Plant chlorophyll determination method and procedure

Methods and Procedures for Determining Plant Chlorophyll Chlorophyll is the most important pigment associated with photosynthesis. Photosynthesis is the process of converting light energy into chemical energy by synthesizing some organic compounds. Chlorophyll actually exists in all organisms that can create photosynthesis, including green plants, prokaryotic blue-green algae (blue fungus) and eukaryotic algae. Chlorophyll absorbs energy from light, and energy is then used to convert carbon dioxide into carbohydrates.
Determination of plant chlorophyll:
(1) Material: fresh (or dried) plant leaves.
(2) Instruments and equipment: 1) Spectrophotometer; 2) Electronic top load balance (thickness 0.01g); 3) Research mortar; 4) Brown volumetric flask; 5) Small funnel; 6) Quantitative filter paper; 7) Absorbent paper 8) Rub the paper; 9) Dropper.
(3) Reagents: 1) 95% ethanol (or 80% acetone) (v 90% aqueous solution of acetone: v ethanol = 2: 1); 2) quartz sand; 3) calcium carbonate powder. Secret 2h, 0.5g, 25ml
(4) Take fresh plant leaves (or other green tissue) or dry materials, clean the surface of the tissue, cut into pieces (remove midribs), and mix.
(5) Weigh 0.2g of freshly crushed samples, total 3, and put them in a mortar, add a small amount of quartz sand and calcium carbonate powder and 2~3ml of 95% ethanol, and research into a homogenate, add ethanol 10ml, Continue grinding until the tissue turns white. Let stand 3~5min.
(6) Take 1 filter paper, set the funnel, moisten with ethanol, pour the extract into the funnel along the glass rod, filter it into a 25ml brown volumetric flask, rinse the mortar, pestle and residue several times with a small amount of ethanol, and finally Pour into the funnel together with the residue.
(7) Using a dropper to suck up the ethanol, 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, make up to 25 ml with ethanol and shake well.
(8) The chloroplast pigment extract was poured into a cuvette with a light path of 1 cm, and 95% ethanol was used as a blank. The absorbance was measured at wavelengths of 663 nm and 645 nm.

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