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High-Throughput Serum 25-Hydroxy Vitamin D Testing with Automated Sample Preparation.

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  • 1Toxicology/Mass Spectrometry Laboratory, Center for Advanced Laboratory Medicine, University of California San Diego, 10300 Campus Point Drive, Room 1314, San Diego, CA, 92121, USA. judy.stone@ucsd.edu.

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Summary

This study details an automated workflow for high-throughput liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis of serum samples. The optimized method significantly increases sample processing capacity for clinical diagnostics.

Keywords:
25-Hydroxy vitamin DAutomated data managementAutomated liquid handlingHigh-throughputLC multiplexing

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Area of Science:

  • Clinical Chemistry
  • Analytical Chemistry
  • Biotechnology

Background:

  • High-throughput analysis of serum samples is crucial for clinical diagnostics.
  • Existing methods often face bottlenecks in sample preparation and analysis.
  • Automation and multiplexing are key to increasing laboratory throughput.

Purpose of the Study:

  • To develop and validate an automated, high-throughput workflow for serum sample preparation and LC-MS/MS analysis.
  • To optimize sample processing for enhanced efficiency and capacity in a clinical laboratory setting.
  • To integrate automated liquid handling with multiplexed LC-MS/MS for streamlined analysis.

Main Methods:

  • Serum samples were processed using automated liquid handlers (ALHs) for reagent addition and precipitation.
  • Solid-phase extraction (SPE) was performed using a 96-channel ALH to remove interfering substances.
  • Samples were analyzed via multiplexed liquid chromatography-tandem mass spectrometry (LC-MS/MS) utilizing dual-channel Shimadzu LC stacks and an AB Sciex API-5000 MS/MS system.

Main Results:

  • The automated workflow demonstrated a high sample extraction capacity of 2500 samples per day by two analysts.
  • LC-MS/MS analysis achieved a throughput of approximately 44 samples per hour per system with dual-channel multiplexing.
  • The integrated system, from automated sample preparation to data analysis, significantly reduced turnaround time.

Conclusions:

  • The developed automated LC-MS/MS workflow provides a robust and efficient solution for high-throughput serum sample analysis.
  • This method enhances laboratory capacity, enabling faster and more comprehensive clinical testing.
  • The integration of ALHs and multiplexed LC-MS/MS represents a significant advancement in clinical laboratory automation.