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Research laboratory at the Department of metabolic care and gerontology is dealing with modern technologies in analytical chemistry.
Modern technologies in analytical chemistry are applied in development of new separation methods for determination of important biological markers. Neopterin is one of the target compounds. Determination of neopterin levels reflect the stage of activation of the cellular immune system which is of important in the pathogenesis and progression of various diseases.
The essential amino acid L-tryptophan is required for the biosynthesis of proteins and is precursor for several biologically important compounds. Tryptophan degradation can be detected in various cells including antigen-presenting cells. Determination of kynurenine (metabolite of tryptophan) is a sensitive laboratory diagnostic tool to detect and monitor Th1-type immune activation in humans. Combination of neopterin, kynurenine, creatinine and tryptophan in serum should predicate micro-inflammatory presence and effectiveness of its treatment. Neopterin levels are elevated during rejection after kidney transplantation, which can make worst graft function. Therefore neopterin in human serum should be important prognostic marker in various diseases such as kidney failure, diabetes mellitus or atherosclerotic or cancer kidney damage and in patients treated by hemodialysis. The same way can be determination of neopterin, kynurenine, tryptophan and creatinine levels of use in gerontology. Determination of neopterin levels in human urine is very often used method, especially for its non-invasive character. For development of method for determination of neopterin in human serum and human urine, monolithic and hybrid stationary phases were used. Sample preparation procedure was focused on micro titration plates which allows handle with 96 samples in one extraction step. Thanks to special auto sampler for micro plates (Rack-Changer) are this method suitable for large sequences of samples (large clinical studies) and these methods should be included as part of routine examination (thank to small volume of samples).
Other target new trend is miniaturization of solid phase extraction procedure for determination of retinol, alpha- and gamma-tocopherol. This method was combined with very fast HPLC separation using monolithic stationary phase. Liposoluble vitamins are determined in human serum, for monitoring of nutrition in gerontology patients and patients with cancer diseases. Pathogenesis of several diseases has been linked to a condition of oxidative stress in aging (Alzheimer's disease, Parkinson's disease, cardiac failure etc.). Antioxidant capacity of cell membrane is monitored by HPLC method for determination of vitamin E in erythrocyte membrane (representative and easy available). Vitamin E is the main scavenger of free oxygen radicals influence cell membrane integrity and protects cell against damage. Therefore knowledge of antioxidant capacity of cell membrane (vitamin E concentration in erythrocyte membrane) is important to predict level of membrane damage and necessity of vitamin E.
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