Lab Report On Chromatography And Lab

In this chromatography lab filter paper was used to examine the different pigments of spinach and red cabbage. There were two hypotheses for both plants. The hypothesis for the red cabbage was that there would be multiple pigments evident. The hypothesis for the spinach was similar in that there would be different pigments present. Both hypotheses were accepted in that there were multiple pigments found on the filter paper of each sample.

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.

In this lab, a few pieces of spinach were grinded to extract all of the pigment molecules from the leaves. The extract was then put through a piece of filter paper to filter out the large bits of spinach. The extract was then put in a microcentrifuge to help separate the pigment molecules from any larger pieces left in the extract. The extract was placed on a single spot on a piece of filter paper. The filter paper was then placed into a chromatography solvent to separate the pigment molecules further. The distance that the pigment molecules went were measured, and then the Rf for each were found. It was found that the leaves of spinach used contained the following pigments; xanthophyll 2, chlorophyll a, and beta-carotene. The pigments xanthophyll 1 and chlorophyll b were not found in this experiment, most likely due to an error in the technique or procedure. Furthermore, the hypothesis was found to be partially accurate due to the fact that chlorophyll a, xanthophyll 2, and beta-carotene were found, but chlorophyll b and xanthophyll 1 were not found. The Rf found experimentally for xanthophyll 2 was 0.15, and the actual Rf is 0.15, which means that there was a zero percent error. The Rf found experimentally for chlorophyll a was 0.52, but the actual Rf is 0.59, which indicates there is twelve percent error. Finally, the Rf found experimentally for beta-carotene was 0.84, but the actual Rf is 0.98, which indicates that there was a fourteen percent error. The Rf

Column chromatography consists of solid-liquid phase partitioning. Several steps are involved in order to perform a separation using this technique. The first step is selecting a column. The size of the sample which will be separated or purified will indicate what size the column should be. The difference in Rf values is also a

Meanwhile, the spinach leaves were being crushed in the crucible into a more liquefied state. Once the spinach leaves were done, a pipette was used to place the spinach extract along the ling drawn around the cylinder. This step was repeated six time, because the darker the line the better the results of the chromatographic separation. After each application of the spinach extract, it was allowed to dry for at least one minute.<p>

Goal: The first goal of today’s laboratory is to separate components of spinach dyes using different eluants. The four eluants will be using are ethanol, chloroform, 9:1 petroleum ether : ethanol, and petroleum ether. The second goal of the lab is to separate fluorine and fluorenone by column chromatography. Thin layer chromatography (TLC) was used to measure the polarity and separate the components in the mixtures. TLC was chosen because of its simplicity, high sensitivity, and speedy separation. For each part of the lab, we measured the retention factor on the TLC plate. To measure the retention factor, we used the formula:

This experiment is concerned with identifying photosynthetic pigments found in spinach plants and determining the spectrum of light each absorbs. The questions being posed are which pigment is the most active in absorbing energy and which wavelengths of light it absorbs. By using paper chromatography, different pigments of spinach leaves can be separated. Based on the results, it can be concluded that chlorophyll is the most active pigment and photosynthetic pigments in plants absorb light across the entire visible spectrum. The knowledge gained in this experiment is relevant to understanding how the process of photosynthesis works. A real-world application for this includes the harvesting of clean energy sources, as scientific advances have

Thin layer chromatography can be used as a physical method to segregate compounds from natural sources. E.g. Spinach leaves are visibly green, but consist of a variety of components that have more colour than others. This experimental procedure uses compounds from spinach leaves that are exposed to chromatography, TLC plate to indicate the different pigments

It can be concluded that column chromatography is an effective way of separating components of a mixture. The crude pigment was sent through the column however three distinctly separate layers were collected (yellow, green-blue and blue). Each coloured layer represented a different pigment. It was made clear by the column chromatography that the beta-carotene (yellow) is the least polar pigment, chlorophyll b (green) is the most polar and chlorophyll a (blue-green) is intermediately polar based on the order the pigments were eluted. In the column, the least polar pigment eluted first as it adhered best to the 8:2 pet ether: acetone mobile phase. The last pigment that passed through was chlorophyll b as it was the most polar and adhered greatly to the silica. The effectiveness of column chromatography was confirmed using thin layer chromatography. On the silica plate, a spot was added for each separated pigment and the crude pigment. A highly effective separation using column chromatography would show corresponding distances travelled for the three separate colours and the three spots the crude separated into on the TLC plate. This confirmed that the column separated the pigments into the correct three layers. On the TLC

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.

After analyzing the results of the paper chromatography, the next step was the determination of the absorption spectrum for the spinach leaf pigments chlorophyll a, chlorophyll b, xanthophyll, and carotenoids. The amount of absorbance was determined with the help of the spectrophotometry, which quantitatively measured the fraction of the light passing through each pigment extract solution, indicated on the absorbance scale. The wavelength was increased every 20-nm, starting with 400-nm, and reaching 740-nm. The results are shown in Table 1 and Table

Objective: The purpose of the experiment is to extract pigments from spinach leaves and separate them by column and thin layer chromatography, determining Rf values for the pigments.

During the column chromatography, frozen spinach was bought from a local grocery store and extracted into acetone to create the “stock spinach extract”. Next, roughly 80mL of the stock spinach extract was placed on a steam bath to remove the acetone. After the acetone is removed, the extract was re-dissolved in about 8mL of 9:1 hexane/acetone.

There are two major types of pigments that are involved in photosynthesis for plants: carotenoids and chlorophylls. Under carotenoids, there are pigments called α and β-carotene and xanthophylls. Chlorophylls include chlorophyll a and chlorophyll b (Pavia et al 208). The main objective of this experiment was to extract these pigments from spinach leaves using chromatography techniques. First, the pigment was extracted from the leaves by column chromatography. However, the column was packed with granular anhydrous sodium sulfate. This first extraction acted as E or the main extract of the pigments.

Chromatography, which was originally discovered and developed by Mikhail Semenovich Tswett in the early 1900s, is a laboratory technique used to separate and distinguish between components of a mixture. The various components of a sample mixture are known as analytes. Chromatography was originally used to separate the various molecules that make up plant pigmentation. Because these components, carotenes, chlorophyll, and xanthophylls, are different colors, the separation of them is responsible for the name of this laboratory technique. Throughout the early to mid-1900s, different styles of chromatography were developed and allowed this technique to be applicable to a wide variety of analyses and separation processes. Chromatography can be applied to analytical process, which aim to examine the presence and relative concentrations of various analytes within a sample. The goal of preparative chromatography is to separate analytes within a mixture for isolation and purification purposes. Chromatography was originally developed by Tswett for the purpose of isolation of particular compounds (10). Moreover, more recent advancements in chromatography are facilitating the separation and distinction of analytes that are extremely comparable to one another and increasing the resolution of the technique.