| Objective: Objective: Saposhnikovia divaricata is a traditional Chinese medicinal material with a long history and extensive application in our country.But in recent years,due to the scarcity of wild wind-proof resources,demand exceeds supply,cultivation wind-proof has become the main market.At present,the quality of cultivated Saposhnikovia divaricata is not uniform in the market,and the picking and processing links of Chinese medicinal materials are random and chaotic,and the storage and transportation links are affected by damp and mildew,which seriously affects the quality and clinical efficacy of Chinese medicinal materials.Therefore,this research takes the Saposhnikovia divaricata from 33 producing areas as the research object,carries out the Saposhnikovia divaricata quality evaluation research,establishes the quality evaluation system,studies the different initial processing and the storage method to the Saposhnikovia divaricata quality influence,the tissue-specific distribution of active components of Saposhnikovia divaricata was analyzed by transcriptome sequencing,which provided a theoretical basis for the exploitation and utilization of Saposhnikovia divaricata.Methods: Based on the methods of Chinese pharmacopoeia(2020 edition),33 batches of Saposhnikovia divaricata from different places were evaluated by 0832 drying method,2302 total ash determination method,2201 hot soaking method and 0512 High-performance liquid chromatography(HPLC).Determination of aflatoxins by High-performance liquid chromatography tandem mass spectrometry.The contents of pesticide residues,heavy metals and harmful elements were determined by the methods in Chinese pharmacopoeia(2020 edition).The different storage methods were evaluated by water content and chromogenin content.The quality formation mechanism of Saposhnikovia divaricata was investigated by transcriptome sequencing and network pharmacology.Results:1.The moisture content of 33 batches of Saposhnikovia divaricata from different producing areas ranged from 5.78% to 9.97%,ash content from 1.95% to 2.99%,alcohol-soluble extract content from 27.88% to 47.59%.The highest content of chromogenic ketones was found in Heilongjiang.A method of thin layer chromatography for extraction of Saposhnikovia divaricata was optimized.75% methanol was used as solvent for ultrasonic extraction for 10 min,the ratio of solid to liquid was 1:5,the sample size was 5 ul,the sample width was 8 mm,the chloroform is ethyl acetate(4:2:1).Two quality markers(4’-O-β-D-glucosyl-5-O-methylvisamminol and Cimifugin)of Saposhnikovia divaricata from different habitats were screened.2.The moisture and ash content of freeze-dried samples were 3.91% and 6.44%,respectively.The contents of prim-O-glucosylcimifugin,4’-O-β-D-glucosyl-5-O-methylvisamminol,Cimifugin,Sec-O-glucosylhamaudol and total chromogenin in the freeze-dried samples were significantly different.The content of chromogenic ketones in freeze-drying was much higher than that in other processing methods.AHP comprehensive evaluation results show that freeze-drying has the highest score.3.The lowest moisture content condition is the carton cool light,the highest is the carton low temperature light.The highest content of chromogenic ketones is the carton low temperature light,the lowest is polyethylene plastic woven bag low temperature light.Grey correlation analysis showed that the retention relationship between carton packaging and chromogenic ketones was the closest.Principal component analysis found that carton packaging was more stable,followed by Gunny bag packaging.AHP comprehensive evaluation and analysis of carton low-temperature light-proof packaging of the highest score.4.The content of prim-O-glucosylcimifugin in the root of Saposhnikovia divaricata was about 2.5 times as much as that in the leaf,and the content of4’-O-β-D-glucosyl-5-O-methylvisamminol in the leaf was about 3 times as much as that in the root.Compared with the roots and leaves of Saposhnikovia divaricata,the expression of C4 H and CYP98 A in the roots was higher than that in the leaves,and the expression of F3 H in the leaves was higher than that in the roots.Conclusion:1.The quality of Saposhnikovia divaricata from different habitats was significantly different.The content of chromogenin in Saposhnikovia divaricata from Heilongjiang was higher.The TLC method was optimized and two differential markers were identified by fingerprint.2.The best initial process is freeze-drying(-80℃): the comprehensive quality is the highest and the active ingredient is the most retained.3.The best storage conditions are low temperature(2-10℃),no light and no aflatoxins detection.4.F3 H,C4H,CYP98 A and other key differential genes were screened based on m RNA sequencing.Therefore,it is inferred that the root and leaf quality differences may be caused by up-or down-regulated expression of these genes. |
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