Presentation Title

High performance liquid chromatographic determination of four biological aminothiols after microwave-enhanced derivatization with SBD-F

Presentation Type

Poster

Keywords

aminothiols, homocysteine, cysteine, glutathione, cysteinyl-glycine, SBD-F, microwave

Department

Chemistry

Major

Chemistry

Abstract

Homocysteine, cysteine, cysteinyl-glycine, and glutathione are significant aminothiols which have been implicated as risk factors in atherosclerosis and other vascular diseases, and therefore rapid determination of these molecules is desirable. Following reduction of the disulfides with tris-carboxyethyl-2-phosphine, a widely used method utilizes derivatization with ammonium 7-fluorobenzo-2-oxa-1,3-diazole-4-sulfonate (SBD-F) as a fluorogenic probe prior to reversed-phase HPLC analysis. We report the results of microwave-enhanced synthesis of the fluorescent derivatives at 90 °C in microcentrifuge tubes. Utilizing microwave heating, we reduced the derivatization time at all temperatures from 1 h to less than 2 min. Standard solutions produced rectilinear calibration curves with correlation coefficients >0.99 for all four aminothiols. We compared the results obtained for the four aminothiols extracted from chicken plasma and liver, and mouse heart using traditional heating and microwave heating. No statistical differences were observed for the aminothiols, except glutathione which was consistently lower with microwave heating. We also report the results of recovery experiments and differences in calibration sensitivities between the two methods.

Faculty Mentor

Dr. David Green

Funding Source or Research Program

Summer Undergraduate Research Program

Location

Waves Cafeteria

Start Date

24-3-2017 2:00 PM

End Date

24-3-2017 3:00 PM

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Mar 24th, 2:00 PM Mar 24th, 3:00 PM

High performance liquid chromatographic determination of four biological aminothiols after microwave-enhanced derivatization with SBD-F

Waves Cafeteria

Homocysteine, cysteine, cysteinyl-glycine, and glutathione are significant aminothiols which have been implicated as risk factors in atherosclerosis and other vascular diseases, and therefore rapid determination of these molecules is desirable. Following reduction of the disulfides with tris-carboxyethyl-2-phosphine, a widely used method utilizes derivatization with ammonium 7-fluorobenzo-2-oxa-1,3-diazole-4-sulfonate (SBD-F) as a fluorogenic probe prior to reversed-phase HPLC analysis. We report the results of microwave-enhanced synthesis of the fluorescent derivatives at 90 °C in microcentrifuge tubes. Utilizing microwave heating, we reduced the derivatization time at all temperatures from 1 h to less than 2 min. Standard solutions produced rectilinear calibration curves with correlation coefficients >0.99 for all four aminothiols. We compared the results obtained for the four aminothiols extracted from chicken plasma and liver, and mouse heart using traditional heating and microwave heating. No statistical differences were observed for the aminothiols, except glutathione which was consistently lower with microwave heating. We also report the results of recovery experiments and differences in calibration sensitivities between the two methods.