高效液相色谱-高分辨四极杆-飞行时间-质谱法测定中成药中非法添加物二乙氨基前他达拉非  

Determination of Diethylaminopretadalafil Illegally Added in Chinese Traditional Patent Medicines by HPLC-Q-TOF-MS

作　　者：舒展 王琤帅 应斌斌 金芩 陈旦 SHU Zhan;WANG Chengshuai;YING Binbin;JIN Qin;CHEN Dan(Jinhua Institute for Drug and Food Control,Jinhua,Zhejiang,China 321000)

机构地区：[1]浙江省金华市食品药品检验检测研究院,浙江金华321000

出　　处：《中国药业》2023年第15期67-71,共5页China Pharmaceuticals

基　　金：浙江省药品监管系统科技计划项目[2020002]。

摘　　要：目的建立检测中成药中非法添加物二乙氨基前他达拉非的高效液相色谱-高分辨四极杆-飞行时间-质谱(HPLC-QTOF-MS)法,并根据他达拉非类化合物的特征碎片离子实现非目标化合物的筛查。方法色谱柱为Agilent EC C_(18)柱(100 mm×3.0 mm,2.7μm),流动相为乙腈-0.1%乙酸溶液(梯度洗脱),流速为0.3 mL/min,柱温为35℃,进样量为1μL;采用Q-TOF-MS作检测器,电喷雾电离(ESI),正离子模式,根据保留时间、精确相对分子质量和二级碎片比进行定性分析;由提取离子流图峰面积与质量浓度关系进行定量分析;基于精确相对分子质量的二级质谱图推测裂解规律;通过提取质荷比(m/z)为135.0441碎片离子的色谱峰识别他达拉非类化合物。结果归纳了常见磷酸二酯酶Ⅴ型抑制剂(PDE5i)他达拉非类化合物的ESI质谱裂解规律。二乙氨基前他达拉非质量浓度在0.11~5.26μg/mL范围与峰面积线性关系良好(r=0.9970,n=5);检测限为0.021μg/mL,定量限为0.053μg/mL;稳定性、重复性、精密度试验结果的RSD均小于4.60%;平均加样回收率大于97.80%(n=9)。1批样品中检出二乙氨基前他达拉非,含量为3.58 mg/g。结论该方法准确、可靠,适用于二乙氨基前他达拉非的定性定量分析,通过特征碎片离子可以实现他达拉非类化合物的快速识别。Objective To establish a high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(HPLC-Q-TOF-MS)method for the determination of diethylaminopretadalafil illegally added in Chinese traditional patent medicines,and to screen non-target compounds according to the characteristic fragment ions of tadalafil derivatives.Methods The chromatographic column was Agilent EC C_(18) column(100 mm×3.0 mm,2.7μm),the mobile phase was acetonitrile-0.1%acetic acid solution(gradient elution),the flow rate was 0.3 mL/min,the column temperature was 35℃,and the injection volume was 1μL.Q-TOF-MS equipped with electrospray ionization(ESI)source was performed in positive ionization mode,and qualitative analysis was carried out according to the retention time,accurate mass and secondary fragment ratio.Quantitative analysis was performed based on the relationship between the peak area of the extracted total ion flow chromatogram and the mass concentration.The fragmentation pattern was inferred based on the second order mass spectrometry with accurate mass.Tadalafil derivatives were identified by the chromatographic peak of fragment ion with extraction mass-to-charge ratio(m/z)of 135.0441.Results The ESI mass spectrometry fragmentation patterns of common phosphodiesterase typeⅤinhibitor(PDE5i)tadalafil derivatives were summarized.The linear range of diethylaminopretadalafil was 0.11-5.26μg/mL(r=0.9970,n=5),the limit of detection(LOD)was 0.021μg/mL,and the limit of quantitation(LOQ)was 0.053μg/mL.The RSDs of stability,repeatability,and precision tests results were all lower than 4.60%,and the average recovery was higher than 97.80%(n=9).Diethylaminopretadalafil was detected in one batch of samples,with a content of 3.58 mg/g.Conclusion The method is accurate,reliable,and suitable for the qualitative and quantitative analysis of diethylaminopretadalafil,and the characteristic fragment ions can be used to quickly identify tadalafil derivatives.

关 键 词：高效液相色谱-高分辨四极杆飞行时间-质谱法 非法添加物 磷酸二酯酶Ⅴ型抑制剂 二乙氨基前他达拉非 非靶向筛查 

分 类 号：R927.2[医药卫生—药学] R286.0