Shimadzu Solutions for Application Notebook - Clinical Research - Application Notebook

Importance of the Topic

Clinical and forensic laboratories face growing demands for rapid, sensitive and reliable analyses of drugs, metabolites and toxic gases in biological fluids. Traditional workflows relying on manual sample preparation and single-analyte methods can limit throughput and introduce variability. Innovative automated sample preparation and high-performance detection strategies are essential for therapeutic drug monitoring, toxicology screening, neonatal metabolite analysis and emergency gas poisoning tests.

Study Objectives and Overview

This collection of application notes presents a unified portfolio of advanced analytical workflows. The goals are to demonstrate how fully automated sample pretreatment modules, on-line SPE, two-dimensional chromatography, direct ionization and bidirectional GC detection enable:

• High-throughput, fully automated LC-MS/MS analysis of immunosuppressants, antiarrhythmics, psychoactive drugs, β-lactam antibiotics, amino acids, acylcarnitines, steroids and NSAIDs.
• Rapid 15-second screening of cyanide in blood serum by probe electrospray ionization mass spectrometry without pretreatment.
• Quantitative multi-target screening (MTS) in whole blood with library-based MS/MS confirmation for over one thousand toxic compounds.
• Sensitive CO measurement in blood by GC with a barrier discharge ionization detector.

Methodology and Instrumentation

The workflows employ Shimadzu’s CLAM-2000 automated sample preparation unit in tandem with triple-quadrupole systems (LCMS-8040, LCMS-8060) and 2D HILIC–LC-MS/MS. Key elements include:

• Online SPE with POROS R1 trapping and biphenyl or C18 analytical columns for clean-up and concentration.
• FIA/LC-MS/MS for integrated amino acid, acylcarnitine and steroid profiling from a single dried blood spot.
• Direct probe electrospray ionization (DPiMS-2020) for rapid blood serum analysis of cyanide derivatives.
• Multi-collision-energy MRM triggered MS/MS library searching for high-confidence forensic toxicology screening.
• Barrier discharge ionization GC detection for ppb-level carbon monoxide quantitation in small blood volumes.

Key Results and Discussion

All methods achieved excellent linearity over three or more orders of magnitude (r2 > 0.995), low limits of quantitation (sub-ppb for steroids, 0.7–6 ng/mL for β-lactams, 10 pg/mL for NSAIDs), and intraday precision <10% RSD. Automated CLAM-2000 protocols eliminated manual variability and increased throughput, achieving cycle times as low as 7–9 minutes per sample. The MTS approach screened over 18–60 analytes with accurate MRM quantitation and library confirmation at therapeutic to toxic ranges. The PESI method detected cyanide in 15 seconds without pretreatment. GC-BID achieved sensitive CO detection at 2 ppm, enabling miniaturized blood sample testing.

Benefits and Practical Applications of the Methods

The integrated platforms deliver:

• Reduced hands-on time and human error via full automation.
• High sample throughput for clinical research, therapeutic drug monitoring and forensic casework.
• Multiplex capability for simultaneous quantitation of diverse analytes in a single run.
• Enhanced specificity and confirmation through MS/MS library matching and multi-ion ratio monitoring.
• Minimal sample consumption for precious specimens and emergency toxicology.

Future Trends and Potential Applications

Advances may include:

• Integration with artificial intelligence and automated reporting for faster clinical decision support.
• Miniaturized point-of-care MS devices with direct sampling probes for emergency diagnostics.
• Expanded library databases and high-resolution MS for emerging designer drugs and novel biomarkers.
• Further development of two-dimensional and multi-dimensional LC-MS to tackle ultra-complex biological matrices.

Conclusion

The application notebook demonstrates that combining fully automated sample pretreatment, innovative ionization approaches and advanced chromatography–MS strategies transforms the analysis of clinical and forensic samples. These workflows deliver the speed, sensitivity and reliability required for modern therapeutic drug monitoring, toxicology screening and emergency diagnostics.

References

1. Shimadzu Application Notes: CLAM-2000 + LCMS-8040 TDM Workflows.
2. Guidance for Industry: Bioanalytical Method Validation. US FDA, 2001.
3. Quantitative Multi-Target Screening using LC-MS/MS and MS/MS Library. Shimadzu, 2016.
4. In-Source CID and Rapid Cyanide Screening with DPiMS-2020. Shimadzu, 2017.
5. Barrier Discharge Ionization GC for Carbon Monoxide in Blood. Shimadzu, 2017.