Messina Maria Anna, Maugeri Ludovica, Forte Giuseppe, Ruggieri Martino, Petralia Salvatore
Expanded Newborn Screening Laboratory, A.O.U Policlinico "G. Rodolico-San Marco", Catania, Italy.
Department of Drug and Health Sciences, University of Catania, Catania, Italy.
Front Chem. 2023 Apr 10;11:1164014. doi: 10.3389/fchem.2023.1164014. eCollection 2023.
The accurate monitoring of phenylalanine concentration plays a prominent role in the treatment of phenylketonuria (PKU). In this study, we present an enzymatic assay based on Phenylalanine Dehydrogenase/NAD and tris (bipyridine) Ruthenium (II/III) as a colorimetric mediator for the detection of Phenylalanine concentration. The amount of amino acid was quantitatively recognized by optical absorption measurements at 452 nm through the conversion of Ru (byp) to Ru (byp) , which is induced by the neoformed NADH. A detection limit of 0.33 µM, a limit of quantification of 1.01 µM, and a sensitivity of 36.6 a.u nM were obtained. The proposed method was successfully tested using biological specimens from patients affected by hyperphenylalaninemia. The proposed enzymatic assay showed a high selectivity, making it a promising alternative for the development of versatile assays for the detection of phenylalanine in diluted serums.
苯丙氨酸浓度的精确监测在苯丙酮尿症（PKU）的治疗中起着重要作用。在本研究中，我们提出了一种基于苯丙氨酸脱氢酶/NAD和三联吡啶钌（II/III）作为比色介质的酶促测定法，用于检测苯丙氨酸浓度。通过新生成的NADH诱导Ru（byp）转化为Ru（byp），在452 nm处通过光吸收测量对氨基酸量进行定量识别。获得了0.33 µM的检测限、1.01 µM的定量限和36.6 a.u nM的灵敏度。使用高苯丙氨酸血症患者的生物样本成功测试了所提出的方法。所提出的酶促测定法显示出高选择性，使其成为开发用于检测稀释血清中苯丙氨酸的通用测定法的有前途的替代方法。
