Chlorophyll is the green pigment that is found in all green plants as well as cynobacteria. Chlorophyll helps plants to absorb light in order to get the energy they need to photosynthesize. Each molecule in chlorophyll has a magnesium atom that is held within a porphyrin ring. Chlorophyll is an intra-membrane chemical found within a thylakoid membrane compartment that lies within the chloroplasts. Since it absorbs blue and red light so strongly but remains transparent to green light, chlorophyll tends to have a strong green color.
In higher plants, thylakoid membranes are primarily made of galactolopids and phospholipids that are arranged systematically across and along the membranes known as chloroplasts. Both have hydrophilic or water-loving heads and hydrophobic or water fearing tails. Within these complexes, chlorophyll plays two major roles which are absorption and transfer of light energy to specific chlorophyll reaction centers of photosystems. Even so, chlorophyll can dissolve in various solvents. Solubility in this context refers to the ability of chlorophyll organic molecules or complex to create an association with water and remain in the solution without being solubilized by the water. Proteins as well as other organic molecules utilize charged ions like sodium and phosphate to form tiny balls of hydrophobic molecules called micelles that are surrounded by charged ion.
In some situations, micelles are created by protein complexes that surround small molecules so they can be transported through water. Chlorophyll is able to combine and form complexes with other molecules to remain in a solution at a biological pH of 7.4. However, its natural environment is not a 7.4 pH, it is 4. At this environment, chlorophyll has a -2 net charge that enables it to form a chemo-gradient that facilitates electron transportation during photosynthesis. Chlorophyll can be extracted from plants using organic solvents that dissolve it and leave other materials intact. Here are the main solvents that dissolve chlorophyll:
A common organic solvent that is often used by scientists to dissolve chlorophyll is known as acetone. The process of dissolving chlorophyll using this solvent starts with dripping hot water on the leaf for several seconds or minutes. This helps stop other chemicals within the leaf from breaking the chlorophyll down. Once this is done, the stems and veins of the leaf are cut and the remaining tissue saved. The leaf should be soaked in pure acetone for a few hours. You know that chlorophyll is dissolving into the solvent when it turns green. The dissolving process can be accelerated by shaking the solution occasionally. Once done, pour the liquid containing the dissolved chlorophyll into a different container and discard the leftover, including the leaf. At this point, the acetone solution has chlorophyll plus other organic compounds and pigments. To separate extracted chlorophyll from the other compounds, paint a line on a paper chromatography strip using concentrated acetone solution and suspend the strip into the solvent. Alternatively, use HPLC to separate chlorophyll from other compounds.
Chlorophyll dissolves easily in alcohol. Chlorophyll molecules have magnesium at the ring center. This makes the molecules ionic and hydrophilic, meaning water loving. Chlorophyll molecules also have rings that are hydrophobic, meaning water fearing with carbonyl groups present near the end, making them polar or hydrophilic. Chlorophyll molecules are held within a water soluble compound known as water soluble chlorophyll binding protein which is soluble in water but largely insoluble in non-polar alkanes and polar alkane alcohol. Chlorophyll itself dissolves easily in alcohol, an aspect that makes it easy to extract using alcohol. However, chlorophyll is not soluble in polar alkanes such as hexane and butane. It has some exceptional relationships with water due to its ionic and polar groups.
- Water and Sodium Chloride
Chlorophyll is not soluble in water. However, the water soluble compound chlorophyll binding molecules can be dissolved in water. This aspect makes it possible to use water to extract and move insoluble chlorophyll molecules. However, extraction of chlorophyll can be done by saturating the water with table salt, which contains sodium chloride. This mixture helps in separating chlorophyll from other extractions. Where membranes have been broken down by enzymes or by a detergent, the only way to extract chlorophyll from alcohol or non-polar solvents is to use lots of water and sodium chloride. The large non-polar areas on chlorophyll molecules make it easy to form a hydrophobic interface that shields the charge at the center. The salt exposes the charge and precipitates chlorophyll into water.
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