Column Chromatography

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Column Chromatography of Plant Pigments
Paul Ibarbia, Gene Paolo Jasmin, Gianpaolo Jimenez and Lorenzo Labicane* Department of Biology, University of Santo Tomas, Manila, Philippines

Abstract
Column chromatography of plant pigments is the separation of plant pigments extracted from Capsicum frutescens (siling labuyo). No slurry is prepared of the eluent; eluent with the stationary phase is set and then carefully poured into the column. The eluent are the solvents which are responsible for the pigments to be divided. The organic material inside the column is advanced by the eluent slowly passing through the inside of the column. Throughout the entire chromatography process, the eluent is collected in series of fractions. The eluent’s flow composition can be observed and each fraction is analysed to what specific color it will show.

Introduction
The Capsicum frutescens (siling labuyo) of the family Solanaceae, is a short-lived perennial with woody stems that reach a height of two meters, glabrous or pubescent leaves, has two or more greenish-white flowers per node, and extremely pungent fruit. This plant is cultivated in the tropics and warmer regions of the United States and is found throughout the Philippines, planted but also thoroughly established in open, waste places in settled areas. Capsicum species are cold sensitive and normally grow best in well-drained, sandy or silt-loam soil. Plantings are established by transplanting or seeding. Hot and dry weather is necessary for fruit ripening. Fruit is generally handpicked as it ripens, and then allowed to dry in the sun. The fruit may be ground intact or after the removal of seeds, placenta parts, and stalks, increasing the fruit color and lowering the pungency. The concentration of capsaicinoids, primarily of capsaicin which is an active component of chili peppers, in the fruit is dependent of the level of its pungency or spiciness. The concentration of capsaicinoids is used to commercially classify Capsicum peppers. The members of the Capsicum species are usually composed of chemicals namely fixed and volatile oils, pungent principles and carotenoids which are mostly capsanthin pigments. Carotenoids are fat-soluble organic pigments which are responsible for many of the red, yellow and orange hues of plants. Carotenoids also functions as an internal biological antioxidant that helps human cells and tissues to be protected from the harmful and damaging effects singlet oxygen and free radicals. Plants used in the production of certain commercial products are recognized for their pungency and color. Capsicum species are used fresh or dried, whole or ground, and alone or in combination with other flavoring agents. It is used in tabasco, tabasco sauce, curry powder and other red chili pepper. The species are the spiciest of their type and is used extensively in Mexican and Italian foods and also as a popular flavoring ingredient in some of our native dishes. As a medicinal plant, the Capsicum species has been used as a carminative or prevention of flatulence, digestive irritant, stomachic effects, stimulant, rubefacient, and tonic. The plants have also been used as folk remedies for dropsy, colic, diarrhea, asthma, arthritis, muscle cramps, and toothache. Prolonged contact with the skin may cause dermatitis and blisters, while excessive consumption can cause gastroenteritis and kidney damage. Having high levels of ground hot pepper has induced stomach ulcers and cirrhosis of the liver. Body temperature, flow of saliva, and gastric juices may be stimulated by capsicum peppers. Capsicum frutescens are generally recognized as safe for human consumption as spices and natural flavorings and as plant extracts and oleoresins. [1] The word chromatography is derived from the Greek word “chromatos” which means “color”. From the work of the Russian chemist Mikhail Tswett, who discovered the technique in 1903 while studying ways to separate mixtures of natural plant pigments. The...
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