| The velvet antler is the young horn with dense hair of the male sika deer(Cervus nippon Temminck)or the red deer(C.elaphus Linnaeus)that is not ossified and.The former is commonly known as "sika deer velvet" and the latter is commonly known as "red deer antler".As a traditional valuable medicine,antler has been widely used for more than 2,000 years for its rich pharmacological effects.According to the statistics of the State Drug Administration,there are 13 kinds of proprietary Chinese medicines containning velvet antler and 65 kinds of health products containing velvet antler.Therefor the supply of antler resources is in short and the price is expensive.The local medicinal herbal standards in Sichuan and Gansu also stipulated that white-lipped deer and sambar could be used as velvet antler in accordance with the local deer antler's prescription habits,resulting in the disorder of the market of velvet antler.Moreover,the reindeer antler,moose antler and others are sold as velvet antler in the market.In addition,the velvet decoction pieces are of a disordered grade and have uneven quality;the commodity specifications of the antler decoction pieces include wax pieces,powder pieces,blood pieces,and bone pieces,occurrence of bleeding pieces from the bone pieces occurs on the market;therefore,it is necessary to conduct overall quality control and evaluation on the authenticity of the velvet antler and the quality of the velvet antler pieces.This paper mainly includes the following three parts:Part ? is the research progress of chemical composition and quality control methods for velvet antler.The chemical composition,standard status,analytical methods,and problems with quality control are mainly outlined.The antler contained more than 20 kinds of amino acids,18 kinds of steroids,26 kinds of mineral elements and other chemical components;The standards of antler were found to be inconsistent when comparing the Chinese Pharmacopoeia,Hong Kong Herbal Standard and local medicinal material standards.Finally,the quality control method of pilose antler was summarized,using ultraviolet(UV),infrared radiation(IR),thin liquid chromatography(TLC),high performance liquid chromatography(HPLC),high performance liquid chromatographymass spectrometry(HPLC-MS),finger-print,gas chromatography(GC),GC-MS and molecular biology as the technical support for quality evaluation of velvet antler medicinal materials and pieces.Part ? is the establishment of the characteristic map of the enzymatic hydrolysis of velvet antler.The transmission voltage and collision energy of the four enzymatic peptide candidate ions were optimized in the selected ion mode and the product ion mode,respectively,and two sub-ions were selected to construct eight ion pairs.The data were collected in the multi-reaction monitoring mode,thus establishing a characteristic map of the enzymatic hydrolysis of velvet antler.Part ? is the establishment of the authenticity identification method of velvet antler and its common mixed varieties.66 batches of samples were collected,including 40 batches of sika deer antler,15 batches of pilose antler,and 11 batches of reindeer antler.Firstly,the protein of velvet antler was extracted at 120? by water,trypsin was used to enzymatically hydrolyze three kinds of velvet antlers and UPLC-QTOF/MS was used to establish the enzymatic peptide map of velvet antler.Then,combining Progenesis QI software with principal component analysis(PCA),the velvet antler was distinguished,and two marker peptides of reindeer antler that have distinctive significance were founded by further analysis using orthogonal partial least-squares discriminant analysis(OPLS-DA).Later,in the product ion pattern,the conditions are optimized to search for two higher-response daughter ions to form four groups of characteristic ion pairs.and multiple reaction monitoring(MRM)mode was used to verify accurate samples and unknown samples;finally,according to the(MS/MS)fragmentation information of the two marker peptides,the sequence of the marker peptides were resolved by Masslynx Biolynx software and the two marker peptides were synthesized to verify the correctness of the characteristic sequences.Part ? is the evaluation of multi-index quality grade of different specifications of velvet antler pieces.Firstly,combined with the status of the change of amino acids content in different specifications of velvet decoction pieces with the growth period,A pre-column derivation of phenylisothiocyanate(PITC)-high performance liquid chromatography was established to simultaneously determine the content of 15 kinds of amino acids in 50 batches of velvet decoction pieces of wax piece,powder piece,blood piece and bone piece and the methodological verification was carried out.The amino acids content was used as an analytical index to evaluate the quality of velvet antler pieces.Secondly,the Chem Pattern software was used to establish fingerprints of different specifications of velvet decoction pieces and perform chemometric analysis.Including similarity analysis,cluster analysis and principal component analysis,which provide a reference for the quality grade evaluation of different specifications of velvet antler pieces.Then,the total nitrogen content of different specifications of velvet decoction pieces was determined by nitrogen analyzer method according to the 2015 edition of the "Chinese Pharmacopoeia",so as to evaluate the quality grade of different specifications of antler and the methodological verification was carried out.Finally,according to the traditional drinking habits of antler pieces,using dilute ethanol as the extraction solvent,the content of the extract under the conditions of hot and cold immersion was test respectively according to the 2015 version of the Chinese Pharmacopoeia alcohol soluble extract determination method(General Regulation 2201).The results showed that the contents of amino acids,total nitrogen and extracts in the four specifications of velvet decoction pieces were the highest in the wax tablets and the lowest in the bone piece,but the amino acids contents of the blood and powder tablets were similar.At the same time,combined with the chemometric analysis results of the fingerprint of antler decoction pieces,the antler pieces of different specifications can be divided into quality grades,and the velvet wax tablets are classified as first-class products,and the antler powder tablets and blood tablets are classified as second-class products,the bone tablets are classified as third-class products;thus,the quality level evaluation of velvet decoction pieces can be carried out to a certain extent,and the phenomenon of velvet decoction pieces is chaotic and shoddy. |
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