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Preparation and activity detection methods of fucoidan components in hizikia fusiformis

A technology of fucoidan sulfate in vegetables and vegetable fucoidan sulfate, applied in the field of preparation and activity detection of fucoidan sulfate components in hijiki, can solve the problem of lack of fucoidan sulfate Efficient preparation process of ester components and other issues

Pending Publication Date: 2019-06-14
WENZHOU UNIVERSITY
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Problems solved by technology

Because hijiki is common in my country's coastal areas and can be cultivated in large quantities, more and more scholars pay attention to the development of hijiki, but at this stage there is a lack of an effective preparation process for the fucoidan sulfate component in hijiki

Method used

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  • Preparation and activity detection methods of fucoidan components in hizikia fusiformis

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Embodiment 2

[0045] An analysis method of fucoidan sulfate ester components in hijiki, including fucoidan sulfate ester components SFF-11, SFF-12, SFF-21, SFF-31, SFF-32, SFF-41 , SFF-42, SFF-51, these eight components were subjected to static cell adhesion experiments.

[0046] Step 1: Prepare Chinese hamster ovary cells CHO and human promyelocytic leukemia cells HL-60, and transfect CHO cells with full-length human P-selectin cDNA to obtain CHO-P cells stably expressing human P-selectin;

[0047] Step 2: Inoculate CHO or CHO-P cells into a 24-well plate, culture overnight to form a monolayer of cells, and then inoculate the cells with blocking monoclonal antibody 9E1, non-blocking monoclonal antibody AC 1.2 or 100 μg / ml The fucoidan sulfate fraction was preincubated at 37°C for 1 hour, and calcein-labeled HL-60 cells were added to the pretreated CHO-P or CHO monolayer cells and incubated for another 1 hour, after washing with PBS, Fluorescence intensity was measured with a Tecan Infinit...

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Abstract

The invention discloses preparation and activity detection methods of fucoidan components in hizikia fusiformis. According to the key points of the technical scheme, the preparation method comprises the steps that the hizikia fusiformis is washed, dried and crushed, then ethanol is added, reflux degreasing and stirring filtration are conducted, degreased algae powder is extracted repeatedly for three times through a calcium chloride solution, and filtrate is merged to obtain a fucoidan extracting solution; the extracting solution is subjected to depressurizing distillation and concentrated toone fifth of the original volume; anhydrous ethanol is added into a concentrated solution, still standing for the night is conducted, and after centrifugal treatment and cleaning through the anhydrousethanol, vacuum drying is conducted to obtain hizikia fusiformis fucoidan; the hizikia fusiformis fucoidan is dissolved in distilled water, separated through DEAE-cellulose ion exchange column chromatography, eluted with the distilled water and then eluted with 0.1 M of NaCl, 0.3 M of NaCl, 0.5 M of NaCl and 1 M of NaCl, an obtained product is purified through Sepharose CL-6B column chromatography and eluted with NaCl, eluant is analyzed through a phenol-sulfuric acid method, and suitable fractions are collected for preparation. The preparation method has the advantage that preparation of hizikia fusiformis fucoidan components is easy and convenient to operate.

Description

technical field [0001] The invention relates to a method for extracting active substances in hijiki, in particular to a method for preparing and detecting the activity of fucoidan sulfate components in hijiki. Background technique [0002] Sargassum fusiforme (Sargassum fusiforme or Hizikia fusiformis) is a marine spore plant belonging to Phaeophyta, Phaeophyceae, Fucusaceae, Sargassum family, and Sargassum genus. Since the 1980s, Dongtou County, Wenzhou City, Zhejiang Province, which is known as the "Hometown of Hijiki in China", has started a large number of artificial breeding and mainly exported to Japan. Hijiki contains a variety of active substances, known as the "longevity vegetable", and has high development and utilization value as a medicinal and edible seaweed. [0003] Fucoidan sulfate is an important active substance in hijiki. Fucoidan sulfate, also known as fucoidan, is a water-soluble heteropolysaccharide. An important molecule in P-selectin-mediated recog...

Claims

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Application Information

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IPC IPC(8): C08B37/00G01N21/64
Inventor 吴明江佟海滨吴思雅张旭高艺恬
Owner WENZHOU UNIVERSITY
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