Clinical workflow solutions

Importance of the topic

Clinical laboratories require sensitive, robust, and high-throughput methods to analyze a wide range of biomarkers, vitamins, hormones, and drugs in complex matrices. Optimized workflows integrating sample preparation, chromatography, and mass spectrometry deliver consistent, high-quality data critical for diagnostics, therapeutic monitoring, forensic investigations, and biomarker research.

Objectives and overview

This document outlines five targeted clinical workflows—metabolomics, vitamin D screening, steroid hormone quantitation (estrone/estradiol), drugs of abuse screening, and alcohol biomarker detection. Each workflow is designed to maximize detection sensitivity and robustness while reducing analysis time and improving throughput.

Methodology and used instrumentation

• Sample Preparation: Protein precipitation (HyperSep plates), solid-phase extraction (SOLA and SOLAµ SPE plates), universal vacuum and positive-pressure manifolds.
• UHPLC Systems: Thermo Scientific Vanquish Flex and Vanquish Horizon UHPLC platforms.
• Columns: Polar embedded mixed-mode C18 (Acclaim PA2), pentafluorophenyl (Hypersil GOLD PFP), polar end-capped C18 (Hypersil GOLD aQ), biphenyl (Accucore Biphenyl), specialized GC columns (TraceGOLD).
• Mass Spectrometry: Thermo Scientific Orbitrap IQ-X Tribrid, TSQ Altis/Altis Plus, and TSQ Quantiva triple quadrupole mass spectrometers.
• Consumables: SureSTART glass vials and caps; WebSeal well plates and sealing tapes for high-throughput workflows.

Main results and discussion

• Metabolomics: Effective retention and baseline separation of highly polar catecholamines in urine on Acclaim PA2, enabling precise quantitation with Orbitrap detection.
• Vitamin D Screening: Baseline resolution of 25(OH)D2, 25(OH)D3, and its epimer using Hypersil GOLD PFP and SOLAµ HRP plates, achieving low-ng/mL sensitivity without evapora­tion steps.
• Steroid Hormones: High-sensitivity analysis of estrone and estradiol in human serum employing Accucore Biphenyl and TSQ Altis, meeting CDC HoSt program requirements.
• Drugs of Abuse: Rapid (<4 min) separation of 23 analytes, including isobaric opioids, on Accucore Biphenyl with SOLA HPR SPE, delivering robust screening in urine.
• Alcohol Biomarkers: Robust quantitation of ethyl sulfate and ethyl glucuronide in urine using Hypersil GOLD aQ and TSQ Quantiva, extending detection windows beyond ethanol measurement.

Benefits and practical applications

• Streamlined protocols reduce sample processing time and manual intervention, increasing daily throughput.
• Enhanced column chemistries and SPE formats improve analyte retention, peak shape, and matrix cleanup for reproducible results.
• Integration with advanced MS detectors ensures low limits of detection and high selectivity for clinical, forensic, and research laboratories.

Future trends and possibilities

• Integration of automated nano- and microfluidic sample processing to further reduce reagent use and increase sensitivity.
• Adoption of data-independent acquisition and multiplexed MS workflows to expand analytical panels in a single run.
• Incorporation of AI-driven data analysis and real-time monitoring to support rapid clinical decision making.
• Development of novel hybrid stationary phases targeting challenging isomers and ultra trace compounds.

Conclusion

Optimized clinical workflows that combine tailored sample preparation, advanced chromatographic separations, and sensitive MS detection achieve robust, high-throughput analyses across a wide range of biomarkers and analytes. These integrated solutions support evolving clinical and forensic demands by delivering reliable, consistent, and actionable data.

References

• Thermo Fisher Scientific application notes and technical briefs on metabolomics, vitamin D, steroid hormones, drugs of abuse, and alcohol biomarker workflows.
• CDC Hormone Standardization (HoSt) program documentation.