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Method for remarkably increasing biomass and active ingredient yield of salvia miltiorrhiza hairy roots and application of salvia miltiorrhiza hairy roots

A technology of active ingredients and hairy roots, applied in botany equipment and methods, medical preparations containing active ingredients, plant/algae/fungus/moss components, etc., to achieve low cost, rapid growth, and easy operation

Inactive Publication Date: 2017-01-11
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, the hairy roots of Salvia miltiorrhiza have been cultured using glucose, fructose and sucrose as carbon sources, and there is no research on using galactose or a series of oligosaccharides composed of it, such as raffinose and stachyose, as carbon sources. precedent

Method used

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  • Method for remarkably increasing biomass and active ingredient yield of salvia miltiorrhiza hairy roots and application of salvia miltiorrhiza hairy roots
  • Method for remarkably increasing biomass and active ingredient yield of salvia miltiorrhiza hairy roots and application of salvia miltiorrhiza hairy roots
  • Method for remarkably increasing biomass and active ingredient yield of salvia miltiorrhiza hairy roots and application of salvia miltiorrhiza hairy roots

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Experimental program
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Effect test

Embodiment 1

[0025] Embodiment 1. The method for efficiently producing salvianolic acid components by cultivating Salvia miltiorrhiza root with galactose

[0026] Experimental Materials:

[0027] The hairy roots of Salvia miltiorrhiza were cultivated by inducing the seeds of Salvia miltiorrhiza by Agrobacterium rhizogenes. The hairy root strain of Salvia miltiorrhiza used in the experiment was cultured on fresh 6,7-V solid medium, cultured statically at 22°C, subcultured every 3-4 weeks, and set aside.

[0028] Experimental reagents: MS medium (Duchefa Biochemie company, product number: M0222); Gamborg's B5 medium (Duchefa Biochemie formula, product number: G0210). Acetonitrile is chromatographically pure (Shanghai Bailingwei Chemical Technology Co., Ltd.); methanol is analytically pure (Shanghai Lingfeng Chemical Reagent Co., Ltd.); trifluoroacetic acid (Sinopharm Chemical Reagent Co., Ltd.); water is deionized water (Hangzhou Wahaha Group Co., Ltd. ).

[0029] Methods and steps:

[0...

Embodiment 2

[0043] Example 2. The preparation method of the salvia miltiorrhiza root to produce the total phenolic acid extract of salvia miltiorrhiza

[0044] Materials and methods: About 2 grams of Salvia miltiorrhiza root seedlings were inoculated into 500 mL of Gamborg’s B5 liquid culture medium containing galactose, and cultured at 22°C for 14 days in the dark with shaking at 120 rpm. In the same way, 10 bottles were cultivated at the same time. Filter out the culture medium, wash with pure water 2 to 3 times, combine and freeze-dry to obtain 580 g of dry hairy roots of Salvia miltiorrhiza.

[0045] Grind the hairy root of Salvia miltiorrhiza, extract twice with 6.0Kg of 70% ethanol, filter, combine the extracts, and recover the ethanol. The concentrated water extract is diluted with 3 times the original volume of deionized water, filtered, and the clarified filtrate is adsorbed by a resin column equipped with 1.0Kg D101 macroporous adsorption resin. After all the adsorption is com...

Embodiment 3

[0047] Embodiment 3. Utilize the preparation method of salvia miltiorrhiza hairy root to produce tanshinone extract

[0048] Materials and methods: About 2 grams of Salvia miltiorrhiza root seedlings were inoculated into 500 mL of Gamborg’s B5 liquid culture medium containing galactose, and cultured at 22°C for 14 days in the dark with shaking at 120 rpm. In the same way, 10 bottles were cultivated at the same time. Filter out the culture medium, wash with pure water 2 to 3 times, combine and freeze-dry to obtain 580 g of dry hairy roots of Salvia miltiorrhiza.

[0049] Grind the hairy root of Salvia miltiorrhiza, extract twice with 6.0Kg of 70% ethanol, filter, combine the extracts, and recover the ethanol. The concentrated water extract is diluted with 3 times the original volume of deionized water, filtered, and the clarified filtrate is adsorbed by a resin column equipped with 1.0Kg D101 macroporous adsorption resin. After all the adsorption is completed, wash once with ...

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Abstract

The invention belongs to the field of biotechnology and medical technology and particularly relates to a method for method for remarkably increasing biomass and active ingredient yield of salvia miltiorrhiza hairy roots and an application of salvia miltiorrhiza hairy roots. According to the method, a culture solution is prepared from galactose and a series of oligose such as raffinose, stachyose and the like consisting of galactose as carbon sources, and the culture solution is used for culturing the salvia miltiorrhiza hairy roots. With the adoption of the method, the biomass and the yield of medicinal active ingredients of the salvia miltiorrhiza hairy roots can be remarkably increased, and an effective approach is provided for effective preparation of the medicinal active ingredients such as salvianolic acids and tanshinone from the salvia miltiorrhiza hairy roots.

Description

technical field [0001] The invention belongs to the field of biotechnology and medicine technology, and in particular relates to a method for significantly increasing the biomass of salvia miltiorrhiza root and the yield of active components, and its use in the field of biomedicine. Background technique [0002] Salvia miltiorrhiza is a labiatae plant Salvia miltiorrhiza ( Salvia miltiorrhiza Bge .) dried roots and rhizomes, have the functions of promoting blood circulation and removing blood stasis, clearing menstrual flow, relieving pain, clearing heart and eliminating troubles, cooling blood and eliminating carbuncles. In traditional Chinese medicine, it is widely used in chest wall pain, wrist, abdomen and hypochondriac pain, irregular menstruation, dysmenorrhea, etc. disease. Its active ingredients are mainly water-soluble salvianolic acid compounds and fat-soluble tanshinone compounds. The former mainly includes danshensu, protocatechuic acid, protocatechuic aldehyde...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K36/537A01H4/00
CPCA01H4/001A01H4/005A61K36/537A61K2236/19A61K2236/35A61K2236/55
Inventor 周国军周铜水
Owner FUDAN UNIV