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Separation and purification method and application of algal inhibition active substances in kelp

A technology for algal inhibitory activity, separation and purification, which is applied in the separation/purification of hydroxyl compounds, chemical instruments and methods, and botanical equipment and methods to achieve the effects of reasonable method design, good algal inhibitory activity and high material purity.

Active Publication Date: 2018-08-17
JIANGSU MARINE RESOURCES DEV RES INST LIAN YUNGANG +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are no reports at home and abroad on the separation and purification of algae-inhibitory active substances that inhibit the growth of red tide microalgae in kelp

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Embodiment 1, the separation and purification method of kelp algae-inhibiting active substance:

[0016] (1) The dry powder of kelp is added with anhydrous methanol, and extracted at 15-20°C for 12 hours to obtain the extract. Repeat the extraction 3 times, and combine the extracts. After filtering and evaporating to dryness under reduced pressure, the kelp extract was prepared. Add 90% methanol aqueous solution to this extract, shake fully, let it stand overnight at 4°C, centrifuge, filter, remove the precipitate, and keep the supernatant. After taking appropriate amount of this supernatant and evaporating to dryness under reduced pressure, be dissolved in methanol, this methanol solution is carried out anti-algae activity detection, the result shows that it significantly inhibits the growth of Karenia militaris;

[0017] (2) Add the rest of the supernatant to ethyl acetate and extract 3 times. The upper layer was evaporated to dryness under reduced pressure to obta...

Embodiment 2

[0019] Embodiment 2, the separation and purification method of kelp algae-inhibiting active substance:

[0020] (1) Dried kelp was crushed into 0.3 mm powder, added with anhydrous methanol, leached at 15-20°C for 12 hours, repeated leached 3 times, and combined the extracts. After filtering and evaporating to dryness under reduced pressure, the kelp extract was prepared. Add 90% methanol aqueous solution to this extract, shake fully, let it stand overnight at 4°C, centrifuge, filter, remove the precipitate, and keep the supernatant. Take an appropriate amount of the supernatant, evaporate to dryness under reduced pressure at 60°C, dissolve it in methanol, and carry out the detection of algae-inhibiting activity (in the present invention, unless otherwise specified, the detection method of algae-inhibiting activity is the same). On a 96-well plate, 1 μL of the component solution to be tested was added to 199 μL of algae liquid, and 1 μL of anhydrous methanol was added to 199 μ...

Embodiment 3

[0024] Embodiment 3, the separation and purification method of kelp algae-inhibiting active substance:

[0025]Weigh 1000 g of dry kelp powder, add 3 L of anhydrous methanol, extract at 15-20 ° C for 12 hours, and pour out the extract. Add fresh anhydrous methanol again, and extract under the same conditions as above. After repeating 2 times, the extracts were combined (3 times in total). After filtering and evaporating to dryness under reduced pressure, 30.16 g of kelp extract was prepared. Add 150 mL of 90% methanol aqueous solution to this extract, shake fully, let stand overnight at 4°C, centrifuge, filter, remove precipitate, and obtain 130 mL of supernatant.

[0026] Take 1 mL of the supernatant, evaporate to dryness under reduced pressure at 60 °C, dissolve in 20 mL of methanol, and prepare a 10 g / L mother solution for the detection of algae inhibitory activity. On a 96-well plate, 1 μL of the component solution to be tested was added to 199 μL of algae liquid, and 1...

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PUM

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Abstract

The invention provides a separation and purification method of algal inhibition active substances in kelp. The method comprises the following steps: extracting kelp dry powder with absolute methanol,and then, performing evaporation to dryness under reduced pressure to obtain extract; performing alcohol precipitation on the extract with a methanol aqueous solution; adding ethyl acetate to supernatant liquid for performing extraction, performing evaporation to dryness under reduced pressure, distilling out ethyl acetate at the lower layer, then adding n-butyl alcohol for performing extraction,and performing evaporation to dryness under reduced pressure respectively to obtain a component B and a component C; performing separation by adopting silica gel column chromatography to obtain B1, B2and C2, performing separation by adopting gel column chromatography, and taking methanol as eluent; performing separation to obtain algal inhibition active components B13, B21 and C21, wherein the silica gel thin layer chromatography is adopted for the B13 and the C21; and finally, obtaining algal inhibition active components B21, B131, B132, C211 and C212 of which the purity reaches the thin layer purity. The separation and purification method of the algal inhibition active substances in the kelp is established on the basis of activity tracking, several algal inhibition active substances areseparated from the kelp, and the kelp can be applied to treatment of red tide microalgae. The method is reasonable in design, and the substances obtained by separation have high purity and good algalinhibition activity.

Description

technical field [0001] The invention relates to a method for separating and purifying algae-inhibiting active substances, in particular to a method for separating and purifying algae-inhibiting active substances in kelp. The present invention also relates to the use of the substance obtained by the aforementioned separation and purification method. The field of marine biochemical engineering. Background technique [0002] Kelp (Laminaiia japonica) is a large marine brown algae with high economic value. Its fresh product has high nutritional value and contains a lot of sugar, protein, vitamins, etc. It can be eaten after cooking, and it is also the main material for abalone farming. one. Kelp is also rich in organic iodine, which can treat and prevent goiter. In addition to edible and medicinal value, kelp is also an industrial raw material with high economic value, and its dry product is an important raw material for extracting alginic acid and mannitol. At present, the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07C33/30C07C35/06C07C35/36C07C29/86C07C29/76C07D307/12A01N31/04A01N31/06A01N43/08A01P13/00
CPCA01N31/04A01N31/06A01N43/08C07C29/76C07C29/86C07D307/12C07C2601/08C07C2602/26C07C33/30C07C35/06C07C35/36
Inventor 孙颖颖周文静徐嘉韩伟孙菁雯王长萍张新
Owner JIANGSU MARINE RESOURCES DEV RES INST LIAN YUNGANG
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