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Determination of Urea in Human Serum and Epithelial Lining Fluid Using LCMS; Benchmarking to a Traditional Diagnostic Colorimetric Kit

GlaxoSmithKline (GSK)
23 Jun 2014

Difficulties are common in estimating the actual concentration of recovered pharmaceutical entities in epithelial lining fluid (ELF) following a bronchoalveolar lavage (BAL) procedure due to uncertainty around existing ELF volume. Therefore, an endogenous biomarker (urea) is typically used to determine the volume of recovered ELF (relative to volume of saline instilled), after a BAL procedure. The most widely used method for urea determination in clinical laboratories utilizes a colorimetric assay kit such as QuantiChrom™, which is designed to measure urea directly in a variety of biological matrices without any sample pretreatment. Current FDA guidance requiring in-lab validation of diagnostic kit-based assays, along with a possible 40% positive bias when analyzing serum samples using a kit, make a selective and accurate LC-MS assay essential and cost-effective to explore.

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LC-MSLC-MS (liquid chromatography-mass spectrometry) systems and equipment are used for separation and quantitative analysis of complex mixtures, combining liquid chromatography and mass spectrometry. Quantify proteins, contaminants, pesticides or screen for drug metabolites with a high level of sensitivity. LC-MS systems and equipment include reverse phase, normal phase and specialized columns integrated with various MS detectors such as time-of-flight (TOF), quadrupole, orbitrap or ion trap mass analyzers. LC-MS/MS instruments equipped with a qTOF or triple quadrupole analyzer give greater sensitivity and resolving power to your analysis. Find the best LC-MS equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.ELISAEnzyme-linked immunosorbent assays (ELISA), also known as enzyme immunoassays (EIA), are used for the detection and quantification of proteins, peptides and antibodies in a sample. ELISA often comes in kit format, with pre-selected antibody pairs to detect specific proteins or biomarkers of diseases such as diabetes and obesity, or cardiovascular and neurological disorders. Detection is made possible with chemiluminescent, fluorescent or colorimetric substrates and detection instruments such as microplate / ELISA readers. Other kits available include: Enzyme-linked immunospot (ELISPOT) assays for the quantification of protein-producing cells and chemiluminescent immunoassays (CLIA) kits. Find the best ELISA kits and products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.ASMSThe American Society for Mass Spectrometry (ASMS) is an organization dedicated to advancing the science and application of mass spectrometry. ASMS supports professionals through events, publications, and research initiatives, promoting innovation in areas like proteomics, metabolomics, and environmental analysis. Human SerumHuman serum is found in blood plasma. The protein is used in a variety of applications including tissue engineering, transplantation and cell therapy.

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Determination of Urea in Human Serum and Epithelial Lining Fluid Using LCMS; Benchmarking to a Traditional Diagnostic Colorimetric Kit