Effects of 105 traditional Chinese medicines on the detection of β-agonists in medicine extracts and swine urine based on colloidal gold immunochromatographic assay

doi:10.1016/S2095-3119(20)63265-8

摘要/Abstract

摘要：

本研究首先建立并验证了一种中兽药的提取方法，该方法能够提取中兽药产品中可能存在的克伦特罗、莱克多巴胺和沙丁胺醇等三种β-受体激动剂及其结构类似的物质。其次，采用该方法对105种批准在动物养殖中使用的中草药进行提取和浓缩，PBS复溶解后采用CGIA筛查存在阳性反应的中草药，出现阳性反应的样品随后经LC-MS/MS（液相色谱-串联质谱法）检测。在动物实验中，将105种中草药分别饲喂三元猪后，接取尿液进行CGIA检测，将有阳性反应的尿液样品进行LC-MS/MS检测。采用乙醇和乙腈（1:9，v/v）为溶剂，对中草药的提取效果良好，在105种受测的中草药的提取物中，青皮、陈皮、木瓜、厚朴和红景天等5种中草药的提取物出现CGIA阳性反应，其他中草药均显阴性反应。105种中草药分别饲喂猪后，喂过青皮和陈皮的猪只尿液出现CGIA检测阳性反应，其他中草药均显阴性反应，在随后的LC-MS/MS检测中喂过青皮和陈皮的猪只尿液未检出克伦特罗、莱克多巴胺或沙丁胺醇。在体外实验中，陈皮、青皮、木瓜、厚朴和红景天能引起克伦特罗、莱克多巴胺和沙丁胺醇CGIA检测阳性现象，该现象属于假阳性；三元猪被超剂量饲喂陈皮或青皮后，猪尿中莱克多巴胺的CGIA检测可出现假阳性。本研究首次提出了中草药的应用可能与临床CGIA检测β-受体激动剂出现假阳性的现象存在一定的关联性，提醒养猪从业者规范中草药添加剂的使用。发现引起猪尿假阳性的青皮和陈皮中存在有一种相同的化学成分——辛弗林，猪尿中存在相对浓度较低的辛弗林可引起莱克多巴胺的CGIA检测呈阳性反应。

Abstract:

Colloidal gold immunochromatographic assay (CGIA) is commonly used for the on-site detection of β-agonists that are sometimes used illegally as feed additives in swine diets. However, few studies have evaluated the causes of false-positive results that sometimes occur when applying CGIA in agricultural settings. In this study, we investigated if this false-positive phenomenon is related to the addition of certain traditional Chinese medicines (TCMs) to swine feed. We established and verified an extraction method for TCMs, and then applied CGIA to detect β-agonists in the extracts of 105 TCMs and in the urine of swine dosed with TCMs, respectively. Liquid chromatography-tandem mass spectrometry was used to validate the results of the urine samples tested positive for β-agonists using CGIA. The results were also verified using TCMs and colloidal gold test strips produced by different manufacturers. The extracts of Citri Reticulatae Pericarpium Viride, Citri Reticulatae Pericarpium, Magnoliae Officinalis Cortex, Chaenomelis Fructus, and Rhodiolae Crenulatae Radix Et Rhizoma were tested positive for β-agonists. Meanwhile, the addition of Citri Reticulatae Pericarpium Viride and Citri Reticulatae Pericarpium to swine feed resulted in false-positive results for β-agonists in swine urine. The results provide a new way to explain false-positive CGIA results and provide valuable information for livestock feeding programs.

Key words: colloidal gold immunochromatographic assay , false positive , traditional Chinese medicine , β-agonists , swine urine

. [J]. Journal of Integrative Agriculture, 2021, 20(6): 1626-1635.

WANG Zi-lin, FENG Ke-ying, GE Xiu-feng, MAI Jia-cheng, WANG Han-chuan, LIU Wen-zi, ZHANG Jia-hui, SHEN Xiang-guang. Effects of 105 traditional Chinese medicines on the detection of β-agonists in medicine extracts and swine urine based on colloidal gold immunochromatographic assay[J]. Journal of Integrative Agriculture, 2021, 20(6): 1626-1635.

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