Method for extracting taurocholic acid in sheep bile through column chromatography isolation method
A technology of taurocholic acid and separation method, which is applied in the field of biomedicine, can solve the problems of low extraction rate, difficult source of cattle bile, high impurity content of products, etc., to reduce extraction cost, shorten process cycle, and high product yield Effect
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Embodiment 1
[0029] The method for extracting taurocholic acid in goat bile by column chromatography separation method comprises the steps:
[0030] (1) Filtration: Take fresh sheep bile and put it in a double-layer gauze for 48 hours to collect the filtrate;
[0031] (2) Protein precipitation treatment: Add 3 times the volume of ethanol to the filtrate in step (1), stir and centrifuge at 15,000 rpm / min, 8°C for 30 minutes, and collect the yellow-green supernatant;
[0032] (3) Pigment removal treatment: Add 10g / 100ml activated carbon to the supernatant in step (2), heat at 40°C for 20 minutes, and after cooling, centrifuge at 15,000rpm / min and 8°C for 45 minutes to collect the colorless supernatant ;
[0033] (4) Concentration: Concentrate the supernatant of step (3) by 15 times in a rotary evaporator;
[0034] (5) Salting out: Add 30 times the volume of water to the concentrated liquid in step (4), adjust the pH value to 6.5 with hydrochloric acid, add sodium chloride until a white pre...
Embodiment 2
[0043] The method for extracting taurocholic acid in goat bile by column chromatography separation method comprises the steps:
[0044] (1) Filtration: Take fresh sheep bile and put it in a double-layer gauze for 48 hours to collect the filtrate;
[0045] (2) Protein precipitation treatment: add 3 times the volume of acetonitrile to the filtrate in step (1), stir and centrifuge at 15,000 rpm / min, 8°C for 30 minutes, and collect the yellow-green supernatant;
[0046] (3) Pigment removal treatment: Add 10g / 100ml activated carbon to the supernatant in step (2), heat at 40°C for 20 minutes, and after cooling, centrifuge at 15,000rpm / min and 8°C for 45 minutes to collect the colorless supernatant ;
[0047] (4) Concentration: Concentrate the supernatant of step (3) by 15 times in a rotary evaporator;
[0048] (5) Salting out: Add 30 times the volume of water to the concentrated liquid in step (4), adjust the pH value to 6.5 with hydrochloric acid, add sodium chloride until a whit...
Embodiment 3
[0057] The method for extracting taurocholic acid in goat bile by column chromatography separation method comprises the steps:
[0058] (1) Filtration: Take fresh sheep bile and put it in a double-layer gauze for 48 hours to collect the filtrate;
[0059] (2) Protein precipitation treatment: Add 3 times the volume of ethanol to the filtrate in step (1), stir and centrifuge at 15,000 rpm / min, 8°C for 30 minutes, and collect the yellow-green supernatant;
[0060] (3) Pigment removal treatment: Add 10g / 100ml activated carbon to the supernatant in step (2), heat at 40°C for 20 minutes, and after cooling, centrifuge at 15,000rpm / min and 8°C for 45 minutes to collect the colorless supernatant ;
[0061] (4) Concentration: Concentrate the supernatant of step (3) by 15 times in a rotary evaporator;
[0062] (5) Salting out: Add 30 times the volume of water to the concentrated liquid in step (4), adjust the pH value to 6.5 with hydrochloric acid, add sodium chloride until a white pre...
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