Establishment Of Evaluation System For Quality Of Velvet Antler And Cervus Nippon Temminck

Velvet antler are the young horn of male deer such as Cervus nippon Temminck or Cervus elaphus Linnaeus which have not ossified. As a precious traditional Chinese medicine, there are many effects for human bodies including enhancing kidney Yang, essence and blood, strenghening bones, and so on. Because of its abundant chemical composition and a variety of pharmacological effects, it has been widely used in the field of food, medical treatment and health care. Due to the different varieties and years of velvet antler,the qualities of velvet antler are different which can not be distinguished easily by the eyes. The velvet antler often mixes the spurious with the genuine and the inferior replaces replacing good ones frequently in the market. Therefore, it is necessary for us to build the methods of distiguising different velvet antler.Infrared spectroscopy technology was used to analyze different kinds of velvet antler. Potassium bromide tablet is used to analyze and evaluate the infrared spectral characteristics of different kinds of velvet antler, and a certain theoretical basis for the establishment of the evaluation system can be provided. The results showed that the characteristic absorption peak of different kinds of velvet antler and C. nippon Temminck varieties at 3500 ~ 3000 cm-1, 2930 cm-1 and 1650 cm-1. For different kinds of C. nippon Temminck, Spiker velvet antler has broad characteristic absorption peak at 3370 cm-1. Three-branch velvet antler was at 3340 cm-1. Regrowth velvet antler has narrower characteristic absorption peak at 3316 cm-1. Two-branch velvet antler has narrowest characteristic absorption peak at 3304 cm-1. Two-branch velvet antler, Three-branch velvet antler and Regrowth velvet antler have a characteristic absorption peak of amide Ⅱ, but Spiker velvet antler was not obvious. Two-branch velvet antler and Regrowth velvet antler have a characteristic absorption peak at 1233 cm-1, but Three-branch velvet antle and Spiker velvet antler were not obvious.For different kinds of velvet antler, Rangifer tarandus has the broadest absorption peak at 3340cm-1 than other velvet antler. C. elaphus Linnaeus was at 3310 cm-1, C. nippon Temminck and New Zealand red deer velvet antler have narrower absorption peak at 3304 and 3322cm-1, respectively. New Zealand red deer velvet antler has a narrower absorption peak at 2938 cm-1. C. nippon Temminck, New Zealand red deer velvet antler and C. elaphus Linnaeus have amide II absorption peak while Rangifer tarandus was not obvious. C. nippon Temminck and Rangifer tarandus have characteristic peaks at 1455,1443 cm-1 while C. elaphus Linnaeus was not. So that we can distinguish different velvet antler and different varieties C. nippon Temminck by peak type, peak number, peak intensity of the characteristic peak. Infrared spectroscopy spectrum is well suited for the rapid identification and quality control of velvet antler for its high resolution, easy operation and strong specialization of absorbance bands.Studies were carried out by basically analyzing the contents of conventional ingredients, protein, polysaccharides, Ca, P and the color index of velvet antler(L、a、b、E、H、H°、C) which can be determined through chromatic meter of seven varieties of velvet antler. The results of principal component analysis(PCA) showed that there are two principal components of the extraction, and the cumulative variance contribution rate is 82.479%. The variance contribution rate of the first principal component was 56.332%, representative variables were water extract, crude protein, moisture and Ca, the variance contribution rate of the second principal components was 26.146%, representative variables were ash and P. Through correlation analysis and stepwise regression analysis of chemical composition content and color indicators of velvet antler, the color of velvet antler had a closer relationship with chemical components. Redness of the color measurement index can be used as the measurement indicator for crude protein which is the first principal component on velvet antler. The content about moisture which was the first principal component of velvet antler influenced on chromatism extremely significantly. The lightness value of L can be used to judge the content of moisture which was the first principal component of velvet antler. The content of Ca which was the first principal component of velvet antler can be judged by tincture. Hue angle H °can be used as the measure indicator of the quantity of velvet antler which was used to determine the content of