Chromatography of Plant Pigments

CHROMATOGRAPHY OF PLANT PIGMENTS Marquez, Ma. Rica Paulene, Moises, Patrisha Kate, Policarpio, Jairus Paolo, Rolda, Zylene Joy Department of Biology, College of Science, University of the Philippines Baguio April 23, 2013 ABSTRACT The objective of this experiment was to apply the technique of opus chromatography as a method for separating individual jell rouges contained in make up tissue extracts containing rouge blends. The extremity of chromatography separates molecules because of the polar solubilities of the molecules in a selected closure.In paper chromatography, paper marked with an un turn inn, such as plant extract, is determined in a beaker covered with a foil containing a specified solvents. The solvent carried the dissolved blushers as it moved up the paper. The pigments were carried at different rates because they were not equally soluble. The around soluble pigment travelled the longest outstrip while the others traveled in a shorter length. The distance o f the pigment traveled was unique for that pigment in set conditions and was used to identify the pigment.The ratio was because used to footprint the Rf (retention factor) value. INTRODUCTION As primary developrs in the food chain with approximately bacteria and algae, plants produce their own food by using the suns cipher to transform carbon dioxide and peeing into glucose. In this process of photosynthesis, plants convert the suns energy into chemical energy that is stored in the bonds of the glucose molecule. Glucose is a simple carbohydrate that provides immediate fuel to cells nevertheless it is in any case a building block for much complex carbohydrates stored by living organisms for future use.For photosynthesis to transform light energy from the sun into chemical energy (bond energy) in plants, the pigment molecules take on light to power the chemical reactions. Plant pigments are macromolecules produced by the plant, and these pigments absorb specified wavelength s of visible light to provide the energy required for photosynthesis. Chlorophyll is necessary for photosynthesis, but supportive pigments collect and transfer energy to chlorophyll. Although pigments absorb light, the wavelengths of light that are not absorbed by the plant pigments are reflected back to the eye.The reflected wavelengths are the colors we see in observing the plant. Plants contain different pigments, and some of the pigments observed include ? chlorophylls (greens) ? carotenoids (yellow, orange red) ? anthocyanins (red to blue, depending on pH) ? betalains (red or yellow) (Plant Traveling Lab. TTU/HHMI at CISER. 2010) As you may know from the popular media, there is currently a substantial research effort in place to look for the potential health benefits of plant pigments to humans. In popular literature, these plant-based immixs are often collectively referred to as phytochemicals most are besides pigments.Flavonoids, anthocyanins, and carotenoids are just som e of the categories of plant pigments known to have antioxidant properties. Antioxidant is a normal term used to describe any substance that has the ability to neutralize free radicals which cause cellular damage by removing electrons from surrounding molecules. Many lines of research suggest that consuming a diet fatty in plant pigments may slow the process of cellular aging and reduce the risks of some types of disease, such as cancer, heart disease, and stroke.The point of this experiment is to look at the oppositeity of some of the common pigments in plant leaves and how that polarity affects their interactions with the cellulose fibers in paper and a few solvents and to apply the technique of paper chromatography as a method for separating individual plant pigments contained in plant tissue extracts containing pigment blends. (https//www. msu. edu) RESULT shelve 1 shows the Rf values of Kangkong (Ipomoea aquatic), Bloodleaf (Iresine herbstii), and Golden bush (Duranta repen s) plant Rf value 50% acetone50% pet. ther 40% acetone, 10% isopropanol, 50% pet. ether 40% acetone, 10% pet. ether, 50% isopropanol 30% acetone70% pet. ether 30% pet. ether 70% acetone Kangkong yellow-bellied 0. 5 embrown 0. 01 Yellow 0. 33Brown 0. 23 Yellow 0. 79Brown 0. 08 Yellow 0. 