COLUMN CHROMATOGRAPHY (MALUNGGAY LEAVES)

Angel Lynn Manalansang, Sheena Patrizzia Martinez, Gilbert John Mascardo Kerwin Jeffrey Maez and Angelo Rafael Mendoza Group 8, 2DMT, Faculty of Pharmacy, University of Santo Tomas ABSTRACT
The main focus of this study was the pigments present in malunggay leaves and the process of chromatography. The result obtained from this experiment was interpreted. The objective of this experiment is to separate the coloured components of malunggay leaves using column chromatography. The qualitative results of the experiment were about the colour and dominance of different pigments in the malunggay leaves. Both yellow pigments and green pigments were seen in the column chromatography set-up. Also, the set-up was only able to extract yellow and green pigments with almost equal amount in drops.

INTRODUCTION Chromatography is a proven method for separating complex samples into their constituent parts, and it is undoubtedly the most important procedure for isolating and purifying chemical. Chromatography relies on the differential solubilities and adsorptive of the components to be separated with respect to two phases; stationary phase and mobile phase. Stationary phase is the part of the chromatographic system though which the mobile phase flows where distribution of the solutes between the phases occurs. Mobile phase, on the other hand, is the part of the chromatographic system which carries the solutes through the stationary phase. MATERIALS AND METHODS In this experiment, a column chromatography setup was used. The components of the setup are: Mortar and Pestel Pasteur pipettes Iron stand Iron Clamp Malunggay Leaves Hexane Acetone Vials Silica gel Cotton

Figure 1. Column Chromatography

First, the malunggay leaves were triturated with the use of a mortar and pestle. Second, 5 mL of hexane: acetone (7:3) was poured to the triturated malunggay leaves. After that, the dropping pipette that will be used for the set up was plugged by cotton and was uniformly packed with silica gel up

to the indented part of the dropping pipette. 3 mL of the extract was placed on the top of the column using a dropping pipette. Then, the pigment mixture was eluted using 2 mL of hexane: acetone (7:3), 2 mL of acetone, and 2 Ml of acetone: methanol (1:1). The solvent systems were introduced in portions and the column was not allowed to run dry. The colourless eluate gotten from the column was discarded while the coloured eluates were collected in different test tubes. Data, like the number of drops per eluate collected in each test tube, were collected and noted.

and green pigments but with different shades. Table 1. Compound and Solvent system used Plant Used Solvent System Used Malunggay (Moringa oleifera) Hexane: acetone (7:3)

Table 2. Column Chromatography Colour of Component 1 2 3 4 5 6 Yellow Light green Green Yellow green Light yellow Yellow

Dried silica gel Cotton Test tube

REFERENCES [1] Bayquen, A.V., Cruz, C.T., de Guia, R.M., Lampa, F.F., Pea, G.T., Sarile, A.S., Torres, P.C. (2010). Laboratory Manual in Organic Chemistry. Manila, Philippines. C& E Publishing Inc. Date: 08/16/2011 [2] Chromatography-online.org. Mobile Phase. http://www.chromatographyonline.org/topics/mobile/phase.ht ml Date: 08/16/2011 [3] Chromatography-online.org. Stationary Phase. http://www.chromatographyonline.org/topics/stationary/phase .html Date: 08/16/2011 [4] Miller, James (2005). Chromatography: Concepts and Contrast. New Jersey: John Wiley & Sons Inc. 08/16/2011

Figure 2. Column Chromatography Set-Up

RESULTS AND DISCUSSIONS Different observations were noted during the experiment. First of all, there were 6 eluetes yielded. Those were yellow, light green, green, and yellow green, light yellow and yellow in respective order. Yellow pigments and green pigments dominance was proven by this observation. This was also proven by the column chromatography set-up which only has extracted yellow