Authors:Aida Begic, Ana Djuric, Borko Gobeljic, Ivana Stevanovic, Vera Lukic, Ivan Stanojevic, Milica Ninkovic, Luciano Saso, Danilo Vojvodic, and Mirjana Djukic
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.
Authors:Ivan Givechev, Dimitar Tanev, and Dancho Danalev
of contamination especially of plants further used as a row materials for production of different good as well as for direct food of animals or humans are described in the scientific literature among which contaminated soil [ 5–7 ], particles from air
Authors:Iskra Boneva, Spaska Yaneva, and Dancho Danalev
possibility for accumulation in the food chain can cause a risk for human health [ 2–4 ]. The presence of these compounds in the food have led to established maximum residue levels (MRLs) in or on food and feed of plant and animal origin are set at Regulation
Authors:Lei Gao, Peng Wang, Zhongxiang Chen, Shizhan Tang, Song Wu, Gang Li, and Dongli Qin
Introduction Fish is a common source of animal protein for people. It also contains long chain ω-3-fatty acids such as eicosapentaenoic acid and docosahexaenoic acid, as well as vitamins and minerals which are beneficial for a healthy diet [ 1
Authors:I. Kowalska, D. Jedrejek, K. Jonczyk, and A. Stochmal
Wheat ( Triticum aestivum L.) is one of the most important crop plants worldwide, an important agricultural commodity, a component of human diet, and a significant source of animal fodder in the world. As wheat
Authors:S.M. Nurulain, S. Ojha, S. Dhanasekaran, K. Kuča, N. Nalin, C. Sharma, A. Adem, and H. Kalász
Distribution of K027, a hydrophilic, positively charged compound is monitored in the body of pregnant mice using high-performance liquid chromatography (HPLC). Intraperitoneal injection was done on the 18th day of pregnancy; the plasma and brains of the mother mice, placentae and the fetuses’ brains were dissected following 5, 15, 30, 60, and 120 min of treatment. Significant incorporation of K027 was found in the placentae and in fetuses’ brains relative to its levels in the mothers’ plasma and brains. This incorporation warns of a possible adjustment of dose of pyridinium aldoxime antidotes in case of pregnancy. Further studies with different gestational periods and animal models are warranted.
Authors:M. Gawlik, R. Skibiński, J. Trawiński, and Ł. Komsta
The number of new drugs launched to the market is constantly increasing; however, the metabolism of many of them is still not fully established. The knowledge of drug metabolism pathways is crucial for the efficacy and safety of therapies and, in classical approach, requires the use of animals as well as human volunteers, but this kind of research is expensive and time-consuming. Therefore, nowadays, more and more biological and chemical in vitro methods are developed for the drug metabolism study. This review is focused on the photocatalytic degradation of chemicals and the application of this process in chromatographic methods of drug metabolism research. A theoretical background of photocatalysis and all its applications in a drug metabolism study were reviewed, and other in vitro methods that are actually used were summarized and discussed. Other analytical methods used in this area were also discussed and compared.
Authors:A. Ahmad, R. M. A. Khan, and K. M. Alkharfy
A rapid, accurate, and sensitive reverse phase high-performance liquid chromatographic method was developed and validated for the simultaneous determination and quantification of glibenclamide and thymoquinone in rat plasma in the presence of internal standard (thymol). Chromatograms were developed with methanol, acetonitrile, and buffer (50:20:30, v/v/v) solvent system on a Symmetry® C18 (5 μm, 3.9 × 150 mm) column, and pH was adjusted to 4.5 with orthophosphoric acid. Mobile phase was pumped at a flow rate of 1.5 mL min−1 with 254 nm ultraviolet (UV) detection. Validation of the method was performed in order to demonstrate its selectivity, linearity, precision, accuracy, limits of detection, and quantification (LOD and LOQ). Standard curves were linear (r2 = 0.996 and 0.999 for glibenclamide and thymoquinone) over the concentration range 0.5–50 μg mL−1. The coefficient of variation (CV) of < 6% and accurate recovery of 87.54–105.19% for glibenclamide and CV of <5% and accurate recovery of 86.08–103.19% for thymoquinone were found to be in the selected concentration range of 0.5–50 μg mL−1. The lower limits of detection and quantitation of the method were 0.109 and 0.332 μg mL−1 for glibenclamide and 0.119 and 0.361 μg mL−1 for thymoquinone, respectively. The within and between-day coefficients of variation were less than 7%. The validated method has been successfully applied to measure the plasma concentrations in a drug interaction study of glibenclamide with thymoquinone in an animal model to illustrate the scope and application of the method.
Authors:M. Crevar-Sakač, Z. Vujić, Z. Vujčić, B. Marković, and D. Vasiljević
A simple and sensitive liquid chromatography—tandem mass spectrometry method was developed for the quantification of atorvastatin, ortho-hydroxyatorvastatin, para-hydroxyatorvastatin, and atorvastatin lactone in rat plasma. Solid-phase extraction was used for preparation of samples. Rosuvastatin was chosen as an internal standard. Chromatographic separation was achieved on ZORBAX Eclipse C18 Analytical, 4.6 × 100 mm (3.5 μm) column with a gradient mobile phase composed of acetonitrile and 0.1% acetic acid, at a flow rate of 400 μL min−1. The column was kept at constant temperature (25 °C), and autosampler tray temperature was set at 4 °C. The following selected reaction monitoring (SRM) transitions were selected: (m/z, Q1 → Q3, collision energy) atorvastatin (559.47 → 440.03, 22 eV), atorvastatin lactone (541.36 → 448.02, 19 eV), ortho-ohydroxyatorvastatin (575.20 → 440.18, 20 eV), para-hydroxyatorvastatin (575.54 → 440.18, 20 eV), and rosuvastatin (482.25 with selected combination of two fragments 257.77, 31 eV, and 299.81, 35 eV) in positive ion mode. The method was validated over a concentration range of 0.5–20 ng mL−1 for ortho-hydroxyatorvastatin and para-hydroxyatorvastatin and 0.1–20 ng mL−1 for atorvastatin and atorvastatin lactone with excellent linearity (r2 ≥ 0.99). This method demonstrated acceptable precision and accuracy at four quality control concentration levels. The detection limits were 0.1 and 0.13 ng mL−1 for orth-ohydroxyatorvastatin and para-hydroxyatorvastatin, respectively, and 0.05 ng mL−1 for atorvastatin and atorvastatin lactone. All analytes were found to be stable at examined conditions. Validated method was applied for determination of atorvastatin and its metabolites in plasma of experimental animals.