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Rapid and efficient extraction method of chlorophylls in large quantity of cyanobacteria samples

An extraction method and chlorophyll technology, applied in chemical instruments and methods, azo dyes, organic dyes, etc., can solve problems such as being unsuitable for large-scale sample extraction, and achieve the effects of high extraction effect, time saving, and experimental error reduction.

Active Publication Date: 2014-09-10
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this method has high chlorophyll extraction efficiency, it is not suitable for large-scale sample extraction

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Take OD 680 5mL of 1.0 Microcystis aeruginosa culture solution, centrifuged at 12000rpm for 5min, and collected Microcystis aeruginosa cells; respectively added the above-mentioned Microcystis aeruginosa cells to 5 centrifuge tubes containing 1mL sterile water, and the 5 centrifuge tubes were respectively Heat in boiling water for 3min, 5min, 10min, 15min, 20min, then cool naturally at room temperature (25°C), add 4mL acetone to 5 centrifuge tubes and mix well, centrifuge at 12000rpm for 5min, the supernatant is chlorophyll extraction liquid. Take each supernatant to measure OD 750 , OD 663 , OD 645 , OD 630 , calculate the chlorophyll content ρ(Chla) (μg / L) by the formula:

[0020] ρ(Chla) (μg / L) =11.64×(OD 663 -OD 750 ) -2.16×(OD 645 -OD 750 )+0.1×(OD 630 -OD 750 ).

[0021] processing method Boiling water for 3 minutes Boiling water for 5 minutes Boiling water 8min Boiling water 10min Boiling water for 15 minutes ρ(Chla) (μg / L) 3...

Embodiment 2

[0023] Take 5ml Microcystis aeruginosa culture solution (OD 680 =1.0), centrifuged at 12000rpm for 5min, and collected the cells; the above-mentioned Microcystis aeruginosa cells were added to two centrifuge tubes containing 1mL sterile water, and the two centrifuge tubes were heated in boiling water for 3min, and the There are two cooling methods: bath cooling and natural cooling at room temperature (25°C). After cooling, add 4 mL of acetone to the two centrifuge tubes, mix well, and centrifuge at 12,000 rpm for 5 min. Take each supernatant to measure OD 750 , OD 663 , OD 645 , OD 630 , the chlorophyll content ρ(Chla) (μg / L) was calculated by the formula. The amount of chlorophyll extracted is shown in Table 2.

[0024] cooling method natural cooling ice bath cooling ρ(Chla) (μg / L) 3.219464 3.134184

Embodiment 3

[0026] Take 5mL Microcystis aeruginosa culture solution (OD 680 =1.0), centrifuge at 12000rpm for 5min, collect the cells, add the above-mentioned Microcystis aeruginosa cells into 7 centrifuge tubes containing 1mL sterile water, heat the 7 centrifuge tubes in boiling water for 3min, (25°C) to cool naturally, after cooling, add 4mL of acetone to the 7 centrifuge tubes and mix well, and then stand in a dark place at room temperature (25°C) for 0h, 0.5h, 1h, 1.5h, 2h, 2.5h, and 6h , centrifuged at 12000rpm for 5min. Take each supernatant to measure OD 750 , OD 663 , OD 645 , OD 630 , the chlorophyll content was calculated by the formula ρ (Chla) (μg / L). The amount of chlorophyll extracted is shown in Table 3.

[0027] extraction time 0h 0.5h 1.0h 1.5h 2.0h 2.5h 6.0h ρ(Chla)(μg / L) 3.169361 3.166472 3.058928 2.670952 2.297768 2.005824 1.749136

[0028] To sum up the above examples 1-3, the heating time in boiling water is 3min, the cooling...

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Abstract

The invention discloses a rapid and efficient extraction method of chlorophylls in a large quantity of cyanobacteria samples. The method comprises the following steps: centrifuging microcystis aeruginosa with the OD (Outer Diameter) 680 of 1.0 in a culture solution and then collecting microcystis aeruginosa strains; respectively treating the microcystis aeruginosa strains in boiling water for 3 minutes and naturally cooling at a room temperature (25 DEG C); adding 4mL of acetone and fully mixing; centrifuging so as to obtain a supernate, i.e., a chlorophyll extracting solution. According to the method, the extraction time of the microcystis aeruginosa is obviously shortened. The method is convenient to operate and high in extraction effect. The method has obvious advantages when being applied to the extraction of a large quantity of cyanobacteria chlorophylls of multiple samples.

Description

technical field [0001] The invention belongs to the field of biochemical industry, and in particular relates to a method for quickly and efficiently extracting chlorophyll in large batches of cyanobacteria samples. Background technique [0002] Chlorophyll is a natural, non-toxic, fat-soluble natural pigment that exists in some cyanobacteria and all green plants and has certain physiological functions. Its products have a wide range of uses, such as in food, pharmaceutical and chemical industries. As a fat-soluble pigment, chlorophyll is insoluble in water but soluble in organic solvents such as acetone and ethanol, so the above reagents are often used as effective solvents for extracting chlorophyll. [0003] Cyanobacteria are the earliest photosynthetic and oxygen-evolving organisms, a type of planktonic algae, and have played a huge role in changing the earth's surface from an oxygen-free atmospheric environment to an aerobic environment. Cyanobacteria are often seen onl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D487/22C09B61/00
CPCC07D487/22C09B61/00
Inventor 孙朋飞赵宇华王冠卢丽玲
Owner ZHEJIANG UNIV
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