评价三种不同的微生物抑制试验检测兔食用组织中磺胺嘧啶残留。
Evaluation of three different microbial inhibition tests for the detection of sulphamethazine residues in the edible tissues of rabbit.
机构信息
Department of Food Hygiene and Technology, University of Veterinary Medicine, Kosice, Slovakia.
出版信息
Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2009 Jul;26(7):978-87. doi: 10.1080/02652030902839715.
The aim of the present study was to evaluate three microbial inhibition tests (MIT) based on inhibition of growth of the test organisms: (a) four plate test (FPT) containing Bacillus subtilis BGA, (b) screening test for antibiotic residues (STAR) containing Bacillus stearothermophilus var. calidolactis_ATCC 10149 and (c) the Premi(R)Test containing Bacillus stearothermophilus var. calidolactis. The tests were used to determine sulphamethazine (SMZ) residues in edible tissues of rabbit after oral administration up to day 15 of the withdrawal period (WP). A solvent extraction procedure was used to enhance the capability of the tests to detect SMZ residues at or below the maximum residue limit (MRL). Para-aminobenzoic acid (PABA) was employed to previously identify SMZ residues in the first stage of the residue screening. The presence of SMZ residues in the samples was confirmed and quantified by a validated HPLC method. The Premi(R)Test detected SMZ residues in the muscle and heart tissue up to day 9 of the WP, and in the liver, lungs and kidneys up to day 10 of the WP. The STAR detected SMZ residues in the edible organs of rabbits up to day 8 of the WP. The kidneys were positive up to day 5 of the WP, the liver until day 4 of the WP and the lungs until day 3 of the WP. No SMZ residues were detected in the muscle and heart. By using the solvent extraction procedure, SMZ residues were detected in the muscle extract up to day 10 of the WP and the muscle was positive until day 6 of the WP. No detection sensitivity was observed using the FPT. After solvent extraction, SMZ residues were detected in the muscle extract until day 8 of the WP and the muscle was positive until day 3 of the WP. No positive results were detected after the addition of PABA into/onto the agar medium. PABA at a concentration of 10 microg ml(-1) completely reversed the inhibitory activity of SMZ and enabled reliable identification of SMZ in the examined samples. Using HPLC, SMZ was detected in the muscle samples until day 10 of WP (0.02 mg kg(-1)) and in the liver until day 12 of the WP (0.09 mg kg(-1)). The results obtained by the HPLC method and the limit of detection (LOD) of screening tests for SMZ (FPT 0.4 microg ml(-1), STAR 0.2 microg ml(-1), Premi(R) Test 0.05 microg ml(-1)) allowed us to state that the most suitable screening tests for the detection of SMZ residues in the edible tissues of rabbits at level corresponding to the MRL of 0.1 mg kg(-1), established for sulphonamides, are the Premi(R)Test and STAR in conjunction with the solvent-extraction procedure.
本研究的目的是评估三种基于测试菌生长抑制的微生物抑制测试（MIT）：（a）含有枯草芽孢杆菌 BGA 的四板测试（FPT），（b）含有嗜热脂肪芽孢杆菌 var 的筛选测试。 calidolactis_ATCC 10149 和（c）含有嗜热脂肪芽孢杆菌 var 的 Premi（R）Test。 calidolactis。 这些测试用于确定兔在停药期（WP）第 15 天之前口服磺胺甲恶唑（SMZ）残留量在可食用组织中。 采用溶剂萃取法提高了这些测试检测 SMZ 残留量的能力，这些残留量低于或等于最大残留限量（MRL）。 对氨基苯甲酸（PABA）用于在残留筛选的第一阶段预先鉴定 SMZ 残留量。 通过验证的 HPLC 方法确认和定量了样品中 SMZ 残留量。 Premi（R）Test 可在 WP 第 9 天检测肌肉和心脏组织中的 SMZ 残留量，在 WP 第 10 天检测肝脏、肺脏和肾脏中的 SMZ 残留量。 STAR 可在 WP 第 8 天检测到兔子可食用器官中的 SMZ 残留量。 肾脏在 WP 第 5 天之前呈阳性，肝脏在 WP 第 4 天之前，肺脏在 WP 第 3 天之前呈阳性。 肌肉和心脏未检测到 SMZ 残留量。 通过使用溶剂萃取程序，在 WP 第 10 天检测到肌肉提取物中的 SMZ 残留量，肌肉在 WP 第 6 天之前呈阳性。 FPT 未观察到检测灵敏度。 经溶剂萃取后，在 WP 第 8 天检测到肌肉提取物中的 SMZ 残留量，肌肉在 WP 第 3 天之前呈阳性。 在琼脂培养基中加入或加入 PABA 后未检测到阳性结果。 PABA 浓度为 10μg ml（-1）完全逆转了 SMZ 的抑制活性，使 SMZ 在检测样品中的可靠鉴定成为可能。 使用 HPLC，在 WP 第 10 天（0.02 mg kg（-1））检测到肌肉样品中的 SMZ，在 WP 第 12 天（0.09 mg kg（-1））检测到肝脏中的 SMZ。 通过 HPLC 方法和磺胺类药物的 SMZ 筛选检测的检测限（LOD）（FPT 0.4μg ml（-1），STAR 0.2μg ml（-1），Premi（R）Test 0.05μg ml（-1））获得的结果允许我们声明，对于在对应于 0.1mg kg（-1）的 MRL 的水平检测兔可食用组织中的 SMZ 残留量，最适合的筛选测试是 Premi（R）Test 和 STAR 与溶剂萃取程序相结合。
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