Study On Extraction And Purification Technology Of Total Amino Acids From Nervilia Fordii(Hance) Schltr And Its Fingerprint

Nervilia fordii (Hance) Schltr., is the traditional export Chinese crude drug as well as a specialty in Guangxi Province. Typhonium Giganteum and Clethra Loosestrife are the aliases of it. Nervilia fordii (Hance) Schltr, cold in nature and bitter in taste, belongs to heart, liver and lung meridian. It has the abilities of watering lung, activating and relieving swelling and pain. It attends Consumptive Lung Disease, cough, stomatitis, sore throat, tuberculosis, carbuncles rash and bones injuries. Nervilia fordii (Hance) Schltr is rich in amino acid, the percentage of which reaches about 10% and makes it one of the primary origin of the production of amino acid.The thesis has studied the relevant literature Nervilia fordii (Hance) Schltr., the method for determination of amino acids and the isolation and purification of amino acids. What's more, it also adopted HPLC Fingerprint of Flavonoids in portation of Nervilia fordii (Hance) Schltr's amino acids.Document Research:The thesis is a review of Nervilia fordii (Hance) Schltr's material resources, biological characteristics, shapes, identification, chemical components, pharmac eutical, the exploiture and applications. Apart from that, the paper also summarizes not only the method for isolation and purification of amino acids but also the pharmacology and Fingerprint of Flavonoids. The above provides the basic theory for the design and operation of the experiment.Research on quality standard:identifiton amino acids in Nervilia fordii (Hance) Schltr by TLC.Drop the solution of sample and reference substance separately on the same silicone G with 4% disodium hydrogen phosphate,butanol-glacial acetic acid-water (4:1:1) is used as developer,develop,dry,5% ninhydrin used as chromogenic reagent,examine by eyes under sunlight,for sample and reference substance separately,there are Leucine,trytophan,valine,,alanine, glutamic,aspartic in the same position.Determination of amino acids:Based on the a-amino acids and ninhydrin color reaction principle, the ninhydrin colorimetry was used to determine quantitatively the amount of the free amino acids in Nervilia fordii (Hance) Schltr. Analysis of effect of PH,temperature,heat time and cool time on the absorbance.The extraction technology for total free amino acids in Nervilia fordii (Hance) Schltr: Employing the content of free amino acids in Nervilia fordii (Hance) Schltr as index,On the base of single factor screening,the effects of the menstruum and temperature,cycles of extracting,the amount of menstruum on the extracting process were investigated. The uniform design was adopted for getting the optimal mixed technique parameters, and was verified by experiment. The optimal condition was 40℃,1.0 h, the solid-liquid ratio was 1:20.The decolorization technology of the extract of Nervilia fordii (Hance) Schltr: Employing the loss of total amino acids and decolorization of the extract of N fordii (Hance) Schltr as index. On the base of single factor screening, the effects of the pH value, the amount of activated carbon, decolorizing temperature and time on the decolorizating process were investigated. The uniform design was adopted for getting the optimal mixed technique parameters, and was verified by experiment. The concentration of pH value of extract solution was 3.0,the activated carbon was 0.1 g/10 mL, tempetature was at 40℃,and decolorizing time lasted 20 min.Purified technology of total amino acids:Amino acids was purified by 732 cation exchange resin.Study the dynamic adsorption propreties of 732 cation exchange resin,research on how these affect the enrichment of total amino acids.The factors are the concentration of liquid filled column,flow velocity of upper samples,divulging curves,elution solvent and elution amout,according to which the optimal technology for enrichment is determined and then to verify it.Use the pre-column derivatization-HPLC to develop fingerprint of Nervilia fordii (Hance) Schltr:Phenylisothiocyanate(PITC) was employed as derivatizing agent.Separation was performed on CAPCELL PAK C18 SG300 S5(250mm×4.6mm) analytical column with mobile phase consisting of pH6.5 sldium acetate buffer solution and acetonitrile.The column temperature was at 40℃,gradient elution with the flow rate of 1.0 mL/min.The Fluorescence detecting wavelength was set at 254 nm; sixteen common peaks on the HPLC fingerprints.There are Aspartic,Glutamic,Serine,Glycine,Threonine,Alanine,Tyrosine, Proline,Viline,Methionine,Isoleucine,Leucine,Phenylalanine,Tryptophane,Lysine and an anknown amino acid.