Process for extracting epigoitrin from isatis root and use thereof
A technology that reports Yichun and Banlangen, applied in the direction of medical preparations containing active ingredients, pharmaceutical formulations, organic active ingredients, etc., can solve the problem of not being able to use large-scale production
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Embodiment 1
[0051] Extraction of main parts
[0052] 1. Materials and Instruments
[0053] Materials: 100 g of isatidis extract prepared by Suzhou Siyuan Pharmaceutical Technology Co., Ltd. by the method of Chinese application 03110306.5.
[0054] Neutral alumina: purchased from Shanghai Ludu Chemical Reagent Factory
[0055] Silica gel H: Qingdao Ocean Chemical Co., Ltd.
[0056] 2. Extraction method:
[0057] Grind the Radix Radix Radix extract to 60 mesh; add it to the neutral alumina column; use chloroform:methanol mixed solvent gradient elution, the gradient is to increase 1ml methanol per 500ml eluent after collecting 50 fractions, the elution speed 6ml / min; collect the fractions containing epichun, and detect the concentration of epichun therein with HPLC; combine the fractions containing epichun; distill off the solvent under reduced pressure at 60°C.
[0058] Dissolve the obtained powder in methanol, add a small amount of silica gel H to the column of silica gel H, and wash w...
Embodiment 2
[0081] Anti-influenza virus experiment of extract
[0082] 1. Samples: a) indirubin, b) quinazolones, c) the epiblast prepared in experiment 3 of Example 1, with a concentration of 0.01 mg / ml, and d) prepared by the applicant according to the method of Chinese application 03110306.5 Radix isatidis extract, the concentration is 0.05mg / ml.
[0083] Control: ribavirin, concentration 0.01mg / ml.
[0084] 2. Detection method:
[0085] (1) Experimental materials:
[0086] Virus strain: influenza type I virus A / shanghai / 26 / 85 (H, N) strain, hemagglutination titer 1:1280 (preserved by the Department of Microbiology and Immunology, Nanjing Medical University).
[0087] Chicken embryos: A total of 22 10-year-old chicken embryos were hatched (Department of Microbiology and Immunology, Nanjing Medical University).
[0088] Chicken erythrocytes: adult males were washed three times with normal saline to prepare a concentration of 0.5%.
[0089] (2) Experimental method:
[0090] a) Grou...
Embodiment 3
[0097] Cytopathic Research
[0098] 1. Test material:
[0099] Sample: Table Yichun prepared in Example 1 Experiment 3
[0100] Virus strain: Influenza virus A / FM1 / 47 / H1N1 (sourced from Department of Microbiology and Immunology, Nanjing Medical University).
[0101] Sendai virus (gifted by the Department of Microbiology and Immunology, Nanjing Medical University).
[0102] Cells: MDCK cells (gifted by the Department of Microbiology and Immunology, Nanjing Medical University).
[0103] 2. Experimental method:
[0104] a) Administer 0.6 μg / ml epiblaston to the cells, and add 100 TCID after 24 hours 50 influenza virus or Sendai virus.
[0105] b) The sample was mixed with 0.6μg / ml and 100TCID 50 Influenza virus or Sendai virus was mixed, placed overnight at 4°C, and the cells were inoculated the next day.
[0106] c) The sample was directly mixed with 0.6μg / ml and 100TCID 50 Influenza virus or Sendai virus and cell mix.
[0107] After 24-48 hours, the cytopathic changes ...
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