Formal Report on Exp 5
COLUMN AND THIN LAYER CHROMATOGRAPHY
Mark Paul P. Pastrana, Mariah Ericka M. Patawaran, Princess Juneire M. Peligro,
Francisco Q. Pua III, Rose Anne L. Quyo and Janille P. Ragpa
Group 8 2B Medical Technology Organic Chemistry Laboratory
ABSTRACT
The main objectives were to separate the colored components of malunggay leaves by means of column chromatography, as well as to determine the purity of the components using thin layer chromatography (TLC) and measure the Rf values of the colored components obtained herein. For column chromatography, the sample prepared was loaded into a Pasteur pipette plugged with cotton and uniformly packed with silica gel. The eluents used were 7 mL hexane:acetone (7:3), 5 mL hexane:acetone (1:1), 5 mL acetone, and 5 mL acetone:MeOH (1:1); the collected eluates were dark green, light green, yellow green, and yellow. For thin layer chromatography, the eluates were applied in spots on a TLC plate pre-coated with silica and placed in a developing chamber lined with filter paper and hexane:acetone (7:3) as the solvent system. The solvent front was measured to be 8.2 cm. The TLC plate was then put under a UV lamp and the Rf values were computed as follows: Dark green—0.5, Light Green—0.49, Yellow Green—0.46, Yellow—0.99, Pink—0.32, and Bluish-violet—0.20.
INTRODUCTION Chromatography is a technique used to separate mixtures into components as a result of differential partitioning behavior between a stationary phase and a mobile phase that percolates through the stationary bed. [1, 2] Column and thin layer chromatography are examples of the solid-liquid method of chromatography, which utilizes an adsorbent solid stationary phase (silica) and a liquid mobile phase. In column chromatography, the solid stationary phase is situated within a tube as the liquid mobile phase is added to the top and allowed to flow down through the tube [3]; in thin layer chromatography, the stationary phase is pre-coated on a plate. Column
Mark Paul P. Pastrana, Mariah Ericka M. Patawaran, Princess Juneire M. Peligro,
Francisco Q. Pua III, Rose Anne L. Quyo and Janille P. Ragpa
Group 8 2B Medical Technology Organic Chemistry Laboratory
ABSTRACT
The main objectives were to separate the colored components of malunggay leaves by means of column chromatography, as well as to determine the purity of the components using thin layer chromatography (TLC) and measure the Rf values of the colored components obtained herein. For column chromatography, the sample prepared was loaded into a Pasteur pipette plugged with cotton and uniformly packed with silica gel. The eluents used were 7 mL hexane:acetone (7:3), 5 mL hexane:acetone (1:1), 5 mL acetone, and 5 mL acetone:MeOH (1:1); the collected eluates were dark green, light green, yellow green, and yellow. For thin layer chromatography, the eluates were applied in spots on a TLC plate pre-coated with silica and placed in a developing chamber lined with filter paper and hexane:acetone (7:3) as the solvent system. The solvent front was measured to be 8.2 cm. The TLC plate was then put under a UV lamp and the Rf values were computed as follows: Dark green—0.5, Light Green—0.49, Yellow Green—0.46, Yellow—0.99, Pink—0.32, and Bluish-violet—0.20.
INTRODUCTION Chromatography is a technique used to separate mixtures into components as a result of differential partitioning behavior between a stationary phase and a mobile phase that percolates through the stationary bed. [1, 2] Column and thin layer chromatography are examples of the solid-liquid method of chromatography, which utilizes an adsorbent solid stationary phase (silica) and a liquid mobile phase. In column chromatography, the solid stationary phase is situated within a tube as the liquid mobile phase is added to the top and allowed to flow down through the tube [3]; in thin layer chromatography, the stationary phase is pre-coated on a plate. Column
References: [1] Brian M. Tissue. Chromatography. http://www.files.chem.vt.edu/chem-ed/sep/chromato.html. Retrieved 8/11/13 [2] Rebecca Carrier and Julie Bordonaro. Intro to Biochemical Engineering http://www.rpi.edu/dept/chem-eng/Biotech-Environ/CHROMO/chromintro.html.Retrieved 8/11/13 [3] University of Colorado at Boulder, Department of Chemistry and Biochemistryhttp://orgchem.colorado.edu/Technique/Procedures/Columnchrom/Columnchrom.html [4] Bayquen, A., Cruz C., de Guia, R., Lampa, F., Peña, G., Sarile, A. and Torres, P. (2009). Laboratory Manual in Organic Chemistry. Manila: C & E Publishing, Inc. [5] University of Colorado at Boulder. Thin Layer Chromatographyhttp://www.ce.gxnu.edu.cn/organic/net_course/content/tlc/Retention_Factor.htm [6] Medical Health Guide. What is Malunggay.http://www.medicalhealthguide.com/articles/malunggay.htm