Extraction of chlorophyte pigment

of these green plants is carried out in chlorophytes, and understanding the composition and properties of chlorophyte pigments is helpful in understanding the nature of photochrome. The content of folicin in leaves is closely related to photolysal intensity and nitrogen nutrition. Therefore, the determination of the content of lustin has become an indispensable means to study photoneculation and nitrogen metabolism, and has been widely used in crop breeding, scientific fertilization and leaf-watching diagnosis.principleprinciple
chlorophyte contains chlorophyte (chlorophylatin a and b) and carotenoids (carotene and chlorophylactin), both pigments are insoluble in water, and dissolved in
organic
solvents containing water, so 95% alcohol and 80% acetone extraction, or with alcohol, acetone and water mixture. Chlorophylic pigment is susceptible to light oxidation, the extraction of pigment should be carried out in low light, and avoid light to preserve pigment.equipment and
reagements
equipment: torque
balance
, measure barrel,
beaker
, plug test tube, research,
centrifuge
, funnel, filter paper and so on.
: alcohol, acetone, calcium carbonate.method and step
1. Chlorophyl pigment extraction for physical and chemical properties identification and color layer separation:
(1) Take fresh plant leaves, such as spinach leaves, first kill at 105 degrees C, then dry at 80 degrees C (not too much at one drying), into a powder,
drying
in the spare, can also be directly extracted with fresh leaves.
(2) said to take leaf powder 2g into a small beost, add 95% alcohol 20-30 ml, for dipping. The immersion process needs to be stirred frequently with a glass rod, such as the temperature is too low can also be appropriate
heating
, when the alcohol is dark green, the solution
filtering
to another beech or triangular bottle in the dark spare.
E.g. with fresh leaves, you can cut the fresh leaves 10g, put in the research, add a little calcium carbonate (the acid in the cell, prevent magnesium from removing from the chlorocerin molecule) and 20 ml of acetone grinding, homogeneous filtration, and then use 10-20 ml of acetone repeatedly wash the pigment in the research and residue, the filter is a pigment extract, preserved in the dark.
2. Chlorophylic pigment extraction for quantitative analysis.
(1) extraction of chlorophyl pigment in leaves: 0.5g fresh leaves cut into pieces into 10ml or 20ml plug test tubes, add 10ml or 20ml of acetone: alcohol : Water: 5:5:1 mixture, covered with plugs, placed in a temperature tank at 40 degrees C immersion (3 hours or more), until the leaves fade white, you can use the test tube solution for quantitative determination.
(2) Chlorophyl pigment extraction of leaf meat cells or chlorophyl suspended fluid: 0.1 ml of leaf meat cells or chlorophyl suspended fluid in the centrifuge tube, plus 4.9 ml 80% acetone, after a moment of rest, with a bench centrifuge 3000 l/min, centrifuge 2 minutes, the upper liquid can be used for chlorophyl content determination.
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