20.500.12556/RUL-152925 Simultaneous determination of free biliverdin and free bilirubin in serum a comprehensive LC-MS approach Background: Prognosis, diagnosis, and treatment of several diseases strongly rely on the sensitive, selective, and accurate determination of specific biomarkers in relevant biological samples. Free biliverdin and free bilirubin represent important new biomarkers of oxidative stress, however, the lack of suitable analytical methods for their determination has hindered progress in biomedical and clinical research. Results: Here, we introduce a first comprehensive approach for robust and simultaneous determination of these bilins in serum using liquid chromatography – mass spectrometry (LC-MS). The developed analytical method exhibits linearity for both analytes within the concentration range of 0.5–100 nM, with limits of detection and quantitation determined at 0.1 nM and 0.5 nM, respectively. Moreover, several analytical pitfalls related to the intrinsic molecular structures of free bilirubin and free biliverdin and their trace concentration levels in biological samples are discussed here in detail for the first time. We have shown that the solubility, chemical stability, and affinity of these bilins to various materials strongly depend on the solvent, pH, and addition of stabilizing and chelating agents. Finally, the validated LC-MS method was successfully applied to the analysis of both bilins in fetus bovine serums, yielding higher free bilirubin/biliverdin ratios compared with previously reported values for human serum. Significance: Failure to recognize and address the challenges presented here often leads to substantial analytical errors and consequently biased interpretation of the obtained results. This pertains not only to LC-MS, but also to many other analytical platforms due to the compound-derived sources of error. biomedical research sensitivity robustness serum stability matrix effects analizna kemija bolezni zdravljenje biomarkerji serum true false true Angleški jezik Slovenski jezik Članek v reviji 2023-12-12 11:16:18 2023-12-12 11:16:18 2023-12-13 03:41:47 0000-00-00 00:00:00 2024 0 0 8 str. Vol. 1287, art. 342073 Jan. 2024 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 543 1873-4324 10.1016/j.aca.2023.342073 176790787 1-s2.0-S0003267023012941-main.pdf 1-s2.0-S0003267023012941-main.pdf 1 F3510607A74C213BAF631BD37945E141 2e238906dd59a97132741b8bb349a4d027b2ab7f8eb2133355752eaf20334b4e cec1ff40-98d7-11ee-b821-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=178662 https://www.sciencedirect.com/science/article/pii/S0003267023012941 1 7dbdf234-98d7-11ee-b821-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=178661 Medicinska fakulteta Fakulteta za farmacijo 0 0 0