Institute of Chemical Biology and Fundamental Medicine SB RAS, Lavrentiev Ave. 8, Novosibirsk 630090, Russia.
Novosibirsk State University, Pirogova Str. 2, Novosibirsk 630090, Russia.
Molecules. 2022 Jun 2;27(11):3577. doi: 10.3390/molecules27113577.
The aim of this work was to develop and validate a sensitive and robust method of liquid chromatography coupled with tandem mass spectrometry to quantitate ST-246 (tecovirimat) in plasma using an internal standard (2-hydroxy-N-{3,5-dioxo-4-azatetracyclo [5.3.2.02.6.08.10]dodec-11-en-4-yl}-5-methylbenzamide). The method was validated in negative multiple reaction monitoring mode following recommendations of the European Medicines Agency for the validation of bioanalytical methods. The calibration curve for the analyte was linear in the 10−2500 ng/mL range with determination coefficient R2 > 0.99. Intra- and inter-day accuracy and precision for three concentrations of quality control were <15%. Testing of long-term stability of ST-246 (tecovirimat) in plasma showed no degradation at −20 °C for at least 3 months. The method was applied to a clinical assay of a new antipoxvirus compound, NIOCH-14. Thus, the proposed method is suitable for therapeutic drug monitoring of ST-246 (tecovirimat) itself and of NIOCH-14 as its metabolic precursor.
本工作旨在开发和验证一种灵敏、稳健的液相色谱-串联质谱法，采用内标（2-羟基-N-[[3,5-二氧代-4-氮杂双环[5.3.2.02,6.08,10]十二-11-烯-4-基]-5-甲基苯甲酰胺]）定量测定血浆中的 ST-246（替科韦瑞玛）。该方法按照欧洲药品管理局对生物分析方法验证的建议，采用负多重反应监测模式进行了验证。分析物的校准曲线在 10-2500ng/mL 范围内呈线性，决定系数 R2>0.99。三种浓度质控品的日内和日间准确度和精密度均<15%。ST-246（替科韦瑞玛）在血浆中的长期稳定性测试表明，在至少 3 个月的时间内，在-20°C 下无降解。该方法已应用于一种新型抗痘病毒化合物 NIOCH-14 的临床检测。因此，该方法适用于 ST-246（替科韦瑞玛）本身以及作为其代谢前体的 NIOCH-14 的治疗药物监测。
