The palm leaf anti-alkali is also called palmatine, and the tetrahydropalmatine sulfate is also called dltetrahydro palmatine sulphate.
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Biochemical Technology Topics: Extraction and Separation of Palmetine and Preparation of Tetrahydropalmatine
Corydalis B is a sedative and tranquilizer used to relieve pain caused by diseases of the stomach system, labor pain, headache, insomnia, etc.
The traditional Chinese medicine, Yanhusuo (the root of Corydalis yanhusuo WT Wang) contains a small amount of tetrahydropalmatine, while its dehydrogenated compound, palmatine, is high in some plants. It contains palmatine in the rhizome of Fibreursa recisa paerre in the south of China, and then hydrogenation reaction to prepare tetrahydropalmatine.
Huang Teng has been included in the "Compendium of Materia Medica", Li Shizhen said that "Teng Shengsheng Lingnan, the shape of the defense has been, but people often serve this vine, the food is poisonous, and naturally not." Modern folk as a heat-clearing anti-inflammatory drug. Commonly used for traumatic infections, tonsillitis, pharyngitis, conjunctivitis, enthusiasm and jaundice.
The purpose of the experiment requires:
1. Master an extraction method for quaternary ammonium alkaloids.
2. Master the general physical and chemical properties of alkaloids and their relationship to their properties.
3. Understand the relationship between the structure and properties of the yellow alkaloids.
4. Familiar with the conditions and methods of alkaloid precipitation reactions.
5. Master the method and purification of preparing alkaloid crystalline salts.
6. Master the preparation method of tetrahydropalmatine.
Physical and chemical properties of known components in yellow vine:
The constituents in the rhizome and root of the yellow vine are mainly palmatine. It also contains a small amount of jatrorrhizine, scutellaria, scutellaria, lactone and sterol. It is also known to contain berberine in roots, roots and bark.
1. Palmetine (Bamatine, Palmatine)
This product is a quaternary ammonium alkaloid, soluble in water, ethanol, almost insoluble in chloroform, ether, benzene and other solvents, palm leaf anti-alkali hydrochloride, palmation Chloride C21H22O4N · Cl · 3H2O is yellow Needle crystals, melting point 205 ° C (decomposition). Its physical and chemical properties are similar to berberine hydrochloride. Palmatine icdide C21H22O4N·I·2H2O is an orange-yellow needle-like crystal with a melting point of 241 ° C (decomposition).
2. Pharmacopoeine (Yato base, Jatrorrhizine)
This product has phenolic hydroxy quaternary ammonium salt, its physical and chemical properties are similar to that of palmatine, but it is more soluble in caustic soda. Its solubility in water is also greater than that of palmatine hydrochloride, which can be separated by this property. . Jatrorrhizine Chloride C20H20O4N·Cl·H2O is a copper needle-like crystal with a melting point of 204-206 ° C. Its bitter acid salt (Jatrorrizine picrate) C20H20O4N·C6H2O7N2 is an orange-yellow columnar crystal with a melting point of 217~ 220 ° C (decomposition).
3. Berberine (Berberine)
This product is a quaternary ammonium alkaloid, its free base is yellow long needle-like crystal, C20H18O4N·OH·5 H2O, melting point 145 ° C, drying at 100 ° C, the loss of crystal water turns brown. Berberine can be slowly dissolved in water (1:20), ethanol (1:100), soluble in hot water, hot ethanol, slightly soluble in acetone, chloroform, benzene, almost insoluble in petroleum ether, small sputum The base can form an adduct with chloroform, acetone and benzene. Berberine Chloride (Cerberine Chloride) C20H16O4N·Cl2H2O, melting point 205 ° C (decomposition), slightly soluble in cold water, more soluble in boiling water, its nitrate and hydroiodide, extremely difficult to dissolve in water (cold water about 1 :2000). The neutral sulfate, phosphate and acetate of berberine are more soluble in water, and the solubility of berberine salts in water:
4. Tetrahydropalmatine:
Tetrahydropalmatine is racemic tetrahydropalmatine, a tertiary amine base, its free base C21H25O4N has a melting point of 146~148 ° C, is insoluble in water, soluble in hot ethanol (smaller solubility in cold ethanol), soluble in Among the chloroform, benzene and diethyl ether, the acid sulfate of tetrahydropalmatine is colorless needle crystal, the melting point is 245~246 °C, the solubility in cold water is small, and it is larger in hot water. The neutral sulfate is long columnar crystal. The melting point is 220 ° C. The solubility in water is larger than that of acidic sulfate, and the hydrochloride is hardly soluble in water.
L-tetrahydropalmatine, Rotundine, has a melting point of 141-142 ° C [α] D16-257.5 ° C (C = 1.1 chloroform). This product is abundant in the roots of the Stephania sinica Diels and the S. rotundalour.
Method for extracting palmatine and preparing fumarate
Palmatine is a quaternary ammonium alkaloid, soluble in water and a highly polar organic solvent (such as methanol, ethanol, etc.) so it can be extracted with methanol, ethanol or water. It is then precipitated by salting out, reducing its solubility in water, and separated from other impurities. Palmatine is an organic base. Although it has a positive charge, its affinity with water is still less than the strong affinity of NaCl, NaCl and water, which reduces the solubility of palmatine in water and causes it to be "squeezed out".
