Development and validation of bioanalytical technique for the LCS-1269 assay by high-performance liquid chromatography

Background. The glycosylated derivative of indolo[2,3-a]carbazole LCS-1269, an DNA intercalator and a chromatin remodulator, synthesized in N.N. Blokhin National Medical Research Center of Oncology (N.N. Blokhin NMRCO), is of great interest for the treatment of malignant tumors. To carry out pharmacokinetic studies of LCS-1269 dosage forms in vivo, it is necessary to develop a method for LCS-1269 assay in biological samples.

Aim. To develop and validate an analytical technique using high-performance liquid chromatography with ultraviolet detection (HPLC-UV) for LCS-1269 pharmacokinetic studies.

Materials and methods. The study used the LCS-1269 substance containing 99.4 % of the active component, blood plasma and liver of healthy cyclic immunocompetent laboratory mice hybrid (C₅₇Bl/6J×DBA2) F₁ of N.N. Blokhin NMRCO breeding. Acetonitrile and sodium chloride were used to prepare biological samples. Reversed-phase HPLC-UV method in the gradient regimen was used for LCS-1269 assay. Statistical processing of the obtained results was carried out in Microsoft Office Excel 2010 software.

Results. The technique based on cooled acetonitrile deproteinization was developed for blood plasma sample preparation; liver samples were prepared using combined cooled acetonitrile and sodium chloride deproteinization. The developed technique validation confirmed its specificity, linearity (r >0.99), correctness (response >80 % at low concentrations) and precision (relative standard deviation <3.5 %) in the entire analytical range; LCS-1269 transfer effect wasn’t observed. It was found that the analyzed samples are stable for 24 hours at room temperature, and the model mixtures are stable during a month when stored at temperatures less than –18 °C.

Conclusion. A bioanalytical technique for LCS-1269 assay in the blood plasma and liver of animals using HPLC-UV has been developed. Has been shown that the technique meets the requirements of the Eurasian Economic Union for the validation of biological samples analysis.

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