Development of a method for the quantitative determination of ritonavir and lopinavir in blood plasma
https://doi.org/10.52420/2071-5943-2023-22-6-50-58
Abstract
Introduction. Therapeutic monitoring of antiretroviral drugs (ARVs) is necessary in routine practice, both to objectively monitor adherence and to ensure a personalized approach to patient treatment. Most methods of quantitative determination of ARVP, in particular, ritonavir and lopinavir, are carried out using high-performance liquid chromatography with a tandem mass-selective detector (HPLC-MS/MS) or in combination with UV detection (HPLC-UV), with a single quadrupole detector (HPLC-MS). However, these methods provided for the use of a large range of expensive solvents of a high degree of purification, a long time of analysis and sample preparation, which prevented the introduction of these methods into routine practice.
The aim of the study to evaluate the effectiveness of the developed technique for quantifying the concentration of Ritonavir and Lopinavir in blood plasma by HPLC-UV.
Materials and methods. The following drugs were used in the study:
Ritonavir and Lopinavir. Efavirenz was used for the internal standard. Randomly selected blood samples with anticoagulant from patients who did not take antiretroviral drugs were used as test samples. Chromatographic separation and detection were performed by HPLC-UV. The chromatographic separation process was controlled and the chromatographic analysis results were processed using the OpenLab CDS Chromatographic Data System software. The calibration curves were calculated using a linear regression mathematical model based on the least square’s method. Statistical data processing was carried out using the software product “Statistica v.12” (StatSoft Russia).
Results. The linear range of plasma concentration determination was 1-50 µg/ml for Ritonavir and 2-100 µg/ml for Lopinavir. Due to the short retention time – less than 7 minutes, the technique allows you to examine up to 7.5 samples per hour on one device.
Discussion. Most of the methods of quantitative determination of Ritonavir and Lopinavir for therapeutic monitoring of ART are carried out using HPLC-MS/MS or HPLC-UV, however, all of them provided for the use of a large range of expensive solvents of a high degree of purification, a long time of analysis and sample preparation, which prevented the introduction of these methods into routine practice.
Conclusion. The developed technique for quantifying the concentration of Ritonavir and Lopinavir in blood plasma using high-performance liquid chromatography with an ultraviolet detector (HPLC-UV) has a sufficiently wide linear range of concentration determination and high performance for further validation and implementation into clinical practice.
Keywords
Supporting agencies: The research was financed by a subsidy for the implementation of research (registration number in the EGISU NIOKTR 121041500042-8) clause 1.2.1 of the sectoral program of Rospotrebnadzor for 2021-2025. “Scientific support of epidemiological surveillance and sanitary protection of the territory of the Russian Federation. Creation of new technologies, means and methods of control and prevention of infectious and parasitic diseases”
About the Authors
P. M. Manakov
Bureau of Forensic Medical Examination
Russian Federation
Russian Federation
Pavel M. Manakov, chemist-expert of a medical organization
Ekaterinburg
O. P. Kolesov
Bureau of Forensic Medical Examination
Russian Federation
Russian Federation
Oleg P. Kolesov, deputy head, forensic medical expert of the highest category, thanatologist
Ekaterinburg
I. L. Tikhonova
Ural State Medical University
Russian Federation
Russian Federation
Irina L. Tikhonova, Candidate of Chemical Sciences, Associate
Professor
Department of General Chemistry
Ekaterinburg
M. V. Piterskiy
Federal Scientific Research Institute of Viral Infections “Virome” Rospotrebnadzor
Russian Federation
Russian Federation
Mikhail V. Piterskiy, researcher
Ural Federal District AIDS Centre
Ekaterinburg
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Review
For citations:
Manakov PM, Kolesov OP, Tikhonova IL, Piterskiy MV. Development of a method for the quantitative determination of ritonavir and lopinavir in blood plasma. Ural Medical Journal. 2023;22(6):50-58. (In Russ.) https://doi.org/10.52420/2071-5943-2023-22-6-50-58
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