| Dendrobium is a precious traditional Chinese medicine,according to the new classification of the National Union of Dendrobium,Dendrobium medicinal divided into 7 categories.Dendrobium medicinal contained in the "Shen Nong's Herbal Classic",as our traditional precious Chinese medicine.It have been widely used in the treatment of weakness after febrile diseases,thirsty,fire excess from yin deficiency,asthenia-syndrome and other diseases.Modern pharmacological experiments show that Dendrobium have anti-tumor,immunomodulatory,anti-oxidation,lower blood sugar and other activities.As people in the pursuit of health,quality of Dendrobium demand is rising.Dendrobium officinale is top grade of medicinal Dendrobium,there are many Chinese medicine's records in books of previous dynasties and it's singled out for the first time in Chinese Pharmacopoeia(2010 edition).Due to the growth of Dendrobium requires a unique natural conditions,Dendrobium officinale growth is very slow.The scarcity of Dendrobium officinale resulting in domestic market supply and demand imbalance.It is seriously restricted the development of Dendrobium candidum industry.In recent years,with the Dendrobium officinale plantation and processing technology breakthroughs,Dendrobium officinale artificial scale planting a "blowout" growth.Dendrobium officinale country now has three major areas:? Guangxi and Yunnan areas;? Danxia areas;?Zhejiang areas.Dendrobium polysaccharides are the effective component of herbs.According to the analysis of research group,different origin of Dendrobium officinale medium polar components with some geographical variation.Mapping technique using feature three areas of Dendrobium polysaccharides,amino acid composition analysis and study,to provide a reliable basis for the identification of different origin Dendrobium officinale.Study on the determination of the content of Dendrobium officinale polysaccharide by HPLC.Objective1.To establish PMP-HPLC characteristic pattern analysis method of Dendrobium officinale polysaccharides,and to provide the basis of the methods to quality control.To compare characteristic spectrum of the different areas of Dendrobium officinale polysaccharides.2.To establish the HPLC method for the determination of mannose and glucose of Dendrobium officinale polysaccharides,to provide reliable means of detection for the determination of Dendrobium officinale polysaccharides.Analysis the composition of Dendrobium officinale polysaccharides,to explore the specificity of different areas of the Dendrobium officinale polysaccharide fingerprint.3.To optimize the method of Dendrobium officinale fingerprint of amino acids,and the analysis of the composition of Dendrobium officinale amino acid composition in different areas and find out the different regions of specific amino acids of Dendrobium officinale.Methods1.Use the HPLC characteristic spectrum analysis method of Dendrobium officinale polysaccharides to analysis 20 batches of samples,specific chromatogram similarity evaluation system software writed by the State Pharmacopoeia Commission Specific chromatogram similarity evaluation system software(2004A version)was adopted to generate a total of mode;using the established method of Dendrobium officinale polysaccharides to analyse the three areas of origin polysaccharide.2.To establish the HPLC determination of mannose and glucose in Dendrobium officinale polysaccharides.Taking glucosamine hydrochloride as an internal standard.The chromatographic column is Agilent ZORBAX Eclipse XDB-C18(4.6×150mm,5?m)Acetonitrile-0.02mol/L ammonium acetate as mobile phase with gradient elution,flow rate is 1ml/min,the detection wavelength is 250nm,the column temperature is 30 ?.3.Using the methods of amino acid column derivatization and the HPLC characteristic spectrum technology to anaylse the composition of the amino acid of different ares of Dendrobium officinale and the similarity evaluation.Results1.Established a PMP-HPLC method for determination of mannose and glucose in polysaccharide of Dendrobium officinale.(1)the Dendrobium officinale polysaccharides are marked out of a total of five common peaks in total control mode,20 batches of samples similarity of 0.986 to 1.000.(2)Analysis of different origin Dendrobium officinale polysaccharide hydrolysis,mannose and glucose peak area and were more than 97%of the total peak area.(3)20 batches of different origin of Dendrobium officinale polysaccharide composition is basically the same.Different regions of mannose and glucose peak area ratio of Dendrobium officinale polysaccharide hydrolysis with a degree of difference,Guangxi,Yunnan area ratio in 3.45?6.62,Danxia producing areas in 2.61?3.92,Zhejiang area in 3.21?5.79.2.Establish the HPLC determination method of content of mannose and glucose in Dendrobium officinale polysaccharide.(1)The HPLC determination method of content of Dendrobium officinale polysaccharide is feasible and reliable and stability.Analysis of 20 batches of Dendrobium officinale,mannose content 17.79%?34.89%content and glucose content 2.92%-14.20%.(2)The content of mannose and glucose in Dendrobium officinale polysaccharide is differences in different areas,but there is no regional differences.(3)The mannose relative peak area percentage content from 72.99%?86.06%,glucose content from 13.07%?25.61%,mannose and glucose content of total polysaccharide content of more than 97%;the sum of mannose and glucose are close to its UV total polysaccharide content,determination of mannose and glucose samples the total amount of HPLC with an average of 1.039 times the amount of UV polysaccharide.(4)The method is accurate,reliable,durable and strong,the stability is good by UV spectrophotometry.3.To analyse the composition of the amino acid of different areas of Dendrobium officinale with the HPLC analytical method.Comparison the similarities and differences of Dendrobium officinale amino acid composition between different areas.The different areas of Dendrobium officinale are basically the same characteristic peaks,the identification of which 15 pairs of amino acids beneficial to the human body(aspartic acid,glutamic acid,serine,arginine,threonine,proline,alanine acid,valine,isoleucine,leucine,histidine,phenylalanine,lysine,tyrosine).All areas composition of amino acid of Dendrobium officinale is similar and no regional differences.Similarity in 19 areas of amino acids are 0.893?0.994;the similarity of Danxia area,Guangxi Yunnan region,Zhejiang region were 0.950,0.993,0.988.Conclusion1.Dendrobium officinale polysaccharides PMP-HPLC characteristic spectrum analysis method is feasible and practical,which can provide the method for the quality control Dendrobium officinale.2.A total of 20 batches of Dendrobium officinale polysaccharide in different producing areas,the hydrolysis monosaccharides composition is basically the same,all containing mannose,glucose,galactose,xylose,arabia,the relative content of mannose and glucose are more than 97%.3.The HPLC content determination method of mannose and glucose in Dendrobium officinale polysaccharide is feasible and reliable,can be used as a means of determination of polysaccharides of Dendrobium candidum.4.The types and proportions of Dendrobium officinale amino acid in different regions are basically the same,but no obvious regularity. |
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