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Method for preparing high-purity chlorophyll a from spirulina platensis

A technology of spirulina platensis and chlorophyll, which is applied in the field of natural pigments, can solve the problem of low purity of chlorophyll a and achieve the effect of high purity

Inactive Publication Date: 2015-11-18
CHINA UNIV OF PETROLEUM (EAST CHINA)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Aiming at the problem of low purity of commercialized chlorophyll-a, the present invention proposes a preparation method of high-purity chlorophyll-a

Method used

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  • Method for preparing high-purity chlorophyll a from spirulina platensis
  • Method for preparing high-purity chlorophyll a from spirulina platensis
  • Method for preparing high-purity chlorophyll a from spirulina platensis

Examples

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Effect test

Embodiment 1

[0025] The concrete steps of the preparation method of high-purity chlorophyll a of the present invention are as follows:

[0026] Take 1 L of Spirulina platensis algae liquid grown to the late logarithmic growth phase, centrifuge and separate the algae, discard the supernatant, enrich the precipitate in a centrifuge tube, and then extract it with 50 mL of ethanol three times, and combine the extracts.

[0027] An appropriate amount of petroleum ether (60-90°C) is added to the extract, water is added until the phases are separated, and the pigment-containing petroleum ether phase is washed twice with water.

[0028] Fill the chromatographic column with 100-200 mesh neutral alumina, first wet the alumina column with 0.3% ethanol petroleum ether solution, then load the sample, and then use 0.3%, 10% ethanol-petroleum ether mixed solvent gradient rinse. The orange-yellow carotenoids are washed out first, followed by the blue-green chlorophyll. The solvent was recovered by rotar...

Embodiment 2

[0030] The concrete steps of the preparation method of high-purity chlorophyll a of the present invention are as follows:

[0031] Take 1 L of Spirulina platensis algae liquid grown to the late logarithmic growth phase, centrifuge and separate the algae, discard the supernatant, enrich the precipitate in a centrifuge tube, and then extract it with 50 mL of ethanol three times, and combine the extracts.

[0032] An appropriate amount of petroleum ether (60-90° C.) was added to the extract, saturated brine was added until the phases were separated, and the pigment-containing petroleum ether phase was washed twice with saturated brine.

[0033] Fill the chromatographic column with 100-200 mesh neutral alumina, first wet the alumina column with 0.3% ethanol petroleum ether solution, then load the sample, and then use 0.3%, 1% ethanol-petroleum ether mixed solvent gradient rinse. The orange-yellow carotenoids are washed out first, followed by the blue-green chlorophyll. The solve...

Embodiment 3

[0035] The concrete steps of the preparation method of high-purity chlorophyll a of the present invention are as follows:

[0036] Take 1 L of Spirulina platensis algae liquid grown to the late logarithmic growth phase, centrifuge and separate the algae, discard the supernatant, enrich the precipitate in a centrifuge tube, and then extract it with 50 mL of ethanol three times, and combine the extracts.

[0037] An appropriate amount of n-hexane is added to the extract, water is added until the phases are separated, and the n-hexane phase containing the pigment is washed twice with water.

[0038] Fill the chromatographic column with 100-200 mesh neutral alumina, first wet the alumina column with 0.3% ethanol-n-hexane solution, then load the sample, and then use 0.3%, 10% ethanol-n-hexane mixed solvent gradient rinse. The orange-yellow carotenoids are washed out first, followed by the blue-green chlorophyll. The solvent was recovered by rotary evaporation to obtain dark green...

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Abstract

The invention discloses a method for preparing high-purity chlorophyll a from spirulina platensis. The method includes the steps that 1, the spirulina platensis cultivated to the later stage of the logarithm growth period serves as raw materials, and platensis bodies are centrifugally collected; 2, the platensis bodies are extracted three times through ethyl alcohol, and extract liquor is combined; 3, an appropriate amount of normal hexane or petroleum ether is added into the extractor liquor, water is added to phase splitting, back extraction is performed, and a solution with pigment is rinsed with water two times; 4, neutral alumina of 100-200 meshes is adopted for column chromatography isolation and purification to obtain chlorophyll a, first, a normal hexane or petroleum ether solution containing ethyl alcohol of 0.3% is used for moistening and rinsing an alumina column, and after sample loading, ethyl alcohol-normal hexane or ethyl alcohol-petroleum ether mixed solvent is used for gradient elution; 5, the solution containing the chlorophyll a is subjected to vacuum-rotary evaporation to recover solvent and obtain the high-purity chlorophyll a. The prepared chlorophyll a does not contain chlorophyll b, wherein the HPLC purity is larger than 99.5%.

Description

technical field [0001] The invention relates to the technical field of natural pigments, in particular to a preparation method of high-purity chlorophyll a. Background technique [0002] Chlorophyll is the main pigment for photosynthesis in higher plants, algae and photosynthetic bacteria. It is a family of lipid-containing pigments located on the thylakoid membrane in chloroplasts. Chlorophyll has multiple uses such as promoting hematopoiesis, supplementing vitamins, strengthening detoxification, and improving disease resistance. For example: Drinking chlorophyll is very beneficial for pregnant women or people who lose blood due to accidents. [0003] The main types of chlorophyll are: magnesium porphyrin compounds (including chlorophyll a, b, c, etc.), prochlorophyll and bacterial chlorophyll. Chlorophyll is not very stable, light, acid, alkali, oxygen, oxidant, etc. can cause its decomposition. Under acidic conditions, chlorophyll molecules can easily lose magnesium io...

Claims

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Application Information

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IPC IPC(8): C07D487/22
CPCC07D487/22
Inventor 于道永李泉张宏玉李富强吴晨露葛保胜黄方
Owner CHINA UNIV OF PETROLEUM (EAST CHINA)
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