Sist Paola, Tramer Federica, Urbani Ranieri, Bandiera Antonella, Passamonti Sabina
Department of Life Sciences, University of Trieste, Italy.
Department of Chemical and Pharmaceutical Sciences, University of Trieste, Italy.
MethodsX. 2024 Dec 20;14:103123. doi: 10.1016/j.mex.2024.103123. eCollection 2025 Jun.
Bilirubin (BR) is the product of cellular heme catabolism and the main bile pigment in animal blood. It is an established biomarker for hemolysis and liver function. Over the last decade, mild hyperbilirubinemia has been shown to be a biomarker for a lower risk of cardiovascular disease, due to its antioxidant and anti-inflammatory effects. In order to use bilirubin as a predictive biomarker, new powerful methods for its direct analysis in human blood are currently being developed. To harmonize the different methods, it is essential to use high-quality BR standard solutions for assay calibration. We present here a protocol for the preparation of stable standard solutions in the range of 10- 10M BR at pH 7.4 that can facilitate a uniform approach for assay calibration without or with a sample preparation step.•The bilirubin standard solutions are prepared in buffered saline solution at physiological pH (not in alkali, not in apolar solvents) added with BSA•The standard solutions are in a wide range of concentrations.•The preparation has a quality control procedure based on direct analysis of bilirubin UV-VIS spectra or fluorescence emission of the its complex with the recombinant fusion protein HELP-UnaG (HUG).
胆红素（BR）是细胞血红素分解代谢的产物，也是动物血液中的主要胆汁色素。它是溶血和肝功能的既定生物标志物。在过去十年中，由于其抗氧化和抗炎作用，轻度高胆红素血症已被证明是心血管疾病风险较低的生物标志物。为了将胆红素用作预测性生物标志物，目前正在开发用于在人血液中直接分析它的新的强大方法。为了使不同方法统一，使用高质量的BR标准溶液进行测定校准至关重要。我们在此介绍一种在pH 7.4条件下制备10 - 10M BR范围内稳定标准溶液的方案，该方案可以促进在不进行或进行样品制备步骤的情况下进行统一的测定校准方法。
•胆红素标准溶液在添加了牛血清白蛋白（BSA）的生理pH缓冲盐溶液中制备（不在碱性溶液中，不在非极性溶剂中）
•标准溶液具有广泛的浓度范围。
•该制备方法有基于胆红素紫外可见光谱直接分析或其与重组融合蛋白HELP-UnaG（HUG）复合物荧光发射的质量控制程序。
