Hložek Tomáš, Křížek Tomáš, Tůma Petr, Bursová Miroslava, Coufal Pavel, Čabala Radomír
Charles University and General University Hospital, First Faculty of Medicine, Institute of Forensic Medicine and Toxicology, Ke Karlovu 2, 121 08, Prague 2, Czech Republic; Charles University, Faculty of Science, Department of Analytical Chemistry, Albertov 6, 128 43, Prague 2, Czech Republic.
Charles University, Faculty of Science, Department of Analytical Chemistry, Albertov 6, 128 43, Prague 2, Czech Republic.
J Pharm Biomed Anal. 2017 Oct 25;145:616-620. doi: 10.1016/j.jpba.2017.07.024. Epub 2017 Jul 26.
High anion gap metabolic acidosis frequently complicates acute paracetamol overdose and is generally attributed to lactic acidosis or compromised hepatic function. However, metabolic acidosis can also be caused by organic acid 5-oxoproline (pyroglutamic acid). Paracetamol's toxic intermediate, N-acetyl-p-benzoquinoneimine irreversibly binds to glutathione and its depletion leads to subsequent disruption of the gamma glutamyl cycle and an excessive 5-oxoproline generation. This is undoubtedly an underdiagnosed condition because measurement of serum 5-oxoproline level is not readily available. A simple, cost effective, and fast capillary electrophoresis method with diode array detection (DAD) for simultaneous measurement of both paracetamol (acetaminophen) and 5-oxoproline in serum was developed and validated. This method is highly suitable for clinical toxicology laboratory diagnostic, allowing rapid quantification of acidosis inducing organic acid 5-oxoproline present in cases of paracetamol overdose. The calibration dependence of the method was proved to be linear in the range of 1.3-250μgmL, with adequate accuracy (96.4-107.8%) and precision (12.3%). LOQ equaled 1.3μgmL for paracetamol and 4.9μgmL for 5-oxoproline.
高阴离子间隙代谢性酸中毒常并发于急性对乙酰氨基酚过量服用，通常归因于乳酸酸中毒或肝功能受损。然而，代谢性酸中毒也可能由有机酸5-氧代脯氨酸（焦谷氨酸）引起。对乙酰氨基酚的毒性中间体N-乙酰-p-苯醌亚胺与谷胱甘肽不可逆结合，其消耗导致随后γ-谷氨酰循环的破坏和5-氧代脯氨酸的过度生成。这无疑是一种诊断不足的病症，因为血清5-氧代脯氨酸水平的测量方法并不容易获得。我们开发并验证了一种简单、经济高效且快速的毛细管电泳方法，该方法采用二极管阵列检测（DAD）同时测定血清中的对乙酰氨基酚（扑热息痛）和五氧脯氨酸。该方法非常适合临床毒理学实验室诊断，能够快速定量对乙酰氨基酚过量病例中导致酸中毒的有机酸5-氧代脯氨酸。该方法的校准依赖性在1.3-250μg/mL范围内被证明是线性的，具有足够的准确度（96.4-107.8%）和精密度（12.3%）。对乙酰氨基酚的定量下限为1.3μg/mL，5-氧代脯氨酸为4.9μg/mL。
