U.S. Food and Drug Administration, Center for Veterinary Medicine, Office of Research, Laurel, MD, USA.
J AOAC Int. 2021 Sep 27;104(5):1213-1222. doi: 10.1093/jaoacint/qsab026.
Antibiotics are used in ethanol production to discourage the growth of bacteria that would lower the yield of the product. Any antibiotic residues remaining in distillers grain (DG) co-product could lead to antimicrobial resistance, which is a public health concern. The U.S. Food and Drug Administration (FDA) Center for Veterinary Medicine (CVM) previously developed an LC-MS/MS analytical method to detect residues of erythromycin A, penicillin G, virginiamycin M1, and virginiamycin S1 in DG to enable regulatory decision making.
The objective of this study was to ensure the method's robustness by carrying out a multi-laboratory validation of the method.
Test portions were extracted with a mixture of acetonitrile and buffer. The extract was cleaned by solid phase extraction. The concentrated eluant was reconstituted and analyzed by LC-MS/MS. Eight laboratories participated in the study.
Average accuracies for the combined three matrixes for all four compounds at all fortification levels ranged from 83->109% with repeatability relative standard deviation (RSDr; within laboratory) ≤17% and reproducibility relative standard deviation (RSDR; between laboratory) ≤21%. The Horwitz ration (HorRat) values ranged 0.4-1.0 indicating that method reproducibility is acceptable.
An interlaboratory study was successfully conducted to evaluate an LC-MS/MS method for the determination of the drugs of interest in DG. The results demonstrate that the method is fit for purpose to determine the drugs in DG and could serve as a regulatory method capable of being used for compliance actions for DG containing these antibiotic contaminants.
The method was posted to the FDA/Foods Program Compendium of Analytical Laboratory Methods.
抗生素被用于乙醇生产中，以抑制会降低产品产量的细菌生长。如果酒糟副产物中残留任何抗生素，都可能导致抗药性，这是一个公共卫生问题。美国食品和药物管理局（FDA）兽医中心（CVM）先前开发了一种 LC-MS/MS 分析方法，用于检测酒糟中红霉素 A、青霉素 G、维吉尼亚霉素 M1 和维吉尼亚霉素 S1 的残留量，以支持监管决策。
本研究旨在通过对该方法进行多实验室验证来确保其稳健性。
用乙腈和缓冲液的混合物提取测试部分。提取物通过固相萃取进行净化。浓缩洗脱液经复溶后通过 LC-MS/MS 进行分析。有 8 个实验室参与了这项研究。
在所有添加水平下，所有四种化合物在三种基质中的平均准确度均在 83%到 109%之间，重复性相对标准偏差（RSDr；实验室内部）≤17%，再现性相对标准偏差（RSDR；实验室之间）≤21%。霍里茨比率（HorRat）值范围为 0.4-1.0，表明方法再现性可接受。
成功进行了一项实验室间研究，以评估用于测定酒糟中关注药物的 LC-MS/MS 方法。结果表明，该方法适合用于测定酒糟中的药物，并且可以作为一种监管方法，用于对含有这些抗生素污染物的酒糟进行合规性行动。
该方法已发布在 FDA/Foods 计划分析实验室方法汇编中。
