关键词: 缬沙坦, 高分辨质谱, 遗传毒性杂质, N-亚硝基-N-甲基-4氨基丁酸, N-亚硝基乙基异丙胺, N-亚硝基二异丙胺, N-亚硝基二丁胺

Abstract:

Objective： To establish an ultra-high performance liquid chromatography coupled with quadrupole/orbitrap high-resolution mass spectrometry （UPLC-HRMS） method for the simultaneous determination of four nitrosamine genotoxic impurities in Valsartan： N-Nitroso-N-methyl-4-aminobutyric acid （NMBA）， N-Nitrosoethylisopropylamine （NEIPA）， N-Nitrosodiisopropylamine （NDIPA）， and N-Nitrosodibutylamine （NDBA）.Methods： Separation was achieved using a Phenomenex Kinetex F5 column （100 mm × 4.6 mm， 2.6 μm） with a mobile phase consisting of 0.1% formic acid in water （mobile phase A） and 0.1% formic acid in methanol （mobile phase B） under a linear gradient elution program. The flow rate was set at 0.6 mL·min⁻¹， and the column temperature was maintained at 40 ℃. Detection was carried out using a mass spectrometer equipped with an electrospray ionization （ESI） source. The instrument was operated in selected ion monitoring （SIM） and parallel reaction monitoring （PRM） modes. The spray voltage was 3.5 kV， the capillary temperature was 350 ℃， the sheath gas flow rate was 40 arb， and the auxiliary gas flow rate and temperature were set at 10 arb and 300 ℃， respectively. Quantification of the four genotoxic impurities was performed using the external standard method.Results： Neither the blank solvent nor valsartan interfered with the detection of NMBA， NEIPA， NDIPA， and NDBA in valsartan. The four nitrosamine genotoxic impurities exhibited good linearity within the concentration range of 0.4-100 ng·mL⁻¹， with correlation coefficients （r） all greater than 0.9995. The limits of detection （LOD） and quantification （LOQ） were 0.2 ng·mL⁻¹ and 0.4 ng·mL⁻¹， respectively. The spike recovery rates at low， medium， and high concentrations were satisfactory， with average recoveries （n=9） ranging from 80.5% to 100.3%. The relative standard deviations （RSD） ranged from 2.9% to 8.9%. The injection precision and repeatability RSD for the four nitrosamine genotoxic impurities were 1.6%-2.9% and 2.1%-9.0%， respectively. The solutions remained stable within 10 hours， and no impurities were detected in the samples.Conclusion： The method is sensitive， simple， specific， and accurate， and can be used for the simultaneous detection of the four genotoxic impurities in Valsartan.

Key words:  , Valsartan； , High-resolution mass spectrometry； , Genotoxic impurity； , N-Nitroso-N-methyl-4-aminobutyric acid； , N-Nitrosoethylisopropylamine； , N-Nitrosodiisopropylamine； , N-Nitrosodibutylamine