THEORY Lycopene (C40H56) and β-carotene (C40H56), two carotenoids, are found in tomato paste. Their chemical structure are shown below. Lycopene and β-carotene are separated using thin layer chromatography (TLC). A mixture of tomato paste containing dissolved lycopene and β-carotene is first spotted on a TLC plate. The technique uses a stationary phase layer, on which the substances stick, and a mobile phase, which migrates upwards the plate and carries the components of the mixture with itself (Chemguide). Depending on their respective polarity, lycopene and β-carotene stick more or less to the stationary phase and so they migrate up the TLC plate at different rates (Chemguide). Thus, as the chromatography is completed, the two components will appear as distinct dots at different heights on the plate. Lycopene has a bright orange color, whereas β-carotene is light yellow. Adsorption chromatography occurs when “one substance form[s] some sort of bonds to the surface of another one”, creating intermolecular forces between the two substances (Chemguide). For thin layer chromatography, the components of the mixture are adsorbed onto the stationary phase that covers the plate (Chemguide). The more polar a substance is, the more strongly adsorbed it is (Chemguide) and the strong intermolecular forces then result in a slower rate of migration with respect to the moving phase. As the solvent touches the TLC plate, the solute (mixture) is allowed to move up the TLC plate
The purpose of this experiment is to practice common organic laboratory techniques inside the lab to get one oriented to the basic methods of procedure that can be used for later experiments. This experiment involves the separation of benzoic acid from a more crude form, consisting of benzoic acid, methyl orange, a common acid/base indicator, and cellulose, a natural polymer of glucose (Huston, and Liu 17-24). The technique that is used to perform this separation is called extraction. Extraction is a systematic process of separating mixtures of compounds, taking advantage of the affinity differences of compounds to separate them (Padias 128-37). This technique recognizes the principle that “like dissolves in like,” that is,
Analysis: In this experiment, every substance tested positive for at least 1 organic compound. Corn oil tested positive for lipids. Honey and oats tested positive for starch. Honey, apple juice, and unknown 1 tested positive for sugar. Egg white, gelatin, and unknown 2 all tested positive for proteins. So, not a single substance didn't test positive for at least 1 organic compound. All the indicators used in this experiment change color when in the presence of a compound; therefore, they chemically react with the compound. No errors were made during this experiment
In the beginning of this experiment, our TA added water, salt, and 75/25 hexane/acetone to spinach leaves to a blender and blended the mixture to assume equal amounts for each group and to avoid erros if each student had to do the blending. The addition of the water to the mixture allowed the it to separate into a distinct organic layer after being run in a centrifuge, which was available to be collected at the top of the centrifuge. Salt reduces solubility, which will force the organic parts of the mixture (the desired pigments for example) to separate into the organic layer at the top. Lastly, 75/25 hexane/acetone is added because this is a moderately polar solvent and will useful for both the non-polar and polar pigments present within the spinach leaves. A mixed solution of hexanes and acetone must be used because acetone is very polar, while hexane in very non-polar, and the spinach leaves contain both non-polar and polar pigments in them that are important in the extraction and for further analysis. The mixture was placed in the centrifuge so the solids in the mixture (mostly cellulose) could be separated from the liquids into separate distinct layers for further extraction and testing. In the tube, the organic substances separated into the top layer, whereas the water layer remains at the bottom of the tube below the solid layer made up of mainly cellulose.
Abstract: The purpose of this lab is to separate and identify pigments and other molecules within plant cells by a process called chromatography. We will also be measuring the rate of photosynthesis in isolated chloroplasts. Beta carotene, the most abundant carotene in plants, is carried along near the solvent front because it is very soluble in the solvent being used and because it forms no hydrogen bonds with cellulose. Xanthophyll is found further from the solvent font because it is less soluble in the solvent and has been slowed down by hydrogen bonding to the cellulose. Chlorophylls contain oxygen and nitrogen and are bound more tightly to the paper than the other pigments.
