Effect of pre-treatment methods of bovine blood on clenbuterol residues by Enzyme-linked Immunosorbent Assay (ELISA)
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摘要: 采用单因素实验和正交实验探讨牛血液样品前处理方式对酶联免疫法（ELISA）检测克仑特罗残留结果的影响。结果表明:通过控制牛血液离心前放置时间、离心后放置时间、抗凝剂添加比例,可以有效提高血清样品的质量,最大程度的减少ELISA法检测牛血样中克仑特罗残留假阳性的结果。牛血液样品前处理的各因素对克仑特罗残留检测结果的影响顺序为:离心前放置时间>抗凝剂添加比例>离心后放置时间。牛血液样品最佳前处理条件为离心前放置30min、抗凝剂添加比例为1:9、离心后放置时间0min。按照牛血液样品最佳前处理条件,用ELISA法检测30份样品,筛选出2份阳性样品,经液相色谱-串联质谱法（LC-MS/MS）确证为阳性,两种方法检测结果一致。在1.0～8.1μg/L线性范围内,ELISA法检测克仑特罗残留结果假阳性率为0%。
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关键词:
- 牛血液; /
- 克仑特罗; /
- 酶联免疫法; /
- 液相色谱-串联质谱法;
Abstract: One-factor-at-a-time and orthogonal array design methods were employed to investigate the effect of bovine blood sample pre-treatment methods on the detection results of clenbuterol residues by Enzyme-linked Immunosorbent Assay (ELISA) .The results indicated that the serum sample quality was effectively improved, ELISA assay for detection bovine blood clenbuterol residual false positive results were reduced at the greatest degree, by controlling standing time of blood sample before centrifugation and after centrifugation, proportion of added anticoagulants into bovine blood.The effect of bovine blood sample pre-treatment methods on detection of clenbuterol residues were in the following order:standing time of blood sample before centrifugation>proportion of added anticoagulants into bovine blood>standing time of blood sample after centrifugation.The optimum bovine blood sample pre-treatment methods were for standing 30min before centrifugation, added anticoagulants in proportion of 1:9, standing 0min after centrifugation.In accordance with the bovine blood sample optimal pre-treatment conditions before analysis, two positive samples were screened out of 30 bovine blood samples using ELISA, which was consistent with the results from LC-MS/MS analysis.Within the linear range of 1.0 ~ 8.1μg/L, the false positive rate of clenbuterol residues was 0%. -
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