PDF(Pubmed)
Abstract：
Various types of milk powder purportedly providing diverse health functions have emerged with the growth of the country\'s elderly population. Some manufacturers illegally add chemical drugs to their products to achieve their reported benefits, which poses a threat to consumer health. The existing standard methods are inapplicable to such complex sample matrices and require testing based on functional claims and classification. These limitations not only consume manpower and resources but also seriously impede daily regulatory efforts to detect unknown risk substances. In this study, a high-throughput method for the screening and quantitative analysis of 300 illegally added chemical drugs in functional milk powder and an identification strategy for unknown structural analogues were established using Zeno SWATH® data-independent acquisition (DIA) ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS) technology combined with a QuEChERS sample purification method. The QuEChERS purification process was developed according to the characteristics of milk powder matrix. The supernatant was separated on a Kinetex F5 column (100 mm×3.0 mm, 2.6 μm) by gradient elution using 5 mmol/L ammonium formate aqueous solution (0.1% (v/v) formic acid, ) and methanol-acetonitrile (1∶1, v/v) as mobile phases. The method was validated in terms of selectivity, linearity, limits of detection and quantification (LODs and LOQs, respectively), matrix effect, accuracy, and precision. Based on a screening database for the 300 target substances, electron-activated dissociation (EAD) fragmentation was applied to obtain rich secondary MS fragmentation information, and unknown structural analogues were identified and confirmed through fragment attribution analysis. The results indicated that all compounds had good linear relationships in certain ranges with correlation coefficients >0.99. The LODs and LOQs were 0.04-2.7 and 0.2-8.0 μg/kg, respectively. The average recoveries at three spiked levels were in the range of 73.1%-125.2%, and the relative standard deviations were ≤14.8% (n=6). When the developed method was applied to detect illegally added chemicals in 60 functional milk powder samples, it detected benzoguanidine and sildenafil and successfully identified ethylphenidate, which is the structural analogue of an amphetamine. The proposed method is simple, sensitive, and accurate; thus, it may have practical application value for the daily supervision and law enforcement of milk powders with reported health functions.
摘要：
随着该国老年人口的增长，据称提供多种保健功能的各种类型的奶粉已经出现。一些制造商在其产品中非法添加化学药物以实现其报告的利益，这对消费者健康构成威胁。现有的标准方法不适用于这种复杂的样本矩阵，并且需要基于功能声明和分类进行测试。这些限制不仅消耗人力和资源，而且严重阻碍了检测未知风险物质的日常监管工作。在这项研究中,使用ZenoSWATH®数据无关采集(DIA)超高效液相色谱-高分辨率质谱(UHPLC-HRMS)技术结合QuEChERS样品纯化方法,建立了功能性奶粉中300种非法添加化学药物的高通量筛选和定量分析方法,以及未知结构类似物的鉴定策略.根据奶粉基质的特点，开发了QuEChERS纯化工艺。上清液在KinetexF5柱(100mm×3.0mm,2.6μm），使用5mmol/L甲酸铵水溶液（0.1％（v/v）甲酸，)和甲醇-乙腈(1∶1，v/v)为流动相。该方法在选择性方面进行了验证，线性度检测和定量限(LOD和LOQ，分别)，基体效应，准确度,和精度。根据300种目标物质的筛选数据库，应用电子激活解离(EAD)片段化以获得丰富的二级MS片段化信息，和未知的结构类似物被鉴定和确认通过片段归因分析。结果表明,各化合物在一定范围内线性关系良好,相关系数>0.99。LOD和LOQ分别为0.04-2.7和0.2-8.0μg/kg，分别。三个加标水平的平均回收率在73.1%-125.2%之间,相对标准偏差≤14.8%(n=6)。将该方法应用于60份功能性奶粉样品中非法添加化学物质的检测，它检测到苯胍和西地那非，并成功鉴定了哌醋乙酯，这是安非他明的结构类似物。该方法简单,敏感,准确；因此，对于报告具有保健功能的奶粉的日常监管和执法可能具有实际应用价值。
