色谱 ›› 2024, Vol. 42 ›› Issue (11): 1077-1086.DOI: 10.3724/SP.J.1123.2024.02018

• 技术与应用 • 上一篇    下一篇

分散固相萃取-高效液相色谱-串联质谱法测定消毒产品中42种抗微生物药物

童嘉琦1,*(), 朱谷焕2, 蓝航镇3, 史红兰2, 朱晓艳1, 孙瑶4   

  1. 1.宁波海关技术中心, 浙江 宁波 315000
    2.宁波中盛产品检测有限公司, 浙江 宁波 315000
    3.宁波大学食品科学与工程学院, 浙江 宁波 315000
    4.大连海关技术中心, 辽宁 大连 116000
  • 收稿日期:2024-03-01 出版日期:2024-11-08 发布日期:2024-10-22
  • 通讯作者: * Tel:(0574)89095049,E-mail:tyrles@yeah.net.
  • 基金资助:
    宁波市公益性科技计划项目(2021S195);国家自然科学基金(32102060)

Determination of 42 antimicrobials in disinfection products by dispersive solid phase extraction-high performance liquid chromatography-tandem mass spectrometry

TONG Jiaqi1,*(), ZHU Guhuan2, LAN Hangzhen3, SHI Honglan2, ZHU Xiaoyan1, SUN Yao4   

  1. 1. Ningbo Customs Technology Center, Ningbo 315000, China
    2. Ningbo Joysun Product Testing Service Company, Ningbo 315000, China
    3. College of Food Science and Engineering, Ningbo University, Ningbo 315000, China
    4. Technological Center of Dalian Customs, Dalian 116000, China
  • Received:2024-03-01 Online:2024-11-08 Published:2024-10-22
  • Supported by:
    Ningbo Science and Technology Program for Public Welfare(2021S195);National Natural Science Foundation of China(32102060)

摘要:

本研究采用分散固相萃取-高效液相色谱-串联质谱(HPLC-MS/MS)测定膏霜、凝胶和水剂等基质类型的消毒产品中42种抗微生物药物(7种磺胺类、10种喹诺酮类、3种林可酰胺类、5种四环素类、3种大环内酯类、8种唑类、3种嘌呤核苷类、1种呋喃类、1种非多烯类及1种类固醇类药物)含量的测定方法。消毒产品样品用水辅助分散,用0.5%甲酸乙腈溶液提取,用EMR-Lipid吸附剂除去提取液中长链烃类化合物等样品基质成分。选用Poroshell 120 EC-C18色谱柱(150 mm×3.0 mm, 2.7 μm)分离,以0.1%甲酸乙腈溶液和0.1%甲酸水溶液为流动相,梯度洗脱。选用电喷雾离子源,正离子(ESI+)扫描模式和多反应监测(MRM)模式检测,保留时间和选择离子定性,外标法定量。在优化的条件下,42种抗微生物药物可以有效分离,基质效应对目标化合物的影响较弱。在0.25~5.0 mg/kg范围内,42种目标化合物线性良好,相关系数(r)>0.99。以色谱峰信噪比(S/N)≥3评估检出限(LOD),在取样量为0.2 g、定容体积为10 mL时,3种基质中42种抗微生物药物的检出限为0.03~0.10 mg/kg。在0.25、1.0和5.0 mg/kg 3个添加水平下,膏霜、凝胶和水剂3种基质中42种抗微生物药物的平均回收率为80.3%~109.8%,相对标准偏差(RSD)≤9.8%。采用本方法检测市售消毒产品中抗微生物药物,共检出2批阳性样品。所建立的方法操作简便,准确性好,精密度高,可适用于膏霜、凝胶和水剂等类型消毒产品中42种抗微生物药物的快速测定。

关键词: 消毒产品, 抗微生物药物, 高效液相色谱-串联质谱, 分散固相萃取

Abstract:

Antimicrobials inhibit the growth and reproduction of microorganisms thereby alleviating skin and other problems caused by microorganisms. Antimicrobials are classified into different categories, including antibacterials, antifungals, and antivirals, among others, and include azoles, sulfonamides, tetracyclines, quinolones, and many other classes of synthetic and natural compound. The inappropriate or excessive use of antimicrobials can damage skin and other human organs and increase antimicrobial resistance. Relevant regulations and standards clearly state that antimicrobials are prohibited for use as ingredients in disinfection products. However, since antimicrobials enhance the disinfection or antibacterial effect of a product, with a significant short-term effect, antimicrobials are occasionally illegally added to disinfectant products, including those intended for human use. Therefore, establishing testing methods that provide technical support for enforcing regulations is an urgent objective.

Herein, a method was established for the analysis of 42 antimicrobials in disinfection products, that is applicable to common types of disinfection-product matrix, including creams, gels, and aqueous solutions, using high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) combined with dispersive solid phase extraction. The 42 antimicrobials comprise antibacterials, antifungals, and antivirals, and include seven sulfonamides, ten quinolones, three lincosamides, five tetracyclines, three macrolides, eight azoles, three purine nucleoside analogs, one furan, one nonpolyene antifungal, and one steroid. Briefly, 0.2 g of a sample was first dispersed in 2 mL of water and then extracted with 10 mL of 0.5% formic acid in acetonitrile, with 3 g of anhydrous Na2SO4 added to remove water. After centrifugation, 5 mL of the supernatant was cleaned using dispersive solid phase extraction with EMR-Lipid as the adsorbent. Lipids, waxes, surfactants, and moisturizing lubricants are commonly used as cream and gel matrices. Matrix substances containing long carbon chains dissolved in acetonitrile were removed using the EMR-Lipid adsorbent. Target analytes were separated on a Poroshell 120 EC-C18 analytical chromatography column (150 mm×3.0 mm, 2.7 μm), with 0.1% formic acid in acetonitrile and 0.1% formic acid aqueous solution used as mobile phases under gradient-elution conditions. The target analytes in the test solution were detected in positive ionization (ESI+) and multi-reaction-monitoring (MRM) modes. Analytes were characterized in terms of their retention times and selected ions, and quantified using the external-standard method. The main factors affecting method response, recovery, and sensitivity, such as the extraction method and solvent, purification method and adsorbent, mobile phase, and MS conditions, were examined during sample pretreatment and instrumental analysis. The 42 antimicrobials were effectively separated under the optimized experimental conditions; the target compounds exhibited linear working curves in the 0.25-5.0 mg/kg concentration range, with correlation coefficients (r) greater than 0.99. Limits of detection (LODs) for the 42 antimicrobials were determined from the signal-to-noise ratios (S/N) of their chromatographic peaks. LODs of 0.03-0.10 mg/kg were determined for the three matrices using 0.2-g samples and 10-mL test solution. Recoveries of 80.3-109.8%, with relative standard deviations (RSDs) of less than 9.8%, were obtained by determining three levels of each target analyte added to the three blank matrices; this process was repeated for six parallel samples. The developed method was used to analyze antimicrobials in commercially available disinfection products, with two sample batches testing positive. The established method is simple, accurate, precise, and suitable for the rapid screening and quantification of antimicrobials in disinfection products. This study provides powerful technical support for regulating the illegal addition of related antimicrobials to disinfection products.

Key words: disinfection products, antimicrobials, high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), dispersive solid phase extraction

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