Abstract
It is challenging to detect 7-aminonitrazepam (7-ANZP) residue in animal tissues simply and sensitively by the enzyme-linked sorbent immunoassay (ELISA) method. This paper demonstrates that utilizing a bioconjugate of gold nanoparticles and enzyme-labeled antibody as a signal probe increases the sensitivity of a traditional ELISA for 7-ANZP by nearly 20 times. The sensitivity of this ELISA for 7-ANZP was 5.6 pg/mL in buffer, and the limit of detection (LOD) of 0.18 μg/kg for 7-ANZP in urine could be achieved after the urine samples were simply hydrolyzed and diluted by buffer. This simple and sensitive method has potential application for improving the sensitivity of ELISA methods against various small molecules.
Highlights
Benzodiazepines are among the most widely prescribed psychotropic drugs, and can eliminate erethism, manic anxiety and have a calming effect on patients by selectively inhibiting unrest and tension
In order to confirm whether 7-aminonitrazepam (7-ANZP) had been obtained, the product obtaining by reducing process of nitrazepam and subsequent preliminary purification was separated by chromatographic solution hexane/acetic acetate (1:1, v/v) on silica TLC plate
It was important to note that the Gold nanoparticles (GNPs) -IgG-HRP conjugate was mono-disperse, which will contribute to the reproducibility of the GNPs-based enzyme-linked sorbent immunoassay (ELISA) method
Summary
Benzodiazepines are among the most widely prescribed psychotropic drugs, and can eliminate erethism, manic anxiety and have a calming effect on patients by selectively inhibiting unrest and tension. Based on fluorescent CdTe nanoparticle bioconjugations and microfluidic chip, we previously reported an immunoassay for the metabolite of clonazepam In this method, the LOD achieved was. The above-mentioned immunoassays have shown very good results, detecting benzodiazepines in urine using simple and sensitive ELISA methods is challenging due to serious matrix interference [11,12,13,14]. A nano-enhanced ELISA was demonstrated for rapid and sensitive detection of 7-ANZP in urine samples. Combining a traditional ELISA format and a GNP-antibody-enzyme bioconjugate as a single probe, this nano-enhanced ELISA method achieved a remarkably low detection limit of 0.18 ng/g 7-ANZP in urine using a simple traditional ELISA protocol.
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