high-throughput-comprehensive-drug-surveillance-using-multisegment

High-throughput and Comprehensive Drug Surveillance Using Multisegment Injection-Capillary Electrophoresis-Mass Spectrometry

New analytical methods are urgently needed to enable high-throughput, yet comprehensive drug screening, given an alarming opioid and prescription drug ...

McMaster University

A collaborative study on the robustness and portability of a capillary electrophoresis-mass spectrometry method for peptide mapping was performed by an international team, consisting of 13 independent laboratories from academia and industry. All participants used the same batch of samples, reagents and coated capillaries to run their assays, whereas they utilized the capillary electrophoresis-mass spectrometry equipment available in their laboratories. The equipment used varied in model, type and instrument manufacturer. Furthermore, different types of sheath-flow capillary electrophoresis-mass spectrometry interfaces were used. Migration time, peak height and peak area of ten representative target peptides of trypsin-digested bovine serum albumin were determined by every laboratory on two consecutive days. The data were critically evaluated to identify outliers and final values for means, repeatability (precision within a laboratory) and reproducibility (precision between laboratories) were established. For relative migration time the repeatability was between 0.05 and 0.18% RSD and the reproducibility between 0.14 and 1.3% RSD. For relative peak area repeatability and reproducibility values obtained were 3-12 and 9-29% RSD, respectively. These results demonstrate that capillary electrophoresis-mass spectrometry is robust enough to allow a method transfer across multiple laboratories and should promote a more widespread use of peptide mapping and other capillary electrophoresis-mass spectrometry applications in biopharmaceutical analysis and related fields.

Interlaboratory study to evaluate the robustness of capillary electrophoresis-mass spectrometry for peptide mapping.

Urinary 1-hydroxypyrene (OH-Pyr) is widely used for biomonitoring human exposures to polycyclic aromatic hydrocarbons (PAHs) from air pollution and tobacco smoke. However, there have been few rigorous validation studies reported to ensure reliable OH-Pyr determination for occupational health and risk assessment. Herein, we report an inter-laboratory method comparison for urinary OH-Pyr when using gas chromatography-high-resolution mass spectrometry (GC-HRMS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) on urine specimens collected from firefighters (n = 42) deployed at the 2016 Fort McMurray wildfire. Overall, there was good mutual agreement in urinary OH-Pyr quantification following enzyme deconjugation with an average bias of 39% with no significant deviation from linearity (slope = 1.36; p > 0.05), whereas technical precision (< 12%) and average recovery (> 85%) were acceptable when using a stable-isotope internal standard. Faster analysis times (4 min) were achieved by LC-MS/MS without chemical derivatization, whereas lower detection limits (0.64 ng/L, S/N = 3) was realized with solid-phase extraction prior to GC-HRMS. A median creatinine normalized OH-Pyr concentration of 128 ng/g was measured for firefighters that were below the recommended biological exposure index due to delays between early stages of emergency firefighting and urine sample collection. Similar outcomes were also measured for 3-hydroxyphenanthrene and 9-hydroxyfluorene that were positively correlated with urinary OH-Pyr (p < 0.05), implying similar uptake, distribution, and liver biotransformation processes. Optimal specimen collection strategies post-deployment together with standardized protocols for OH-PAH analysis are critical to accurately evaluate smoke exposure in firefighters, including experimental conditions to ensure quantitative enzyme hydrolysis of urine samples. Graphical abstract.

Urinary hydroxypyrene determination for biomonitoring of firefighters deployed at the Fort McMurray wildfire: an inter-laboratory method comparison.

Expanded newborn screening of inborn errors of metabolism (IEM) based on tandem mass spectrometry (MS/MS) technology is one of the most successful preventative healthcare initiatives for presymptomatic diagnosis and treatment of rare yet treatable genetic diseases in the population. However, confirmatory testing of presumptive screen-positive cases is required using high efficiency separations for improved specificity in order to improve the positive predictive value (PPV) for certain classes of IEMs. Here, we describe recent advances using capillary electrophoresis-mass spectrometry (CE-MS) for reliable second-tier screening or confirmatory testing based on targeted analysis of amino acids, acylcarnitines, nucleosides, and other classes of polar metabolites associated with IEMs. Additionally, nontargeted metabolite profiling enables the identification of unknown biomarkers of clinical significance for other genetic diseases that are currently screened by bioassays and/or mutation panels, such as cystic fibrosis (CF). Noteworthy, CE-MS allows for resolution of isobaric/isomeric interferences without complicated sample handling that is ideal when analyzing volume-restricted biospecimens from neonates/infants, including dried blood spots and sweat specimens. New developments to improve concentration sensitivity, as well as enhance sample throughput and quality control for unambiguous confirmatory testing of IEMs will also be discussed when using multiplexed separations based on multisegment injection-CE-MS.

Article	Year
Interlaboratory study to evaluate the robustness of capillary electrophoresis-mass spectrometry for peptide mapping. Journal of separation science\| PubMed ID: 26147246	2015
Urinary hydroxypyrene determination for biomonitoring of firefighters deployed at the Fort McMurray wildfire: an inter-laboratory method comparison. Analytical and bioanalytical chemistry\| PubMed ID: 30683964	2019
New Advances for Newborn Screening of Inborn Errors of Metabolism by Capillary Electrophoresis-Mass Spectrometry (CE-MS). Methods in molecular biology (Clifton, N.J.)\| PubMed ID: 30847789	2019

Meera Shanmuganathan

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