A Study of Bromine Speciation in Human Serum and Ambroxol Determination in Rat Plasma by Liquid Chromatography–Inductively Coupled Plasma Mass Spectrometry
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In this study, we employed liquid chromatography–inductively coupled plasma mass spectrometry (LC–ICP-MS) to explore bromine bioforms in human serum. Three bromine species were found by size exclusion chromatography–inductively coupled plasma mass spectrometry (SEC–ICP-MS), and one of the most abundant species was identified as Br− by anion exchange chromatography–inductively coupled plasma mass spectrometry (IC–ICP-MS).The quantitative detection of Br− was performed by SEC–ICP-MS, and the total bromine was detected by ICP-MS. Results for the determination of Br− and total bromine in human serum revealed that Br− is the major form of bromine in human serum, with a ratio ranging from 57.3 to 68.8%. Furthermore, we developed a method for the determination of ambroxol based on reversed phase high-performance liquid chromatography–inductively coupled plasma mass spectrometry (RP-HPLC–ICP-MS), and then applied it in rat plasma analysis. After optimization of the RP-HPLC–ICP-MS parameters and pre-treatment procedures, the retention time of ambroxol was 4.6 min and the detection limit was 7.0 μg L−1. The average recovery was 85.4%, with relative standard deviation of 8.4%. The above results showed that the RP-HPLC–ICP-MS method possessed the advantages of low matrix interference and time efficiency, which were better than those reported for traditional RP-HPLC methods. Therefore, this new method for the detection of ambroxol based on RP-HPLC–ICP-MS was certainly a good tool for ambroxol analysis.
KeywordsBromine speciation Human serum Rat plasma Ambroxol Liquid chromatography–inductively coupled plasma mass spectrometry
We gratefully acknowledge the financial support from the Jiangsu Provincial Medical Youth Talent (QNRC2016540), and the Key Project and Open Research Fund of State Key Laboratory of Bioelectronics, Southeast University. This study was also supported by the Jiangsu Provincial Commission of Health (CXTDB2017012), the National Natural Science Foundation of China (NSFC91743205, NSFC81773479), and the Jiangsu Social Development Project (BE2018745).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no competing interests.
All biological sample experiments in this study were carried out in accordance with institutional and national guidelines for the care and use of human serum and laboratory animals. The human serum experiment was a project of the Chinese National Bio-monitoring, and the ethical review was approved by the National Institute of Environmental Health, Chinese Center for Disease Control and Prevention, No. 201814. The ethical review of the in vitro and in vivo rat plasma experiments was approved by the Jiangsu Center for Disease Control and Prevention, No. 2018013.
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