Chromatography

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COLUMN AND THIN LAYER CHROMATOGRAPHY Maria Janine B. Abarientos, Kuia B. Allarde, Aliana Keshia P. Andino Mary Viadelle E. Andrada and Nina Marian Robelea G. Ang Group 1 2C Pharmacy Organic Chemistry Laboratory ABSTRACT In this experiment, the techniques column and thin layer chromatography was used to separate and determine the purity of the colored components of siling labuyo and malunggay leaves. The results obtained relied on differential solubilities and adsorptivities of the components to be separated with respect to the mobile and stationary phases, wherein the stationary phase/s are silica gel and silica gel G, while the mobile phase/s are the extracts on different solvent system. INTRODUCTION This experiment aims to make…show more content…
EXPERIMENTAL A. Sample Used Sample Used: Siling Labuyo (Capsicum frutesceus) B. Procedure 1. Column Chromatography The pigments of the red siling labuyo were extracted using DCM-hexane (1:1). Portions of the extracts were set a side for Thin Layer Chromatography. In preparing the column chromatography set up, the column was plugged with cotton and uniformly packed with silica gel up to the intended part of the dropping pipette. 0.5 mL of the extract was then placed on top of the column using a pipette. The pigment mixture was eluted using 10 mL of DCM-hexane (1:1) and afterwards introduced in portions. The members were instructed not to allow the column to run dry. The colorless eluates were discarded while the colored eluates were collected in separate test tubes. The number of drops of the eluate collected in each test tube was noted. Figure 1. Column Chromatography Set-up 2. Thin Layer Chromatography In the Thin Layer Chromatography (TLC), the eluates were applied on a 5 cm x 8 cm per coated TLC plate by spotting 10 times. The member performing the application allowed each spot to dry before applying the next and made sure the spots are small as possible. Meanwhile, another member prepared the developing chamber by placing an approximate amount of the solvent system, DCM-hexane (1:1). The inner wall of the chamber with filter paper was lined, covered with a watch glass, and was then allowed to
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