Role of Electron-Donating Withdrawing Character, Ph, and Stoichiometry on the Chemiluminescent Reaction of Tris(2,2'-Bipyridyl)ruthenium(iii) with Amino-Acids
by Bruñe, Stephen Newell; Bobbitt II, Donald R.
A postcolumn chemiluminescent technique for the detection of underivatized amino acids using electrogenerated tris-(2,2'-bipyridyl)ruthenium(III) is described. The reaction chemistry has been investigated, and it has been shown that the electron-withdrawing/donating character of the R group attached to the alpha-carbon of the amine influences the chemiluminescent efficiency of the reaction. Stoichiometric studies conducted on four amines produced a mole ratio of 2:1, ruthenium: amine. At optimum reaction conditions the relative intensities of the primary amino acids tested varied by a factor of 55, with serine yielding the lowest chemiluminescence and leucine yielding the highest chemiluminescence. Detection limits for serine and leucine have been calculated to be 135 and 3 pmol, respectively, at a S/N of 3. Linearity has been demonstrated over 2 orders of magnitude for Leu. The ability to implement this system for postcolumn chemiluminescence detection of amino acids following a protein digest has been demonstrated.
- Analytical Chemistry
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