Using thin-layer chromatography, it is possible to see what pigments plant leaves contain. For example, I can test whether there are chlorophyll a or b, carotenoids, or anthocyanins.

What kind of technique I would need to quantify the amount of these pigments in the leaves?

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Once you've found a suitable mobile phase in analytical TLC, you could upscale the separation and transfer the results to preparative TLC or column chromatography.

In the case of column chromatography, collect the eluted material in test tubes, check the content by analytical TLC and/or UV-VIS spectroscopy, unite identical fractions, evaporate the solvent and determine the weight of the respective fractions.

In the case of preparative TLC, scratch the separated bands off the plate, elute with a solvent of your choice and proceed as above.

If necessary, repeat the chromatographic separations for the different compound classes with other solvent mixtures.

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  • $\begingroup$ How should the results from spectroscopy be interpreted in this case? Is it a confirmatory step or does it serve some other purpose? $\endgroup$ – mmh Feb 15 '15 at 14:46
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    $\begingroup$ @mmh One spot on tlc indicates at least one compound. A UV spectrum gives additional Information on which fractions to pool and which not. $\endgroup$ – Klaus-Dieter Warzecha Feb 15 '15 at 19:32

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