Research On The Quality Of Antler Medicinal Materials

Purpose:Through the qualitative identification and chemical composition analysis of the antlers and antlers of the two medicinal materials of the antler medicinal materials,the differences and commonalities of the two were analyzed.The deer bones of the same stagnation were used as the control to identify and control the quality of antlers.Provide evidence.Materials and methods:Materials:The samples were 5 batches of antler medicinal materials,4 batches of antlers and medicinal materials,and 1 batch of deer bone medicinal materials.methods:1 Qualitative identification of antler medicinal materials1.1 Microscopic identification The microstructures of antlers,antlers and deer bones were observed and their characteristics（number,shape and area of bone lacunae in bone fragments）were measured.Inter-group T test was used to compare antlers with deer bones.The difference in the microstructure of the antlers.1.2 Study on Fingerprint of Amino Acids Fingerprints of amino acid components of antler medicinal materials were analyzed by pre-column derivatization high performance liquid chromatography with 18 amino acid controls such as aspartic acid.2 Quantitative analysis2.1 Analysis of total polysaccharide components2.1.1 Analysis of total polysaccharide content The phenol-sulfuric acid method and the fluorenone-sulfuric acid method were used for color development,and D-anhydrous glucose was used as a control to determine the total polysaccharide in the sample by spectrophotometry.2.1.2 Determination of monosaccharide composition The type and content of monosaccharides were determined by pre-column derivatization-HPLC method using mannose,glucosamine hydrochloride,ribose,glucuronic acid,galactose hydrochloride,glucose and galactose as controls.2.2 Phospholipid composition analysis2.2.1 Determination of phosphatidylcholine content The color of the total phospholipids in the samples was determined by spectrophotometry using molybdenum blue color reagent to develop color and potassium dihydrogen phosphate as control.2.2.2 analysis of phosphatidylcholine content The content of phosphatidylcholine in the sample was determined by HPLC method using phosphatidylcholine as a control.2.3 Determination of water soluble protein content Coomassie Brilliant Blue G-250 was used for color development,and bovine serum albumin was used as a control to determine the content of water-soluble protein in the sample by spectrophotometry.2.4 Determination of amino acid components and content The content of 18 amino acids in the sample was determined by pre-column derivatization high performance liquid chromatography with 18 amino acid controls such as aspartic acid.2.5 Determination of nucleoside components and content The contents of four nucleoside components in the samples were determined by HPLC method using uridine,hypoxanthine,xanthine and uracil as controls.2.6 Determination of putrescine content The content of putrescine component in the sample was determined by cation exchange resin column chromatography,benzoyl chloride pre-column derivatization,and HPLC method.3 Data processing and statistical analysis3.1 Cluster Analysis Based on the measured bone area ratio of bone filling and the content of chemical components,the SPSS 19.0 statistical software was used to systematically cluster the samples..3.2 Discriminant analysis Based on the measured bone area ratio of bone filling and the content of each chemical component,the SPSS 19.0 statistical software was used to discriminate the sample.Results:1 Qualitative identification results of antler medicinal materials1.1 Microscopic identification study results The area of bone filling in the antlers was35.5%⁴4.2%;the area of talion powder bone filling area was 46.9%⁵6.3%;the area of bone filling in the deer bone accounted for the area of bone fragments.71.9%.Therefore,the proportion of bone filling nests is from small to large,followed by antlers,antlers and deer bones.The ratio of the length of the bone filling in the antlers is 2.04:1².38:1;the ratio of the length of the bone filling in the antlers is 1.69:1².00:1;the ratio of the length of the bone filling in the deer bone is 1.54:1.Therefore,the length and the short diameter of the bone filling nest are from the big to the small,and the antlers are detached,the antlers and the deer bones.There is a significant difference between the area of the staghorn and the antlers,the area of the bone fragments,and the length and diameter ratio of the bone filling.1.2 Amino acid composition fingerprint study results The fingerprints of amino acid components of staghorn medicinal materials were analyzed.There were 21 common peaks,including 2 solvent peaks,18 known peaks and 1 unknown peak（No.3 is unknown peak）.The similarity calculation was performed on the common peaks,and the