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<p>The study aimed to develop and validate an isotope-dilution liquid chromatography–tandem mass spectrometry (ID-LC–MS/MS) method as a candidate reference measurement procedure (RMP) for the determination of theophylline in human serum. Analytical performances (selectivity, matrix effects, linearity, trueness and precision, etc.) of the ID-LC–MS/MS method were rigorously evaluated. Sample preparation was performed using a simple protein precipitation (PPT) approach, and chromatographic separation was achieved on a pentafluorophenyl column. Under optimal conditions, theophylline showed excellent linearity over the concentration range from 3.43 to 336.98&#xa0;μmol/L (correlation coefficient: 0.9998). The average spiked recoveries ranged from 98.2 to 101.2%, and the measurement bias was less than 1.3% compared with existing reference measurement procedures using the same RELA samples. The method demonstrated high sensitivity with a limit of detection (LOD) of 0.10&#xa0;μmol/L and a limit of quantification (LOQ) of 0.31&#xa0;μmol/L. The intra-day and inter-day coefficients of variation (CV) were 0.37 to 1.63% and 1.19 to 1.93%, respectively. The established ID-LC–MS/MS method is accurate, precise, robust, and features simple sample preparation and short analysis time. It shows promise as a candidate RMP for theophylline determination and is applicable for measurement traceability and calibration of routine detection systems.</p>

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An isotope-dilution liquid chromatography–tandem mass spectrometry as a candidate reference method for the measurement of theophylline in human serum

  • 期刊论文

  • 发表日期：

    2026年7月24日

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  • Jing Lin,
  • Xiaodong Yang,
  • Yongnan Jin,
  • Jing Zhou,
  • Minmin Tu,
  • Qijun Wang,
  • Min Shen

摘要

The study aimed to develop and validate an isotope-dilution liquid chromatography–tandem mass spectrometry (ID-LC–MS/MS) method as a candidate reference measurement procedure (RMP) for the determination of theophylline in human serum. Analytical performances (selectivity, matrix effects, linearity, trueness and precision, etc.) of the ID-LC–MS/MS method were rigorously evaluated. Sample preparation was performed using a simple protein precipitation (PPT) approach, and chromatographic separation was achieved on a pentafluorophenyl column. Under optimal conditions, theophylline showed excellent linearity over the concentration range from 3.43 to 336.98 μmol/L (correlation coefficient: 0.9998). The average spiked recoveries ranged from 98.2 to 101.2%, and the measurement bias was less than 1.3% compared with existing reference measurement procedures using the same RELA samples. The method demonstrated high sensitivity with a limit of detection (LOD) of 0.10 μmol/L and a limit of quantification (LOQ) of 0.31 μmol/L. The intra-day and inter-day coefficients of variation (CV) were 0.37 to 1.63% and 1.19 to 1.93%, respectively. The established ID-LC–MS/MS method is accurate, precise, robust, and features simple sample preparation and short analysis time. It shows promise as a candidate RMP for theophylline determination and is applicable for measurement traceability and calibration of routine detection systems.