| Objectives:Trichonephila clavata spider,as the raw material of some classic ancient prescriptions,is a kind of animal with a wide distribution range and high medicinal value,but it is not included in the Chinese pharmacopoeia and local standards of Chinese medicinal materials.There were no reports on the pharmacognosy system of T.clavata.In this subject,a relatively systematic pharmacognostic study was conducted on T.clavata in different places of Yunnan Province according to the relevant requirements of Chinese Pharmacopoeia(2020 edition),the textbook of Pharmacognosis(5th edition)and the technical guidelines for quality standard research of Chinese medicinal materials(ethnic medicinal materials)in Yunnan Province(released in 2017).The research contents included the identification method,the determination of examination items and the content determination of main components of the medicinal materials.The basic data of pharmacognosis of T.clavata were obtained,which provided reference for the further development and utilization of the medicinal materials of T.clavata.Methods:The characters and microscopic characteristics of T.clavata were observed and described by magnifying glass and microscope.Amino acid was used as control substance for TLC identification.The possible chemical constituents in T.clavata were preliminarily determined by systematic pretest method.DNA barcode of COI sequence was established by molecular biology technology to identify the authenticity of medicinal materials.Water content,ash content and extracts of different solvents in ten batches of samples of medicinal materials were studied and determined according to the inspection items stipulated in the 2020 edition of Chinese Pharmacopoeia.The contents of heavy metals and other microelements were determined by atomic spectrophotometer and atomic fluorescence spectrophotometer.The molecular weight distribution of proteins and peptides in the samples were investigated by SDS-PAGE.The lipopolysaccharides in samples were extracted by Soxhlet extraction method,the types and structures of lipopolysaccharides were identified by gas chromatography-mass spectrometry,and the relative percentage of each compound was determined.The contents of water-soluble protein and total polysaccharide in 10 batches of samples were determined by ultraviolet spectrophotometer,and the contents of nucleosides in samples were determined by high performance liquid chromatography.Results:In this paper,the characters and microscopic characteristics of T.clavata were described systematically,and a TLC identification method with three amino acids as the control was established.The color spots were rose red,and there were no less than 6chromogenic spots.The pre-test results of the system showed the chemical composition of proteins,sugars,alkaloids and phenols in the samples,and provided the corresponding picture data.The phylogenetic tree constructed based on COI gene sequence can distinguish T.clavata from other spider species,and the calculated inter-specific and intraspecific genetic distances can also distinguish different species.According to the relevant requirements of Chinese Pharmacopoeia,the test items of 10 batches of T.clavata samples were determined: the moisture content was 4.06%～5.73%,with an average content of 5.09%.The total ash content was 8.88%～13.81%,and the average ash content was 11.58%.The acid insoluble ash content is 1.63%～2.55%,and the average content is 1.98%.In the determination of extractives,the content of extractives obtained by hot water solvent leaching method was the highest,ranging from 25.24%～34.44%,with an average content of 32.08%.The content of volatile ether extract was the lowest,ranging from 12.14%～16.27%,and the average content was 13.78%.The contents of eight metal elements in 10 batches of samples were determined.The average contents of arsenic,mercury,copper,antimony,selenium,iron,manganese and zinc were 1.805mg/kg,0.028 mg/kg 116.75 mg/kg,0.087 mg/kg,0.539 mg/kg,659.89 mg/kg,58.09mg/kg and 737.02,respectively mg/kg.The results of SDS-PAGE analysis of molecular weight distribution of protein polypeptides showed that most of the molecular weight was about 75 KDa,and more oligopeptides and polypeptides were distributed around 11 KDa and 35 KDa.The content of fat-soluble components in 10 batches of T.clavata samples ranged from 12.25% to 16.10%,with an average content is 13.81%.The content of watersoluble protein was determined by biuret method in the range of 196.18～287.34 mg/g,and the average content was 240.24 mg/g.The content of water-soluble protein was determined by Coomassie brilliant blue method in the range of 18.91～40.75 mg/g,and the average content was 34.19 mg/g.The total polysaccharide content was 1.331～1.722mg/g,and the average content was 1.509 mg/g.The total contents of uracil,guanine,uridine,inosine and guanosine were 4.017～6.473mg/g,and the average content was 5.495mg/g.Conclusions:In this project,a systematic and comprehensive pharmacognosy study was carried out on ten batches of samples of T.clavata collected.The authenticity identification method of T.clavata was established,and the data of moisture,ash,extract,heavy metal and trace elements were determined.The contents of main substances in T.clavata were determined,including lipid solubility,protein,total polysaccharide and nucleoside,and the quality control method was established.This study further improved the pharmacognosy research content of T.clavata and provided important scientific basis and reference materials for the sustainable development of T.clavata and other arthropod resources and the subsequent exploitation and utilization of potential medicinal value. |
PDF Full Text Request