Scientific article
Open access

Internal calibration as an emerging approach for endogenous analyte quantification: Application to steroids

Published inTalanta, vol. 240, 123149
Publication date2022-04
First online date2021-12-15

The use of mass spectrometry methods with triple quadrupole instruments is well established for quantification. However, the preparation of calibration curves can be time-consuming and prone to analytical errors. In this study, an innovative internal calibration (IC) approach using a one-standard calibration with a stable isotope-labeled (SIL) standard version of the endogenous compound was developed. To ensure optimal quantitative performance, the following parameters were evaluated: the stability of the analyte-to-SIL response factor (RF), the chemical and isotopic purities of the SIL, and the instrumental reproducibility. Using six clinically important endogenous steroids and their respective SIL standards, we demonstrated that RFs obtained on different LC-MS platforms were consistent. The quantitative performance of the proposed approach was determined using quality control samples prepared in depleted serum, and showed both satisfactory precision (1.3%–12.4%) and trueness (77.5%–107.0%, with only 3 values outside ±30%). The developed method was then applied to human serum samples, and the results were similar to those obtained with the conventional quantification approach based on external calibration: the Passing-Bablok regression showed a proportional bias of 6.8% and a mean difference of −5.9% between the two methodologies. Finally, we showed that the naturally occurring isotopes of the SIL can be used to provide additional calibration points and increase the accuracy for analytes with low concentrations.

  • Internal calibration
  • Multiple isotopologue reaction monitoring
  • Quantification
  • Response factor
  • Steroids
Citation (ISO format)
VISCONTI, Gioele et al. Internal calibration as an emerging approach for endogenous analyte quantification: Application to steroids. In: Talanta, 2022, vol. 240, p. 123149. doi: 10.1016/j.talanta.2021.123149
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Article (Published version)
ISSN of the journal0039-9140

Technical informations

Creation01/14/2022 6:09:00 AM
First validation01/14/2022 6:09:00 AM
Update time03/16/2023 2:22:45 AM
Status update03/16/2023 2:22:45 AM
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