Experiments on the extraction, isolation, properties and quantitative determination of chloroplast pigments
Experiments on the extraction, isolation, properties and quantitative determination of chloroplast pigments
Chloroplast pigment is an important substance for plants to absorb sunlight energy for photosynthesis. This experiment is to learn the extraction and separation of chloroplast pigment in plants, to understand the important physical and chemical properties of chloroplast pigment, the principle and method of quantitative analysis.
Operation method
Experiments on the extraction, isolation, properties and quantitative determination of chloroplast pigments
Principle
1. solubility: chlorophyll and carotenoids are insoluble in water but soluble in organic solvents, commonly used in ethanol or acetone extraction. 2. separation of pigments in a variety of methods, such as paper chromatography. Column chromatography, etc. Paper chromatography is one of the simpler methods. Paper chromatography is one of the simpler. When the solvent continuously flows through the chromatographic filter paper due to the various components of the mixed pigments in the two phases (i.e., mobile and stationary phases) have different partition coefficients between them, they move at different speeds, so that the various components in the sample to be separated.3. Chlorophyll by light excitation, can emit red fluorescence, reflecting the light visible red fluorescence.4. Saponification reaction: chlorophyll is a dicarboxylic acid ester, and react with a strong base, the formation of a green Soluble chlorophyll salt, it can be separated from the carotenoids in organic solvents. 5. substitution reaction: in the acidic and heating conditions, chlorophyll porphyrin ring in the Mg2 + can be replaced by H + and Cu2 + to form a brown de-magnesium chlorophyll and green copper chlorophyll. 6. quantitative analysis, chlorophyll absorbs red and blue-violet light, the red region of the light can be used in the quantitative analysis, which is used in the quantitative analysis of 645 and 663 Chlorophyll a, b.
Materials and Instruments
Wheat Plant leaves Move I. Materials and equipment: Common Problems Result Calculation: For more product details, please visit Aladdin Scientific website.
Electronic balance Spectrophotometer Chromatography cylinder
1, experimental materials: leaves of wheat, spinach and other plants
2, Instruments: (1) electronic balance (2) spectrophotometer (3) chromatography cylinder
Second, the method:
Take 3g of fresh leaves + 100% acetone 10ml, grind into homogenate 
Method of pigment separation: take a round filter paper with a diameter slightly larger than that of the Petri dish, suck the sample with a pipette to drop a drop at the center of the filter paper, air-dry or bake dry, and repeat seven times at the original position. Take another long strip of filter paper about 1cm wide, roll it into a paper core and pass it through the center of the sample point, so that the core is perpendicular to the circular filter paper. Put a small aluminum box in the petri dish, flat on the petri dish, the box add appropriate amount of gasoline and add 1-2 drops of benzene, has been treated with the lower end of the filter paper core immersed in gasoline, quickly cover the petri dish lid, at this time, gasoline through the capillary gravity along the paper diffusion of the paper to the round filter paper, and chlorophyll along the filter paper to the surrounding area to promote, soon you can see the separation of various pigments of the concentric rings.