results showed that the similarity was greater than 0.9.2 Principal component analysis results2.1 Determination of total polysaccharide content The results of the anthrone-sulfuric acid method showed that the total polysaccharide content of the antlers was 0.53%¹.6%,the total polysaccharide content of the antlers was 0.37%⁰.42%,and the total polysaccharide content of the deer bone was 0.35%.The results of the phenol-sulfuric acid method were used to remove the antlers.The total polysaccharide was 0.34%⁰.96%,the total polysaccharide content of antler was 0.060%⁰.098%,and the total polysaccharide content of deer bone was 0.020%.In the two polysaccharide determination methods,the total polysaccharide content from high to low is:antlers,antlers,deer bones.There was a significant difference in the total polysaccharide content measured by the two methods of antlers and antlers.The results of determination of total polysaccharides by phenol-sulfuric acid method were higher than those of phenol-sulfuric acid method.2.2 Determination of the type and content of monosaccharides The antler polysaccharide is mainly composed of 7 kinds of monosaccharides such as mannose,glucosamine hydrochloride,ribose,glucuronic acid,galactose hydrochloride,glucose and galactose.D-galacturonic acid is not detected,and the highest glucose content is followed by mannose.And ribose is less.The total content of seven monosaccharides in the antlers was 0.65 mg·g^-10.96 mg·g^-1,and the monosaccharide composition ratio was 4.02:2.58:1:3.73:4.24:28.71:5.15;seven kinds of monosaccharides in the antlers The total content is 0.31 mg·g^-10.40 mg·g^-1,the monosaccharide composition ratio is 4.41:4.1:1:3.51:3.56:6.21:4.56;the seven monosaccharides in deer bone are 0.14 mg·g^-1 The monosaccharide composition ratio is4.82:2.65:1:3.73:2.05:5.25:3.43.The contents of the seven monosaccharides from high to low were:antlers,antlers,deer bones,antlers and antlers,and the total monosaccharide content was significantly different,which was consistent with the determination of total polysaccharide content.The ratio of glucose content in antlers and antlers is the largest and significant difference,so the total monosaccharide and total polysaccharide content can be identified by glucose content.2.3 Total phospholipid content determination results The total phospholipid content in antlers ranged from 9.99 mg·g^-1 to 21.6 mg·g^-1,and the total phospholipid content in antlers was 2.21 mg·g^-1 to 7.47 mg·g^-1.The total phospholipid content in deer bone was 32.1.mg·g^-1,the total phospholipid content from high to low in order:deer bone,antlers,antlers off the plate.There is a significant difference in the total phospholipid content of antlers and antlers.2.4 Determination of phosphatidylcholine content The content of phosphatidylcholine in antlers was 1.04 mg·g^-12.15 mg·g^-1,and the content of phosphatidylcholine in antlers was0.18 mg·g^-11.13 mg·g^-1.The choline content was 3.33 mg·g^-1.The content of phosphatidylcholine from high to low is:deer bone,antler,antlers,antlers and antlers.There is a significant difference in the content of phosphatidylcholine.The results are consistent with the determination of total phospholipids in the sample,so phospholipids can be used.The choline content determination results identify the total phospholipid content.2.5 Determination of water-soluble protein content The content of water-soluble protein in antlers was 0.20%⁰.30%.The determination of water-soluble protein content in antlers was 0.27%⁰.43%,and the content of water-soluble protein in deer bone was 0.22%.There was no significant difference in the water-soluble protein content between the antlers and the antlers.2.6 Determination of amino acid type and content Ten kinds of free amino acids were detected in antlers,antlers and deer bones.The free amino acid content of antlers was 0.62%⁰.86%,and the free amino acid content of antlers was 0.73%⁰.88%.The result was 0.8%.Only one of the essential amino acids was detected,which was lysine,and the content was more than 0.1%.18 kinds of hydrolyzed amino acids were detected in antlers,antlers and deer bones.The content of hydrolyzed amino acids in antlers was 26%³0.84%,and the amino acid content in antlers was 29.33%³0.81%.It is 52.04%.Among them,Gly glycine content is the highest,both exceeding 4.5%.Eight essential amino acids were detected and the lysine content was the highest.There was no significant difference in the types and contents of amino acids in the antlers and antlers,which was consistent with the determination of water-soluble protein content.2.7 nucleoside component types and content determination results The content of nucleosides in antlers and antlers was compared.The total content of four nucleosides in antlers was 20.71μg·g^-1117.1μg·g^-1,and the total content of four nucleosides in antlers was31.33.