Method for producing high-quality extract by rapidly fermenting ginkgo leaf powder

A high-quality technology of ginkgo biloba powder, applied in the directions of medical raw materials, pharmaceutical formulations, plant raw materials, etc. derived from Ginkgo biloba Compare and improve the effect of product quality

Active Publication Date: 2018-06-22
SHAANXI SCI TECH UNIV
2 Cites 0 Cited by

AI-Extracted Technical Summary

Problems solved by technology

[0004] The existing production methods of Ginkgo biloba extract include water extraction, organic solvent extraction, supercritical fluid extraction, ultrasonic-assisted extraction, etc., but they all have defects such as low extraction efficiency and high cost.
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Abstract

The invention discloses a method for producing a high-quality extract by rapidly fermenting ginkgo leaf powder. A strain used for fermentation is trichoderma reesei. The method comprises the steps that dried ginkgo leaves are smashed to 20-40 mesh and soaked in deionized water according to the material-to-liquid ratio of 1:5-1:8 to adjust the pH value to be 3-6, the mixture is sucked into a fermentation tank to be sterilized for 3-10 min at 116-125 DEG C and then cooled to 25-30 DEG C, a trichoderma reesei seed solution with the mass accounting for 1-10% of the dry weight of the ginkgo leaf powder in fermentation broth is added, aerated fermentation is conducted for 2-10 days at 25-30 DEG C, and the rotation speed during stirring is 180-300 r/min; after fermentation, a 20-95% ethanol solution is used for reflux extraction for 1-3 h for 2-3 times according to the material-to-liquid ratio of 1:7-1:12, extraction solutions are combined, filtered and subjected to vacuum concentration, thenthe product is purified by using LX-5 macroporous adsorption resin, the product is washed with deionized water and a 10-20% ethanol solution in sequence and finally eluted by using a 65-85% ethanol solution, and eluant is subjected to vacuum concentration to be free of alcohol smell; after filtering is completed, the concentration of the product is adjusted to be proper with deionized water, thenginkgolic acid is removed by using LXD-200 macroporous adsorption resin, and a deacidification extraction solution is subjected to concentration, drying and smashing to obtain the ginkgo leaf extract. The ginkgo leaf extract prepared through the method is high in extraction efficiency, good in product quality and suitable for industrial large-scale production.

Application Domain

Ginkgophyta medical ingredientsPlant ingredients

Technology Topic

Fermentation brothReflux extraction +6

Examples

  • Experimental program(4)
  • Effect test(1)

Example Embodiment

[0030] Example 1
[0031] (1) Take 50.0g of dried Ginkgo biloba powder from 20-40 mesh, dilute with water at a material-to-liquid ratio of 1:5 (m/v), stir well, adjust the pH of the solution to 3.0 with 2mol/L hydrochloric acid, and fill it into a 500mL triangle Sterilize the bottle in an autoclave at 116°C for 10 minutes.
[0032] (2) After the sterilization, the temperature was lowered to 25°C, and 1 mL of Trichoderma reesei seed liquid was added at a 2% inoculum amount in a sterile environment, and fermented on a constant temperature shaker at 25°C for 10 days at a speed of 180r/min.
[0033] (3) After the fermentation, add 200mL 95% ethanol solution for reflux extraction, the extraction temperature is 40℃, the extraction time is 3h, after the extraction, filter, add 500mL 20% ethanol to the filter residue and extract once according to the above conditions, filter, combine the filtrate twice, and concentrate To 300mL.
[0034] (4) Purify the extract with LX-5 macroporous adsorption resin, the column bed volume is 100mL, the sample rate is 1BV/h, and then the impurities are washed with 3BV of deionized water and 10% ethanol solution in sequence, and the flow rate is controlled to 3BV/ h, finally eluted with 4BV 55% ethanol solution at a speed of 1BV/h. The eluent was concentrated under reduced pressure at 50℃ and -0.1Mpa to 1/3 of the original volume, and adjusted with deionized water after filtration. To 2 times the volume of the concentrate.
[0035] (5) The above solution is selected from LXD-200 macroporous adsorption resin to remove ginkgolic acid, and the loading rate is controlled at 1BV/h. The effluent is collected, concentrated under reduced pressure to an extract, and then dried in a vacuum drying cabinet at 70°C to constant weight , Ginkgo biloba extract is obtained after crushing.
[0036] (6) Take appropriate amount of Ginkgo biloba extract to determine the content of total flavonoids and total lactones in accordance with the method shown in the Chinese Pharmacopoeia. The results showed that 3.08 g of Ginkgo biloba extract was obtained, the extraction rate was 6.16%, the total flavonoid content was 30.19%, the total lactone content was 7.96%, and the total ginkgolic acid content was 3.73 ppm.

