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Preparation of a Broadly Specific Monoclonal Antibody-Based Indirect Competitive ELISA for the Detection of Benzodiazepines in Edible Animal Tissues and Feed
- Source :
- Food Analytical Methods. 9:3407-3419
- Publication Year :
- 2016
- Publisher :
- Springer Science and Business Media LLC, 2016.
-
Abstract
- Incorrect use of benzodiazepines could result in serious health problems. To monitor the illegal use of benzodiazepine compounds in animals, a group-specific monoclonal antibody (mAb) was prepared in this study. The obtained 3D7 mAb, which is an IgG1 isotype mAb, displayed an IC50 value of 8.9 ng mL−1 for diazepam and exhibited cross-reaction for diazepam (100 %), nitrazepam (49 %), nordiazepam (140 %), temazepam (32 %), oxazepam (17 %), estazolam (7.5 %), and alprazolam (2.4 %). Based on this mAb, for the first time in this study, an optimized indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) protocol that did not require complicated sample preparation and clean-up was developed. The detection limits of this ic-ELISA for benzodiazepines ranged from 1.2 to 3.3 μg kg−1 in muscle matrix and from 25.2 to 55.4 μg kg−1 in feed matrix. The recoveries ranged from 70.9 to 111.3 % with coefficients of variation below 15.0 %. Good correlations (r > 0.9494) between the results of the ic-ELISA and high-performance liquid chromatography were also observed. This simple method reduced the time required for sample preparation, ensured greater throughput, and met the requirements for benzodiazepine residue analyses. In conclusion, the proposed method is a sensitive and rapid multi-residue technique that offers a cost-effective alternative to current published procedures without any concession in the ability to detect benzodiazepine sedative misuse.
- Subjects :
- Benzodiazepine
Chromatography
Nitrazepam
Chemistry
Temazepam
medicine.drug_class
010401 analytical chemistry
Pharmacology
030226 pharmacology & pharmacy
01 natural sciences
Applied Microbiology and Biotechnology
Estazolam
0104 chemical sciences
Analytical Chemistry
Matrix (chemical analysis)
03 medical and health sciences
0302 clinical medicine
Alprazolam
Oxazepam
medicine
Safety, Risk, Reliability and Quality
Safety Research
Diazepam
Food Science
medicine.drug
Subjects
Details
- ISSN :
- 1936976X and 19369751
- Volume :
- 9
- Database :
- OpenAIRE
- Journal :
- Food Analytical Methods
- Accession number :
- edsair.doi...........c1747f7c8a8c076ad03a080bcb16c541