Quantitative determination of a new sidnonymine derivative in blood plasma and organs of mice using HPLC-MS/MS

Introduction. Sydnonymines are a class of heterocyclic chemical compounds with a wide range of biological activity and promising in terms of the search and development of new drugs. One of these compounds is the candidate molecule BBP2023, N-(ethoxycarbonyl)-3-(4-methylhexane-2-yl)sydnonymine. It has been determined that BBP2023 accumulates in the brain tissues of laboratory animals and causes a cerebral vasodilating effect, which makes it possible to consider this compound as a promising drug used to treat cerebral circulatory disorders.

Goal – development and validation of a bioanalytical method for the determination of a new sidnonymine derivative in various tissues of laboratory mice with a view to further application in experimental pharmacokinetic research.

Material and methods. Quantitation of BBP2023 in blood plasma and organ homogenates of mice was performed by HPLC-MS/MS. The technique of precipitation of proteins with methanol was used as the basis for biomaterial sample preparation. Chromatographic separation was performed with a mixture of deionized water and acetonitrile in a gradient mode on a Zorbax Eclipse XDB-C18 4,6×50 mm, 5 µm column. Detection was performed in the MRM mode.

Results. As a result of the experiment, a technique for sample preparation of biological material of mice was developed, optimal conditions for chromatography and detection of the studied compound were selected. The bioanalytical technique has been validated according to the following parameters: selectivity, matrix effect, recovery rate, carry-over, linearity of the calibration range, lower limit of quantitative determination, intra- and inter-run accuracy and precision, dilution factor, as well as stability of the analyte and the internal standard at the analysis stages.

Conclusion. As a result of the experiment, a method was developed for the quantitative determination of a new derivative of sydnonymine BBP2023 in blood plasma and internal organs of mice. The analytical range of the method ranged from 5 to 5,000 ng/ml for blood plasma and from 25 to 25,000 ng/g for liver, kidneys, heart, lungs, brain, spleen, skeletal muscles and adipose tissue. The bioanalytical method fully meets the validation requirements and can be used to perform the analytical part of an experimental study of the pharmacokinetics of BBP2023, including an assessment of the distribution of the studied compound across organs and tissues.