Isolation of Chlorophyll and Beta-Carotene from Plant Leaves
Yokoyama Yuri B1 (A0142349E)
1. Aim:
a) To separate beta-carotene and chlorophyll via column chromatography.
b) To analyse the contents of the plant extract as well as the separated beta-carotene and chlorophyll via thin layer chromatography (TLC)
c) To analyse the absorption peaks of both beta-carotene and chlorophyll using UV-Vis spectroscopy
2. Results:
Fig 1. UV-Vis spectroscopy results for S2 (Beta-Carotene extract)
Wavelength at peak/ nm Peak obtained/ A
448.00 0.4699
472.00 0.3993
649.00 0.0030
3. Discussion:
3.1 Data analysis
On inspection of Fig 4., a spectroscopic reading of beta-carotene should give three peaks of absorbance. The first peak would range between 200nm - 300nm, the approximate value nears that of 280nm. The next two peaks are found between 400nm - 500nm, the maxima of the major peak would be close to approximately 450nm while the second, minor peak would have a reading at about 480nm. The data procured …show more content…
This allows polar substances to have a greater affinity with the stationary phase while the non-polar substance will have greater affinity with the mobile phase. The initial eluting solvent used was hexane, a non-polar solvent, which allows the non-polar molecules to elute first and allowing compaction of the column. This resulted in beta-carotene eluting first, implying non-polar characteristics of beta-carotene. Chlorophyll is seen to have a better affinity with the stationary phase due to its polar characteristics. This is further proven when a change in eluting solvent was used, with 1:1 hexane-ethyl acetate. Ethyl acetate contributes polarity to the solvent, allowing the polar substances such as chlorophyll to be eluted. The change in eluting solvent is necessary in order to successfully remove
Yokoyama Yuri B1 (A0142349E)
1. Aim:
a) To separate beta-carotene and chlorophyll via column chromatography.
b) To analyse the contents of the plant extract as well as the separated beta-carotene and chlorophyll via thin layer chromatography (TLC)
c) To analyse the absorption peaks of both beta-carotene and chlorophyll using UV-Vis spectroscopy
2. Results:
Fig 1. UV-Vis spectroscopy results for S2 (Beta-Carotene extract)
Wavelength at peak/ nm Peak obtained/ A
448.00 0.4699
472.00 0.3993
649.00 0.0030
3. Discussion:
3.1 Data analysis
On inspection of Fig 4., a spectroscopic reading of beta-carotene should give three peaks of absorbance. The first peak would range between 200nm - 300nm, the approximate value nears that of 280nm. The next two peaks are found between 400nm - 500nm, the maxima of the major peak would be close to approximately 450nm while the second, minor peak would have a reading at about 480nm. The data procured …show more content…
This allows polar substances to have a greater affinity with the stationary phase while the non-polar substance will have greater affinity with the mobile phase. The initial eluting solvent used was hexane, a non-polar solvent, which allows the non-polar molecules to elute first and allowing compaction of the column. This resulted in beta-carotene eluting first, implying non-polar characteristics of beta-carotene. Chlorophyll is seen to have a better affinity with the stationary phase due to its polar characteristics. This is further proven when a change in eluting solvent was used, with 1:1 hexane-ethyl acetate. Ethyl acetate contributes polarity to the solvent, allowing the polar substances such as chlorophyll to be eluted. The change in eluting solvent is necessary in order to successfully remove