Preparation of dry powder water soluble extract of ground beetle and application thereof in medicine
A technology of water extraction and soil turtle, which is applied in the field of biomedicine, can solve the problems of weakening the therapeutic effect, reducing the amount of medicine taken, and large dosage, and achieves the effect of safe polypeptide components and safe clinical application
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Embodiment 1
[0030] The preparation of embodiment 1 soil beetle water extraction polypeptide coarse powder
[0031] Suspend 15 g of dry wood beetle powder (obtained from grinding dried wood beetle) in 500 ml of distilled water. The resulting suspension was stirred at a constant temperature of 37.5°C for 40 minutes. The suspension was filtered naturally under normal pressure, and the filtrate was freeze-dried to obtain 2.25g (15%) of the ground beetle water-extracted polypeptide coarse powder (W 0 ), stored at 4°C until use.
Embodiment 2
[0032] Example 2 Separation and Purification of Water-extracted Polypeptide Coarse Powder of Wood Beetle
[0033] Sephadex G25 was processed and packed into a column (2.6×60cm), and 0.005mol / L ammonium acetate was filtered through a 0.45nm microporous membrane as a buffer balance. Dissolve 2.25g of ground beetle water-extracted polypeptide coarse powder in 1ml of distilled water, and centrifuge at 10,000 rpm for 10 minutes. The supernatant was applied to a Sephadex G25 chromatography column, eluted with 0.005mol / L ammonium acetate buffer solution, the flow rate was 0.5mL / min, and one tube was collected in 8 minutes. Each tube was measured by a UV spectrophotometer with a wavelength of 280nm, and the data was recorded. Curves were made with Microsoft Excel, fractions were pooled according to peak shape, and freeze-dried. yielded 0.24 g (10.5%) of W I , 0.54g (24%)W II , 0.74g (33%)W III and 0.19g (8.5%) W IV Four kinds of water-extracted peptide components.
[0034] Examp...
Embodiment 5
[0038] Example 5W IV The sequence of the polypeptide in
[0039] Measure W according to the method of Example 4 IV After the HPLC-ESI-MS, select suitable peak to measure ESI-MS, analyze collection of graphs according to complementary principle ( Figure 4 ), the amino acid sequence is as follows: H-Ser-Cys-Leu(Ile)-Ser-Pro-Val-Ser-Leu(Ile)-Thr-Arg-Pro-Trp-OH.
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