| 丁晨红，黄晓梅，王威利，万 凯，苏秋权，李亚菲.HPLC 法测定饲料中 7 种青霉素类药物含量[J].广东农业科学,2021,48(1):126-133 |
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HPLC 法测定饲料中 7 种青霉素类药物含量 |
| Determination of seven Penicillins in Feed by High Performance Liquid Chromatograph |
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| DOI：10.16768/j.issn.1004-874X.2020.07.016 |
| 中文关键词: 高效液相色谱 饲料 青霉素 衍生化 |
| 英文关键词: high performance liquid chromatograph feed penicillin derivatization |
| 基金项目:国家重点研发计划项目（2018YFD0500300）；国家农业行业标准制定与修订项目（农财发【2018】
46号）；国家农产品质量安全风险评估项目（GJFP201700702）；广东省现代农业产业技术体系创新团队项目
（2019KJ130） |
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| 中文摘要: |
| 【目的】建立饲料中 7 种青霉素药物（阿莫西林、氨苄西林、青霉素 G、青霉素 V、苯唑西林、
氯唑西林和双氯西林）含量的高效液相色谱测定方法。【方法】配合饲料、浓缩饲料、精料补充料样品采用
20% 乙腈溶液提取，添加剂预混合饲料样品经 EDTA-Mcllvaine 缓冲液提取，HLB 小柱净化，衍生化后，用 C18
（250 mm×4.6 mm, i.d 5 μm）色谱柱，以磷酸盐缓冲溶液－乙腈二元流动相梯度洗脱，325 nm 紫外波长检测，
基质匹配标准工作液校准，外标法定量。【结果】配合饲料、浓缩饲料和精料补充料中 7 种青霉素类药物在 0.25~10
μg/mL 浓度范围内线性关系良好，添加剂预混和饲料中 7 种青霉素类药物在 0.10~10 μg/mL 浓度范围内线性关
系良好；加标回收率为 69.6%~103.7%，RSD 值 <15%。配合饲料、浓缩饲料、精料补充料的检测限为 2.5 mg/
kg，定量限为 5 mg/kg；添加剂预混合饲料的检测限为 1.0 mg/kg，定量限为 2.0 mg/kg。【结论】该方法具有快速、
准确、专属性和重现性好的特点，适合日常饲料样品中 7 种青霉素类药物含量测定。 |
| 英文摘要: |
| 【Objective】The study was aimed to establish a HPLC method for the determination of amoxicillin,
ampicillin, penicillin G, penicillin V, oxacillin, cloxacillin, and dicloxicillin in feeds.【Method】Firstly, the samples of
formula feed, concentrated feed and concentrate supplement were extracted with 20% acetonitrile solution, and the premixed
feed samples were extracted by EDTA-Mcllvaine buffer solution and purified by HLB solid-phase extraction column, and
then derivated by the reagents. Eventually, the samples were separated on a C18（150 mm×4.6 mm, i.d 5 μm）column by
gradient elution with acetonitrile - phosphate buffer solution as mobile phase, detected by UV detecor at 325 nm, calibrated
by matrix matching standard working solution and quantified by external standard method.【Result】There were good linear
relationships among the seven kinds of penicillins of formula feed, concentrated feed and concentrate supplement in the
concentration range of 0.25-10 μg/mL, and there were good relationships among the seven kinds of penicillins of premixed additive feed in the concentration range of 0.10-10μg/mL. The average recovery rates were in the range of 69.6%-103.7% with
RSD value less than 15%. The limit of detection（LOD）of compound feed, concentrated feed and concentrate supplement
was 2.5 mg/kg , and the limit of quantification（LOQ）was 5 mg/kg. The LOD of premixed additive feed was 1.0 mg/kg and the
LOQ was 2.0 mg/kg.【Conclusion】The method was proved to be specific, rapid, accurate and reproducible for determination of
seven penicillin antibiotics in daily feed . |
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