中药成分辛弗林对胶体金免疫层析法检测β-受体激动剂的影响

Effect of Traditional Chinese Medicine Compound Synephrine on the Detection ofβ-agonists Using Colloidal Gold Immunochromatographic Assay

旨在探究陈皮、青皮、木瓜、厚朴、红景天等5种中药材提取液以及饲喂了陈皮和青皮的猪尿液胶体金免疫层析法检测结果呈阳性的原因。建立测定猪尿及中药提取物中辛弗林的LC-MS/MS检测方法,测定陈皮、青皮的提取物和饲喂陈皮、青皮后猪尿中的辛弗林浓度。用小鼠肝微粒体孵育CGIA检测呈阳性的陈皮、青皮、木瓜和厚朴4味中药提取物。饲喂陈皮和青皮后的猪尿液检出辛弗林,浓度分别为1.36和1.65μg·mL^(-1);在陈皮和青皮的提取复溶液中检出辛弗林,浓度分别为132.6和312.7μg·mL^(-1)。厚朴和木瓜两种中药提取物经过肝微粒体孵育后,在2~5 h逐渐由CGIA检测阳性转为阴性,陈皮和青皮孵育后,0~6 h的结果均为CGIA检测阳性;阴性中药组CGIA检测均呈阴性,而添加辛弗林的阳性对照组0~6 h CGIA检测均呈阳性。猪较大剂量使用陈皮和青皮会引起猪尿β-受体激动剂CGIA检测出现假阳性,该假阳性现象跟中药成分辛弗林有关,体外试验中辛弗林不易被小鼠肝微粒体代谢。

This study was conducted to explore the reason why traditional Chinese medicine extractives and swine urines from pigs fed with some herbal were tested positive by CGIA.A LC-MS/MS method was established to determine synephrine in pigs’urine and traditional Chinese medicine extractives.Citri Reticulatae Pericarpium Viride and Citri Reticulatae Pericarpium’s extractives were determined,along with urine after herbal feed.In vitro mouse liver microsomes model was used to incubate the extractives of Citri Reticulatae Pericarpium Viride,Citri Reticulatae Pericarpium,Magnoliae Officinalis Cortex,and Chaenomelis Fructus.Synephrine was detected in pig urines after feeding with Citri Reticulatae Pericarpium and Citri Reticulatae Pericarpium Viride,and the concentration was 1.36 and 1.65μg·mL^(-1)respectively.The concentration of synephrine in Citri Reticulatae Pericarpium and Citri Reticulatae Pericarpium Viride's extractive was 132.6 and 312.7μg·mL^(-1)respectively.Extractives of Magnoliae Officinalis Cortex and Chaenomelis Fructus were tested gradually from positive to negative during 2-5 h incubation.Extractives of Citri Reticulatae Pericarpium Viride and Citri Reticulatae Pericarpium were tested positive during 0-6 h incubation.The experiment group of negative herbs were tested negative during incubation,and control group of synephrine matched were tested positive during 0-6 h incubation.Great dosing of Citri Reticulatae Pericarpium Viride and Citri Reticulatae Pericarpium in swine can cause false positive results ofβ-agonists’CGIA test in swine urine,and this phenomenon is related to synephrine.Synephrine is hard to be metabolized by mouse liver microsomes in vitro.