Lab Report Column Chromatography

Multiple extractions with smaller volumes are more efficient than a single extraction at a one large volume. When an organic solvent is used to extract a compound from the aqueous solvent, smaller volumes will result in a better extraction. The success upon the collection of the crude material is depended on how well the water is absorbed by the anhydrous drying material. The presence of the drying material correlates with the vapor pressure of the other compound. When the vapor pressure is low, there is a smaller amount of moisture in the gas produced. Once the anhydrous material is added and clumping was avoided, the evaporation of the added organic substance can proceed. The final material collected can be physically identified by the final color of the precipitate. A greenish- white precipitate is most likely identified to be pure, and a brownish color indicates that the collected material is wet, and not

Chromatography is used to separate mixture of chemicals to figure out what compound are present in the chemical, the chemical that are suitable to separate is by chromatography that include inks, dyes and colouring agents in foods. The experiment is carried out on paper where you put a spot of the chemical near the bottom of a chromatography piece of paper and the paper is in a solvent which is usually water. When the solvent (water) soaks up the paper it carries the chemical up with it. Different components of mixtures will move at a different rate as other, some would move faster or slower than others which separates the mixtures from each other. They all have a stationary phase that is a solid or a liquid supported on a solid. A mobile phase is a liquid or a gas.

Paper chromatography is used to separate mixtures of substances into their components. There are different types of chromatography but they are all based on the same principal. Paper chromatography is an analytical method that is used to separate colored chemicals or substances, especially pigments. They all have a stationary phase and a mobile phase. The moving substance is called the mobile phase while the stationary phase stays put. The mobile phase flows through the stationary phase and carries the components of the mixture with it. The stationary phase is motionless and is the actual medium that performs the separation. Ninhydren reacts with amino acids to give colored compounds and detect the location of the amino acids. This is used because amino acids are colorless. Different components travel at different rates. Each one undergoes adsorption in a slightly different way and spends more or less time in either the solid or the liquid phase. Components of the samples will separate readily according to how strongly they absorb on the stationary phase vs. how readily they dissolve in the mobile phase.

A liquid-liquid extraction is used when separating two different liquids that are immiscible. Often the two different liquids will be soluble in water and a solution that is organic. These two types of liquids will create two distinct layers of isolated solution. This is caused because one of the liquids will dissolve into the water and the other liquid will dissolve into the organic solution and since water is a polar molecule and the organic solution is a non-polar molecule the two layers will not mix at all. Therefore, creating two distinct layers that can then successfully be separated. In this experiment, the top layer of solution in the separatory funnel consists of the ether solution layer containing the unknown neutral compound and

Chromatography can be defined as a series of steps used to identify, analyze and separate compound. It is a method used to obtain components from a non-volatile mixture (Preethi, Harita & Rajesh, 2017). There are various types of chromatography separations methods. These include: column, gas, supercritical fluid chromatography and Thin layer chromatography. All these separation techniques operate under the same procedure;

Column one was a 30m x 0.25mm-i.d coated with 0.5 μm 5% diphenyl and 95% dimethyl polysiloxane (Rtx®-5). Column two was a 30m x 0.25mm-i.d coated with 0.5 μm midpolarity phase consisting of 50% phenyl and 50% dimethyl polysiloxane (Rxi®-17sil). Column three was a 30m x 0.25mm-i.d, capillary coating with 0.5 μm film of 100% trifluoropropyl methyl polysiloxane (Rtx®-200). The separation was performed using the same temperature program on all these stationary phases. The temperature program used for separation was consisted of an initial temperature hold at 80 °C for 1.0 min, ramped up to 300 °C at a rate of 30 °C/min, held at 300 °C for 0.5 min then ramped to 340°C at a rate of 5.0 °C/min and held at 340 °C for 5.0 min with a total run 21 min. (TP1). Column four was a 30m x 0.25mm-i.d, capillary coated with 0.5 μm film of midpolarity phase consisting of 35% phenyl and

