Vědět a znát, to mnohý by chtěl, ale učit se nechce.

Walther Von Der Vegelweide

Výzkum

Isolation of biogenic amines using paramagnetic microparticles as an auxiliary analytical tool

Biogenic amines are formed as products of normal metabolic activities of plants, animals and microorganisms [1]. They arise from the effects of amino acid decarboxylase, which contain pyridoxal phosphate as the cofactor [2] in process - decarboxylation. Decarboxylase enzyme cleaves the carboxyl carbon dioxide to form basic amine [3]. The polyamines spermine (Spm) and spermidine (Spd) and diamines putrescine (Put) and cadaverine (Cad) are constituents of eukaryotic and prokaryotic cells, having multiple functions in living organisms [4]. High polyamine levels are toxic to cells and facilitate cell death mainly by oxidative mechanisms [5, 6]. Thus their pathological presence in various matrices may be utilized for doing diagnostic more precise. In particular, urine was previously shown to be closely linked with increased levels of biogenic amines associated with various types of tumors [7, 8], and recovery of urinary diacetylpolyamines to the normal levels, accompanied with remission of cancer was described [9]. For diagnostic, namely putrescine, spermidine, spermine and cadaverine may be interesting, because of their presence in many malignant diseases [10]. On the other hand histamine is an active amine complex, and may affect the reactions of many cells as a sort of chemical conductive substance in many states, such as anaphylaxis, dermatosis, phlogistic reactions, rheumatoid arthritis and many others [11]. Moreover biogenic amines dopamine (Da), norepinephrine (Ne), and serotonin (5-HT) are important neurotransmitters in central and peripheral nervous system [12]. Concentrations of norepinephrine and dopamine in urine specimens above 80 and 400 µg per 24 h have been reported to be linked with pheochromocytoma [13].

In order to determine the concentrations of biogenic amines in complicated biological matrices like urine, techniques providing high resolution and sensitivity are highly demanded. In our study, we decided to use ion-exchange liquid chromatography (IEC) with post-column ninhydrin derivatization and visible (VIS) light range for biogenic amines separation and detection, which minimize the sample pre-cleaning requirement [14]. Main aim of the present study consisted of synthesis and characterization of two types of paramagnetic microparticles (PMPs), composed of nanomaghemite (?-Fe2O3) core, modified with chitosan, or tetraethyl orthosilicate (TEOS) covered with Dowex. Subsequently, our prepared microparticles were tested for their ability to establish a binding with biogenic amines (Spermine, Spermidine, Putrescine, Cadaverine, Histamine, Tyramine,), and finally they were applied as a first part of our 2D separation approach, consisting of PMPs and IEC for determination of biogenic amines in sample without complicated pre-treatment, thus facilitating biogenic amines analysis.

Práce je spojená s projektem SIX CZ.1.05/2.1.00/03.0072.

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