Porous graphitic chromatography in urine analysis

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Authors

ŠMAK Pavel GREGOROVÁ Jana NOVOTNÁ Lucie KUBINYIOVÁ Lenka PEŠ Ondřej

Year of publication 2021
Type Conference abstract
MU Faculty or unit

Faculty of Medicine

Citation
Description Urine is rich in the products of metabolism, some of which are linked to different conditions, such as inflammation or metabolic disorders. The traditional analytical procedure involving reversed-phase liquid chromatography (LC) is unsuitable for analysis of polar compounds unless ion pair reagents are used. Hydrophilic interaction chromatography often requires high ionic strength to overcome the broad tailing of basic compounds and is limited to pH < 8. Porous graphitic chromatography (PGC) in connection to mass spectrometry (MS) may encompass analysis of polar and nonpolar compounds at the same time using a broad range of conditions such as pH and ionic strength of MS compatible buffers and additives. Reliable quantitation of analytes in urine demands knowing either daily diuresis or the concentration of creatinine, which is often done separately (e.g., Jaffe's reaction, or direct injection analysis). Here we aimed to establish the background for the development of routine methods for urine compounds separation and quantitation. Selected compounds from the class of catecholamines and creatinine have been analyzed using LC MS on a PGC column and various conditions affecting the final separation have been tested. The ability to quantify selected analytes and creatinine in a single run could significantly simplify the analytical process (and potentially lower the costs of the analyses). Nevertheless, such a method is not currently available for routine analyses in clinical practice.
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