. Author manuscript; available in PMC: 2012 Nov 21.

Published in final edited form as: Chem Res Toxicol. 2011 Sep 12;24(11):1924–1936. doi: 10.1021/tx200273z

Table 2.

Regioselectivity of the reaction between GSH-BDA and ornithine, lysine or spermidine

	Amine mixture^a		Individual amines^b
Amine	Buffer	Media	Buffer	Media	Hepatocytes
N^α/N^δ ornithine (7a/7b)	0	0	0.47	0.55	0.51 ± 0.17
N¹/N⁸ spermidine (8a/8b)	2.5 ± 0.3	6.9 ± 0.6	2.6	7.8	5.5 ± 1.8
N^α/N^ε lysine (2b/2a)	0.45 ± 0.05	0.26 ± 0.06	0.57	0.37	0.083 ± 0.021^c

GSH (5 mM) was combined with 100 μM BDA and an equimolar mixture of amines (100 μM each putrescine, cadaverine, spermine, spermidine, ornithine, and lysine) in either 150 mM sodium phosphate, pH 7.4 (buffer) or RPMI 1640 media, containing 10 mM HEPES, pH 7.4 (media). The reaction mixture was analyzed by LC-MS and the resultant peak areas were corrected for the ionization efficiency of each reaction product (Supplemental Table 1) prior to calculating the ratio of the two possible regioisomers.

GSH (5 mM) was combined with 100 μM BDA and 100 μM ornithine, lysine or spermidine in either 150 mM sodium phosphate, pH 7.4 (buffer) or RPMI 1640 media, containing 10 mM HEPES, pH 7.4 (media). The reaction mixtures were analyzed as described above.

Previously published.¹¹