Example Embodiment

[0037] Example 2
[0038] (1) Take 100.0g of dried Ginkgo biloba powder from 20-40 mesh, dilute with water at a material-to-liquid ratio of 1:6 (m/v), stir well, adjust the pH of the solution to 4.0 with 2mol/L hydrochloric acid, and divide into 4 In a 500mL Erlenmeyer flask, sterilize in a high-pressure steam sterilizer at 118°C for 8 minutes.
[0039] (2) After the sterilization, the temperature was lowered to 27°C, and 1 mL of Trichoderma reesei seed liquid was added to each bottle at a 4% inoculum amount in a sterile environment. Fermentation was carried out on a constant temperature shaker at 27°C for 8 days at a speed of 220r/min.
[0040] (3) After the fermentation, add 250mL of 95% ethanol solution to each bottle for reflux extraction. The extraction temperature is 55℃, and the extraction time is 2h. After the extraction is completed, filter, add 800mL of 40% ethanol to the filter residue and extract twice under the above conditions, filter, and combine the three filtrates. , Concentrated to 1000mL.
[0041] (4) Purify the extract with LX-5 macroporous adsorption resin, the column bed volume is 150mL, the sample rate is 1.5BV/h, and then use 3BV of deionized water and 15% ethanol solution to wash impurities, and control the flow rate to 3BV /h, and finally eluted with 4BV 65% ethanol solution at a speed of 1BV/h. The eluent was concentrated to 1/3 of the original volume under reduced pressure at 60℃ and -0.09Mpa, and filtered with deionized water Adjust to 2 times the volume of the concentrate.
[0042] (5) The above solution is selected from LXD-200 macroporous adsorption resin to remove ginkgolic acid, and the loading rate is controlled at 1BV/h. The effluent is collected, concentrated under reduced pressure to an extract, and then dried in a vacuum drying cabinet at 70°C to constant weight , Ginkgo biloba extract is obtained after crushing.
[0043] (6) Take appropriate amount of Ginkgo biloba extract to determine the content of total flavonoids and total lactones in accordance with the method shown in the Chinese Pharmacopoeia. The results showed that 6.35 g of Ginkgo biloba extract was obtained, the extraction rate was 6.35%, the total flavonoid content was 29.88%, the total lactone content was 8.04%, and the total ginkgolic acid content was 5.24 ppm.

Example Embodiment

[0044] Example 3
[0045] (1) Take 200.0g of dried ginkgo biloba powder with 20-40 mesh, dilute with water at a material-to-liquid ratio of 1:7 (m/v), stir well, adjust the pH of the solution to 5.0 with 2mol/L acetic acid, and divide into 7 Sterilize in a 1000mL Erlenmeyer flask at 121°C for 6 min in an autoclave.
[0046] (2) After the sterilization, the temperature was lowered to 28°C, and 1.7 mL of Trichoderma reesei seed liquid was added to each bottle at a 6% inoculum amount in a sterile environment, and fermented on a constant temperature shaker at 28°C for 6 days at a speed of 250r/min.
[0047] (3) After the fermentation, add 300mL of 95% ethanol solution to each bottle for reflux extraction. The extraction temperature is 70℃, the extraction time is 1.5h. After the extraction is completed, filter, add 2000mL of 60% ethanol to the filter residue and extract 2 times under the above conditions, filter, and combine 3 times The filtrate was concentrated to 2000mL.
[0048] (4) Purify the extract with LX-5 macroporous adsorption resin, the column bed volume is 300mL, the sample rate is 1.5BV/h, and then use 3BV of deionized water and 15% ethanol solution to wash impurities, and control the flow rate to 3BV /h, finally eluted with a 4BV 75% ethanol solution at a speed of 1BV/h. The eluent was concentrated to 1/3 of its original volume under reduced pressure at 70°C and -0.08Mpa. After filtration, deionized water was used. Adjust to 2 times the volume of the concentrate.
[0049] (5) The above solution is selected from LXD-200 macroporous adsorption resin to remove ginkgolic acid, and the loading rate is controlled at 1BV/h. The effluent is collected, concentrated under reduced pressure to an extract, and then dried in a vacuum drying cabinet at 70°C to constant weight , Ginkgo biloba extract is obtained after crushing.
[0050] (6) Take appropriate amount of Ginkgo biloba extract to determine the content of total flavonoids and total lactones in accordance with the method shown in the Chinese Pharmacopoeia. The results showed that 12.77 g of Ginkgo biloba extract was obtained, the extraction rate was 6.38%, the total flavonoid content was 30.26%, the total lactone content was 8.12%, and the total ginkgolic acid content was 8.69 ppm.

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