Institute of Clinical Pharmacy and Pharmacotherapy, Heinrich Heine University, Duesseldorf, Germany.
Institute of Clinical Pharmacy and Pharmacotherapy, Heinrich Heine University, Duesseldorf, Germany.
Steroids. 2019 Aug;148:73-81. doi: 10.1016/j.steroids.2019.04.009. Epub 2019 May 8.
As part of the renin-angiotensin-aldosterone system (RAAS), aldosterone is key to the pathology of cardiovascular and renal diseases, leading to end-organ damage and cardiovascular death. Because of different aetiology and metabolism, pharmacotherapy in adults shows only limited transferability to children. Comprehensive investigations of humoral parameters, their precursors, and metabolites are necessary to establish a more rational and safe therapy in children. The LENA (Labeling of Enalapril from Neonates up to Adolescents) project aims to generate these missing data in neonates up to adolescents and provide insight into the maturing RAAS.
A HRMS (high-resolution mass spectrometry) assay was developed, utilizing blank serum depleted of the endogenous aldosterone, its precursor, 18-hydroxycorticosterone, and its main metabolite, tetrahydroaldosterone. A TOF-MS (time-of-flight-mass spectrometry) scan run in parallel with the simultaneous determination of all three analytes enriches the acquired data. Validation of aldosterone was conducted according to EMA and FDA bioanalytical guidelines.
Using the Sciex TripleTOF 6600, a reliable determination in 50 µL serum was successfully shown. Appropriate calibration ranges from 19.53 pg/mL for aldosterone, 39.06 pg/mL for 18-hydroxycorticosterone, and 78.13 pg/mL for tetrahydroaldosterone to 2500 pg/mL were established to ensure the applicability in diseased paediatric patients. Between-run accuracy and precision for aldosterone ranged between -1.21 and -6.99 % and 2.07 and -10.22 %, respectively, confirming compliance with international guidelines.
A simultaneous bioanalytical LC-HRMS assay for the determination of the biomarker aldosterone, its precursor, and main metabolite, utilizing 50 µL serum, was successfully established. This assay facilitates insight into the maturing RAAS from neonates up to adolescents.
醛固酮作为肾素-血管紧张素-醛固酮系统（RAAS）的一部分，是心血管和肾脏疾病病理生理学的关键，导致终末器官损伤和心血管死亡。由于发病机制和代谢不同，成人的药物治疗仅在有限程度上可转移到儿童身上。全面研究体液参数、其前体和代谢物对于在儿童中建立更合理和安全的治疗方法是必要的。LENA（从新生儿到青少年的依那普利标签）项目旨在生成新生儿至青少年的这些缺失数据，并深入了解不断成熟的 RAAS。
开发了一种 HRMS（高分辨率质谱）测定法，利用空白血清去除内源性醛固酮、其前体 18-羟基皮质酮及其主要代谢物四氢醛固酮。与同时测定所有三种分析物的 TOF-MS（飞行时间质谱）扫描并行，可丰富获得的数据。根据 EMA 和 FDA 生物分析指南对醛固酮进行验证。
使用 Sciex TripleTOF 6600，成功地在 50 µL 血清中进行了可靠的测定。建立了适当的校准范围，从醛固酮的 19.53 pg/mL、18-羟基皮质酮的 39.06 pg/mL 和四氢醛固酮的 78.13 pg/mL 到 2500 pg/mL，以确保在患病儿科患者中的适用性。醛固酮的批间精密度和准确度范围分别为-1.21%至-6.99%和 2.07%至-10.22%，证实符合国际指南。
成功建立了一种同时测定生物标志物醛固酮、其前体和主要代谢物的 LC-HRMS 分析方法，利用 50 µL 血清，该方法有助于深入了解从新生儿到青少年的不断成熟的 RAAS。
