Preparation of wood louse powder extract and uses thereof
A technology of extracts and soft-shelled turtles is applied in the application field of preparing analgesic drugs, and can solve problems such as inconvenience of patients
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Embodiment 1
[0048] Example 1 Preparation of water-extracted polypeptide coarse powder of soft-shelled turtle
[0049] 15g of dry soil turtle powder (obtained by grinding dry soil turtle) was suspended in 500 ml of distilled water. The resulting suspension was stirred at a constant temperature of 50°C for 60 minutes. The suspension is naturally filtered under normal pressure, and the filtrate is freeze-dried to obtain 3.0 g (20%) of the crude soft-shelled turtle water-extracted polypeptide powder, which is stored at 4°C until use.
Embodiment 2
[0050] Example 2 Separation and Purification of Polypeptide Coarse Powder from Soft-shelled Turtle
[0051] Sephadex G25 is treated and packed into a column (2.6×60cm), 0.007mol / L ammonium acetate is filtered through a 0.45nm microporous membrane and used as a buffer balance. 3.0g of water-extracted polypeptide coarse powder of soft-shelled turtle was dissolved in 0.5ml of distilled water and centrifuged at 10,000 rpm for 10 minutes. The supernatant was applied to Sephadex G25 chromatography column, eluted with 0.007mol / L ammonium acetate buffer, flow rate 0.8mL / min, 8min collected in one tube, each tube was measured by UV spectrophotometer, wavelength 280nm, recorded data, Use Microsoft Excel to make a curve, combine the fractions according to the peak shape, and freeze-dry. Obtained 0.20g (6.7%) W I , 0.50g (17%) W II , 0.99g (33%) W III And 0.30g (10.0%) W IV Four water-extracted peptide components.
Embodiment 3WI
[0052] Example 3W I , W II , W III And W IV Analgesic activity determination
[0053] Divide male ICR mice (22±2g) into a control group (W 0 ), and W I , W II , W III And W IV Treatment group, 10 mice in each group. W I , W II , W III And W IV Dissolve them with 0.3ml of saline. W I , W II , W III And W IV The gavage dose is calculated according to the dose (mg / kg) = 9.01×38mg / piece×yield rate, that is, 0.4mg / 0.3ml / piece W I , 9.1mg / 0.3ml / W II , 12.5mg / 0.3ml / W III With 3.2mg / 0.3ml / only W IV . W 0 Group 0.3ml / only normal saline. The mouse is placed in a special plastic cylinder, the tail is exposed outside the cylinder, and the indoor temperature is kept at (20±1℃). After the animal is stable for 30 minutes, the pain is measured by the radiant heat tail flick method, and a special lampshade is used to make it emit A light beam with a diameter of 2mm is irradiated at a distance of about 2-3cm from the end of the tail, and the voltage of the light source is adjusted to make the late...
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