| Purpose:In recent years,the planting area of A.melanocarpa has gradually increased in China,but due to the different climatic environment and geographical location,the quality of the medicinal material has some differences.At present,the fruit quality evaluation system of A.melanocarpa has not been established,and its quality control is not good,so that its functional characteristics of medicine and food are limited.Therefore,in this experiment,11 batches of A.melanocarpa fruit of the same kind and different origins were selected as the research object,and their traits,microscopic identification,moisture,ash,and extract content were determined,and their polyphenols,flavonoids and their contents were determined.Carrying out fingerprint research to establish a fruit quality evaluation system of A.melanocarpa,providing an effective reference basis for its in-depth research and further application and promotion in the future,and at the same time laying a foundation for subsequent pharmacodynamic research.Material and method:1.Materials: The experimental materials of this subject are 11 batches of A.melanocarpa fruit,which was identified by Professor Kang Tingguo of Liaoning University of Traditional Chinese Medicine as the fruit of the genus A.melanocarpa.The voucher specimen is kept in the dark in the laboratory 304 of Chinese Medicine Innovation Engineering Technology Center of Liaoning University of Traditional Chinese Medicine.2.Methods:2.1 In this experiment,the direct observation method and vernier caliper measurement method in the first part of the 2015 edition of the Chinese Pharmacopoeia were used to identify the traits of the fruit and seeds of A.melanocarpa from different places.Relying on the 2015 edition of the Chinese Pharmacopoeia,the fourth chapter of the general rule,the powder preparation method was used to observe the microscopic characteristics of its fruits.The contents of water,ash,acid-insoluble ash,water-soluble extracts,alcohol-soluble extracts of the fruits were determined using the drying method,ash determination method,acid insoluble ash determination method,and cold soaking method.2.2 Taking the polyphenol extraction rate as the evaluation index,design five single factors of extraction solvent,absolute ethanol extraction fraction,extraction time,material-liquid ratio,and extraction temperature,screen out the main factors based on the single factor inspection,and adopt L9(3)4 positive The cross experiment optimized the extraction process of total polyphenols in the fruits of A.melanocarpa.Taking the total flavonoid extraction rate as the evaluation index,the four single factors of absolute ethanol extraction fraction,extraction time,material-liquid ratio,and extraction temperature were designed,and the main factors were screened according to the single factor in spection,and the L9(3)4 orthogonal test was used The extraction process of total flavonoids in the fruits of A.melanocarpa was optimized.At the same time,the content of total polyphenols and total flavonoids in 11 batches of A.melanocarpa fruit samples was determined by ultraviolet spectrophotometry.In addition,HPLC method was used to determine the content of cyanidin-3-O-glucoside,the index component of 11 batches of A.melanocarpa.2.3 The HPLC method was used to study the fingerprints of 11 batches of A.melanocarpa fruit samples.The similarity evaluation software of the 2012 version of the traditional Chinese medicine fingerprint spectrum similarity evaluation software was used fo rcluster analysis.Results:1.The Aronia melanocarpa fruit is a berry when it is fresh,it is round,and its trans verse diameter is 0.7-1.3 cm,the vertical diameter is 0.8-1.2 cm.The surface,flesh and seeds are black-purple or dark red,with luster,flat top,style and brown.The top is oblate,with astyle,brown,and the top contains a protruding style base.The base has brown-red fruit-stalk marks,often retaining fruit stems,1-2.5 cm,slightly airy,slightly sour,the Aronia melanocarpa fruit is oval,dark brown,with irregular vertical wrinkles,hard texture,seeds,kidney-shaped,tan,4-8 capsules,horizontal diameter 3-4.5 mm,vertical diameter after drying 0.1-0.2 mm.2.The Aronia melanocarpa fruit powder reddish brown,thin exocarp,reddish brown orpurple brown,more common reddish brown blocks,with wooden plug cells,clearly visibleoil spots,neatly arranged,keratinous lines on the surface,cell wall.It is thickened with rosary,so it is also called rosary thickened cells;thin-walled cells,closely arra nged together;stone cells in the outer layer of the seed coat are slightly longer or polygonal,the walls are slightly thick,the cell cavity is slightly smaller,and the cavity contains yellow;the inner layer of the seed coatis round,oval,etc,the wall