CN103948654A - Method of extracting, purifying and inhibiting alpha-glucosaccharase active ingredient from chestnut shell - Google Patents

Abstract

The invention relates to a method of extracting, purifying and inhibiting an alpha-glucosaccharase active ingredient from a chestnut shell. The method comprises the following steps: firstly, cleaning the chestnut shell, drying, crushing, refluxing and extracting by 75% (v/v) alcohol, concentrating under reduced pressure, adding water and statured n-butyl alcohol liquor to extract; and finally, carrying out adsorption elution by using macroporous resin AB-8 to obtain polyphenol extract with effect of inhibiting activity of alpha-glucosaccharase.

Description

A kind of method of extracting purification Inhibiting α-glucosidase active component from chestnut shell

Technical field

The invention belongs to natural product chemistry field, particularly a kind of method of extracting purification Inhibiting α-glucosidase active component from chestnut shell.

Background technology

Chestnut shell is the acorn-cup of Fagaceae chestnut (Castanea mollissima Blume), is deciduous tree, general high 15～20 meters, suitable plantation on the sandy loam of mountain region endroit, and extensively cultivate various places, China north and south.Fruit acorn-cup is spherical, above has acupuncture, and the close pubescence of being close in thorn ftractures and the nut that sheds when ripe.

That the pharmacological action of chestnut shell mainly contains is antibacterial, antiinflammatory, anticoagulation, treatment chronic bronchitis and treatment bacillary dysentery etc., now the research of chestnut shell is mainly concentrated on the aspect such as extraction of pigment and bioactive substance.In chestnut shell, contain the chemical analysis such as polyphenol, organic acid, polysaccharide (or glycoside), flavone (or Saponin class), plant sterol (or triterpene), lactone, coumarin (or its glycoside) and tannin.Brown pigment in chestnut shell belongs to flavone compound, has good fungistatic effect; The polyphenol extracting from chestnut shell has higher oxidation resistance (Li Jinfeng etc., the extraction of polyphenol and antioxidation in vitro Journal of Sex Research in chestnut shell, chemistry of forest product and industry, 2010,30 (1): 54-58).

Alpha-glucosidase and diabetes and obesity are closely related, and Inhibiting α-glucosidase can reduce the sickness rate of diabetes effectively.Alpha-glucosidase inhibitor, by the alpha-glucosidase of competitive inhibition small intestinal epithelial cells, delays the absorption of intestinal to carbohydrate, reaches the treatment to diabetes thereby slow down post-prandial glycemia rising.Develop taking alpha-glucosidase inhibitor as basic medicine is as Acarbose and Miglitol etc. at present.Plurality of Chinese extract also shows has obvious inhibitory action to alpha-glucosidase, there is research to point out the substance activity of Inhibiting α-glucosidase effectively such as flavonoid, alkaloids, terpenoid, phenols, report and from chestnut shell, isolate the material with Inhibiting α-glucosidase activity but not yet find that there is so far.

Chestnut shell belongs to the garbage in food processing, therefrom extract the chemical substance with Inhibiting α-glucosidase activity, can be used for the treatment of diabetes and obesity, this further processing and utilization to chestnut shell and the exploitation of related drugs and functional food, have certain directive significance.

Summary of the invention

The object of this invention is to provide a kind of method of extracting purification Inhibiting α-glucosidase active component from chestnut shell, first chestnut shell is cleaned, dries, pulverized, with 75% (v/v) alcohol reflux, concentrating under reduced pressure, add water-saturated n-butanol solution extraction, finally carry out absorb-elute with macroporous resin AB-8, obtain having the polyphenol extract of Inhibiting α-glucosidase activity.Concrete steps are as follows:

(1) chestnut shell is cleaned, dries, pulverized, add 75% (v/v) ethanol according to solid-liquid ratio 1: 10 (w/v), boiling reflux extracts 2～3 times, each 2～4 hours;

(2) merge chestnut shell extracting solution, 50 DEG C of concentrating under reduced pressure 2～4 hours, obtain concentrated extract;

(3) concentrated extract water is deployed into the solution of 10mg/mL, then with the water-saturated n-butanol extraction of 4 times of volumes 3～5 times, each 2～4 hours;

(4) merge water-saturated n-butanol extract, in 70～80 DEG C of concentrating under reduced pressure 2～4 hours, obtain concentrated extract, then in-70～-90 DEG C of lyophilizations 10～24 hours, obtain powdered extract;

(5) powdered extract water is made into 0.5mg/mL solution, with macroporous resin, AB-8 adsorbs, and the flow velocity eluting with 60% (v/v) ethanol with 1BV/h is collected eluent;

(6) measure suppression ratio and the polyphenol content of eluent to alpha-glucosidase, collect the enzyme higher eluent of suppression ratio of living, get final product to such an extent that there is the chestnut shell component that suppresses this enzymatic activity.

Brief description of the drawings

Fig. 1 is polyphenol content in chestnut shell extract macroporous adsorbent resin AB-8 eluent and to alpha-glucosidase activity suppression ratio curve.

Detailed description of the invention

The following examples will be further explained the present invention, but the present invention is not limited only to these embodiment, the scope that these embodiment do not limit the present invention in any way.

