Multidimensional chromatography amino acids

Figure 4.8. MuDPIT platform. The MuDPIT or multidimensional gel protein identification system is an multidimensional liquid chromatography (LC)-based system of separation prior to tandem mass spectrometry (MS/MS). It is not necessary to purify proteins prior MuDPIT, although a reduction in protein complexity by some prior purification is helpful in obtaining interpretable spectra. Protein lysates are digested into peptides that are loaded onto a strong cation exchange (SCX) support. Peptides are sequentially eluted onto a reverse-phase column for a second separation. Eluted peptides from the reverse-phase column are electrosprayed into a tandem mass spectrometer for amino acid sequencing and identification of proteins in the sample.

Multidimensional Liquid Chromatography of Proteins Separation of Amino Acids—Amino Acid Analysis References... 

The basis for developing multidimensional chromatographic approaches to separate complex mixtures is the work of Martin and his coUeagues, using paper chromatography to separate wool hydrolysate. The hydrolysate was applied at one corner of the square filter paper and developed in collidine for 3 days, taken out and dried, turned 90, and then developed in phenol for 27 hr. A similar approach was also used to separate a mixture of the hydrochlorides of 22 amino acids. 

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