5种β受体拮抗剂类药物中的N-亚硝基类杂质的含量研究OA北大核心CSTPCD

OBJECTIVE To determine the contents of N-nitroso impurities in raw materials/formulations of propranolol,metoprolol,atenolol,esmolol and bisoprolol,and clarify the attention threshold.METHODS Ultra-high performance liquid chromatography-quadrupole/electrostatic field orbitrap high-resolution mass spectrometry(UPLC-Q/Orbitrap HRMS)was adopted.An ACE Excel 3 C18-AR column was used for the separation and a mixture of 0.2%formic acid solution with 0.01 mol/L ammonium acetate and methanol was employed as the mobile phase by gradient elution,at a flow rate of 0.60 mL/min.The column temperature was set at 40℃,and the sample size was 5 μL.The heated electrospray ionization source was employed in the positive full mass spectra-selected ion monitoring mode.The contents of N-nitroso impurities in raw materials/formulations of 15 batches of β-blockers from 10 manufacturers were determined by this method.Discovery Studio software was applied to predict the toxicity of the impurities and estimate the attention threshold.RESULTS Among 5 kinds of β-blockers,the linear ranges of N-nitroso propranolol,N-nitroso metoprolol,N-nitroso atenolol,N-nitroso esmolol and N-nitroso bisoprolol were 1.01-503.38,1.02-508.38,0.97-483.63,1.11-554.27 and 1.05-523.92 ng/mL,respectively(r＞0.999).The limits of quantitation were 1.04,0.25,0.05,0.55 and 1.05 ng/mL,and the limits of detection were 0.52,0.08,0.02,0.17 and 0.52 ng/mL,respectively.RSDs of precision,reproducibility,recovery,stability and durability tests were all lower than 7.5%(n=6 or n=5).Among the 15 batches of samples,except for 1 batch,N-nitroso propranolol(1.07-8.91 ng/mg),N-nitroso metoprolol(1.43-3.37 ng/mg),N-nitroso atenolol(1.33 ng/mg),N-nitroso esmolol(0.19 ng/mg)and N-nitroso bisoprolol(1.27 ng/mg)were detected in all other batches.According to predictions,the above 5 impurities had varying degrees of reproductive toxicity,mutagenicity and carcinogenicity,with attention thresholds of 1.0,0.4,4.3,0.2 and 46.7 ng/mg,respectively.CONCLUSIONS The established method is simple,rapid,sensitive and specific,the estimated attention thresholds are clear,which can be used for the control of N-nitroso impurities in variousβ-blockers.