The extraction procedure isolates the pigments in spinach that will be used in the TLC analysis by leaving the insoluble properties behind. Each step serves a purpose in doing so, for example, the spinach is blended into a puree in order facilitate the centrifuge process. 75/25 hexane/acetone solution is used in the blending process because hexane alone is not enough and acetone alone is miscible in water. Anhydrous sodium sulfate is used to dry the organic solution after it has been through the centrifuge tube and separated into a test tube. The purpose of the alumina column is to filter unwanted inorganic chemicals while letting the desired organic chemicals to pass through. The pigments tested in this experiment are carotenes, pheophytin a and b, …show more content…
Carotene is a yellowish/orange pigment that comes in two forms, a and b, which differ in their double bond position in the cyclohexene rings. The remaining carotene consists of methyl groups and single and double bonds, it is the least polar out of all of the pigments tested. Carotene resulted with an Rf factor of 0.77 cm, the highest out of the pigments tested. Chlorophyll is the green pigment which also consist of two structures, a and b, it is found in the plants chloroplasts. Both chlorophyll structures have a porphyrin ring the difference is that chlorophyll a has a (-CH3) bond while chlorophyll b has a (-CH=0) bond, making b more polar. Chlorophyll was the second least polar pigment tested, its Rf factor resulted at 0.18 cm, significantly more polar than carotene. Pheophytin is a grey pigment which also has two structures, a and b. Pheophytin a also has a
The objective of this experiment was to perform extraction. This is a separation and purification technique, based on different solubility of compounds in immiscible solvent mixtures. Extraction is conducted by shaking the solution with the solvent, until two layers are formed. One layer can then be separated from the other. If the separation does not happen in one try, multiple attempts may be needed.
The experiment was conducted using carbon dioxide to see how it affected the rate of photosynthesis in spinach leaves. Carbon dioxide should increase the rate of photosynthesis because there will be more carbon dioxide, a reactant in the photosynthesis formula.
Experiment #3: The purpose of this experiment to test the chromatography of plant pigments the alcohol test strip test will be used.
The pigment line of the sample leaf was extracted by repeatedly rolling a coin along a ruler edge that held the leaf 1.5-2cm from the bottom of Whatman #1 chromatography paper. Subsequently, a saturated environment was created to ensure that the solvent was separated by placing the beaker containing the rolled Whatman paper with the sample line on the outside into a mason jar containing the separation solvent, and sealing both compartments.
Summary Paragraphs: In three dishes, paper was placed in the bottom to absorb the moisture. Twenty-five spinach seeds were added to each using tweezers. Two milliliters of different solutions were then added to each. The solutions were deionized water, 1 mL water and 1 mL vinegar, and 1 mL water and 1 mL Borax. The water was used to form a neutral environment, the vinegar to form an acidic environment, and the Borax to form a basic environment. The petri dishes were then labeled, taped shut, and placed on the windowsill for seven days.
Paper chromatography is a useful technique for separating and identifying pigments and other molecules from cell extracts that contain a mixture of molecules. As solvent moves up the paper, it carries along any substances dissolved in it. The more soluble, the further it travels and vice versa. The purpose of this experiment was to separate plants pigments.
The test is to see if spinach leaves discs can be effected by photosynthesis. I predicted that more disc would float in the light than in the dark. After writing down my hypothesis, I put the discs in water on a petri dish and waited 15 minutes. I then recorded if the in the light were floating or not. I also found out that the disc will also change color when they are in the light or in the dark. The hypothesis is supported when the light does have more disc floating in the petri dish than the dark did. I also determined other things as well such as the the dependent and independent variables, and the control treatments. The independent variable is the amount of oxygen in the discs. The dependent variable is the light, darkness, and the amount
Materials used in the experiment included 5-7 g of the potato tissue, 50ml of 2.0M phosphate buffer coffee filter and guaiacol dye.
Human pigmentation is influenced by hemoglobins within blood vessels in the skin, carotene and melanins. Melanin, the basis of pigmentation, can be found in the forms of eumelanin and phaeomelanin. Eumelanin is the brown-black pigment located in the skin, hair, and eyes. Phaeomelanin is a yellow to reddish-brown pigment found in small quantities within the skin, eyes, and red hair. Because of these two pigments, to a greater or lesser degree, we have the variation in human pigmentation that is seen today.
Starting this experiment, we knew that the extraction was going to form varies layers due to the density differences. When placing three different substances, we saw that two layers formed because the Clove Oil is soluble in MTBE, but not in water. In order, to get the organic layer we used separatory funnel to take out the excess substances and leave the oil layer. Then we transferred to a beaker and dried with Magnesium Sulfate. Lastly, we filtered the liquid using funnel; we placed the liquid to boil, let it cool to room temperature. The purpose of drying and evaporation is to help us with the Gas Chromatography analysis of the product.
The structure of chlorophyll involves a hydrophobic tail embedded in the thylakoid membrane which repels water and a porphyrin ring which is a ring of four pyrrols (C4H5N) surrounding a metal ion which absorbs the incoming light energy, in the case of chlorophyll the metal ion is magnesium (Mg2+.) The electrons within the porphyrin ring are delocalised so the molecule has the potential to easily and quickly lose and gain electrons making the structure of chlorophyll ideal for photosynthesis. Chlorophyll is the most abundant photosynthetic pigment, absorbing red and blue wavelengths and reflecting green wavelengths, meaning plants containing chlorophyll appear green. There are many types of chlorophyll, including chlorophyll a, b, c1, c2, d and f. Chlorophyll a is present in all photosynthetic organisms and is the most common pigment with the molecular formula C55H72MgN4O5. Chlorophyll b is found in plants with the molecular formula C55H70MgN4O6, it is less abundant than chlorophyll a. Chlorophyll a and b are often found together as they increase the wavelengths of light absorbed. Chlorophyll c1 (C35H30O5N4Mg) and c2 (C35H28O5N4Mg) are found in algae, they are accessory pigments and have a brown colour. Chlorophyll c is able to absorb yellow and green light (500-600nm) that chlorophyll a
HPLC technology works on the principle of conventional chromatography where in there is a stationary phase and a mobile phase. The sample containing the mixture of components is introduced in a column packed
Materials and Methods: An ion exchange chromatography column was obtained and set up for purification with the addition of 0.5 ml ion exchange matrix. 1 ml
Hue is the common name for the colours in the spectrum which are red, orange, yellow, green, blue, and violet. A pigment is a colouring ag...
Analyze each fraction by spotting 10 times with capillary tubes on a TLC plate, which is exposed to iodine vapor for 15 minutes.