Method for separating and extracting abscisic acid from fermented liquid by ionic exchanging and reversed phase chromatography

A technology of reverse phase chromatography and ion exchange, applied in the separation/purification of carboxylic acid compounds, organic chemistry, etc., can solve the problems of high price of natural abscisic acid, affecting the research of natural abscisic acid, affecting the application of natural abscisic acid, etc., to achieve The extraction method is simple and reasonable, the method is simple, and the effect of easy recycling

Inactive Publication Date: 2007-04-11
CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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Problems solved by technology

Therefore, until now there is no simple and effective method for the preparation of high-purity natural abscisic acid. The higher price of high-purity natural abscisic acid has greatly affected the research on the physiological effects of natural abscisic acid on plants. Thus affecting the application of natural abscisic acid, an endogenous plant hormone with comprehensive physiological functions, in agricultural production

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  • Method for separating and extracting abscisic acid from fermented liquid by ionic exchanging and reversed phase chromatography
  • Method for separating and extracting abscisic acid from fermented liquid by ionic exchanging and reversed phase chromatography

Examples

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Embodiment 1

[0032] 1000mL fermentation medium composition: 5.0% dextrin, 0.3% wheat bran, 0.02% ethylene glycol, 2.0% glucose, 3.0% sucrose, 1.0% bean cake powder, 0.01% dipotassium hydrogen phosphate, 0.05% magnesium sulfate.

[0033] Divide 1200mL of the above medium into four 1000mL Erlenmeyer flasks, 300mL in each bottle, sterilize at 120°C for 30 minutes, inoculate the spore suspension of the activated abscisic acid high-yielding strain Botrytis cinerea TBI-9 after cooling, and inoculate at 23-29°C under fermentation. Oscillation speed 150rpm, amplitude 20cm. The fermentation time is 8-12 days, and the concentration of natural abscisic acid reaches 0.14%.

[0034] Take 1000 grams of fermentation broth (the content of natural abscisic acid is 1.431 grams), add 3N sulfuric acid solution to adjust the Ph to 2.0-5.0, and carry out acidification treatment for 30 minutes. The fermented broth after acidification is centrifuged at room temperature for 10 minutes at a centrifugal speed of 5...

Embodiment 2

[0040] The preparation method of fermented liquid is the same as Example 1, get 1000 grams of fermented liquid (natural abscisic acid content is 1.410 grams), add 9.0 liters of water, make the concentration of natural abscisic acid in the liquid be 0.014%, replace original fermented liquid with this solution and extract separate. Add 3N hydrochloric acid solution to adjust the Ph to 2.0-5.0, centrifuge at 5000-9000 rpm for 10 minutes at room temperature, and collect the supernatant. The centrifuged supernatant was in contact with 200-1000 grams of 101 macroporous cation exchange resin for 1 hour, and then washed with 1000-5000 mL of 40-85% methanol for 30 minutes, and the methanol solution containing natural abscisic acid was collected. The abscisic acid content is 1.387 grams.

[0041] The eluting methanol liquid was distilled under reduced pressure on a Buchi R-3000 rotary distillation device, the vacuum degree was maintained at 0.06-0.09Mpa, the temperature of the water ba...

Embodiment 3

[0044] The preparation method of fermented liquid is the same as embodiment 1, gets 3000 grams of fermented liquid, and natural abscisic acid content is 4.248 grams in fermented liquid. Concentrate under reduced pressure to 1000 g, so that the concentration of the fermented liquid is 0.423%, and use this solution to replace the original fermented liquid for extraction and separation. The extraction and separation operation is basically the same as in Example 1. Finally, 1.006 grams of natural abscisic acid with a purity of 95.8% and 2.623 grams of natural abscisic acid crystals with a purity of 99.1% were obtained.

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Abstract

The ion exchange and reversed phase chromatographic separation process for extracting abscisic acid from fermented liquid includes the following steps: the first macroporous resin adsorption of abscisic acid from fermented liquid to extract and concentrate abscisic acid in fermented liquid initially; the subsequent silica gel chromatography to eliminate much of fermentation side products and obtain natural abscisic acid product with relatively high purity (higher than 90 %); and final C18 silica gel reversed phase chromatographic separation to obtain natural abscisic acid product with purity higher than 98 %. The process can obtain high purity abscisic acid product suitable for plant physiology research.

Description

technical field [0001] The invention belongs to the field of biochemical industry, in particular to the field of extraction and separation of natural substances. Background technique [0002] Abscisic acid is an endogenous plant hormone with comprehensive physiological functions. With the in-depth research on the physiological function of abscisic acid, it has been confirmed that it plays a variety of roles in the growth and development of plants, such as the development and maturation of seeds, the response to environmental inverse factors, and the regulation of gene expression, etc. It has an important regulatory function. It can promote the maturity and development of fruits, grains, beans, etc., can greatly increase their yield and quality, and can greatly enhance their cold resistance, drought resistance and salt-alkali resistance. However, the source of natural abscisic acid has been greatly limited, especially to obtain high-purity natural...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07C59/90C07C51/47
Inventor 周金燕谭红杨杰
Owner CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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