Methods used to separate miscible solvents include thin layer chromatography (TLC) as well as column chromatography. A method used to separate a mixture of miscible solvents is column chromatography, which is used to purify and separate compounds. The particular speed of the solvents used in the experiment depended upon the properties, being polar or nonpolar, as well as the properties of the prepared column (INSERT SOURCE HERE). With column chromatography, there are two phases that include mobile and stationary phases. In the experiment performed, the glass column used was first packed with a piece of cotton followed by a layer of sand in order to keep the silica gel in place and also to prevent the gel from flowing out of the tube when the particular solvent was added. Based upon whether the

In this lab, paper chromatography will be used to separate the components of known and unknown mixtures and then used to identify those components. Paper chromatography is a technique where a drop of solution containing a mixture is placed on a piece of filter paper. One end of the filter paper is then placed into a liquid solvent. The mixture will separate into its different components as the solvent moves up the filter paper. The filter paper is known as the stationary phase. The solvent is known as the mobile phase because it uses capillary action to move up the paper. The mixture will be separated into its different components because each component will have a unique chemical affinity for the paper and for the solvent. Theses affinities

The solvents that were used in the column chromatography were hexanes, 75/25 hexanes/acetone solvent, and acetone. The adsorbent used in the column chromatography was alumina, which is polar. All of the pigments moved through the column based on their polarity. Hexanes were used because they are nonpolar due to their hydrocarbon chain and was used as the initial solvent before the extract was placed in the Pasteur pipette. Carotene, which is nonpolar hydrocarbon chain, was the first pigment extracted. The nonpolar solvent was used because it is soluble in the nonpolar carotene, thus moving it through the column quickly. To elute the next pigment of chlorophyll the solvent was the hexane acetone mix. This mixture is slightly more polar because

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Afterward, PTEN purified using affinity chromatography. Since PTEN fused with His6 tag, Ni-NTA affinity chromatography would work to purify PTEN. Ni-NTA is a type of immobilized metal affinity chromatography that contain coupled Ni2+ nickel chelate forming ligand (imiondiacetic acid) that immobilized to agarose. The science behind using Ni-NTA affinity chromatography is that histidine bind specifically and reversibly to the nickel chalet thus washing step will remove other contaminate and then His-PTEN eluted using imidazole. Bradford assay and SDS-PAGE performed to quantify and analyze the progress toward PTEN purification. The calculated concentration of the samples presented in table 8. The Concentration of samples make since having the

Background information: Four categories of contaminants have been identified by environmental scientists and have said to be existent in municipal water supplies. These pathogens can cause disease that can lead to cancer and acute poisoning. Public health officials have taken note of the issue and are now attempting to take action to counter the hazard.

The function of Columnar epithelial tissue is to surround glands and ducts, which then help in absorption, secretion of mucus, enzymes, and other substances. The columnar epithelial tissue is made of large epithelial cells that are specialized for absorption, and are commonly found in areas where there is a lot of wear and tear, such as in the digestive tract. The larger shape of the columnar cell allows this tissue to perform its function in two ways. First, it increases the number of organelles to synthesize needed material, and second, it increases surface area. Simple columnar tissue commonly surrounds glands and ducts, therefore the increased amount of organelles aid in the production of material that is secreted. Columnar epithelial

Chromatography is the term used to describe all the separation methods based on the distribution of compounds between two separate phases. TLC, an abbreviation for thin layer chromatography, is a chromatographic technique which is used to separate components of a mixture using a thin stationary which has an inert backing, this occurs as one phase is fixed on a plate (stationary phase) and the other phase is mobile and migrates through the stationary phase (mobile phase). During the chromatographic development process, the mixture to be separated is distributed between the stationary and mobile phases. (Spangenberg Bernd et al 2011). It is mostly used because of its simplicity, relatively low cost, high sensitivity and relatively fast speed of separation. TLC also functions on the same principle as all other chromatography techniques. A compound has different affinities for the mobile and stationary phases, and this affects the speed at which it migrates. The goal of TLC is to obtain well defined, well separated spots. (O. H. Lowry, 1951). In contrast to HPLC, many samples can be run at the same time on TLC. (Journal of Chromatography, vol 19 (2011)).

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