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The simple isocratic HPLC-UV method for the simultaneous determination of reduced and oxidized glutathione in animal tissue

Begic A., Djuric A., Gobeljic B., Stevanovic I., Lukic V., Stanojevic I., Ninkovic M., Saso L., Vojvodic D., Djukic M.

Acta Chromatographica

2017

Vol. 29, no. 1

67--84

The aim of our work was to optimize and apply simple high-performance liquid chromatography method with ultraviolet detection (HPLC-UV) for simultaneous determination of reduced (GSH) and oxidized (GSSG) glutathione in biological matrix (specifically, the rat liver tissue was used herein), since the ratio between oxidized and reduced glutathione forms (GSSG-GSH) has been recognized as an important biological marker of oxidatively depleted GSH in oxidative stress (OS)-associated diseases and poisonings. An isocratic chromatographic separation of GSH and GSSG (2.8 min and 6.3 min, respectively) was performed with the mobile phase consisted of sodium perchlorate solution (pH adjusted to 2.8) at flow rate of 1 mL min−1, detection set at 215 nm, and column temperature of 40 °C. The method offers short run time, linearity in the range of 0.01-200 μM concentration for both compounds (R2 = 1), low limits of detection and quantification (GSH: 0.18 μM and 0.56 μM, GSSG: 0.52 μM and 1.58 μM, respectively), precision, accuracy (bias < 2%), and high reproducibility. Through suitable sample handling, an overestimation of GSSG was prevented. High recovery (>99%) was achieved. The method was successfully applied for the analysis of GSH and GSSG in liver homogenates of Wistar rats intraperitoneally exposed to cadmium (Cd) (1 mg kg−1 CdCl2/21 days). Regardless of other Cd-mediated hepatotoxicity mechanisms, herein, we have exclusively interpreted/emphasized oxidative GSH depletion. The presented method is acceptable for a routine analysis of GSH and GSSG in biological matrix, while the calculated ratio GSSG-GSH is considered as a valuable OS marker.

Cable-suspended CPR-D type Parallel Robot

Filipovic M., Djuric A.

Journal of Theoretical and Applied Mechanics

2016

Vol. 54 nr 2

645--657

This paper deals with the analysis and synthesis of a newly selected Cable-suspended Parallel Robot configuration, named CPR-D system. The camera carrier workspace has the shape of a parallelepiped. The CPR-D system has a unique Jacobian matrix that maps the relationship between internal and external coordinates. This geometric relationship is a key solution for the definition of the system kinematic and dynamic models. Because of the CPR-D system complexity, the Lagrange principle of virtual work has been adapted. Two significant Examples have been used for the CPR-D system analysis and validation.