Thin Layer Chromatography (TLC) works in relation to the polarity of chemicals. A plate is first covered with aluminum foil or silica etc. and has solutions of varying polarities placed upon the aluminum foil or silica. When placed in a in a puddle of solvent that moves up the plate, the different inks i.e. the solution will move up the place based on their Rf values. Adherence increases with increasing polarity, so the less polar compounds will be carried farther by the solvent. Eventually the dyes will separate into their compontnets, which can be visibly seen. This is then used to determine who the ink of the unsigned note belonged to along with the pen that it belonged
On a thin chromatography plate, five spots were placed ( as shown in table 2) and the plate was developed using chloroform/methanol. This was later visualized with dragendorff’s reagent under the UV light. All separated components were observed, identified and recorded.
During the separation of the pigments by chromatography paper, chlorophyll b traveled the shortest distance, chlorophyll a went above it, and the highest went beta carotene. This
Used of lycopene have been studied extensively through epidemiological and biochemical investigations of its properties and its bioavailability. No adverse events have been reported in association with the consumption of lycopene-containing foods (Kalai, 2011). Lycopene is synthesized by plants and microorganisms, but not by animals. It is a red open-chain unsaturated carotenoid, acyclic isomer of beta-carotene, and longer than any other carotenoid. This highly unsaturated hydrocarbon contains 11 conjugated and 2 unconjugated double bonds, predisposing lycopene to isomerization and degradation upon exposure to light, excessive heat, and air (Lee and Chen, 2002; Yang et al.,2006).
Chromatography is a separation technique in which the mixture to be separated is dissolved in a solvent and the resulting solution, often called the mobile phase, is then passed through or over another material, the stationary phase. The separation of the original mixture depends on how strongly each component is attracted to the stationary phase. Substances that are attracted strongly to the stationary phase will be retarded and not move alone with the mobile phase. Weakly attracted substances will move more rapidly with the mobile phase.
The purpose of this experiment was to take spinach leaves and extract the chlorophyll and carotenoid pigments by using acetone as the solvent. The chlorophyll and carotenoid pigments were extracted by using column chromography and alumina was used as the solvent. Solvents of different polarities were used, starting with the least polar, to extract the certain components from the leaves. They were then analyzed by using thin- layer chromatography.
Vitamin C is an organic compound that is also known as ascorbic acid. The molecular formula for Vitamin C is C6O8H6. This structure contains a 5-membered ring. Vitamin C contains 4 alcohol functional groups, an alkene, and an ester. Within the ring it contains one double bond, the carbon=carbon bond in the ring structure, this is an alkene functional group. The Vitamin C structure also contains an ester which helps form the five-membered ring.
Chromatography Investigation Chromatography is a highly regarded technique used to separate the components of a mixture. It is based on the principle that each component possesses a unique affinity for a stationary phase and a mobile phase. The components that are more inclined to enter the mobile phase will migrate further on the chromatogram and distinguish themselves from the other components. The type of solvent used in chromatography is known to directly affect the separation of the mixture. In this experiment, thin-layer and column chromatography will be utilized to separate the numerous chlorophyll and carotenoid pigments of a spinach extract.
Gel-Filtration Chromatography is a commonly used method used in order purify a protein from a mixture, by means of separations. Different biomolecules differ in size, or their molecular weight. In the gel matrix inside the chromatography column, there are gel beads which are porous to allow certain sized molecules to enter. The molecules that are able to enter the pores of the gel, are held in stationary phase and will elute from the column later on, these are usually smaller, to medium sized molecules. Larger molecules that are not able to fit in the pores will elute out of the column first, they are involved in mobile phase where they just go straight through the column without interacting with the gel beads. Smaller molecules will have a higher elution volume, while the larger molecules will have a lower elution volume. The volume to elute the protein is inversely proportional to the molecules size.
AIM : Thin-Layer Chromatography can show many different characteristics of a mixture. It is recognized for isolation , separation ,identification, and anaylsis of the mixture’s components. The purpose of this experiment is to separate carbohydrates into its pure components such as mixtures of monosacrides by TLC. TLC is used to identify sugars in normal and pancreatic disease urine, the procedure is easy and reproducible .
38 photosynthesis and prevention from photo oxidative stress [7]. Carotenoids are utilized as natural colorants and 39 value addition for juices as well as in animal feeds like poultry and fish [8]. Carotenoids applications have also 40 reached its arm in cosmetic industries [9]. The