1. Take fresh plant leaves (or other green tissue) or dry material, wipe the surface dirt, cut (remove the midrib), and mix.

2. Weigh 0.2g of freshly chopped fresh sample into a mortar, add a small amount of quartz sand and calcium carbonate powder and 2~3ml of 95% ethanol, grind into a homogenate, add 10ml of ethanol, continue grinding To the organization turned white. Allow to stand for 3 to 5 minutes.

3. Take 1 piece of filter paper, place it in a funnel, moisten with ethanol, pour the extract into the funnel along the glass rod, filter into a 25ml brown volumetric flask, rinse the mortar, pestle and residue several times with a small amount of ethanol, and finally The residue was poured into the funnel together.

4. Pipette the ethanol with a dropper and wash all the chloroplast pigments on the filter paper into a volumetric flask. Until there is no green in the filter paper and residue. Finally, dilute to 25 ml with ethanol and shake well.

5. Pour the chloroplast pigment extract into a cuvette with a diameter of 1 cm. The absorbance was measured at a wavelength of 665 nm and 649 nm with 95% ethanol as a blank. Calculation of experimental results:

The measured absorbance values ​​were substituted into the following formula: Ca = 13.95 A665 - 6.88 A649; Cb = 24.96 A649 - 7.32 A665. According to this, the concentrations of chlorophyll a and chlorophyll b (Ca, Cb: mg/L) can be obtained, and the sum of the two is the concentration of total chlorophyll. Finally, according to the following formula, the content of chlorophyll in plant tissues can be further determined:

Chlorophyll content (mg/g) = [concentration of chlorophyll x volume of extract × dilution factor] / fresh weight (or dry weight) of the sample.

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