Embodiment 1:

(1) 100g chestnut shell is cleaned, dries, pulverized, add 75% (v/v) ethanol boiling reflux 2 times, each 2 hours according to solid-liquid ratio 1: 10 (w/v);

(2) merge chestnut shell extracting solution, concentrating under reduced pressure 2 hours, obtains concentrated extract at 50 DEG C;

(3) concentrated extract water is deployed into the solution of 10mg/mL, then with the water-saturated n-butanol extraction of 4 times of volumes 3 times, each 2 hours;

(4) merge water-saturated n-butanol extract, in 70 DEG C of concentrating under reduced pressure 2 hours, obtain concentrated extract, then in-70 DEG C of lyophilizations 10 hours, obtain 0.5g powdered extract;

(5) powdered extract water is made into 0.5mg/mL solution, with macroporous resin, AB-8 adsorbs, and the flow velocity eluting with 60% (v/v) ethanol with 1BV/h is collected eluent;

(6) measure the polyphenol content of eluent in different eluting pipes and the suppression ratio (Fig. 1) to alpha-glucosidase, the peak value plyability of polyphenol eluting peak and suppression ratio is better, illustrate that polyphenol is the material of effective Inhibiting α-glucosidase activity, collect enzyme live suppression ratio higher, the eluent at elution volume 1.0-1.5BV place, get final product to such an extent that there is the chestnut shell component of this enzymatic activity of inhibition.

Embodiment 2:

(1) 10kg chestnut shell is cleaned, dries, pulverized, add 75% (v/v) ethanol boiling reflux 3 times, each 4 hours according to solid-liquid ratio 1: 10 (w/v);

(2) merge chestnut shell extracting solution, concentrating under reduced pressure 4 hours, obtains concentrated extract at 50 DEG C;

(3) concentrated extract water is deployed into the solution of 10mg/mL, then with the water-saturated n-butanol extraction of 4 times of volumes 5 times, each 4 hours;

(4) merge water-saturated n-butanol extract, in 80 DEG C of concentrating under reduced pressure 4 hours, obtain concentrated extract, then in-90 DEG C of lyophilizations 24 hours, obtain 50g powdered extract;

(5) powdered extract water is made into 0.5mg/mL solution, with macroporous resin, AB-8 adsorbs, and the flow velocity eluting with 60% (v/v) ethanol with 1BV/h is collected eluent;

(6) measure suppression ratio and the polyphenol content of eluent to alpha-glucosidase, collect the enzyme higher eluent of suppression ratio of living, get final product to such an extent that there is the chestnut shell component that suppresses this enzymatic activity.

Embodiment 3:

(1) 100kg chestnut shell is cleaned, dries, pulverized, add 75% (v/v) ethanol boiling reflux 2 times, each 3 hours according to solid-liquid ratio 1: 10 (w/v);

(2) merge chestnut shell extracting solution, concentrating under reduced pressure 3 hours, obtains concentrated extract at 50 DEG C;

(3) concentrated extract water is deployed into the solution of 10mg/mL, then with the water-saturated n-butanol extraction of 4 times of volumes 4 times, each 3 hours;

(4) merge water-saturated n-butanol extract, in 75 DEG C of concentrating under reduced pressure 3 hours, obtain concentrated extract, then in-80 DEG C of lyophilizations 12 hours, obtain 450g powdered extract;

(5) powdered extract water is made into 0.5mg/mL solution, with macroporous resin, AB-8 adsorbs, and the flow velocity eluting with 60% (v/v) ethanol with 1BV/h is collected eluent;

(6) measure suppression ratio and the polyphenol content of eluent to alpha-glucosidase, collect the enzyme higher eluent of suppression ratio of living, get final product to such an extent that there is the chestnut shell component that suppresses this enzymatic activity.

Claims (1)

1. a method of extracting purification Inhibiting α-glucosidase active component from chestnut shell, its step is as follows:

(1) chestnut shell is cleaned, dries, pulverized, add 75% (v/v) ethanol according to solid-liquid ratio 1: 10 (w/v), boiling reflux extracts 2～3 times, each 2～4 hours;

(2) merge chestnut shell extracting solution, 50 DEG C of concentrating under reduced pressure 2～4 hours, obtain concentrated extract;

(3) concentrated extract water is deployed into the solution of 10mg/mL, then with the water-saturated n-butanol extraction of 4 times of volumes 3～5 times, each 2～4 hours;

(4) merge water-saturated n-butanol extract, in 70～80 DEG C of concentrating under reduced pressure 2～4 hours, obtain concentrated extract, then in-70～-90 DEG C of lyophilizations 10～24 hours, obtain powdered extract;

(5) powdered extract water is made into 0.5mg/mL solution, with macroporous resin, AB-8 adsorbs, and the flow velocity eluting with 60% (v/v) ethanol with 1BV/h is collected eluent;

(6) measure suppression ratio and the polyphenol content of eluent to alpha-glucosidase, collect the enzyme higher eluent of suppression ratio of living, get final product to such an extent that there is the chestnut shell component that suppresses this enzymatic activity.