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Extraction and content determination methods for natural astaxanthin of thick-wall microalgae

A technology of natural astaxanthin and an extraction method, which is applied in the field of extraction and content determination of natural astaxanthin from thick-walled microalgae, can solve the problems of limited sensitivity of thin-layer scanning, complicated operation, low accuracy, etc., and avoid high temperature decomposition. Or oxidative deterioration, high extraction efficiency, and complete wall-breaking effects

Active Publication Date: 2014-03-12
QINGDAO XUNON BIOLOGICAL ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] This method has the following obvious shortcomings and deficiencies: (1) The sample pretreatment and determination process cannot effectively eliminate the interference of other pigments such as chlorophyll; (2) The detection process can only obtain the information of the total astaxanthin content, and cannot distinguish the Other β-carotene and various composition forms of astaxanthin contained in it; (3) The absorbance is measured by the ultraviolet method, and the accuracy is low; (4) NaOH and acetic acid are used as the alkali and acid to break the wall during the detection process. Increased risk of compositional changes and oxidative degradation of astaxanthin and other carotenoids
Although this method can detect the concentration and composition of different astaxanthin and other carotene respectively, the method of this invention is cumbersome to operate, the detection process takes a long time and needs to be carried out at room temperature, which easily causes the oxidation of astaxanthin and other carotenoids Decomposition and photodegradation, therefore, less accurate
In addition, the reproducibility of the sampling process is poor, the sensitivity of TLC scanning detection is very limited, and the accidental errors caused by subjective accidental factors are also relatively large, which will have a certain impact on the detection results of astaxanthin and carotenoids and other pigments

Method used

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  • Extraction and content determination methods for natural astaxanthin of thick-wall microalgae
  • Extraction and content determination methods for natural astaxanthin of thick-wall microalgae
  • Extraction and content determination methods for natural astaxanthin of thick-wall microalgae

Examples

Experimental program
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Effect test

Embodiment 1

[0042] Example 1 Preparation and Content Detection of Haematococcus Pluvialls Fresh Algae Liquid Extract

[0043] (1) Extraction of Astaxanthin

[0044]1) Determination of dry weight of algae cells: take 50mL of fresh Haematococcus pluvialis algal liquid and filter it with qualitative filter paper (the qualitative filter paper has been baked in an oven at 80°C to constant weight), and then put the filtered filter paper in an oven at 80°C Baked to constant weight, weighed Haematococcus pluvialis dry weight 31.5mg, 31.3mg (parallel test).

[0045] 2) Astaxanthin extraction: take another 50mL of fresh algae liquid and centrifuge at a speed of 8000r / min, remove the supernatant, transfer the centrifuged Haematococcus pluvialis algal cells into a mortar, add appropriate amount of quartz sand in turn, and Pour liquid nitrogen into the mortar and freeze. After the algal cells are fully frozen, add 0.5 mL of extraction solvent (dichloromethane-methanol (v:v=1:3)) and grind thoroughly...

Embodiment 2

[0056] Example 2 Preparation and Content Detection of Haematococcus Pluvialls Dry Algae Powder Extract

[0057] (1) Extraction of Astaxanthin

[0058] 1) Astaxanthin extraction: Take 50.0 mg of dried Haematococcus pluvialis powder in a mortar, add appropriate amount of quartz sand in turn, and pour liquid nitrogen into the mortar to freeze. After the algal cells are fully frozen, add 1 mL of extraction solvent (dichloromethane-methanol (v:v=1:1)) and grind thoroughly, transfer the algal cells together with the extraction solvent to a centrifuge tube, and rinse the mortar with the extraction solvent , wash until there is no residue, centrifuge the residue and supernatant at a speed of 3000r / min, and collect the supernatant. Then repeat the above liquid nitrogen grinding and extraction steps until the centrifuged supernatant is colorless, and combine the supernatants.

[0059] 2) Preparation of astaxanthin test solution: Concentrate the above-mentioned astaxanthin sample by pu...

Embodiment 3

[0064] Example 3 Preparation and Content Detection of Haematococcus Pluvialls Fresh Algae Liquid Extract

[0065] (1) Extraction of Astaxanthin

[0066] 1) Determination of dry weight of algae cells: take 100mL of fresh Haematococcus pluvialis algae liquid and filter it with qualitative filter paper (the qualitative filter paper has been baked in an oven at 80°C to constant weight), and then put the filtered filter paper in an oven at 80°C Baked to constant weight, weighed Haematococcus pluvialis dry weight 62.9mg, 62.7mg (parallel samples).

[0067] 2) Astaxanthin extraction: Take another 100mL of fresh algae liquid and filter it with qualitative filter paper, then use the extraction solvent (chloroform-methanol (v:v=1:5)) to wash the algae cells on the filter paper into the mortar, Until no algal cells remain on the filter paper, add appropriate amount of quartz sand in turn, and pour liquid nitrogen into the mortar to freeze. After the algae cells are fully frozen, add 0....

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Abstract

The invention discloses extraction and content determination methods for natural astaxanthin of thick-wall microalgae. According to the extraction method for the natural astaxanthin of the thick-wall microalgae, the astaxanthin is obtained by utilizing an extraction solvent composed of dichloromethane or trichloromethane and methanol at a low temperature through low-temperature liquid nitrogen protection. According to the extraction method, the astaxanthin is effectively prevented form being decomposed at a high temperature or oxidized so as to avoid going bad; the operation is simple and walls are completely broken; the extraction efficiency is high; the extraction method is applicable to living organisms including the thick-wall microalgae and the like. The extraction method for the natural astaxanthin of the thick-wall microalgae is combined with a high performance liquid chromatography to detect the content of the astaxanthin, namely the content of total carotenoids can be obtained, namely, content information of free astaxanthin, astaxanthin monoester, astaxanthin diester, xanthophyll, zeaksantyna and the like can also be obtained; an important methodology is provided for developing correlational researches on the microalgae astaxanthin and other carotenoids. The high performance liquid chromatography is used for detecting the content of the astaxanthin; the determination accuracy is high and the reliability is high.

Description

technical field [0001] The invention relates to a method for extracting and determining the content of natural astaxanthin from thick-walled microalgae. Background technique [0002] Carotenoids such as lutein and astaxanthin are commonly found in green algae. Haematococcus (Haematococcus, also known as Haematococcus) is a green alga of the Volvox family Haematococcus. The astaxanthin content accumulated by this microalgae can be as high as 1.5% to 6.0%, which is regarded as a "concentrated product" of natural astaxanthin. A large number of studies have shown that the accumulation rate and total production of astaxanthin in Haematococcus pluvialis are higher than those of other green algae, and the ratio of astaxanthin and its esters contained in Haematococcus pluvialis (about 70% monoester, 25% diester and 5% monomer) are very similar to astaxanthin in aquaculture animals (shrimp, crab, salmon, etc.). The structure of astaxanthin in Haematococcus pluvialis is mainly 3S-3...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
Inventor 陈伟曹腾腾李康徐瑶刘永梅曹磊李西清
Owner QINGDAO XUNON BIOLOGICAL ENG
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