POTĚŠIL, David, Jitka PETRLOVÁ, Josef ZEHNÁLEK, Vojtěch ADAM, Jana ZELENÁ, Bořivoj KLEJDUS and René KIZEK. Determination of thiol compounds using high-performance liquid chromatography with electrochemical detection. In Vitamins 2004 Targeted Nutritional Therapy. Pardubice: Univerzita Pardubice, 2004, p. 179-180, 292 pp. ISBN 80-7194-644-3.
Other formats:   BibTeX LaTeX RIS
Basic information
Original name Determination of thiol compounds using high-performance liquid chromatography with electrochemical detection.
Name in Czech Určení thiolových sloučenin pomocí HPLC-ED
Name (in English) Determination of thiol compounds using high-performance liquid chromatography with electrochemical detection.
Authors POTĚŠIL, David (203 Czech Republic, guarantor), Jitka PETRLOVÁ (203 Czech Republic), Josef ZEHNÁLEK (203 Czech Republic), Vojtěch ADAM (203 Czech Republic), Jana ZELENÁ (203 Czech Republic), Bořivoj KLEJDUS (203 Czech Republic) and René KIZEK (203 Czech Republic).
Edition Pardubice, Vitamins 2004 Targeted Nutritional Therapy. p. 179-180, 292 pp. 2004.
Publisher Univerzita Pardubice
Other information
Original language Czech
Type of outcome Proceedings paper
Field of Study 10406 Analytical chemistry
Country of publisher Czech Republic
Confidentiality degree is not subject to a state or trade secret
RIV identification code RIV/00216224:14310/04:00011125
Organization unit Faculty of Science
ISBN 80-7194-644-3
Keywords in English GSH; HPLC-ED; PC
Tags GSH, HPLC-ED, PC
Changed by Changed by: prof. RNDr. Vojtěch Adam, Ph.D., učo 63820. Changed: 31/5/2005 13:15.
Abstract
Thiol compounds such as cystein (Cys), reduced (GSH) and oxidized (GSSG) glutathione and phytochelatins (PCs) play an important role in heavy metal detoxification in plants. These thiols are biological active compounds whose function is elimination of oxidative stress in plant cells. The aim of our work was to optimise sensitive and rapid method of high-performance liquid chromatography coupled with electrochemical detector (HPLC-ED) for determination of abovementioned thiol compounds in biological samples. An HPLC-ED system was consisted of two delivery pumps, chromatographic column Zorbax XDB (150.0 x 2.1 mm, 5 um particle size) and CoulArray detector containing eight working electrodes made from porous graphite. The most suitable isocratic mobile phase for the determination and separation of Cys, GSH, GSSG and PC2 consisted of methanol (MET) and trifluoroacetic acid (TFA). In addition, the influence of concentrations of TFA and ratio of MET:TFA on chromatographic separation and detection of the thiol compounds were studied. The mobile phase consisting from methanol and 0.05% TFA in ratio (97:3) (%; v/v) was found the most suitable for the thiol compounds determination. Optimal flow rate of the mobile phase was 0.18 ml.min-1 and the column temperature was 35 C. For the selection of the most effective working electrodes potentials, hydrodynamic voltamograms of all studied compounds was obtained. From the results we selected two detection potentials 700 mV (for the determination of GSSG and PC2) and 680 mV (Cys and GSH). HPLC-ED full scan of studied thiols. Moreover, we were able to determine femtomole levels of studied compounds by optimised HPLC-ED method. The optimised HPLC-ED method was applied to the study of influence of different cadmium concentrations (0, 10 uM and 100 uM Cd concentration) on content of Cys, GSH, GSSG and PC2 in maize (Zea mays L.) kernels. According to the increasing time of Cd treatment, content of GSH, GSSG and PC2 in maize kernels increased but content of Cys decreased. Decreasing concentration of Cys probably relates with the increasing GSH and phytochelatins synthesis.
Abstract (in English)
Thiol compounds such as cystein (Cys), reduced (GSH) and oxidized (GSSG) glutathione and phytochelatins (PCs) play an important role in heavy metal detoxification in plants. These thiols are biological active compounds whose function is elimination of oxidative stress in plant cells. The aim of our work was to optimise sensitive and rapid method of high-performance liquid chromatography coupled with electrochemical detector (HPLC-ED) for determination of abovementioned thiol compounds in biological samples. An HPLC-ED system was consisted of two delivery pumps, chromatographic column Zorbax XDB (150.0 x 2.1 mm, 5 um particle size) and CoulArray detector containing eight working electrodes made from porous graphite. The most suitable isocratic mobile phase for the determination and separation of Cys, GSH, GSSG and PC2 consisted of methanol (MET) and trifluoroacetic acid (TFA). In addition, the influence of concentrations of TFA and ratio of MET:TFA on chromatographic separation and detection of the thiol compounds were studied. The mobile phase consisting from methanol and 0.05% TFA in ratio (97:3) (%; v/v) was found the most suitable for the thiol compounds determination. Optimal flow rate of the mobile phase was 0.18 ml.min-1 and the column temperature was 35 C. For the selection of the most effective working electrodes potentials, hydrodynamic voltamograms of all studied compounds was obtained. From the results we selected two detection potentials 700 mV (for the determination of GSSG and PC2) and 680 mV (Cys and GSH). HPLC-ED full scan of studied thiols. Moreover, we were able to determine femtomole levels of studied compounds by optimised HPLC-ED method. The optimised HPLC-ED method was applied to the study of influence of different cadmium concentrations (0, 10 uM and 100 uM Cd concentration) on content of Cys, GSH, GSSG and PC2 in maize (Zea mays L.) kernels. According to the increasing time of Cd treatment, content of GSH, GSSG and PC2 in maize kernels increased but content of Cys decreased. Decreasing concentration of Cys probably relates with the increasing GSH and phytochelatins synthesis.
Links
GP525/04/P132, research and development projectName: Studium obranných mechanismů rostlin při stresu způsobeném těžkými kovy
LN00A081, research and development projectName: Signální dráhy u rostlin
PrintDisplayed: 12/5/2024 17:37