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Method for preparing quantitative standard substance of photosynthetic pigments

A quantitative standard product, photosynthetic pigment technology, applied in the field of quantitative standard product preparation of phytoplankton photosynthetic pigment, can solve the problems of high price, low concentration, incomplete variety, etc., achieve high accuracy and reliability, single component, low cost cheap effect

Inactive Publication Date: 2014-05-14
HUBEI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] 1. The types are incomplete, and the standard photosynthetic pigments provided are limited, which is often difficult to meet the qualitative and quantitative requirements for photosynthetic pigments of phytoplankton in water bodies;
[0010] 2. The concentration is not high, and the price is relatively expensive;
[0011] 3. When the biomass of phytoplankton in the environment is high, especially in the state of water bloom, the concentration of photosynthetic pigments in the field samples will fall outside the working curve, resulting in quantitative errors;
[0013] 5. The quantitative standard of photosynthetic pigment must be stored at -20°C to ensure that it will not degrade. However, for products that need to be imported from abroad, in the process of transnational transportation, it is necessary to pay a high price to ensure this condition. cost
[0014] At present, the production method of a series of quantitative standards for photosynthetic pigments has not been published

Method used

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  • Method for preparing quantitative standard substance of photosynthetic pigments
  • Method for preparing quantitative standard substance of photosynthetic pigments
  • Method for preparing quantitative standard substance of photosynthetic pigments

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1. Separation and extraction of Microcystis aeruginosa zeaxanthin standard pigment

[0054] 1) Experimental algae:

[0055] The monocultured Microcystis aeruginosa (Microcystis sp.7806) was selected from the Freshwater Algae Species Bank of the Institute of Hydrobiology, Chinese Academy of Sciences.

[0056] 2) Preparation of quantitative standards for separation and extraction of pigments and photosynthetic pigments:

[0057] A. Cultivation method of experimental algae: the above-mentioned experimental algae with BG11 medium was cultivated in the Laboratory of Aquatic Biodiversity and Environmental Resource Effects of Hubei Normal University under the light intensity of 1000lux and the temperature of 25°C.

[0058] B. Extraction of mixed pigments: Take 300ml of algae liquid in the logarithmic growth phase and collect it through GF / F membrane filtration. The negative pressure of suction filtration is 0.02MPa. The filter membrane is stored at -20°C. Liquid chro...

Embodiment 2

[0067] Example 2. Separation and extraction of Microcystis aeruginosa zeaxanthin standard pigment

[0068] Compared with Example 1, the rest of this example is the same, except that the culture medium of the algae used in the experiment is a CSI medium, and the extraction steps of the mixed pigment are as follows: after accurately counting 300 mL of cultured algae cells, centrifuge them at 5000 rpm / 5 min and collect them in In a 2mL centrifuge tube, after discarding the excess water in the supernatant, add 2.0mL DMF, shake well, and then extract in the refrigerator for 40min, shaking twice during the period. After the pigment is extracted, filter it with a syringe with PTEF, take 0.5mL into the chromatographic bottle, and then add 0.5mL of 1.0M ammonium acetate. This operation is carried out before loading the sample on the chromatograph, so as not to cause flocculent precipitation in the sample .

Embodiment 3

[0069] The making of embodiment 3 zeaxanthin standard pigment working curves

[0070] 1), material:

[0071] From Example 1.

[0072] 2), method:

[0073] A, methods such as pigment extraction and separation see embodiment 1.

[0074] B, the zeaxanthin obtained by separating is measured the absorbance value under its maximum wavelength 450nm under the ultraviolet spectrophotometer, deducts the impurity absorption value at 750nm place, calculates according to the following formula, draws its accurate concentration:

[0075] Carotene: C = A λ max 0.1 × E 1 cm 1 % × d × P c 100 ...

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Abstract

The invention discloses a method for preparing a quantitative standard substance of photosynthetic pigments. The method comprises the following steps: carrying out xenicculture on limnetic algae, extracting mixed photosynthetic pigments in the limnetic algae under dark condition, and performing separating and purification, thus obtaining target photosynthetic pigments; and concentrating to obtain the enriched target photosynthetic pigments, quantifying the enriched target photosynthetic pigments, thus obtaining the quantitative standard substance of photosynthetic pigments. The method can be used for rapidly and economically preparing the quantitative standard substance at least including nine common target diagnosis photosynthetic pigments in all limnetic algae photosynthetic pigments; the quantitative standard substance is convenient to apply, good in repeatability, and high in accuracy and reliability, and can be widely applied to the field of researching phyto-group formation of phytoplankton or ultra-miniature phytoplankton.

Description

technical field [0001] The invention relates to the technical field of environmental monitoring, in particular to a method for preparing quantitative standard substances of phytoplankton photosynthetic pigments. Background technique [0002] Phytoplankton are the main contributors to primary productivity in aquatic ecosystems, and they are also the culprits of ecological disasters such as algae blooms. The biomass and community structure characteristics of phytoplankton are an important indicator of water environment assessment. Common analysis methods for phytoplankton biomass and community structure characteristics include: ultraviolet-visible spectrophotometry, fluorescence spectrophotometry, and microscopy. Although microscopic analysis can obtain detailed information on the composition of phytoplankton species, it has the disadvantages of heavy workload and long time consumption; and the analyst needs strong knowledge and background of phytoplankton taxonomy, and the a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28G01N33/00
Inventor 侯建军胡俊李家园贺云彦周婷郑和龙毕永红
Owner HUBEI NORMAL UNIV
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