Validation of an LCMS Hybrid Assay with EVOSEP Cleanup for the Quantitation of Islet Amyloid Polypeptide in Human Plasma
Islet Amyloid Polypeptide (IAPP) is a peptide hormone produced by the pancreas’ beta cells that regulates blood glucose. Research on IAPP and its role in diabetes is ongoing, and there is a need for a reliable method to accurately detect this hormone at clinically relevant levels and possibly the different forms of the peptide. We set out to validate a hybrid LCMS assay for this biomarker that could be validated to an appropriate level to support clinical studies.
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Validated Biomarker Assay for the Analysis of Coproporphyrin I and Coproporphyrin III in Human Plasma
Coproporphyrin I (CP‐I) and Coproporphyrin III (CP‐III) are potential endogenous biomarkers for hepatic organic anion transporting polypeptide (OATP)1B1/1B3 function. We developed and validated a bioanalytical assay for monitoring these biomarkers to assess OATP1B1/1B3 inhibition in place of a standalone prospective clinical drug-drug interaction (DDI) study. Currently, investigational drugs that alter the pharmacokinetics of other medications are subject to additional testing to understand how to manage a DDI risk safely. By monitoring the effect of an investigation drug on the levels of these endogenous substrates of OATP1B1/1B3 in early clinical development, the potential need for a dedicated clinical DDI study could be avoided.
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Development of a Mitra Tip Extraction Assay Coupled with LC-MSMS
Biomarkers such as L-Citrulline, L-Arginine, and L-Argininosuccinic Acid play crucial roles in various physiological processes and pathological conditions. However, their accurate quantitation presents challenges due to sample complexity and low concentrations. Here, we present a novel approach utilizing Mitra tip extraction coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the simultaneous quantitation of these biomarkers. Mouse models are known to produce small blood volumes while Mitra tips offer advantages in sample collection, allowing for convenient and reproducible extraction from biological matrices.
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A Derivatization-free LCMS Assay for 4β-hydroxycholesterol
In clinical studies, 4β-hydroxycholesterol is an important biomarker for assessing the activity of CYP3A. Numerous quantitative methods have been reported for analyzing the total amount of 4β-hydroxycholesterol (free and conjugated) after saponification and derivatization. Derivatization is often required to achieve clinically relevant assay sensitivity when utilizing LCMS for detection, but this additional step can be complex and time-consuming.
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