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12 cze 2019 · Learn how to use thin-layer chromatography to separate and identify the pigments in different leaf extracts. The best solvent for chlorophyll chromatography is a combined mixture of hexane, acetone and trichloromethane (3:1:1).
Using a non-polar solvent in the chromatography of chloroplast pigments is critical because the pigments themselves are non-polar compounds. Non-polar solvents effectively dissolve these pigments, allowing them to travel up the chromatography paper or TLC plate.
Chlorophyll a is slightly soluble in a 3:1:1 mixture of petroleum ether, acetone, and water. Carotenoids are very soluble in this solvent system. These solubility differences will allow the separation of chlorophyll a from the carotenoids and chlorophyll b on a paper chromatogram.
Try this class practical using paper chromatography to separate and investigate the pigments in a leaf. Most leaves are green due to chlorophyll. This substance is important in photosynthesis (the process by which plants make their food).
The solvents must be HPLC grade or vacuum filtered through a nylon membrane of 0.2 μm pore size before use (as for the Tris buffer in solvent A) in order to avoid problems in the accuracy of the system.
18 sie 2018 · High performance liquid chromatography (HPLC) is one of the best options to characterise the exact pigment composition in a plant tissue, and it is the standard method to quantify individual carotenoids.
8 lut 2020 · Reversed-phase supercritical fluid chromatography (RP-SFC), which can be achieved with a non-polar C18-bonded phase, is an interesting alternative for the separation of this type of non-polar compounds. With regards to the organic solvents used in NARP-LC, carbon dioxide has many advantages: ecological, economic and physico-chemical.
