We want to prepare pyridine for solvent system to be runned for chromatography for carbohydrates. To teach post graduate students.
Dear Miss Shah,
Personally, I go out of my way to avoid pyridine whenever possible. It stinks, is extremely flammable and may be genotoxic or teratogenic. I also understand that you may not have many options. I suggest that you read the article, Thin-layer chromatography of carbohydrates; M. Ghebregzabher, S. Rufini, B. Monaldi, and M. Lato, Journal of Chromatography A, Volume 127(2):133-162, 18 August 1976. This article goes through the various different solvent mixtures that can be used for carbohydrate separations. That said, I would strongly urge you to consider looking at other chromatographic methods than TLC. While illustrative, few people actually use TLC methods today because they are time-consuming, hard to quantify, offer poor resolution of many isomers, and expose the user to dangerous chemicals. I would suggest that you consider HPAEC-PAD (high pH anion exchange chromatography with pulsed amperometric detection) of non-derivatized carbohydrates, or GC or CE (capillary electrophoresis) of derivatized sugars. HPAEC provides phenomenal resolution while PAD allows detection of extremely small amounts. Best yet, carbohydrates do not need to be derivatized for separation or detection. [Background science. Carbohydrates like glucose and sucrose are very weak organic acids. At high pH they are easily separated. It is even possible to separate 2-deuteroglucose from glucose. PAD oxidizes a small amount of the carbohydrate to generate signal.] Gas chromatography requires derivatization of the carbohydrate to eliminate all hydroxyl and amino groups. Many of the derivatizing reagents can also derivatize the hydroxyl and amino groups of the researcher so care is required in the procedures but they are very easy. Capillary electrophoresis requires derivatization for detection. Usually the reducing position of the sugar is reacted with an amine to generate a Schiff base which is then reduced to the amine. There are dozens of derivatization tags and many ways to separate the modified sugars. I suggest the somewhat dated. "Carbohydrate analysis by capillary electrophoresis." In the Handbook of Capillary Electrophoresis, Olechno, J. D., and J. A. Nolan ed. JP Landers, CRC Press, Boca Raton, FL (1997): 297-345.
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