ORIGINAL RESEARCH article
Front. Chem.
Sec. Analytical Chemistry
Volume 13 - 2025 | doi: 10.3389/fchem.2025.1631203
Development and validation of a novel UPLC-Q-Orbitrap HRMS method for determining p-Nitroaniline and its metabolites in blood
Provisionally accepted
- Zhejiang Center for Disease Control and Prevention (Zhejiang CDC), Hangzhou, China
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An accurate, reliable, and sensitive method was successfully developed to simultaneously determine p-Nitroaniline (p-NA) and its metabolites in blood based on the hyphenated technology of quadrupole-orbitrap high resolution mass spectrometry (Q-Orbitrap HRMS) and ultra-performance liquid chromatography (UPLC). After the blood sample was extracted with ethyl acetate, the organic layer was taken for injection and analysis. Within the concentration range of 1 -100 μg/L, the calibration curves of p-NA and its metabolites had a good linearity, with correlation coefficient(r) values greater than 0.999. This method has excellent precision and accuracy. The intra-and inter-day coefficients of variation (CVs) were less than 9.9 % and 8.7 % respectively, and the analytical accuracy ranged from 83.1 % to 101.3 %. The lower limits of detection (LLODs) for all target analytes were between 0.6 μg/L and 2.2 μg/L, and the lower limits of quantification (LLOQs) were between 2.0 μg/L and 7.4 μg/L. In addition, the application potential of this method was verified by analyzing the blood samples of workers exposed to p-NA. The results indicated that this method was accurate, reliable and sensitive, and was applicable to the detection of p-NA and its metabolites in the blood.
Keywords: Occupational Exposure, p-Nitroaniline, Blood, UPLC-Q-Orbitrap HRMS, metabolite
Received: 19 May 2025; Accepted: 18 Aug 2025.
Copyright: © 2025 Wang, Wang, Xu, Zou, Gao, Ren and Cao. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Peng Wang, pwang@cdc.zj.cn
Yiyao Cao, yycao@cdc.zj.cn
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