Development of a multi-matrix LC–MS/MS method for urea quantitation and its application in human respiratory disease studies
| Autor: | Tracy Staton, Meire Bremer, Stephanie Cape, Yang Gao, Dennis Milanowski, Xiao Ding, Fang Cai, Jianshuang Wang, Brian Dean, Drew Dorshorst |
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| Rok vydání: | 2017 |
| Předmět: |
Analyte
Bioanalysis Respiratory Tract Diseases Clinical Biochemistry Pharmaceutical Science Buffers Tandem mass spectrometry 01 natural sciences Analytical Chemistry 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Tandem Mass Spectrometry Drug Discovery Humans Urea Carbon Radioisotopes Chromatography High Pressure Liquid Spectroscopy Active ingredient Chromatography Nitrogen Isotopes Elution Hydrophilic interaction chromatography 010401 analytical chemistry Extraction (chemistry) Reproducibility of Results Culture Media 0104 chemical sciences 030228 respiratory system chemistry |
| Zdroj: | Journal of Pharmaceutical and Biomedical Analysis. 133:96-104 |
| ISSN: | 0731-7085 |
| Popis: | In human respiratory disease studies, liquid samples such as nasal secretion (NS), lung epithelial lining fluid (ELF), or upper airway mucosal lining fluid (MLF) are frequently collected, but their volumes often remain unknown. The lack of volume information makes it hard to estimate the actual concentration of recovered active pharmaceutical ingredient or biomarkers. Urea has been proposed to serve as a sample volume marker because it can freely diffuse through most body compartments and is less affected by disease states. Here, we report an easy and reliable LC-MS/MS method for cross-matrix measurement of urea in serum, plasma, universal transfer medium (UTM), synthetic absorptive matrix elution buffer 1 (SAMe1) and synthetic absorptive matrix elution buffer 2 (SAMe2) which are commonly sampled in human respiratory disease studies. The method uses two stable-isotope-labeled urea isotopologues, [15N2]-urea and [13C,15N2]-urea, as the surrogate analyte and the internal standard, respectively. This approach provides the best measurement consistency across different matrices. The analyte extraction was individually optimized in each matrix. Specifically in UTM, SAMe1 and SAMe2, the unique salting-out assisted liquid-liquid extraction (SALLE) not only dramatically reduces the matrix interferences but also improves the assay recovery. The use of an HILIC column largely increases the analyte retention. The typical run time is 3.6min which allows for high throughput analysis. |
| Databáze: | OpenAIRE |
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