Chen Guoying, Smith Emily, Qin Feng, Liu Linshu
Eastern Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, Wyndmoor, Pennsylvania 19038, USA.
J Agric Food Chem. 2006 May 3;54(9):3225-30. doi: 10.1021/jf053243p.
Analyses of chemical residues in animal tissue matrices require multistep sample preparation. To simplify this process, a methodology was developed that combines sorbent extraction and solid-matrix time-resolved luminescence (TRL); it was applied to tetracycline screening in milk. Reported here is an effort to extend its application to tissue matrices, illustrated by oxytetracycline (OTC) screening in catfish muscle. Extraction and enrichment are accomplished by immersing small C18 sorbent strips into tissue homogenates for 20 min, followed by a 3 min rinse in water and a 2 min dip in a reagent solution. After desiccation, TRL is measured directly on the sorbent surface. Tissue particulates no longer interfere via attenuation or scattering, rendering centrifugation and filtration unnecessary. The integrated TRL intensity shows a linear dependence on OTC concentration in the 0-8 microg/g range (R2 = 0.9992) with a 0.026 microg/g limit of detection. To screen OTC at 2 microg/g, the U.S. regulatory tolerance level, a threshold is established at x2-3sigma2, where x2 and sigma2 are the mean and standard deviation, respectively, of the TRL signals from 15 samples fortified at 2 microg/g. Among 45 blind samples randomly fortified at 0-4 microg/g, 41 were screened correctly and 4 negative samples were presumed positive. This method has the potential to improve throughput and save assay costs by eliminating acids, organic solvents, centrifugation, and filtration.
对动物组织基质中的化学残留物进行分析需要多步骤的样品制备。为简化这一过程,开发了一种将吸附剂萃取与固体基质时间分辨发光(TRL)相结合的方法;该方法应用于牛奶中的四环素筛查。本文报道了将其应用扩展到组织基质的研究工作,以鲶鱼肌肉中的土霉素(OTC)筛查为例进行说明。萃取和富集是通过将小的C18吸附剂条浸入组织匀浆中20分钟来完成的,随后在水中冲洗3分钟,再在试剂溶液中浸泡2分钟。干燥后,直接在吸附剂表面测量TRL。组织颗粒不再通过衰减或散射产生干扰,因此无需离心和过滤。积分TRL强度在0 - 8微克/克范围内显示出与OTC浓度呈线性关系(R2 = 0.9992),检测限为0.026微克/克。为筛查美国监管耐受水平2微克/克的OTC,在x2 - 3sigma2处设定一个阈值,其中x2和sigma2分别是来自15个添加2微克/克的样品的TRL信号的平均值和标准偏差。在45个随机添加0 - 4微克/克的盲样中,41个被正确筛查,4个阴性样品被误判为阳性。该方法有可能通过省去酸、有机溶剂、离心和过滤来提高通量并节省检测成本。