Discussion on Some Problems in the Determination of Walnut Chlorophyll Content in Luodian Millet
Chlorophyll is one of the most important substances in photosynthesis of plant leaves. The content of chlorophyll is an important indicator of plant photosynthesis ability. For the measurement of chlorophyll content, many domestic and foreign related research, such as extraction of chlorophyll, chlorophyll meter measurement method using acetone, direct extraction of plant chlorophyll blade dimethyl sulfoxide and the like. Each method has its own advantages and disadvantages, but in practical applications, the chlorophyll meter is most frequently used for measurement at this stage because it is the quickest and simplest method. And the accuracy is extremely high.
The chlorophyll content determined by the Arnon filtration method is quite different from other methods. The main reason may be that the Arnon method requires filtration when determining the chlorophyll content, and the filtration process causes a partial loss of chlorophyll content. Arnon centrifugation is slightly more efficient than Arnon filtration. The efficiencies of the chlorophyll content determination by extraction method were as follows: 80% acetone extraction method <95% ethanol extraction method <three mixture solution extraction method <two mixture solution extraction method.
In addition, with the increase of the proportion of ethanol in the mixture, the synergistic effect increases, and the content reaches the maximum when the ratio of ethanol to acetone reaches 1:1; subsequently, ethanol is added to acetone, and the synergistic effect becomes smaller. It can be seen that when the ratio of ethanol to acetone reaches 1:1, the extraction of chlorophyll from Luodianxiaomi walnuts is the most abundant. Experiments with different proportions of synergistic effects showed that when the ratio of anhydrous ethanol to anhydrous acetone was 1:1, the amount of extraction was the largest. Therefore, the single factor test used the above solution as the experimental extraction solution. In the related literature of chlorophyll extraction, three main factors affecting the extraction of chlorophyll were proposed: extraction temperature, extraction time and extract-to-liquid ratio. At the same extraction temperature and extract liquid-liquid ratio, when the extraction time increased from 2 h to 5 h, the chlorophyll content of Luodian walnut groats gradually increased and reached its maximum at 5 h, and then followed by leaching. As time increases, the chlorophyll content continues to decline. This shows that the optimum extraction time of chlorophyll is 5 h, and less than or more than 5 h, the extraction of chlorophyll is not sufficient. This is due to the fact that after leaching begins, there is a process of outward diffusion of chlorophyll from the tissues, so the extracted chlorophyll increases continuously. When the extraction time was 5 h, the diffusion of chlorophyll into the extraction solvent reached the equilibrium and the chlorophyll content in the solution reached the highest. After the leaching time exceeds 5 h, the chlorophyll content in the solution begins to decrease. The probable reason is that with the increase of extraction time, the chlorophyll leakage of Luodian millet walnuts will take longer under light, and the instability of light will cause the chlorophyll content to decrease continuously.
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