If the pigments were found in the storma rather than in the thylakoid membrane how would this affect their movement on the tlc plate ? Background and Procedure:Photosynthetic plants utilize a variety of light-absorbing pigments. Due to the large quantity and intensely colorfulnature of the chlorophylls (green), under many circumstances it is not possible to notice the presence of the otherpigments.Differences in chemical structure of these pigments result in differing amounts of polarity, allowing us to separatethem from one another using Thin Layer Chromatography (TLC). Most of these pigments are found inmembranes of the thylakoid and are predominantly hydrophobic. Therefore, we will use organic solvents, such asisopropyl alcohol ("rubbing alcohol"), as the solvent and petroleum ethers for the mobile phase or "developingsolution" for the chromatography.1. Weigh approximately 3-4 grams of fresh, dry spinach leaves with the stems completely removed.2. Using a pair of scissors cut the leaves into very small pieces into the mortar. Using the pestle grind andpound the leaves into a mash. This will take several minutes and is an essential step, so don't rush it.3. Add 10 mL of anhydrous isopropyl alcohol to the mortar and carefully, without splashing, continue to mixand crush the slurry for several more minutes. If some of the alcohol has evaporated you can add acouple more milliliters to maintain the original volume. Then place plastic wrap, a beaker, or other objectover the top of the mortar to prevent evaporation of the alcohol. Wait five minutes.4. Stir and crush the mixture for another thirty seconds. The liquid should be a dark green color. Use atransfer pipet to move the isopropyl-pigment solution into a glass test tube. Try to avoid the spinachsolids. Keep in mind that this tube of pigment/alcohol solution will also be used for Activity #3.5. Carefully cut the TLC plate lengthwise, to create two, more narrow strips. One half of the plate will beused per group. Using a ruler draw a thin line 1.5 cm from one end (this will be the bottom) using astandard pencil.6. Using the glass capillary tube aspirate a small amount of the isopropyl solvent containing the leafpigments. Carefully make a small dot on the center of the pencil line on the TLC plate. Allowapproximately 1 minute to dry.7. Repeat step 6 at least five times. You want a very dark and concentrated spot of pigments at one placeon the thin pencil line. Be certain the spot is dry before continuing to the next step.8. In a clean glass test tube add 0.75 mL of chromatography developing solution. Caution: this is a mixtureof petroleum ethers. This solvent is highly flammable and mildly toxic. Be sure to dispose of this mixtureproperly (as outlined by your instructor) and take care with handling.9. Carefully place your TLC strip into the test tube containing the developing solution. The dot containing theofpigments should NOT come in direct contact with the solution. Using a piece of parafilm seal the topthe test tube and place in a test tube rack.10. Observe the TLC plate for the next few minutes to watch the developing solution front move up the platewhile the pigments separate.11. When the developing solution is within 1 cm of the top of the plate remove it from the tube with a pair offorceps and place it on a paper towel to dry. Properly dispose of the remaining developing solution in theorganic waste receptacle and dispose of your test tube in the glass waste.Documentation and Retardation Factor (R):Since the developing solution is extremely nonpolar, more hydrophobic pigments will travel easier with the solventfront nearer the top. The more polar (less hydrophobic) pigments will stick to the silica further down on the plate.4 of 10 updated 7/19yellowgreendarkenlighterFinisyellowravelsLightpeend name of pigment

Introduction :- Plants performs the process of photosynthesis with the use of photosynthetic pigments ( mainly the Chlorophyll pigments) . Chloroplast is the cell organelle which consist of thylakoids and stroma , and different photosynthetic pigments are present here . Chromatography is a bio-physical technique which is used to separate the different pigments from one another . TLC ( Thin layer Chromatography) is a type of Chromatography , which is advantageous to the paper chromatography because of less time taken in the separation of pigments and also due to clear separation .Answer :- If the pigments were found in the stroma rather than in the thylakoid membrane then there would reversal of the effect , on the movement of pigments on the tlc ( thin layer Chromatographic) plate takes place . And it will lead to the appearance of dark colour ( Dark yellow , dark light and dark green) . Because Chlorophyll pigments are present in the thylakoid and absorbs differ wavelength of light and gives the green colour . But when it is present in the stroma , then we know that other pigments are also present in the stroma . So , in this case , firstly Chlorophyll will show the colour and then the other pigments . So, the effect will gets reversed due to changing the location of pigments. And the main reason behind this is that different pigments absorbs light at different wavelengths.Explanation :- Chromatography is the physical method for separation of compounds . In its first use as an experiment , this technique was used to separate the various plant pigments such as Chlorophyll and xanthophyll by passing solution of these compounds through a glass column , packed with finely divided calcium carbonate , by Russian botanist Tswet. Chromatography is based on the principle of mobile and stationary phase. The sample that we want to examine is allowed to interact with them . Thin layer Chromatography is a type of Chromatography and the biophysical technique , used to separate the molecules , compounds .