04Brown 0. 96 Yellow 0. 31Brown 0. 76 Bloodleaf (purple leaf) colour0. 05 gullible0. 11Yellow0. 11 Purple0. 06Green0. 13 Purple0. 13Green0. 58 Purple0. 03Green0. 58Yellow0. 38 Purple0. 097Green0. 86 Golden bush Green0. 5Brown0. 5 Green0. 47Brown0. 09 Green0. 6Brown0. 067 Green0. 3Brown0. 96 Green0. 094Brown0. 96 DISCUSSION The extracts of kangkong, golden bush, and purple leaves were applied to a horizontal line about ? of an butt against from the bottom of a filter paper using a capillary tube. The filter paper then was soaked one by one on a beaker with a mixture of 40% acetone, 10% petroleum ether and 50% isopropanol. These solvents are used because they are capable of separating mixtures that contain both polar and non-polar compounds, or to increase separation of mixtures of compounds that have similar behavior with a single solvent.The beaker was covered with aluminium foil to make sure that the atmosphere in the beaker is saturated with the solvents vapor. Saturating the atmosphere in the beaker with the solvents vapor stops the solvent from evaporating as it rises up the filter paper. As the solvent slowly travels up the paper, the different components of the extract travel at different rates and the extracts are disordered into different colors. After 3-5 minutes, the distance traveled by each pigment and solvent were measured. Plants extracted Color/ pigment Rf values Kangkong Yellow (solvent) 0. 9 mm Brown (solute) 0. 08 mm Purple leaves Green (solvent) 0. 58 mm Purple (solute) 0. 13 mm Golden bush Green (solvent) 0. 6 mm Brown (solute) 0. 067 mm Table 1 Pigments and Rf values for each plant extract The distance traveled relative to the solvent is called the Rf value, or the Retardation value. It can be computed with the formula Rf = distance travelled by the solute Distance travelled by solvent We had the following computations A. For Kangkong Rf= 0. 08 mm 0. 79 mm = 0. 1012 B. For purple leaves Rf= 0. 13 mm 0. 58 mm = 0. 241 C. For Golden bush Rf= 0. 067 mm 0. 6 mm = 0. 1117 These values imply that the larger Rf value a compound has, the larger the distance it travels. It also means that it is less polar because it interacts less strongly with the polar hygroscopic on the filter paper. So similarly, the smaller the Rf value a compound has, the shorter the distance it travelled. It also means that is is more polar because it interacts more strongly with the polar absorbent on the filter paper. comparability to the other groups results, there were similar colors that sprung up.The distances travelled by the pigments were significantly different than ours because they used different percentages of solvents. CONCLUSION Paper chromatogra phy proved to be an accurate method of separating and observing the various colors of plant pigments. The pigments dissolved in the solvent and migrated upward. The colors were observed and their migration distances measured & recorded. TheRfvalueof each pigment was determined by dividing its migration by the migration of the solvent. We have always understood chlorophyll, a pigment that is very alpha in photosynthesis, to be green.However, through this experiment we have discovered that many other pigments are also present in the leaves. For example the kangkong leaf also contains different pigments even though the leaf is prevail by the color green. We have observed that the kangkong leaf not only carries a green pigment, but that it also carries yellow and brown pigment through the chromatography LITERATURE CITED * Mcmurry, John. 2010. Foundations of Organic Chemistry. Pasig City, Philippines Cengage Learning Asia Pte Ltd. * Thompson, R. 2008. Illustrated guide to alkali Chemi stry experiments. Canada O Reilly media.Page 109 * Clark, J. 2007. http//chemguide. co. uk/analysis/chromatography/paper. html. Retrieved April 22, 2013 * Unknown. PlantTravelingLab. TTU/HHMI at CISER. 2010 . http//www. ciser. ttu. edu. * 2010. 04-20-13. * Unknown. separation of pigments by paper chromatography. http//www. hsu. edu/pictures. aspx? id=1653, n/a. 04-20-13 * Unknown. plant pigment analysis. https//www. msu. edu. n/a. 04-20-13 * http//apnewtechbiology. wordpress. com/lab-6-plant-pigments/ * http//www. biologyjunction. com/chromatography_of_simulated_plan. htm ANSWERS TO QUESTIONS