different varieties of velvet antler in the second principal component P and the first principal component of the alcohol extract. They could reflect the inner chemical quality of velvet antler by using the chromatism parameters, and provide a new method and reference for the traditional evaluation methods for velvet antler.High intensity pulsed electric fields method(PEF) which were united with the extraction of proteins and polysaccharides from velvet antler were established and were optimized.The results were as follows: the extraction rate was the highest under the condition of water as solvent, solid-liquid ratio of 1:30 g·mL-1, 20 KV·cm-1 in the electric field intensity and pulse number 10 in the high intensity pulsed electric fields, protein extraction rate reached 12.32% and polysaccharides extraction rate could reach 3.25%.The extraction rate of proteins and polysaccharides by PEF is the higest, compared with ultrasonic extraction, heat reflux extraction about water-soluble components. Therefore, the extraction technique of PEF which were using in combining with the extraction of proteins and polysaccharides from velvet antler were described as practicable reasonably.The study focused on the effect of the aqueous extract of two branched and three branched velvet antler(C. nippon Temminck) on cisplatin-induced nephrotoxicity in mice. The model of renal injury in mice was induced by intragastrical administration of CDDP at the dose of 15 mg/kg. After treatment, kidney index(KI), the levels of creatinine(CRE) and blood urea nitrogen(BUN) in serum, the activity of superoxide dismutase(SOD) and Glutathione peroxidase(GSH-Px), the content of malonic dialdehyde(MDA) in kidney were determined, HE-staining was employed to observe renal pathologise changes. The results shows that each dose group markedly decreased the serum levels of BUN and CRE, and the content of MDA(P<0.05), elevated the activity of SOD and GSH-Px in mice pretreated with CDDP(P<0.05). Kidney pathology morphology was improved obviously, moreover, the aqueous extract of two branched velvet antler has the better protect effect than three branched at the same concentration. The aqueous extract of both two branched and three branched velvet antler has a certain protective effect on cisplatin-induced nephrotoxicity, which may be associated with which increases anti-oxidative capability of mice kidney tissue.To investigate the antioxidant activity of different kinds of velvet antler in vitro,the actibity of SOD, CAT, POD and the inhibitory activity of tyrosinase were measure. The results showed that in the same concentration, the order of the activity of SOD about different kinds of velvet antler is : C. nippon Temminck > Rangifer tarandus > New Zealand red deer > C. elaphus Linnaeus. The order of the activity of CAT is : Rangifer tarandus > C. nippon Temminck > C. elaphus Linnaeus > New Zealand red deer. The order of the activity of POD is : C. nippon Temminck > C. elaphus Linnaeus > New Zealand red deer > Rangifer tarandus. The order of inhibitory activity of tyrosinase is : C. nippon Temminck > C. elaphus Linnaeus > New Zealand red deer > Rangifer tarandus. SOD, POD actibity and the and inhibition rate of tyrosinase activity of C. nippon Temminck is the highest compared with other velvet antler. The actibity of CAT of Rangifer tarandus is higher than other velvet antler, and SOD actibity is higher than New Zealand red deer velvet antler and C. elaphus Linnaeus, but POD actibity and and inhibition rate of tyrosinase activity are both the lowest. CAT, POD actibity of C. elaphus Linnaeus is higher than New Zealand red deer velvet antler. The order of the activity of SOD about different kinds of C. nippon Temminck is : Spiker velvet antler > Two-branch velvet antler > Regrowth velvet antler > Three-branch velvet antler. The order of the activity of CAT is : Two-branch velvet antler > Spiker velvet antler > Three-branch velvet antler > Regrowth velvet antler. The order of the activity of POD is : Three-branch velvet antler > Two-branch velvet antler > Regrowth velvet antler > Spiker velvet antler. The order of inhibitory activity of tyrosinase is : Regrowth velvet antler > Three-branch velvet antler > Two-branch velvet antler > Spiker velvet antler. For different kinds of C. nippon Temminck, each kind has different activity and its own advantages in different aspects. Thus, we can evalute different kinds of velvet antler and C. nippon Temminck through SOD, CAT, POD activity and inhibition rate of tyrosinase activity, respectively, which provides scientific basis for the development of velvet antler as a natural antioxidant.