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Method for determining content of carotenoids in microalgae

A carotenoid and a determination method are applied in the field of content determination of carotenoid components in microalgae, which can solve the problems of long time consumption, large instrument dependence, large amount of solvent and the like, and achieve low cost, strong sensitivity, and reagent dosage. less effect

Inactive Publication Date: 2019-06-04
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, carotenoids are mostly extracted with organic solvents, which requires a large amount of solvent and takes a long time
Methods such as supercritical fluid extraction, solvent extraction, and countercurrent chromatography have also been gradually applied to the extraction of carotenoids, but these methods rely heavily on instruments and are costly

Method used

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  • Method for determining content of carotenoids in microalgae
  • Method for determining content of carotenoids in microalgae
  • Method for determining content of carotenoids in microalgae

Examples

Experimental program
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Effect test

Embodiment 1

[0027] (1). Matrix-dispersed solid-phase extraction: Weigh 100 mg Nannochloropsis and 200 mg C8 bonded silica filler into an agate mortar, grind thoroughly with an agate pestle, and put the mixture into an empty SPE with a PTFE gasket Fill the plastic column tube (6mL specification) evenly, and put a polytetrafluoro gasket on the mixture. Place the packed SPE column on the SPE device, first rinse the SPE column with 2mL of 30% methanol-water to wash away the highly polar impurities, then elute with 4mL of methanol (containing 0.1% BHT by mass fraction), and collect the eluted After blowing with nitrogen, dilute to 5mL with methanol, and analyze by high performance liquid chromatography.

[0028] (2). High performance liquid chromatography method

[0029] 1) High performance liquid chromatography analysis method

[0030] The high-performance liquid chromatograph is HITACHI Chromaster from Japan, including quaternary solvent manager, automatic sampler, column thermostat, DAD d...

Embodiment 2

[0053] Change the Nannochloropsis in the (one) section in embodiment 1 into Chrysophylla, and others are the same as in embodiment 1.

[0054] (3) Determination of the actual Chrysophylla sample content

[0055] Process the Nannochloropsis sample according to the above-mentioned matrix dispersion solid-phase extraction method, and detect through high-performance liquid chromatography, and the analytical spectrum is shown in image 3 , as can be seen from the figure, this method can extract and enrich the fucoxanthin in the golden algae, after adopting the external standard method to quantify, the content of the fucoxanthin in the golden algae is 1.13mg / g (n=3, RSD = 8.78%). .

Embodiment 3

[0057] Change the Nannochloropsis in the (one) section in embodiment 1 into flat algae, and others are the same as in embodiment 1.

[0058] (3) Determination of content of actual flat algae samples

[0059] Process the flat algae sample according to the above-mentioned matrix dispersion solid-phase extraction method, and detect through high-performance liquid chromatography, and the analytical spectrum is shown in Figure 4 , as can be seen from the figure, this method can extract and enrich the lutein in the flat algae. After quantification by the external standard method, the content of lutein in the flat algae is 153.1 μg / g (n=3, RSD= 4.77%).

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Abstract

The invention relates to a method for determining the content of carotenoids in microalgae, which simultaneously carries out content determination on three free carotenoids such as fucoxanthin, astaxanthin and lutein under the same chromatographic condition by adopting matrix solid phase dispersion-extraction pretreatment and high performance liquid chromatography. The method comprises the steps of 1) weighing 0.1-5g of a reversed-phase solid phase extraction filler and microalgae powder with the mass ratio with the filler ranging from 1:2 to 1:20, and grinding the filler and the microalgae powder in an agate or glass mortar; 2) loading the uniformly grinded mixture into an empty SPE tube with a polytetrafluoroethylene gasket, carrying out drip washing by water and methanol with the volumeratio ranging from 100% to 70%, and carrying out elution with an organic solvent; 3) and concentrating the eluant to a constant volume and then adopting high performance liquid chromatography analysis. The method has the characteristics of simplicity, rapidness, low consumption of reagents, accurate and reliable result, good stability and the like, and is applicable to qualitative and quantitative analysis for the free carotenoids in the microalgae.

Description

technical field [0001] The invention relates to a method for determining the content of carotenoids in microalgae. It is characterized in that: three kinds of free fucoxanthin, astaxanthin and lutein carotenoid components in microalgae are detected under the same chromatographic condition by matrix dispersion solid phase extraction pretreatment and high performance liquid chromatography. Simultaneously carry out content determination. The method has the characteristics of simplicity, rapidity, less reagent consumption, accurate and reliable results, and good stability, and is suitable for qualitative and quantitative analysis of free carotenoids in microalgae. technical background [0002] Carotenoids are a class of naturally occurring fat-soluble pigments that are commonly found in algae, plants, fungi and bacteria. They are not only a good food additive, but also widely used in cosmetics and health products. Humans and animals cannot synthesize carotenoids and must obtai...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
Inventor 梁鑫淼金高娃郭志谋闫竞宇
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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