高分辨质谱“诊断离子”结合核磁共振表征检测食品中伐地那非药品去-N-乙基衍生物

Determination of de-N-ethyl derivative of vardenafil drug in foods based on high-resolution mass spectrometry “diagnostic ion” coupled with nuclear magnetic resonance characterization

中文关键词:  HRMS“诊断离子”  核磁波谱  非靶向筛查  伐地那非衍生物  定量检测

英文关键词:high-resolution mass spectrometry “diagnostic ion”  nuclear magnetic resonance spectroscopy  noN-targeted screening  vardenafil derivatives  quantitative determination

中文摘要:

      目的 建立一种基于高分辨质谱“诊断离子”策略结合核磁共振波谱法分析的食品中伐地那非药品去-N-乙基衍生物的分析方法。方法 待测样品经甲醇水溶液溶解, 超声提取后, 注入高分辨质谱采样, 分析确定未知物的那非类型。阳性样品中的未知物经提取、富集、净化、浓缩后, 进行核磁共振波谱表征。采用超高效液相色谱-三重四极杆质谱法(ultra performance liquid chromatography-coupled triple quadrupole mass spectrometry, UPLC-MS/MS)定量检测食品基质中的伐地那非衍生物。结果 未知物经核磁共振波谱表征被确证为伐地那非药品去-N-乙基衍生物。UPLC-MS/MS分析表明: 伐地那非药品去-N-乙基衍生物在4种基质中的质谱响应在2~100 ng/mL质量浓度范围内线性良好, 4个加标浓度下的平均回收率为74.07%~110.05%, 相对标准偏差为0.36%~5.09%。检出限(S/N=3)和定量限(S/N=10)分别为0.05 mg/kg和0.10 mg/kg。采用本方法对50批次样品进行测定, 在5批次样品中检出伐地那非药品去-N-乙基衍生物, 含量为722~1548 mg/kg。结论 该方法无需复杂样品前处理、灵敏度高、准确度好, 经6家检验检测机构复验, 可满足常规实验室检测食品中伐地那非药品去-N-乙基衍生物的要求。

英文摘要:

      Objective To establish an analytical method for detecting of de-N-ethyl derivative of vardenafil drug in foods by high-resolution mass spectrometry “diagnostic ion” coupled with nuclear magnetic resonance spectroscopy. Methods The samples to be tested were dissolved in methanol/H2O, extracted by ultrasound and injected into high-resolution mass spectrometry to analyze and determine the nafil-type of the unknown compound. After extraction, enrichment, purification and concentration of the targeted analyte in the positive sample, it was identified through nuclear magnetic resonance spectroscopy analysis. A quantitative method for detecting of vardenafil derivative in foodstuffs was developed by ultra performance liquid chromatography-coupled triple quadrupole mass spectrometry (UPLC-MS/MS). Results The unknown compound was characterized as de-N-ethyl derivative of vardenafil drug. UPLC-MS/MS analysis showed that the mass spectrometric response of de-N-ethyl derivative of vardenafil drug in 4 kinds of food matrix were well linear in the mass concentration range of 2–100 ng/mL. The spiking average recovery rate in 4 kinds of food matrix were ranged from 74.07%–110.05% with relative standard deviations of 0.36%–5.09% in 4 different concentrations. The limits of detection (S/N=3) and limits of quantitation (S/N=10) was 0.05 mg/kg and 0.10 mg/kg, respectively. This method was developed and employed to screen 50 batches of samples and de-N-ethyl derivative of vardenafil drug was detected in 5 batches of samples along with a content between 722 mg/kg to 1548 mg/kg. Conclusion The established detection method does not require complex sample pretreatment, and has high sensitivity and good accuracy, which can meet the requirements for detecting de-N-ethyl derivative of vardenafil drug in food in routine laboratory tests after being re-examined by 6 inspection and testing institutions.