-5.3 times that of the corresponding shell， and the content of bornyl acetate in the seed mass was 8.8-62.1 times that of the corresponding shell， but there was little difference in the extract content. ConclusionBased on the above research， it is considered that the content of bornyl acetate in A. longiligulare contained in the 2020 edition of Chinese Pharmacopoeia remains to be discussed. It is tentatively determined that the total ash content of Amomi Fructus should not be more than 10.0%， and the extract content should not be less than 15.0%. At the same time， it is suggested that when Amomi Fructus is used as medicine， the dosage of Amomi Fructus should be calculated according to the removal rate of 20%-30% of shell， and it should be crushed regardless of whether it is used in shell or not.
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Chinese medicines of Doukou includes Amomi Fructus Rotundus， Alpiniae Katsumadai Semen， Galangae Fructus and Myristicae Semen. They have a long medicinal history and are also commonly used in cooking and seasoning. Due to the similar names and limited to the traffic conditions in ancient times， the records of Doukou in ancient literature are often confused with many plants in the same family， and there are still many kinds of confused products. In order to promote the development of famous classical formulas containing the medicinal materials， the ancient literature of Doukou in the past dynasties was comprehensively combed from the aspects of name， origin， genuine area， medicinal parts， harvesting and processing and processing methods. It has been found that the basic original plants of Amomi Fructus Rotundus are Amomum kravanh and A. compactum， the original plant of Alpiniae Katsumadai Semen is Alpinia katsumadai and it often confused with Tsaoko Fructus. The main source of Galangae Fructus recorded in the ancient materia medica is the fruit of A. officinarum， while the 2020 edition of Chinese Pharmacopoeia stipulates that the original plant is A. galanga. Myristica fragrans is the original plant of Myristicae Semen. It was found that except M. fragrans， the other three kinds of medicinal origin of Doukou had changed， there are many other plants confused with each other. The four kinds of Doukou are produced in Southeast China and Southeast Asia， and most of Amomi Fructus Rotundus and Myristicae Semen are imported. The Chinese medicines of Doukou have clear medicinal parts and simple processing methods， the main methods in the past dynasties are cleaning， stir frying and simmering， and the processed products are selected according to the needs of different diseases. It is suggested to use the dry mature seeds of A. katsumadai in Houpo Wenzhongtang， which is from Guangxi， Guangdong， Hainan， Fujian and Yunnan and so on， among which Wanning in Hainan province is genuine producing area. The fruits should be harvested in summer and autumn， and dried to 90% dry in the sun， or slightly scalded with water and dried to half dry in the sun， and removed the peel， taken out the seed group， dried in the sun and then be used as medicine.
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Objective: To study the powder properties and powder modification technology of Shenling Baizhu Pulvis (SBP), so as to lay a foundation for the study of the suitability of powder modification technology in the solid preparation of traditional Chinese medicine. Methods: The characterization and evaluation methods of powder in the field of materials science and pharmaceutical science were used for reference to evaluate the grouping of single medicinal materials in the particle design and the process rationality of composite particles. The preparation process of composite particles of SBP was optimized by L9(34) orthogonal test, and the surface properties of the composite particles were evaluated by SEM, IR and XRD. Results: The study on the powder properties of prescription raw materials showed that there was a good correlation between the grinding time and the particle size. Finally, the best process for composite particles was as following: the pulverization temperature for powders of Ginseng Radix et Rhizoma, Dioscoreag Rhizoma, Nelumbinis Semen, Lablab Semen Album, Coicis Semen and Platycodonis Radix was -10 ℃ for 45 min, and then pulverization for another 4 min after adding with Atractylodis Macrocephalae Rhizoma, Poria, Amomi Fructus and Glycyrrhizae Radix et Rhizoma. The results showed that the composite particles were well formed and the preparation process was stable and feasible. Conclusion: The powder modification technology solves the powder defects in the preparation process of traditional powder, which provides experimental basis for powder modification technology to improve the quality of traditional Chinese medicine solid preparation and promote the development and upgrading of powder, pill and other traditional dosage forms.
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Objective:To study the change rule of decocting quantity of the effective components in Amomi fructus and Amomi fructus rotundus with decocting time to determine whether or not decocted later and optimal decocting time. Methods:The herbs were extracted by the traditional water decoction, and at different time points, sampling was carried out. Using camphor and eucalyptol as the index components, the change rule of decocting quantity of the effective components with the decoction time under the condition of single and combined decoction was investigated. Results:When the decoction time of Amomi fructus was within the range of 3-6 min, the total amount of camphor in the decoction reached relatively high value, and the total amount lost more than 45% when the decoction time exceeded 10 min. Amomi fructus rotundus boiled for a short time below 2 min, and when the decoction time was more than 5 min, more than 50% eucalyptol lost. Conclusion:Amomi fructus and Amomi fructus rotundus should be decocted later with decocting time within 3-6min and below 2 min, respectively. The analytical method is reliable and precise in the quality control of relative decoction.
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Objective To establish a GC method for simultaneous determine of camphor, borneol and bornyl acetate in Shenling Baizhu Powder. Methods Capillary column was used with 100% methyl polysiloxane as stationary phase. The temperature-programmed was as follows:starting temperature of 60 ℃, 5 ℃/min, rose to 130 ℃, kept for 5 minutes, followed by 20 ℃/min up to 230 ℃, and kept for 5 minutes. Results The average recovery rate of camphor was 99.87%, RSD was 1.3%, and good linear relationship was showed in the range of 0.003 77-0.150 8 μg (r=0.999 9). The average recovery of borneol was 100.71%, RSD was 2.1%, and good linear relationship was showed in the range of 0.002 411-0.096 4 μg (r=0.999 9). The average recovery rate of bornyl acetate was 101.95%, RSD was 1.3%, and good linear relationship was showed in the range of 0.003 02-0.121 1 μg (r=0.999 9). Conclusion The method is simple, reliable, accurate, and can effectively control the quality of Amomi Fructus in Shenling Baizhu Powder.
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Objective: To evaluate the genetic diversity of germplasm resources for Amomi Fructus from various habitats in different phenotypes. Methods: Inter-simple sequence repeats (ISSR) markers were used to analyze the genetic diversity and the genetic relationship among 21 samples Amomi Fructus collected from Yunnan, Hainan, Guangdong, and Fujian Provinces; At the same time, the phenotypic characters of 16 Amomum villosum samples were measured in height, stem diameter, blade number, and blade size, etc. Then cluster analysis on all populations of Amomi Fructus was carried out based on the above data. Results: Eleven primers selected from 60 ISSR-primers were used for amplification and a total of 54 DNA bands were obtained,including 22 polymorphic bands. At species level, the average percentage of polymorphic bands (PPB) was 40.74%, Nei's gerne diversity (H) was 0.116 1, Shannon's information index (I) was 0.184 2. There was a little difference of morphological characteristics. Conclusion: The genetic diversity of germplasm resources in Amomi Fructus is lower.