Wang Yi-Ming, Li Xiao-Tong, Chu Kun, Wang Qian-Qian, Wu Shuai, Chen Chen
Yantai Testing Center for Food and Drug,Yantai 264000,China.
Se Pu. 2025 Sep;43(9):1034-1044. doi: 10.3724/SP.J.1123.2024.10026.
The consumption of agricultural products has increased in recent years owing to abundant production and improved living standards. Veterinary drugs are highly commercialized and widely used in animal husbandry to ensure animal health and production performance. Moreover, pesticides can become enriched during animal breeding, resulting in animal-derived food pollution through foraging, drinking, and environmental disinfection that can potentially damage human health. Consequently, food-safety issues associated with pesticide and veterinary-drug residues have attracted considerable attention. However, few reports on the multi-residue analysis of livestock and poultry meat using ultra performance liquid chromatography-quadrupole time of flight mass spectrometry (UPLC-Q-TOF/MS) have been published. Therefore, developing high-throughput and efficient screening methods for monitoring various illegal and/or restricted drugs in animal-derived foods is imperative. In this study, we developed a protocol for simultaneously examining seven pesticide and veterinary-drug types that uses an accurate mass-spectral library. UPLC-Q-TOF/MS was then employed to screen 111 such compounds, including quinolones, macrolides, cephalosporins, and antiviral drugs. The developed protocol was subsequently used to establish a method for quantitatively analyzing more than 90 compounds in livestock and poultry meat. The formula, theoretical exact mass, experimental exact mass, and retention time of each analyte were recorded and used for identification purposes. The main factors affecting the response and sensitivity of the method, such as the LC separation conditions (chromatographic column and mobile phases) and MS parameters, were optimized during instrumental analysis. Pork and chicken samples were extracted with an 80% acetonitrile aqueous solution, after which the supernatant was purified using an Oasis PRiME HLB solid-phase extraction column. The 111 target analytes were separated on a Waters HSS T3 analytical chromatographic column (100 mm×2.1 mm, 1.8 μm) after being blown with nitrogen and redissolved, with gradient elution performed using mobile phases composed of 0.1% formic acid aqueous solution and methanol. The analysis process included a flow rate of 0.4 mL/min, a column temperature of 40 ℃, and an injection volume of 5 μL, with positive electrospray ionization (ESI) and time of flight mass spectrometry full scan information-dependent acquisition-product ion (TOF MS-IDA-Product Ion) scanning modes used. The method was validated in terms of linearity, limits of screening and quantification (SDLs and LOQs, respectively), matrix effects, accuracy, and precision. Quantification was performed using matrix-matched external-standard calibration. All target compounds exhibited good linearities in their corresponding concentration ranges, with all correlation coefficients () above 0.99. The SDLs of all analytes were in the range of 0.5-10 μg/kg, and the proportion of LOQs within the range of 0.5-10 μg/kg were 88.3% and 86.5%, respectively. The compounds quantified in pork and chicken exhibited recoveries of between 60.2% and 100.2%, and 61.1% and 116.7%, respectively, at spiked levels of LOQ, 2×LOQ and 10×LOQ, with relative standard deviations (RSDs) ranging from 1.1% to 13.9% and 1.0% to 14.1%, respectively. Simulated positive samples and commercial livestock and poultry meat samples were screened using an in-house-constructed mass spectrometry database. Commercial samples were screened while enrofloxacin was detected in two pork samples and tilmicosin was detected in one chicken sample, with content in the range of 4.94-29.1 μg/kg. The method developed in this study is advantageous because it involves simple sample processing and is less time consuming than existing methods; consequently, it is suitable for the rapid and high-throughput screening of pesticides and veterinary residues in livestock and poultry meat.
近年来,由于产量丰富和生活水平提高,农产品的消费量有所增加。兽药高度商业化,广泛应用于畜牧业,以确保动物健康和生产性能。此外,在动物养殖过程中,农药可能会富集,通过觅食、饮水和环境消毒导致动物性食品污染,进而可能损害人类健康。因此,与农药和兽药残留相关的食品安全问题引起了广泛关注。然而,关于使用超高效液相色谱 - 四极杆飞行时间质谱(UPLC - Q - TOF/MS)对畜禽肉进行多残留分析的报道很少。因此,开发高通量、高效的筛选方法来监测动物源性食品中的各种违禁和/或受限药物势在必行。在本研究中,我们开发了一种使用精确质谱库同时检测七种农药和兽药类型的方案。然后采用UPLC - Q - TOF/MS筛选111种此类化合物,包括喹诺酮类、大环内酯类、头孢菌素类和抗病毒药物。随后,所开发的方案被用于建立一种定量分析畜禽肉中90多种化合物的方法。记录了每种分析物的分子式、理论精确质量、实验精确质量和保留时间,用于鉴定目的。在仪器分析过程中,对影响该方法响应和灵敏度的主要因素,如液相色谱分离条件(色谱柱和流动相)和质谱参数进行了优化。猪肉和鸡肉样品用80%乙腈水溶液提取,然后上清液用Oasis PRiME HLB固相萃取柱纯化。111种目标分析物经氮气吹干并重新溶解后,在Waters HSS T3分析色谱柱(100 mm×2.1 mm,1.8 μm)上分离,使用由0.1%甲酸水溶液和甲醇组成的流动相进行梯度洗脱。分析过程包括流速0.4 mL/min、柱温40℃和进样量5 μL,采用正电喷雾电离(ESI)和飞行时间质谱全扫描信息依赖采集 - 产物离子(TOF MS - IDA - Product Ion)扫描模式。该方法在线性、筛选限和定量限(分别为SDLs和LOQs)、基质效应、准确度和精密度方面进行了验证。使用基质匹配外标校准进行定量。所有目标化合物在其相应浓度范围内均表现出良好的线性,所有相关系数()均高于0.99。所有分析物的SDLs在0.5 - 10 μg/kg范围内,LOQs在0.5 - 10 μg/kg范围内的比例分别为88.3%和86.5%。在猪肉和鸡肉中定量的化合物在LOQ、2×LOQ和10×LOQ加标水平下的回收率分别为60.2%至100.2%和61.1%至116.7%,相对标准偏差(RSDs)分别为1.1%至13.9%和1.0%至14.1%。使用自行构建的质谱数据库对模拟阳性样品和市售畜禽肉样品进行筛选。对市售样品进行筛选时,在两个猪肉样品中检测到恩诺沙星,在一个鸡肉样品中检测到替米考星,含量在4.94 - 29.1 μg/kg范围内。本研究开发的方法具有优势,因为它涉及简单的样品处理,比现有方法耗时少;因此,它适用于畜禽肉中农药和兽药残留的快速高通量筛选。