کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10974715 | 1108028 | 2015 | 8 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
A rapid HPLC method for indirect quantification of β-lactamase activity in milk
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کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری
علوم کشاورزی و بیولوژیک
علوم دامی و جانورشناسی
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چکیده انگلیسی
To circumvent the strictly regulated limits of antibiotics in milk, illegal addition of β-lactamase to lower the antibiotic levels in milk has been reported recently in China. Herein, we describe a fast, sensitive, and robust HPLC-UV method for the determination of β-lactamase activity in milk, based on an indirect quantification strategy. The test milk sample was mixed with a known amount of penicillin G, a specific substrate of β-lactamase. After incubation, an aliquot of the mixture was injected into the HPLC-UV system to quantify the remaining penicillin G in less than 10 min. Comparative analysis of the amount of penicillin G before and after incubation was used to indirectly deduce the activity of β-lactamase in the test sample. This method was successfully applied to milk products with different fat percentages, resulting in a detection limit of 0.6 U/mL. Good recoveries, ranging from 94 to 105%, were obtained from blank milk samples spiked with β-lactamase at levels of 2 to 50 U/mL, with relative standard deviations <6%. A good correlation was demonstrated between the HPLC method and the conventional culture-based assay. Using this method, the activity changes in β-lactamase during milk pasteurization, sterilization, and storage were investigated. The advantages of low-cost, accurate quantification and easily accessible instrumentation make the proposed method an ideal alternative for high-throughput routine analysis in the dairy industry.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Dairy Science - Volume 98, Issue 4, April 2015, Pages 2197-2204
Journal: Journal of Dairy Science - Volume 98, Issue 4, April 2015, Pages 2197-2204
نویسندگان
Shuang Zhou, Dan Wang, Yunfeng Zhao, Yongning Wu,