Quality Evaluation Of Commercial Deer Velvet

Objective: In order to provide the theoretical basis for the quality control of velvet antler,this paper summarized the quality status of velvet antler in China,and carried out quality analysis and composition study of pilose antler.To study the effect of the antler extract on proliferation of human renal tubular epithelial cells(HK-2)and the protective effect of cisplatin on the kidney injury.Methods: Physical and chemical identification and TLC identification were carried out according to the fourth part of Chinese Pharmacopoeia in 2015,and water,ash and extract were examined.The content of amino acid in velvet antler was determined according to agricultural industry standard: NY/T1618-2008.The content of hypoxanthine in velvet antler was determined by high performance liquid chromatography(HPLC).The mobile phase was 0.07% glacial acetic acid aqueous solution-methanol(96:4).The column temperature was 28 ℃,the detection wavelength was 254 nm,and the flow rate was 1mL/min.Besides the method of the content determination,we developed the specific fingerprint chromatograms of nucleic acid.And then the biological activity of pilose antler was performed.The extract of velvet antler was prepared into three concentrations: low,medium and high.The mice were given velvet antler until the end of experiment,and cisplatin was given in the seventh day to establish the renal injury model.The blood samples were used to detect the contents of urea nitrogen and creatinine on the 3th day after the administration of cisplatin.After the mice were sacrificed,kidney tissue was used for histochemistry to evaluate the protection effect of velvet antler on the kidney injury.Furthermore,cell viability was measured by CCK-8 assay.The effects of different concentrations of velvet antler extract on cell viability of HK-2 cells and protection effect of cisplatin-induced kidney injury were investigated.Results: 1.The color reaction of velvet antler was observed by the physical and chemical identification.By thin layer chromatography(TLC),the main spots of the same color were found in the chromatogram of the samples,and the spots of the same color were found in the corresponding position of the chromatogram of the sample and that of the control medicinal material,and the spots of the same color were found in the correspondingposition of the chromatogram of the sample.The amino acid content in velvet antler was determined.The moisture content of velvet antler collected in the market was tested,and moisture content should not be more than 13.0%,and the total ash content of the collected velvet antler medicinal material in the market was detected,and the total ash content should not exceed 40%.The content of water-soluble extracts and alcohol-soluble extracts of velvet antler from the market was determined.Moreover,water-soluble extracts and alcohol soluble extract should not be less than 5.0% and 3.0%,respectively.Hypoxanthine,as the main component of velvet antler,the content should not be less than 0.02%.Hypoxanthine used to develop the HPLC specific chromatogram of velvet antler.2.Pathological sections of kidney were observed.The renal tubule degeneration was irregular in cisplatin model group.With the increase of concentration,the degree of renal tubular necrosis decreased and the serum urea nitrogen and creatinine contents decreased.Velvet antler extract could promote the proliferation of HK-2 cells.With the increase of velvet antler concentration,we observed the more obvious proliferation effect.When the drug concentration reached 1mg/mL,the effect was more obvious.The damage model of HK-2 cells induced by cisplatin was established successfully.The optimal concentration of cisplatin was 40 μ m ·ml-1.Velvet antler extract could inhibit cisplatin-induced HK-2damage,and the cell survival rate increased with the increase of concentration in a certain range.The pathological sections of the kidney were observed.Conclusion: The method is practical and reproducible,and can be used to develop the quality standards of velvet antler.It can improve the quality standard system of velvet antler.Importantly,the extract of velvet antler has protective effect on cisplatin-induced nephrotoxicity,promote the proliferation of HK-2 cells,and protect the cells damaged by cisplatin and inhibit renal tissue necrosis.