I need analytical methods for sample preparation for analysis particularly for Alkaloids from plant material
Dear Suleiman Abubakar Garba , in order to fractionnate your extract you can use a CPC (centrifugal partition chromatography) this method can be coupled to UV to give, more information. for the preparative methods you can see prep-MPLC too, then a purification with Column chromatography and NMR analysis with H and or C, but in order to identify a molecule you need FTIR, UV, H-NMR and C-NMR data, best regards
Preparative HPLC is the most common technique used to separate, fractionate and collect samples. For some sample types, CCC (counter current chromatography) or CPC work well. Your specific sample and goals will determine which techniques are most applicable to start with.
"TLC" is not generally used for sample preparation.
"Normal Column chromatography" ? I think you may mean Normal phase chromatography (NP) which is a mode of liquid chromatography. For samples that are fully soluble in NP liquids, this mode uses volatile solvents which makes it easy to remove the solvents leaving the material. Good for preparative separations and fraction collection.
Please review a basic text on analytical methods, specific to what your own needs are. There are many techniques available for purification and it is important that you learn the terminology and also review techniques which are specific to your sample type of interest. Books are a great resource as are general articles on the web. Invest your time in researching these first to expand your knowledge.
Additionally, many basic bench extraction techniques may be useful too. Review the properties of the plant materials you wish to extract for the simplest initial approach.
Dear Suleiman Abubakar Garba
Good question. All depends on the purpose of your analysis. By purification, do you mean pre-purification? After this step (pre)-purification, which technique (s) do you intend to use for the analysis? This information might help you to select techniques among the already mentioned techniques.
Regards,
JPM.
When it was introduced in the early 1980s HPLC tended to eclipse CCC and related methods. CCC was slower and (like TLC and paper chromatography) needed knowledge and experience. For a time, there was a tendency to think that chromatographic efficiency is a substitute for selectivity; this was unfortunate given the limited range of selectivities provided by the dominant HPLC reversed-phase techniques (and the annoying tendency of lipids and other metabolites not to give nice sharp peaks). Nowadays, convenient CCC equipment is available and, although I don't have practical experience, I think it should be preferred at least for the preliminary stages of isolations. For solvent systems it would be worth consulting older textbooks such as Separation methods in biochemistry by Morris an Morris, which are easily available on the new-fangled internet. You could also get a feel for the subject by looking up the 1950s literature on particular projects such as the isolation and identification of the hormone vasopressin.
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