Thin Layer Chromatography Introduction Thin Layer Chromatography or TLC is a technique used as a separation and identification technique. There are many forms of chromatography‚ but one thing that remains constant throughout all of the types of chromatography is that there is a stationary phase and a mobile phase. In the case of TLC the stationary phase is the silica gel on the TLC tray. Procedure Chromatograph method is a method of separating mixtures of two or more compounds. Two phases
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Figure 1: Affinity chromatography of fumarase with the Ni2+-NTA-agarose column. Extract (9.9 mL) containing yeast (3.76g) in extraction buffer containing 0.1% Igapel CA-630 and protease inhibitors were pumped through Ni2+-NTA-agarose column. Fractions were collected by 1.5 mL portions by use of wash buffer (20.0 mL)‚ imidazole elution buffer (26.3 mL)‚ and wash buffer (10.0 mL)‚ again. Absorption readings were taken for all fractions with a Cary50 set at 280nm. The fumarase activity was determined
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Abstract By separating chemicals out of a mixture and purifying them through sublimation and recrystiliation‚ changes in the substances physical properties can be shown. A salicylic acid-naphthalene mixture was mixed together with diethyl ether and saturated sodium bicarbonate. Two layers separated and were extracted after acidifying with HCl. This precipitated the crude salicylic acid. The left over mixture was combined with water and the two layers separated where crude naphthalene was recovered
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Thin Layer Chromatography of the Unknown Analgesic Jessica Bajao*‚ Phoebe Abalos‚ Kevin Antiga‚ Carmelus Aseneta 3-Biology 2 College of Science‚ University of Santo Tomas‚ Manila‚ Philippines Abstract The group used five different analgesics in this experiment: aspirin‚ acetaminophen‚ ibuprofen‚ caffeine‚ and mefenamic acid. Six analgesics were spotted on the TLC plate including the unknown. After the development of the TLC plate‚ it was placed under ultraviolet light for the spots resulted
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Affinity chromatography technique is used to separate proteins found in a mixture of solution. Affinity chromatography uses the strong interaction between a given protein and its corresponding molecule. In today’s lab‚ affinity chromatography was used to purify L-lactate dehydrogenase‚ which contains histidine-tagged protein. The histidine- tagged protein forms a strong interaction with the Ni-NTA column due to the presence of nickel ions. Varying concentration of imidazole was added to the column
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Separation and Identification of Plant Pigments by Paper Chromatography Biology 1107L Introduction All living organisms require energy for their chemical processes. The ultimate source of this energy is the sun. Plants convert light energy into the chemical energy of sugars. During photosynthesis pigments are used to capture light energy. Pigments of green plants can easily be separated and identified using a technique called paper chromatography. The purpose of this
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understand that because substances have different properties‚ such as mass‚ that can be separated by chromatography. In our experiment‚ we chose 4 different color source to separate in water. We made sure that all the sources were water soluble because only polar substances will dissolve in water. We chose a black wet-erase marker‚ a red marker‚ a green marker‚ and mixed food coloring to test. The chromatography paper was split into 4 sections‚ about 2 cm above the bottom edge and evenly spaced out. Each
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Separation techniques LIQUID CHROMATOGRAPHY ‘THE ART OF SEPARATION’ CHROMATOGRAPHY – AN INTRODUCTION Chromatography is a technique through which a mixture of chemical components are separated‚ identified and determined accurately. This technique while provides a way for analytical separations‚ also useful for preparative techniques by which pure compounds can be obtained. Detector Signal Blue Compound Sample Injection + Mobile Phase Retention Time Red Compound It is i defined d fi d as a
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Column and Thin Layer Chromatography Beverly Abstract: Plant pigments were separated and concentrated from a crude spinach extract through the use of column chromatography and an eluatropic series of hexanes‚ hexane/acetone‚ and methanol. The pigments were analyzed using thin layer chromatography with a 30% ethyl acetate/hexane developing solvent. Introduction: Chromatography is a technique used to separate a mixture of two or more components based on
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Purification of Recombinant Green Fluorescent Protein (rGFP) From E. coli strain‚ BL21(DE3)‚ Using Ni2+-Agarose Affinity Chromatography Abstract: The purpose of these series of experiments was to express and purify recombinant Green Fluorescent Protein (rGFP) from the E. coli strain‚ BL21(DE3) by beginning with its purification via a Ni2+-agarose affinity chromatography column. The His6 tag of the rGFP bound to the Ni2+-agarose column and washes and elutions were obtained‚ with elution 3 containing
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