Quantification of a dimeric dipeptide mimetic with antidepressant-like activity in rat blood plasma by high performance liquid chromatography-mass spectrometry

A new dimeric dipeptide mimetic with antidepressant-like activity has been developed at the FSBSI "FRC of Original and Promising Biomedical and Pharmaceutical Technologies". A specific, sensitive, fast and reproducible method for the determination of bis-(N-monosuccinyl-L-asparaginyl-L-asparagine hexamethylenediamide (GTS-301) in rat blood plasma using high-performance liquid chromatography tandem mass spectrometric detector has been developed and validated. The lower limit of quantification in blood plasma was 50 ng/ml. All calibration curves were described by a quadratic equation (correlation coefficient averaged 0.996, n = 3) in the concentration range of 50÷10000 ng/ml. The accuracy values for the quality control samples varied throughout the day from an average of 93.2 to 113.3 %. The precision ranged from 1.1 to 11.4 %. During the validation period, the accuracy values averaged 101.1–108.8 % and accuracy — 5.4–7.3 %. For the calibration standards, the values of the validation parameters were also within acceptable values (accuracy — 92.2–106.8 %, accuracy — 0.1–11.5 %). The results of the dilution study of experimental blood plasma samples with intact biomaterial in 10and 100-fold lie within the acceptable values of 85–115 %. It has been established that GTS-301 is stable in the biological material at ambient temperature for short-term storage (3 hours), exposure to a thermostatically controlled autosampler (5 °C) during an analytical experiment (up to 20 hours), and prolonged storage at -50 °C (for 4 weeks), and also if subjected to several freezing/defrosting cycles (3 cycles). Thus, the above-mentioned characteristics of the developed technique make it suitable for use in preclinical pharmacokinetic studies. The pharmacokinetics of GTS-301 in rat blood plasma after a single intraperitoneal injection at a dose of 150 mg/kg was described.

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