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27.11.2022 | Original Article

Rapid quantification of phenobarbital and barbital in human whole blood by liquid–liquid extraction combined with DART-orbitrap-HRMS

verfasst von: Shi Ke, Ru Lian, Rong Wang, Yulan Rao, Chen Liang, Jianying Liang, Yurong Zhang

Erschienen in: Forensic Toxicology | Ausgabe 1/2023

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Abstract

Purpose

This study aims to develop and validate a rapid, simple, and efficient bioanalytical method for the simultaneous quantification of phenobarbital and barbital in human whole blood using liquid–liquid extraction combined with direct analysis in real time (DART) and high-resolution mass spectrometry (HRMS).

Method

Phenobarbital-d5 and aprobarbital were selected as internal standards (ISs) of phenobarbital and barbital, respectively. A mixed solvent of o-xylene and ethyl acetate at a ratio of 1:6 was used to extract analytes of interest and ISs from 100 μL of human whole blood samples. Phenobarbital and barbital were detected by DART-HRMS. The proposed method has been validated in accordance with United States Food and Drug Administration Guidelines for Bioanalytical Method Validation in terms of selectivity, linearity, accuracy, precision, matrix effect, recovery, stability, and dilution integrity.

Results

The lower limits of quantification (LLOQs) of phenobarbital and barbital were both 10 ng/mL. The linearities were in the range of 10–1000 ng/mL (R2 ≥ 0.99). The mean recovery values of phenobarbital and barbital were 99.7% and 88.1%, respectively. The interday and intraday precision values were less than 10.4%, and the interday and intraday accuracy values ranged from 87.6 to 106.7%. Furthermore, the validated method was applied to four cases of phenobarbital poisoning at the Shanghai Institute of Forensic Science.

Conclusion

The developed and fully validated method enabled the simultaneous quantification of phenobarbital and barbital in human whole blood and was successfully applied to authentic cases.
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Metadaten
Titel
Rapid quantification of phenobarbital and barbital in human whole blood by liquid–liquid extraction combined with DART-orbitrap-HRMS
verfasst von
Shi Ke
Ru Lian
Rong Wang
Yulan Rao
Chen Liang
Jianying Liang
Yurong Zhang
Publikationsdatum
27.11.2022
Verlag
Springer Nature Singapore
Erschienen in
Forensic Toxicology / Ausgabe 1/2023
Print ISSN: 1860-8965
Elektronische ISSN: 1860-8973
DOI
https://doi.org/10.1007/s11419-022-00650-w

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