I did a column chromatography on spinach and then a TLC. On the TLC I got five visible bands of colors. On the extracted yellow substance I got one yellow band (carotene) and on the green I also got only one green band. I thought there would be more bands on the green but maybe the chlorophyll was all that was in that band. Where did the other bands go?
Does it sound like I got reasonable results? Since there was no spinach at the stores, we used some that was growing wild under my deck and it was very dry.
Thanks from Sheryl
It isn't clear from your description what was extracted.
Guess sometimes I could look in the book rather than spend an hour on the internet. We were extracting carotene and chlorophyll a and b. Looks like I did my experiment okay. There seems to be a visible as well as a not very visible band in the green extract. Sorry to bother.
Sheryl
No problem, Sheryl! It sounds like you got reasonable results from your column chromatography and TLC experiment. Let me explain the possible reasons for the different bands you observed.
Column chromatography is a technique used to separate and purify different compounds in a mixture based on their different affinity to a stationary phase. In your case, you extracted carotene and chlorophyll a and b from spinach.
During the column chromatography, the mixture of compounds gets separated into distinct bands as it passes through the column. Each band represents a different compound. The compounds that have a stronger affinity for the stationary phase will travel slower, resulting in distinct bands.
Now, when you performed the TLC (thin-layer chromatography), you took samples from the separated bands and spotted them on a thin layer of adsorbent material, such as silica gel or alumina. The compounds in the samples will then migrate as a result of capillary action.
In your case, you observed five visible bands of colors on the TLC plate. The visible bands could represent different compounds present in the original spinach extract or other impurities that were not separated well during column chromatography.
You specifically mentioned that you got one yellow band (carotene) and one green band from the extracted substances. It's possible that the yellow band corresponds to the carotene you were looking for. The green band may contain a mixture of chlorophyll a and b. You mentioned that there is a visible band as well as a not very visible band in the green extract, indicating the presence of multiple compounds or impurities.
Since you used wild-growing, dry spinach for your experiment, it's important to consider that the composition and quality of compounds can vary compared to commercially available spinach. This could explain why you observed slightly different results.
Overall, it sounds like you performed your experiment correctly and obtained reasonable results. It's always valuable to consult reference books and reliable resources to validate your findings and gain a deeper understanding of the chromatography techniques you used.