ELISA detection of clonazepam and 7-aminoclonazepam in whole blood and urine.
ABSTRACT The ability of five commercially available enzyme-linked immunosorbent assay (ELISA) benzodiazepines to detect clonazepam and 7-aminoclonazepam in blood and urine was investigated. To determine the cross-reactivity of various ELISA assays, drug free blood and urine were fortified with clonazepam and 7-aminoflunitrazepam at concentrations of 1, 2.5, 5, 10, and 25microg/dl. The cross-reactivity, with respect to oxazepam, for clonazepam was 16, 37, 80, 93, and 109% with Immunalysis, Diagnostix, Neogen, OraSure, and Cozart, respectively; for 7-aminoclonazepam, none of the five ELISA assays showed any cross-reactivity above 10%.
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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.International Journal of Molecular Sciences 01/2013; 14(10):19474-83. · 2.46 Impact Factor
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ABSTRACT: SUMMARY: In recent years, there has been a notable increase in the number of reports of drug-facilitated crimes (DFCs). Usually, individuals report that they were robbed or assaulted while incapacitated by drugs. Most often, these cases have involved drugs that have the ability to produce an effect that leaves the victim in a semiconscious or unconscious state. It is reasonable to assume that the purpose of drug-induced incapacitation is probably largely unchanged with time. This covers the full range of property offences (particularly theft) and crimes against the person. What have changed are the drugs themselves: the number; type; their accessibility; effects and detection. The purpose of this review is to explore the different aspects related to the involvement and use of ethanol, sedative-hypnotics drugs, gamma-hydroxybutyrate (GHB) and ketamine in DFCs or offences, which may help people working in this field to expand their knowledge for better understanding of the nature of these crimes or offences.Annales Pharmaceutiques Françaises 09/2010; 68(5):275-85.
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ABSTRACT: A class selective enzyme-linked immunosorbent assay (ELISA) was developed for the detection of benzodiazepines, such as nitrazepam, clonazepam, 7-aminonitrazepam, 7-amidoclonzepam, chlordiazepoxide, delorazepam and nordazepam. Class selective artificial antigens were prepared by haptens of 7-aminonitrazepam and 7-aminoclonazepam conjugating to the carrier protein bovine serum albumin (BSA). Class selective polyclonal antibodies (PcAbs) were generated by immunising male New Zealand white rabbits with the class selective immunogens. Characterisation studies of the PcAbs showed that PcAbs raised by 7-aminonitrazepam–BSA had high affinity and specificity to the six relative benzodiazepines, while PcAbs raised by 7-amidoclonazepam–BSA had high affinity and specificity to five. So, PcAbs raised by 7-aminonitrazepam–BSA were selected to establish an indirect competitive ELISA for multi-residue determination. The IC50s of the optimised immunoassay were 2.50 µg/l, 3.13 µg/l, 1.92 µg/l, 2.78 µg/l, 3.13 µg/l and 4.55 µg/l for nitrazepam, clonazepam, 7-aminonitrazepam, 7-aminoclonazepam, delorazepam and nordazepam, respectively. Assay performance was validated using spiked pig urine by adding nirazepam. Recoveries were between 75 and 103%, and the coefficient variation was below 14%. This assay can be applied to detect multi-residues of benzodiazepines in urine.Food and Agricultural Immunology 01/2009; 20(4):281-293. · 0.73 Impact Factor