Department of Clinical Pharmacy and Pharmacology, University of Groningen, University Medical Center Groningen, Hanzeplein 1, 9713 GZ Groningen, The Netherlands; Department of Nephrology, University of Groningen, University Medical Center Groningen, Hanzeplein 1, 9713 GZ Groningen, The Netherlands.
ICON Bioanalytical Laboratories, Amerikaweg 18, 9407 TK Assen, The Netherlands.
J Chromatogr B Analyt Technol Biomed Life Sci. 2024 Oct 15;1247:124329. doi: 10.1016/j.jchromb.2024.124329. Epub 2024 Sep 28.
A novel liquid chromatography-tandem mass spectrometry method is described for the quantitative determination of the kidney function markers iothalamate and hippuran in human serum and urine. It is based on protein precipitation with methanol followed by dilution of the supernatant for serum and simple dilution for urine. The polar analytes are chromatographically separated by a 6.5-min gradient on a low-ligand density reversed-phase column; detection is performed by electrospray ionization tandem mass spectrometry in the positive ion mode against stable-isotope labeled internal standards. The results of a thorough method validation show that iothalamate and hippuran can be simultaneously quantified in the concentration ranges 0.500-30.0 ng/mL and 10.0-5000 ng/mL for serum and urine, respectively, with values for CV and absolute bias not exceeding 10 %, and with sufficient stability in all relevant matrices and solvents. The method was successfully applied for the analysis of serum and urine samples of multiple individuals who received both iothalamate and hippuran.
一种新的液相色谱-串联质谱法被描述用于定量测定人血清和尿液中的肾功能标志物碘海醇和 hippuran。它基于用甲醇进行蛋白质沉淀，然后对血清进行上清液稀释，对尿液进行简单稀释。极性分析物在低配体密度反相柱上通过 6.5 分钟梯度进行色谱分离；通过电喷雾电离串联质谱在正离子模式下针对稳定同位素标记的内标进行检测。彻底的方法验证结果表明，碘海醇和 hippuran 可以在血清和尿液中分别为 0.500-30.0ng/mL 和 10.0-5000ng/mL 的浓度范围内同时定量，CV 和绝对偏差值不超过 10%，并且在所有相关基质和溶剂中都具有足够的稳定性。该方法成功地应用于接受碘海醇和 hippuran 的多个个体的血清和尿液样本的分析。
