Clinical workflow solutions Chromatography columns and consumables

Significance of the Topic

Clinical research and diagnostic laboratories require analytical workflows that combine low detection limits, high specificity, and rapid turnaround times. Accurate quantitation of endogenous and xenobiotic compounds in complex biological matrices underpins decisions in toxicology, endocrinology, metabolomics, and therapeutic monitoring. Streamlined methods that integrate sample preparation, chromatographic separation, and mass spectrometric detection yield more consistent data and higher throughput, ultimately supporting improved patient care and more reliable research findings.

Objectives and Overview

This summary presents five complementary clinical workflows developed to address distinct analytical challenges:

• Metabolomics of highly polar compounds (e.g., catecholamines) in urine
• Vitamin D screening, including separation of 25-hydroxyvitamin D2, D3, and epimers
• High-sensitivity measurement of estrone and estradiol in serum
• Rapid screening of 23 drugs of abuse, with resolution of isobaric opioids
• Quantitation of alcohol biomarkers (ethyl sulfate and ethyl glucuronide) in urine

Each workflow illustrates an optimized combination of sample cleanup, chromatographic chemistry, and mass spectrometric detection tailored to its target analytes.

Methodology and Instrumentation

All workflows rely on two core operations: efficient sample preparation and UHPLC–MS(/MS) analysis.

• Sample Preparation: Protein precipitation and solid-phase extraction (SPE) using HyperSep protein crash plates, SOLA SPE plates, and SOLA HRP 96-well formats. These minimize matrix effects and reduce evaporation steps.
• Chromatography: Thermo Scientific Vanquish Flex and Horizon UHPLC systems coupled to core-enhanced columns (Accucore Biphenyl, Acclaim PA2, Hypersil GOLD PFP/aQ) to achieve high efficiency, complementary selectivity, and robustness under extended pressure.
• Detection: High-resolution Orbitrap IQ-X Tribrid MS for untargeted metabolomics; triple quadrupole platforms (TSQ Altis Plus, TSQ Quantiva™) for targeted quantitation with mass accuracy ≤5 ppm and run times under 4 minutes.
• Consumables: Direct-connect guard holders (Uniguard™), fritless SPE design, 0.2 µm UHPLC filters, and SureSTART® vials/caps to ensure sample integrity and reproducibility.

Main Results and Discussion

• Metabolomics: Polar-embedded mixed-mode C18 columns (Acclaim PA2) achieved baseline separation of catecholamines in urine with robust retention and peak shape.
• Vitamin D: SOLAµ HRP plates enabled SPE elution in 25 µL volumes. PFP columns (Hypersil GOLD) separated isobaric 25(OH)D3 and its epimer, supporting 1 ng/mL quantitation.
• Estrone/Estradiol: Biphenyl chemistry on Accucore columns provided exceptional sensitivity and specificity for low-abundance steroids in serum, compliant with CDC HoSt program criteria.
• Drugs of Abuse: Accucore Biphenyl columns resolved 23 analytes, including isobaric opioids, within a 4-minute gradient, demonstrating high throughput and minimal co-elution.
• Alcohol Biomarkers: Polar endcapped C18 (Hypersil GOLD aQ) separated ethyl sulfate/glucuronide isotopically labeled pairs, enabling robust urine testing with minimal run times.

Benefits and Practical Applications

These workflows deliver:

• Enhanced robustness and reproducibility across complex biological samples.
• Shortened sample preparation times and elimination of evaporation steps.
• High throughput with fast gradients (<4 minutes) without sacrificing resolution.
• Lower detection limits suitable for trace-level endogenous and exogenous compounds.
• Cost-efficient consumables and universal manifolds compatible with automation.

Applications span clinical research, forensic toxicology, therapeutic drug monitoring, endocrinology studies, and high-throughput metabolic profiling.

Future Trends and Potential Applications

Emerging developments include integration of microflow and nano-LC to further reduce solvent consumption, automated SPE-UHPLC platforms for unattended high-throughput screening, and expanded use of hybrid high-resolution/triple quadrupole workflows for multiplexed panels. Advances in stationary phase chemistries will continue to address ever more challenging isomeric separations, while machine-learning software may optimize method development and data processing workflows.

Conclusion

Thermo Fisher Scientific’s complete clinical workflow solutions demonstrate a cohesive strategy to tackle diverse analytical needs in clinical and forensic laboratories. By harmonizing advanced sample preparation, innovative column chemistries, and sensitive mass spectrometry, these methods achieve rapid, reliable, and high-sensitivity analyses. Adoption of these optimized protocols can elevate laboratory throughput, data quality, and confidence in decision-making processes across multiple application areas.