Homocysteine is coupled to folate metabolism and has been implicated in reduced cognitive function associated with Down Syndrome. Other low molecular weight aminothiols, cysteine, cysteinyl-glycine, and glutathione, have been identified as markers for risk factors in atherosclerosis and other vascular diseases. Thus, rapid and accurate determination of these aminothiols is desirable. A widely-used method for the determination of these biological aminothiols utilizes RP-HPLC with fluorescence detection after reduction of the disulfides and derivatization with ammonium 7-fluorobenzo-2-oxa-1,3-diazole-4-sulfonate (SBD-F). We report the results of a new microwave-assisted synthesis (MAS) of the fluorescent derivatives, resulting in reduced reaction times from 60 minutes for the traditional method to less than 2 minutes with MAS. Standard solutions produced rectilinear calibration curves with correlation coefficients &gt;0.99 for all four aminothiols. In addition to MAS, we also explored different disulfide reducing agents to minimize the use of hazardous reagents. We compared the results obtained for the four aminothiols in chicken plasma and liver and mouse heart after derivatization with traditional heating and MAS. No statistical differences were observed for the aminothiols, except glutathione which was inconsistently lower after MAS. We also report the results of recovery experiments and differences in calibration sensitivities between the two methods.

Microwave-assisted synthesis of SBD-F derivatives for the HPLC-FLD determination of four biological aminothiols

253rd National Meeting (2017)

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