is relatively thin,the cellcavityis relatively large,and there are densely packed thin-walled holes.It also has endosperm cells and threaded catheters.3.The moisture content of 11 batches of samples ranged from 78.49% to 82.65%,with an average moisture content of 81.03%.The ash content is 2.59%-4.66%,the averagec ontent is 3.72%,the acid insoluble ash content is 0.45%-1.18%,the average contentis0.78%,the water soluble extract content is 49.71%-61.19%,the average content is 57.08%,alcohol The soluble extract is 55.71%-69.99%,the average content is 63.83%.4.In this experiment,the best method for extracting the total polyphenols in the fruit of Aronia melanocarpa was obtained by orthogonal test.The ethanol fraction was 70%,the material-liquid ratio was 1:50(g/m L),and the extraction temperature was 55?,time was 75 min.The average extraction rate of total polyphenols measured by this me thodwas 17.88%,and the RSD was 1.27%.The content of total polyphenols in the fruits of Aronia melanocarpa from 11 batches of production areas was found to be different,and the content of total polyphenols was different between different production ar eas.Overall,the range was 17.47±1.17%-19.10±1.56%,of which Shenyang,Liaoning The averagecontent is the highest at 19.10%,and Anshan in Liaoning is the lowest at17.47%.5.The results of orthogonal experiments in this experiment showed that the best extrac tion process of total flavonoids in Aronia melanocarpa fruit was ethanol fraction 60%,material-liquid ratio 1:40(g/m L),extraction temperature 55?,extraction time 80 min.The above-mentioned optimal extraction method was used to conduct three parallel determinations of confirmatory experiments,and the average extraction rate of flavonoids was 0.102% and RSD was 0.09%.The content of total flavonoids in the fruits of Aronia melanocarpa in 11 batches of production areas was found to be different,and thecontent of total flavonoids was different between different production areas.Overall,the range of flavonoids was 0.04±0.42%-0.14±0.49%.The average percentage content is highest at 0.14%,and Dalian in Liaoning is the lowest at 0.04%.6.The cyanidin-3-O-glucoside content in the fruit of Aronia melanocarpa from different places has a certain difference,in the range of 0.34%-0.87%,of which the highest content in Shenyang,Liaoning is 0.87%,the content of Baishan in Jilin The lowest is0.34%.7.In this experiment,there are 17 peaks in 11 batches of Aronia melanocarpa fruit samples.Peaks 3,9,10,12 and 13 are identified by the standard products as gallic acid,chlorogenic acid,caffeic acid,Cyanidin-3-O-glucoside,epicatechin.The similarity of their fingerprints is above 0.90.The results show that according to the method under this experiment,fingerprint analysis of the fruit samples of Aronia melanocarpa has good similarity.Conclusion:1.This experiment was the first time to conduct microscopic identification and researc on the determination of moisture,ash,and extractables content.A preliminary evaluation system for the quality of A.melanocarpa fruit was established to fill the gaps in its quality evaluation and to provide research and popularization A valid reference basis.2.Optimize the extraction process of polyphenols and flavonoids by ultrasonic method,add 0.2% hydrochloric acid solution to the extraction solvent,the content of polyphenols is greatly increased.The results show that the method is simple,efficient and reproducible,and can be applied to the extraction of its active ingredients.3.The content of polyphenols,flavonoids,and anthocyanins,which are representative co mponents of cyanidin-3-O-glucoside,were the highest in 11 batches of A.melanocarpa fruits.You can use Shenyang,Liaoning as a reference.This may be related to Shenyang's unique geographical location,soil conditions,and climate,but the specific reasons need tobefurther studied.4.An HPLC fingerprint analysis method for the fruit of A.melanocarpa was established for the first time,and its precision,stability,repeatability,and similarity were all within thespecified range.The quality of the fruit has important guiding significance.5.Studies with other reported literature have shown that the polyphenol content and the anthocyanin representative component of cyanidin-3-O-glucoside in the fruit of A.melanocarpa are higher than other domestic fruits.It may be one of the reasons for its strong antioxidant capacity.Therefore,the fruit of A.melanocarpa has important significance for further research,which can lay a solid foundation for its further development and promotion.6.The quality evaluation system of A.melanocarpa has been preliminarily established,which is of great significance for its standardized use. |
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