Study On Quality Evaluation Of The Zaocys Dhumnades(Cantor) Based On The Method Of Micro-morphological Identification

Objective: Chinese Animal Medicine is an important part of Chinese Traditional Medicine,which has been recorded in 《Shen nong ben cao jing》.Due to the shortage of wild resources of Chinese Animal Medicine and immature breeding technology,the price of Chinese Animal Medicine is expensive.There are a large number of animals of the same family in the market to pretend to be authentic,so it is difficult to identify them.Rapid identification of Chinese Animal Medicines has become an urgent problem to be solved.In this study,the method of Micro Character identification is mainly used to identify the Zaocys dhumnades(Cantor)and its adulterants,and 3D high-definition feature map is taken,and then the results of Micro Character identification are verified by DNA bar code identification.In addition,the identification and microstructure characters,heavy metal concentrations,alcohol-solube extract content and relative amino acid content of the Zaocys dhumnades(Cantor)were systematically studied to ensure the safety and effectiveness of clinical medication.Method:1.Micro-morphological identification: In this study,the scales,nasal bones,parietal bones and vertebrae of the head and body of the snake were identified by3 D technology.The ratio of height of spinous process to that of the whole bone was measured.Spss22.0 software was used for statistical analysis.2.DNA barcode identification: The DNA of the samples was extracted by PCR,amplified and sequenced by cytb primer,analyzed by mega5.1 software,and the NJ cluster tree was constructed by adjacency method。3.Pharmacognostical study on the Zaocys dhumnades(Cantor): The identification and microstructure characters of the Zaocys dhumnades(Cantor)and its adulterants were identified;the contents of Pb,Cd,Hg and As were determined by ICP-MS;the alcohol soluble extracts of the Zaocys dhumnades(Cantor)were determined by hot soak;the relative contents of amino acids of the snake and its adulterants were determined by GC-MS.Result:1.There are 36 samples,12 samples are authentic Zaocys dhumnades(Cantor)and the others are all confusions.Among them,7 were the Ptyas mucosa(Linnaeus),8 were the Ptyas korros(Schlegel),3 were the Naja atra Cantor,1 was the orthriophis taeniurus cope,1 was the Elaphe carnata(cunther),3 were the xenopeltis unicolor reinward,1 was the Cerberus Rynchops(Colubridae),which were different from the previously reported forgeries.By identifying buccal scale,body scale type,end socket of scale,longitudinal stripe of capsule,fingerprint like texture of base,morphology of nasal bone and parietal bone,height of spinous process of vertebrae,etc.,we can distinguish the genuine and the confused products of black top snake.The results of statistical analysis showed that the height of spinous process was the highest in the range of 2.82 ～5.80 mm.There is a significant difference between the ratio of spike height to the whole vertebra height of the snake and its adulterants.The spike height is the highest,which corresponds to the word "sword ridge" in traditional character identification.2.Through DNA bar code identification and cluster analysis,different snakes are divided into different kinds,each of which has good single lineage.It is proved that the accuracy and reliability of Micro-morphological identification can achieve the purpose of rapid identification.3.The results showed that the content of arsenic in some samples was 5 times higher than the specified value.The alcohol-soluble extract content basically conforms to the regulations of 2015 edition of Pharmacopoeia of the people’s Republic of China.16 amino acids were detected by GC-MS,including leucine,proline,valine,D-isoleucine,D-proline,glycine,serine,threonine,alanine,aspartic acid,hydroxyproline,glutamic acid,phenylalanine,lysine and tyrosine.The highest content was glycine,the lowest was isoleucine.The relative content of alanine was significantly different between the two products.Conclusion:1.Through DNA molecular verification,the identification characteristics of the method of Micro-morphological identification are stable and not easy to change with the changes of the outside world.The above identification characteristics can be used to accurately identify the Traditional Chinese medicine of Zaocys dhumnades(Cantor).2.The content of arsenic in Zaocys dhumnades(Cantor)is easy to exceed the standard,and the alcohol-soluble extract content basically conforms to the provisions of Pharmacopoeia of the people’s Republic of China in 2015.There was significant difference in the relative content of alanine between the Zaocys dhumnades(Cantor)and its mixture,but there was no significant difference in the relative content of other amino acids.3.Micro-morphological identification using 3D depth of field synthesis technology and 3D big picture splicing technology can take the 3D high-definition characteristic map of the authentic and confused products of the Zaocys dhumnades(Cantor),which provides a strong basis for the identification of the Zaocys dhumnades(Cantor).The method is easy to operate,convenient to use,low cost,suitable for large-scale promotion and use,and is an important means of on-site rapid inspection.