Method for extracting taurocholic acid in sheep bile through column chromatography isolation method

Abstract

The invention discloses a method for extracting taurocholic acid in sheep bile through a column chromatography isolation method. The method comprises the following steps: filtering, performing protein precipitation treatment, performing depigmentation treatment, salting out, extracting, dewatering concentration and performing column chromatography isolation. According to the method, the sheep bile serves as an extracting raw material, the extraction cost of the taurocholic acid is reduced, the extraction process is simple, and the income is higher. The taurocholic acid is extracted through the column chromatography isolation, the column chromatography isolation is good in purification effect, the extracted taurocholic acid does not have a fishy smell and is high in purity, the product is high in yield, the extraction process is simple, and industrial production is promoted.

Description

[0001] technical field

[0002] The invention belongs to the technical field of biomedicine and relates to a method for extracting taurocholic acid, in particular to a method for extracting taurocholic acid in goat bile by a column chromatography separation method. Background technique

[0003] Bile acid is the main component of medicinal bile and is a general term for various bile acids. Various bile acids are combined with glycine or taurine in the liver and secreted into the gallbladder in the form of sodium salt. Sodium glycocholate and sodium taurocholate, as the main components of bile salts, have an important relationship with the digestion and absorption of fat in the body. They can not only enhance the activity of lipase, catalyze the decomposition of fat, and increase the absorption of fat-soluble vitamins, It can also promote the secretion of bile and stimulate the movement of the small intestine.

[0004] Taurocholic acid is widely found in the bile of cattle, ...

Examples

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...

[0001] technical field

[0002] The invention belongs to the technical field of biomedicine and relates to a method for extracting taurocholic acid, in particular to a method for extracting taurocholic acid in goat bile by a column chromatography separation method. Background technique

[0003] Bile acid is the main component of medicinal bile and is a general term for various bile acids. Various bile acids are combined with glycine or taurine in the liver and secreted into the gallbladder in the form of sodium salt. Sodium glycocholate and sodium taurocholate, as the main components of bile salts, have an important relationship with the digestion and absorption of fat in the body. They can not only enhance the activity of lipase, catalyze the decomposition of fat, and increase the absorption of fat-soluble vitamins, It can also promote the secretion of bile and stimulate the movement of the small intestine.

[0004] Taurocholic acid is widely found in the bile of cattle, ...

Examples

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...