Pheretima injection preparation and its preparing method
A technique for injection and preparation of Dilong, which is applied in the directions of anti-inflammatory agents, pharmaceutical formulations, antiviral agents, etc., can solve the problems of low activity, difficult industrialization, complicated operation, etc., achieve good pharmacological effects, easy industrial production, and technological operation. Simple and actionable effects
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Embodiment 1
[0071] (1) Take earthworm, add 6% sodium chloride to soak, wash 3 times, remove yellow mucus substance, add water for injection, wash away impurities, fully mash with electric meat grinder to make a rough homogenate, add Bacterial saline, shake well, place in 4°C refrigerator overnight, centrifuge the soaked saline, and centrifuge at 5000 rpm for 30 minutes at 4°C to obtain the supernatant;
[0072] (2) The supernatant is ultrafiltered with a hollow fiber column with a molecular weight cut-off of 8000, and the circulating fluid is retained, and concentrated to a concentrated solution with a relative density of 1.05 at 20° C.;
[0073] (3) Adjust the pH value of the ultrafiltration concentrated liquid to 5.0 with phosphate buffer, and separate and purify the ultrafiltration concentrated liquid through a CM-sephadex C25 ion-exchange gel chromatographic column and a sephadex G50 ion-exchange gel chromatographic column. -The mobile phase of sephadex C25 ion-exchange gel chromatogr...
Embodiment 2
[0080] (1) Take earthworm, add 8% sodium chloride to soak, wash 5 times, remove yellow mucus substance, add water for injection, wash away impurities, fully mash with electric meat grinder to make a rough homogenate, add Bacterial saline, shake well, place in 4°C refrigerator overnight, centrifuge the soaked saline, and centrifuge at 5000 rpm for 30 minutes at 4°C to obtain the supernatant;
[0081] (2) The supernatant is ultrafiltered with a hollow fiber column with a molecular weight cut-off of 10,000, the circulating fluid is retained, and concentrated to a concentrated solution with a relative density of 1.10 at 20° C.;
[0082] (3) The ultrafiltration concentrate is adjusted to a pH value of 7.0 with tris-hydrochloric acid buffer solution, and the ultrafiltration concentrate is passed through a CM-sephadex C25 ion-exchange gel chromatographic column and a sephadex G50 ion-exchange gel chromatographic column Separation and purification: the mobile phase of the CM-sephadex ...
Embodiment 3
[0089] (1) Take earthworm, add 7% sodium chloride to soak, wash 4 times, remove yellow mucus substance, add water for injection, wash away impurities, fully mash with electric meat grinder to make a rough homogenate, add Bacterial saline, shake well, place in 4°C refrigerator overnight, centrifuge the soaked saline, and centrifuge at 5000 rpm for 30 minutes at 4°C to obtain the supernatant;
[0090] (2) The supernatant is ultrafiltered with a hollow fiber column with a molecular weight cut-off of 8000, and the circulating fluid is retained, and concentrated to a concentrated solution with a relative density of 1.08 at 20° C.;
[0091] (3) Adjust the pH value of the ultrafiltration concentrate to 6.0 with a phosphate buffer buffer, and separate the ultrafiltration concentrate through a CM-sephadex C25 ion-exchange gel chromatographic column and a sephadex G50 ion-exchange gel chromatographic column Purification: The mobile phase of CM-sephadex C25 ion-exchange gel chromatograph...
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