The solubility of palmatine hydrochloride in cold water (slightly larger than that of berberine hydrochloride) can be separated by this property.
method one)
2. Process
(1) Leaching: Take 100g of yellow vine powder, cold soak with 1% acetic acid (about immersion raw material, about 500ml) for 1-2 days, filter with nylon cloth, add slag and add 15 acetic acid for one day, and combine the filtrate ( Leave 1 ml for the alkaloid qualitative reaction).
(2) Salting out, neutralization: The filtrate is adjusted to pH 9 with 40% NaOH solution and 7% refined salt is added, that is, yellow insoluble matter is precipitated, and the precipitate is incubated at 80 ° C to allow the precipitate to coagulate, and the supernatant is decanted. The chrysanthemum filter paper was filtered to obtain crude palmatine free base and dried.
(3) Refining: Crude palmatine, add 80ml of 80% ethanol, heat and reflux to dissolve, about ten minutes, heat filtration, residue and a small amount of ethanol to treat once, suction filtration and finally take the dropper to take the ethanol Insoluble in the Buchner funnel, the filtrate was combined and 6N hydrochloric acid was added dropwise to the filtrate to place it in PH2, that is, golden yellow needle crystals were precipitated, suction filtered, and then recrystallized once with ethanol, and the method was to thicken palmatine chloride. In the crystallized conical flask, 95% ethanol was added to the dropping tube to dissolve the crystal (heated on a water bath), and the clear liquid was obtained by suction filtration, and the crystal was deposited by stoppering. If the filtrate has precipitated crystals during suction filtration, it can be heated and crystallized on a water-soluble solution and then placed in a stopper, so that the crystal form precipitated is good. If the filtrate volume is too large, it can be melted and condensed until the edge of the bottle wall is slightly solid. The mother liquor is properly concentrated and a portion of the palmatine chloride can be precipitated.
(4) Reduction: 1 g of palmatine chloride purified product was taken. Add 50ml of distilled water to the 50~100ml round bottom flask, concentrate 0.5ml of sulfuric acid, 0.7g of zinc powder (industrial production should be added zinc powder in batches), keep boiling for direct heating, and react for 4~5 hours (the color of the solution is gradually increased during the reaction) Lighten until colorless), after the reaction is completed, pour out the supernatant liquid, and then wash the reaction bottle and zinc slag slightly with distilled water (1~2ml) for 1~2 times. The combined solution is cooled and there is tetrahydropalmatine acid. Sulfate precipitates. After suction filtration, the crystals were placed in a 25 ml Erlenmeyer flask and dissolved by heating with 10 ml of 70% ethanol. After hot filtration, wash the container with a small amount of ethanol (1~2ml), add ammonia water to PH9 to the hot solution (about 60 ° C), precipitate scaly crystals immediately, filter by suction, wash with distilled water (the crystal obtained here adds water) Can it dissolve? Can chloroform dissolve?) Dry the fumarate free base. drying.
(5) salt formation: weighed 0.5 g of tetrahydropalmatine free base. Place in a 25 ml Erlenmeyer flask (or small flask) and add 4 ml of 90% ethanol, heat on a water bath to dissolve, and add 5% sulfuric acid ethanol solution to the Congo red test paper with a crude capillary to light blue (pH about 5). Cooling and crystallization. The yellowish columnar crystals are dried by suction filtration and dried at 50 ° C or lower, that is, tetrahydropalmatine neutral sulfate. It can be recrystallized once with water, and the melting point is measured after drying. Melting point - °C, yield -.
Extracted with a small sample of acetic acid water, the general yield is 1-2%, the refluxing rate of ethanol is 3~4%, 100g of yellow vine powder is taken, and the ethanol is refluxed with 95% ethanol for three or four times to about 120ml. It can be operated under (3) refining, and hydrochloric acid is added to obtain palmatine chloride.
3. Extraction process description
(1) The polysaccharides contained in the yellow vine are soluble in water, which often affects the salting out of the water immersion liquid and the filtration after neutralization. Therefore, it is preferred to pour most of the clarified liquid and then use a cloth bag to collect the precipitate after standing. The medicinal materials should not be smashed too fine.
(2) Palmatine chloride is a gold needle crystal with strong yellow fluorescence. Tetrachloropalmatine is a colorless flaky crystal with no fluorescence. Therefore, the color of the reaction liquid can be judged as the end point of the reduction. At the beginning of the reduction, the reaction solution is orange-yellow. As the reduction reaction proceeds, the color of the reaction solution gradually recedes, and the light beige or colorless color can be used as the end point of the reaction.
(3) Tetrahydropalmatine is more easily oxidized to palmatine, so it should be continuously operated as soon as possible during the preparation process.
(4) Tetrahydropalmatine hydrochloride and acid sulfate have low solubility in water, so it is generally made into a water-soluble neutral sulfate for use in formulating injections.