μg·g^-1137.9μg·g^-1,the total content of four nucleosides of antlers was 36.93μg·g^-1.Among them,the highest content of nucleosides was astragalus and hypoxanthine,and the content of uridine was the lowest.There was no significant difference between the determination results of nucleosides in antlers and the antlers.2.8 Determination of putrescine content The content of putrescine in the antlers ranged from 7 mg·g^-1 to 47.9 mg·g^-1,the content of staghorn detoxification was 31.2 mg·g^-1 to51.9 mg·g^-1,and the content of deer-putrescine was 28.4 mg·g^-1.The results of the determination of the content of putrescine in the two were analyzed by SPSS.There was no significant difference,and there was no significant difference in the content of antler and antlers.3 data processing and statistical analysis results3.1 Cluster Analysis Results The ratio of bone fragmentation area and eight chemical components（18 kinds of hydrolyzed amino acid content,water-soluble protein content,total polysaccharide content,putrescine content,total phospholipid content,total monosaccharide content,nucleoside component）The content of phosphatidylcholine and the content of phosphatidylcholine are data sources.Cluster analysis can be used to aggregate antler medicinal materials into one class,and antler detached medicinal materials（except 8#samples）are grouped into one group,and deer bones are clustered separately.3.2 Discriminant analysis results The sample medicine bone filling nest accounts for the area ratio of bone fragments and eight chemical components（18 kinds of hydrolyzed amino acid content,water soluble protein content,total polysaccharide content,putrescine content,total phospholipid content,total monosaccharide content,nucleoside content）And the content of phosphatidylcholine)has a typical discriminant function of 0.001,which is statistically significant.Through stepwise discriminant analysis,the area ratio of bone filling to bone fragments was obtained;total hydrolyzed amino acid;water-soluble protein;total polysaccharide;total monosaccharide component;putrescine;total phospholipid;nucleoside was a typical discriminant function coefficient,The typical discriminant function values are calculated.The eigenvalues of the six chemical components such as the bone filling area and the total hydrolyzed amino acid are:Y=0.004 X1+4.257 X2+186.871 X3+71.232 X4-179.25 X5+173.873 X6+3182.981 X7-183.713（Y>0,judged as Category 1,Y<0,judged as Category 2）.In the nine samples,the discriminant model obtained by discriminant analysis of seven principal components such as total amino acids can completely separate the antler and antler disk knockout samples.This method correctly classifies all the samples in the initial grouped samples and discriminates the classification effect better.Conclusion:1 In the qualitative identification,from the perspective of microscopic identification,there is a significant difference in the number of antlers and deer bone filling nests and the length and diameter ratio of bone filling.The bone filling nest accounts for the area of the bone fragments in the range of 35%to 45%.The plate is staghorn in the range of 45%to 60%,and more than 60%is deer bone.With the deer bone as the control,the ossified bone filling space accounted for an increase in the area ratio of the bone fragments,and the antlers were more ossified than the antlers.Measuring the area ratio of bone filling to bone fragments can be an important basis for identifying antlers and antlers,which can be identified by microscopic measurement.There is no difference in the amino acid fingerprints of the antlers and antlers.2 In the content analysis,the antlers and antlers were significantly different in the angles of polysaccharides and phospholipids:the total polysaccharide content in the antlers was higher than the total polysaccharide content in the antlers（the total polysaccharide content could be identified by the glucose content）;The content of phospholipids is higher than that of antlers（the total phospholipid content can be identified by phosphatidylcholine content）,which can be used to distinguish the antlers from the antlers.With deer bone as the control,with the increase of ossification degree,the total polysaccharide content decreased and the total phospholipid content increased.There were no significant differences in water-soluble proteins,amino acids,nucleosides,and putrescine.3 Through cluster analysis and discriminant analysis,the typical discriminant function is obtained,which can distinguish antlers,dislocation of antlers and bones of deer significantly.The clustering discriminant effect is very good.4 In clinical application,when nourishing effect is used in clinic,the dislocation of antler and antler can be combined.In the treatment of breast carbuncle,the dislocation of antler and antler should be distinguished.