Establishment Of Immunofluorescence Chromatography Test Strip For Drug Metabolites And Its Multi-detection Method

Objective: Establish immunofluorescence chromatography test strips for single and multiple quantitative detection of 4 drug metabolites.It is used to detect the four most common drug metabolites tetrahydrocannabinic acid,methamphetamine,morphine,and ketamine in urine,hair and sewage.To achieve the rapid judgment,traceability and treatment monitoring of the drug situation.Method: Fluorescent probes were prepared by labeling tetrahydrocannabinic acid(THCA),methamphetamine(MET),morphine(MOR),ketamine(KET)monoclonal antibodies and goat anti-chicken Ig Y with Europium nanoparticles(Eu NPs)respectively.Chicken Ig Y and THCA-BSA,MET-BSA,MOR-BSA,KET-BSA are coated on the NC membrane as the control line(C line)and text line(T line).Based on the principle of competitive immunochromatography,immunofluorescence chromatography test strips for single and multiple quantitative detection of drug metabolites were established.After the assembly of single and multiple quantitative test strips,a series of conditions have been optimized.Correspondingly evaluate the linear range,sensitivity,specificity and accuracy of each test strip under the best conditions.The detection results of actual samples are compared with Liquid chromatography-mass spectrometry(LC-MS).Results:(1)A better THCA detection material was selected,and the linear range of the optimized THCA immunofluorescence test strips is 10-500ng/m L.The test strip test results of the actual urine sample are compared with LC-MS,and the correlation between the two methods is good.(2)The linear ranges of the established MET/MOR/KET single detection fluorescent test strips are MET: 1-200 ng/m L,MOR:0.05-5 ng/m L,and KET: 0.1-100 ng/m L.Comparing the results of detecting hair with standard samples and LC-MS,the recoveries are between 81%-114%.(3)The linear ranges of the(MET+MOR+KET)Multi-detection immunofluorescence test strips(TRF-MLFIA)are MET: 0.5-50ng/m L,MOR: 0.02-5ng/m L,and KET: 0.02-50ng/m L.The results of the actual samples of hair and sewage have a good correlation with LCMS.Conclusion: In this study,single-detection and multiple-detection immunofluorescence test strips were prepared for the detection of four drug metabolites of tetrahydrocannabinic acid,methamphetamine,morphine,and ketamine.Among them,the results of the actual urine samples and hair-standard samples detected by singledetection immunofluorescence test strips are compared with LC-MS,and the accuracy rate is well.Compared with LC-MS,the results of multiple-detection immunofluorescence test strips for detecting hair and sewage samples have a good correlation.TRF-MLFIA simplifies the detection process,reduces testing costs and sample usage,and meets the testing needs of anti-drug agencies to the greatest extent.It also provides a new idea for the one-step method to determine the concentration of multiple analytes in a single sample.