Method for preparing pig-bile-combination-type total bile acid and application thereof
A combined type, pig bile technology, applied in the treatment of colonic inflammation, the field of preparation of combined total bile acid, to achieve the effect of blocking damage
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Embodiment 1
[0023] Taking fresh pig bile method 100mL as an example, the preparation method of pig bile-bound cholic acid is as follows:
[0024] 1. Ethanol extraction
[0025] Take fresh pig bile, add 500mL of ethanol with a mass fraction of 95%, let it stand for 12-14 hours, filter, recover the solvent, and obtain a brown-black extract;
[0026] 2. Separation and purification
[0027] Take 2g of the above-mentioned extract, add 4g of 100-160 mesh column chromatography silica gel to mix the sample, evaporate the solvent, and grind into 80 mesh fine powder; dry the obtained fine powder and neutral alumina according to the mass ratio of 1:15 In the chromatographic column, elute with ethanol with a mass fraction of 95%, collect the eluate, and spot the sample on a silica gel G thin-layer plate with a capillary tube. The volume ratio of ethyl acetate to methanol, water, and glacial acetic acid is 10 : 2: 1: 0.1 mixed developing agent, developed with vanillin sulfuric acid test solution col...
Embodiment 2
[0031] Taking 100 mL of fresh pig bile as an example, the preparation method of pig bile-bound cholic acid is as follows:
[0032] In the separation and purification step 2, take 2 g of the above extract, add 4 g of 100-160 mesh column chromatography silica gel to mix the sample, evaporate the solvent, and grind into 80 mesh fine powder; the obtained fine powder and neutral alumina are mixed according to the mass ratio of 1 The ratio of : 10 is installed in the chromatographic column, and the ethanol with a mass fraction of 95% is used for eluting, and the eluate is collected, and the sample is spotted on a silica gel G thin-layer plate with a capillary tube, and ethyl acetate, methanol, water, The volume ratio of glacial acetic acid is 10:2:1:0.1 and the mixed developing agent is developed, and the color is developed with the vanillin sulfuric acid test solution color reagent, and the eluents are combined when the spots of conjugated hyodeoxycholic acid appear on the TLC plate...
Embodiment 3
[0035] Taking 100 mL of fresh pig bile as an example, the preparation method of pig bile-bound cholic acid is as follows:
[0036] In the separation and purification step 2, take 2 g of the above extract, add 4 g of 100-160 mesh column chromatography silica gel to mix the sample, evaporate the solvent, and grind into 80 mesh fine powder; the obtained fine powder and neutral alumina are mixed according to the mass ratio of 1 The ratio dry method of : 20 is installed in the chromatographic column, eluting with 95% ethanol with mass fraction, collecting the eluate, spotting the sample on the silica gel G thin-layer plate with a capillary, and using ethyl acetate, methanol, water, The volume ratio of glacial acetic acid is 10:2:1:0.1 and the mixed developing agent is developed, the color is developed with the vanillin sulfuric acid test solution, and the eluents are combined when the spot of conjugated hyodeoxycholic acid appears on the TLC plate , until the spots disappear.
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