Clinical Physiology of Circulation

Chief Editor

Leo A. Bockeria, MD, PhD, DSc, Professor, Academician of Russian Academy of Sciences, President of Bakoulev National Medical Research Center for Cardiovascular Surgery


Application of high performance liquid chromatography and mass spectrometry for the quantitative determination of local anesthetics in blood plasma

Authors: Chichanovskaya L.V., Popov N.S., Federyakin D.V., Belevskiy Е.V., Mayorov М.О., Zatsepin А.G.

Company:
Tver State Medical University, Tver, Russian Federation

E-mail: Сведения доступны для зарегистрированных пользователей.

DOI: https://doi.org/10.24022/1814-6910-2022-19-2-177-185

UDC: 615.382:543]:616-072

Link: Clinical Physiology of Blood Circulaiton. 2022; 2 (19): 177-185

Quote as: Chichanovskaya L.V., Popov N.S., Federyakin D.V., Belevskiy Е.V., Mayorov М.О., Zatsepin А.G. Application of high performance liquid chromatography and mass spectrometry for the quantitative determination of local anesthetics in blood plasma. Clinical Physiology of Circulation. 2022; 19 (2): 177–85 (in Russ.). DOI: 10.24022/1814-6910-2022-19-2-177-185

Received / Accepted:  04.02.2022 / 10.03.2022

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Abstract

Objective: development and validation of a method for the quantitative determination of local anesthetics in blood plasma using high performance liquid chromatography and mass spectrometry.

Material and methods. The objects of the study were lidocaine, bupivacaine and ropivacaine. Mepivacaine was used as an internal standard. The quantitative determination of drugs in blood plasma was carried out using an Agilent Technologies 1260 Infinity II high performance liquid chromatograph and an AB Sciex 3200 QTrap MD mass spectrometer. A Phenomenex Synergi 4 μm Fusion-RP 50 ×2 mm chromatographic column was used. A mixture of deionized water and acetonitrile with the addition of 0.1% formic acid was used as the mobile phase. The choice of the sample preparation method was based on the obtained data on the degree of extraction of analytes and the matrix effect. At the development stage, protein precipitation methods with acetonitrile and methanol were compared, as well as liquid extraction.

Results. Local anesthetics were detected by the corresponding MRM transitions: bupivacaine – m/z 289.1→140.2, ropivacaine – m/z 275.1→126.2, lidocaine – m/z 235.1→86.2, mepivacaine (internal standard) – m/z 247.1→98.1. The following chromatographic parameters were used: mobile phase – 70% aqueous solution of acetonitrile with the addition of 0.1% formic acid, flow rate – 300 μl/min, chromatographic column temperature – 40 °C, sample injection volume – 10 μl, total analysis time – 5 minutes. The retention times of bupivacaine, ropivacaine and lidocaine averaged 1.1 minutes. Extraction of local anesthetics from blood plasma was carried out with hexane. The analytical range of the developed method was from 1 to 1000 ng/ml for bupivacaine, ropivacaine and lidocaine. The developed technique is characterized by speed of execution, selectivity, reproducibility, and accuracy.

Conclusion. The developed method for the quantitative determination of bupivacaine, ropivacaine, and lidocaine in blood plasma fully complies with the requirements of domestic and foreign regulatory documents and can be used to conduct therapeutic drug monitoring of local anesthetics and establish individual pharmacokinetic characteristics of drugs.

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About Authors

  • Lesya V. Chichanovskaya, Dr. Med. Sci., Professor, Chief of Chair of Neurology, Rehabilitation and Neurosurgery; ORCID
  • Nikita S. Popov, Cand. Pharm. Sci., Head of Research Laboratory; ORCID
  • Denis V. Federyakin, Dr. Med. Sci., Chief of Chair of Surgery and Anesthesiology-Intensive Care; ORCID
  • Еvgeniy V. Belevskiy, Cand. Med. Sci., Assistant Professor of Chair of Surgery and Anesthesiology-Intensive Care, Head of Department of Anesthesiology-Intensive Care;
  • Мaksim О. Mayorov, Postgraduate, Anesthesiologist-Intensivist;
  • Аleksandr G. Zatsepin, Postgraduate

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