δ13C Determination of Benzoic Acid by High-Resolution Orbitrap MS: How Sample Introduction Strategy Governs Accuracy and Precision.
- 1Chemistry Institute, Federal University of Goiás, Goiânia74690-900, Brazil.
- 2UNIFAL, Universidade Federal de Alfenas, Alfenas37130-001, Minas Gerais, Brazil.
- 3PETROBRAS Research and Development Center (CENPES), PETROBRAS , Rio de Janeiro21941-915, Brazil.
Abstract
Stable carbon isotope ratio measurements (δ13C) are traditionally performed by isotope ratio mass spectrometry (IRMS), but recent advances in high-resolution Orbitrap mass spectrometry (Orbitrap MS) enable molecular-level isotope ratio analyses without conversion to simple gases. Here, we systematically evaluate and compare three sample-introduction strategies for Orbitrap-MS-based δ13C determination of benzoic acid─dual-inlet (DI), single-inlet (SI), and automated HPLC flow-injection (column-free mode)─assessing how each governs measurement accuracy and precision. Analyses were performed using paired injections of an isotopically certified benzoic acid reference (IAEA-601) and a benzoic acid sample previously characterized by elemental analysis-IRMS (EA-IRMS), following a seven-block bracketing sequence (Ref-Smp-Ref-Smp-Ref-Smp-Ref) to minimize temporal drift and to generate within-run triplicates. Across all configurations, high analytical precision was achieved, within-block relative standard error (σRSE) consistently below approximately 0.6‰. Under DI conditions, replicate means closely approached the EA-IRMS benchmark (-27.77‰), with residuals of -0.40 to +0.50‰ on the Exploris 240 and -0.58 to -2.73‰ on the Q Exactive. SI analysis enabled the acquisition of a higher number of individual scans per block, contributing to lower σRSE (approximately 0.50-0.65‰) but exhibited larger accuracy offsets (residuals up to approximately 1-3‰), indicating greater susceptibility to systematic bias. Automated HPLC injection on the Exploris 240, optimized for concentration and AGC (50 μmol L-1 and 2.5 × 105, respectively), delivered δ13C values of 28.1‰, -27.19‰, and -27.79‰ (residuals of approximately 0.20-0.45‰) with no significant differences among replicate means (ANOVA/Tukey, 95% confidence level). Collectively, these findings confirm Orbitrap-MS as a reliable alternative to IRMS for δ13C analysis of benzoic acid, and reveal that the choice of injection strategy is the primary determinant of the accuracy-precision-throughput balance achievable with this platform.
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BibTeX
@article{bevilaqua2026δ13c,
title = {{δ13C Determination of Benzoic Acid by High-Resolution Orbitrap MS: How Sample Introduction Strategy Governs Accuracy and Precision.}},
author = {Giovanni B Bevilaqua and Hugo G Machado and Júlio C O Ribeiro and Nerilson Marques Lima and Alexandre A Ferreira and Ygor S Rocha and Gabriel F Dos Santos and Boniek Gontijo},
journal = {Journal of the American Society for Mass Spectrometry},
year = {2026},
month = oct,
doi = {10.1021/jasms.6c00221},
url = {https://europepmc.org/article/MED/42916763},
note = {CC BY},
}
RIS
TY - JOUR
TI - δ13C Determination of Benzoic Acid by High-Resolution Orbitrap MS: How Sample Introduction Strategy Governs Accuracy and Precision.
AU - Giovanni B Bevilaqua
AU - Hugo G Machado
AU - Júlio C O Ribeiro
AU - Nerilson Marques Lima
AU - Alexandre A Ferreira
AU - Ygor S Rocha
AU - Gabriel F Dos Santos
AU - Boniek Gontijo
PY - 2026
DA - 2026/10/01
JO - Journal of the American Society for Mass Spectrometry
DO - 10.1021/jasms.6c00221
UR - https://europepmc.org/article/MED/42916763
AB - Stable carbon isotope ratio measurements (δ13C) are traditionally performed by isotope ratio mass spectrometry (IRMS), but recent advances in high-resolution Orbitrap mass spectrometry (Orbitrap MS) enable molecular-level isotope ratio analyses without conversion to simple gases. Here, we systematically evaluate and compare three sample-introduction strategies for Orbitrap-MS-based δ13C determination of benzoic acid─dual-inlet (DI), single-inlet (SI), and automated HPLC flow-injection (column-free mode)─assessing how each governs measurement accuracy and precision. Analyses were performed using paired injections of an isotopically certified benzoic acid reference (IAEA-601) and a benzoic acid sample previously characterized by elemental analysis-IRMS (EA-IRMS), following a seven-block bracketing sequence (Ref-Smp-Ref-Smp-Ref-Smp-Ref) to minimize temporal drift and to generate within-run triplicates. Across all configurations, high analytical precision was achieved, within-block relative standard error (σRSE) consistently below approximately 0.6‰. Under DI conditions, replicate means closely approached the EA-IRMS benchmark (-27.77‰), with residuals of -0.40 to +0.50‰ on the Exploris 240 and -0.58 to -2.73‰ on the Q Exactive. SI analysis enabled the acquisition of a higher number of individual scans per block, contributing to lower σRSE (approximately 0.50-0.65‰) but exhibited larger accuracy offsets (residuals up to approximately 1-3‰), indicating greater susceptibility to systematic bias. Automated HPLC injection on the Exploris 240, optimized for concentration and AGC (50 μmol L-1 and 2.5 × 105, respectively), delivered δ13C values of 28.1‰, -27.19‰, and -27.79‰ (residuals of approximately 0.20-0.45‰) with no significant differences among replicate means (ANOVA/Tukey, 95% confidence level). Collectively, these findings confirm Orbitrap-MS as a reliable alternative to IRMS for δ13C analysis of benzoic acid, and reveal that the choice of injection strategy is the primary determinant of the accuracy-precision-throughput balance achievable with this platform.
SN - 1044-0305
SN - 1879-1123
ER -