Derivatization-based quadruplex LC/ESI-MS/MS method for high throughput quantification of serum...

Area of Science:

• Analytical Chemistry
• Clinical Chemistry
• Mass Spectrometry

Background:

• Dehydroepiandrosterone sulfate (DHEAS) quantification aids in diagnosing various diseases.
• Immunoassays lack specificity for DHEAS quantification, while LC/ESI-MS/MS offers high specificity but requires throughput improvement.

Purpose of the Study:

• To develop a quadruplex LC/ESI-MS/MS method for simultaneous quantification of DHEAS in four serum samples.
• To enhance the analysis throughput for DHEAS measurement using sample-multiplexing.

Main Methods:

• Developed a quadruplex LC/ESI-MS/MS method involving deproteinization and derivatization of four serum samples with Girard reagents.
• Mixed the derivatized samples and analyzed them in a single LC/ESI-MS/MS run.
• Evaluated method performance using 36 serum samples from healthy subjects across nine batches.

Main Results:

• Achieved a limit of quantitation of 0.050 μg/ml, suitable for clinical laboratory use.
• Demonstrated high precision (intra- and inter-assay RSDs ≤ 3.6%) and accuracy (94.4-108.1%).
• The method proved robust against matrix effects and reduced analysis time by approximately 60% compared to conventional methods.

Conclusions:

• The developed quadruplex LC/ESI-MS/MS method is sensitive, precise, accurate, and robust for DHEAS quantification.
• Sample-multiplexing significantly enhances analysis throughput, making it advantageous for clinical laboratory applications.
• This method offers a valuable tool for efficient and reliable DHEAS measurement in diagnosing diseases.