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individually packed milk samples collected from 5 retail stores in 3 cities. Some of these samples had extremely high βlactamase activity. ? The ministry of health of the People’s Republic of China included βlactamase into its newly published “black list” of illegal food additives. 25 L U N D U N I V E R S I T Y β 內(nèi)酰胺酶檢測 Methods HPLC Cylinder plate method chromogenic method Disadvantages Sample preparation Time consuming: 24 hr/test Time consuming: 30min24hr/test Modified cylinder plate method: betalactamase activity is defined as a difference between the inhibitory zones of the test samples and control samples. Chromogenic method: hydrolytic activity of bacterial fllactamases was estimated by the measurement of absorbance changes of nitrocefin and piridinium2azopdimethylamiline chromophase (PADAC). 26 L U N D U N I V E R S I T Y 乳品快速檢測裝置 β內(nèi)酰胺 酶響應(yīng) 待測奶樣品 27 L U N D U N I V E R S I T Y 乳品中 β 內(nèi)酰胺酶檢測 β 內(nèi)酰胺酶檢測分析是基于這一原理 : 即 , 將特異性的酶催化反應(yīng)與精確的流動量熱分析結(jié)合起來 , 從而實現(xiàn)獨一無二的 、 快速的乳品分析 。其檢測原理如下圖所示 。 28 L U N D U N I V E R S I T Y 測量方法和結(jié)果 1. Calibration 29 L U N D U N I V E R S I T Y 1. Effect of flowrate 2. Effect of sample volume 3. Effect of milk contents 30 L U N D U N I V E R S I T Y 牛 奶中 β 內(nèi)酰胺 酶的檢測響應(yīng) Control y = R178。 = 01230 1 2 3 4 5 6ΔT (m ℃) penicillinase activity (IU/ml) 0 10 20 30 40 50 60 70 80 Time( min) Penicillinase activity determined in milk with 3% fat and 50% (v/v) dilution. 1mM penicillin G, flow rate of ml/min and the sample volume of 350 μl were applied. 31 L U N D U N I V E R S I T Y 不同方法優(yōu)缺點對比 cost per sample Time(min/sample) Continuity Detection limit Modified ET method ≈$ 6 min Semi Cylinder plate method[5] n/a 24hr No 4u/ml Chromogenic method[7] n/a 30min~24hr n/a 32 L U N D U N I V E R S I T Y ET特點 分析方法簡單 、 儀器操作方便 待測奶樣品無需特殊預處理 測量快速 ( 約 57分鐘 /每樣品 ) 檢測成本低 ( 約 1元人民幣 /每樣品 ) 靈敏度高 ( β 內(nèi)酰胺酶的檢測低于 U 酶活性 /ml 奶樣品) 適合原料奶 、 奶制品和其他液態(tài)食品中的 β 內(nèi)酰胺酶的檢測 可用于其它酶的檢測或酶底物的分析 33 L U N