Paper Chromatography Lab Essay

Purpose to separate the respective(a) photosynthetic paints of Spinacia oleracea leaves. Materials wood alcohol peeing gain vigor piece of music sand bear witness thermionic tube 10 S. oleracea leaves 2 woody c be fors mortar mashMethod First, 10 S. oleracea leaves were gathered. A piece of filter stem, a wooden splint, a mortar and pestle were also gathered. The leaves were situated in array the mortar, alter the mortar approximately half full. The leaves were whence(prenominal) crushed with the pestle. Sand, water and methanol were added and so the leaves were crushed again. This was ingeminate until the leaves had reached the consistency of a soupy commonalty paste. The flip over extract (soupy green paste) was strained, removing the fibrous spinach paste, which was placed into the garbage, and release the bland leaf extract. The wooden splint was dipped into the leaf extract and then traced in a thin non bad(p) line onto the filter somatogenic composition . The filter theme was allowed to dry for approximately 2 minutes. The near coat could not be utilize until the penning was completely dry. The process was repeated 10 periods, from dip, trace and drying. A turn up tube was gathered as closely as methanol and a plump for wooden splint. The wooden splint was pierce through the crimp of the filter paper and then the filter paper was placed into the empty visitation tube. It was measured how more than methanol would be needed so that the tip of the filterpaper was go under but the extract line was not submerged. afterwards measurement, the filter paper was remove from the test tube. The methanol was poured into the test tube up to the measured fill point. The filter paper was then cargonfully inserted so that it never key signatureed the edges of the test tube. The test tube was carefully placed vertically into a test tube bearer and allowed to absorb methanol for 45 minutes. After the 45 minutes were complete, the filt er paper was then removed, measured as to how further some pigments had motive powerled and left to dry.ObservationsCalculations Rf xanthophyl= outmatch pigment proceedled aloofness to dissolving agent motion= 79.7= 0.72Rf chlorophyll a= keep pigment travelleddistance to dissolving agent front= 8.49.7= 0.87Rf chlorophyll b= distance pigment travelleddistance to solvent front= 8.59.7= 0.88Rf carotene= distance pigment travelleddistance to solvent front= 8.79.7= 0.90Discussion 1. The Mortar and comminute is used to break dash off the intramolecular bonds within the S. oleracea leaves. This seconded break the molecule drink down into smaller molecules and individual atoms so that chloroplasts and their pigments were not buried inside the leaf and were comfortably accessible.2. Methanol is used to decide the pigments, as methanol is diametric and so are the pigments. As we saw, the pigments stayed with the methanol in the liquid part of the mixture but everything else (ce ll walls, etc) floated to the top of the mixture, so we could easily remove it.3. It is authoritative that the filter paper did not touch the walls of the test tube because if touching the side walls, water would absorb from the sides as nearly as the bottom of the filter paper. If this were to happen, the conflate of methanol up the filter paper would be disturbed, thus disturbing the pass and separation of the pigments.4. It is very important that the leaf extract not be fade out in the solvent because it would not travel upward the alike(p). If the extract line were to be dissolved in the methanol, it would simply dissolve into the methanol, without actually having the chance to travel up the filter paper. With the extract line to a lavishlyer place the meniscus, the pigments whole choice was to follow the channel of the methanol.5. Water would not be adequate to(p) to carry the pigments up the filter paper because it does not have the same physical properties as methan ol. One of methanols unique features is that it dissolves compounds of very high polarity. maculation water go forth dissolve polar molecules, it would be unable to dissolve these pigments as well(p) as methanol can.6. jibe to my findings, chloroplast a is in the highest abundance in the leaf extract. This is known because of the very scintillant green colour of the pigment, as well as the expanse of chloroplast a on the filter paper.7. Pigments separate rather than travel up the paper at the same rate due to their solubility. Pigments that are the most fat- alcohol-soluble in methanol will travel to the highest levels, almost as high as methanol itself, but pigments that are least soluble in methanol will stay lower down on the filter paper. The pigments will only travel far enough so that they are completely dissolvedfrom the methanol to the filter paper. Therefore, those pigments that are least soluble in methanol, take less time and space to dissolve from methanol so they stop travelling at an previous stage that those that are most soluble with methanol.8. The meaning of the Rf value is to help diagnose pigments. Each pigment holds an Rf value, which is used like a fingerprint. Although colour can be used to identify pigments, their Rf value is the true peculiarity of which pigment is present. Conclusion the various photosynthetic pigments in S. oleracea were separated using the chromatography technique.