Determination of umbilical cord and maternal plasma concentrations of fentanyl by using novel spectrophotometric and chemiluminescence enzyme immunoassays |
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Authors: | Yannis DotsikasYannis LLoukas Ioanna Siafaka |
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Institution: | a Department of Pharmaceutical Chemistry, School of Pharmacy, University of Athens, Panepistimioupoli Zografou 15771, Athens, Greece b Department of Anaesthesia, Pain Therapy and Palliative Care, Aretaieion Hospital, Vas. Sofias 76, Medical School, University of Athens, Athens, Greece |
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Abstract: | Two novel enzyme immunoassays of fentanyl have been developed using Horseradish Peroxidase (HRP) as an enzyme, 3,3′,5,5′tetramethylbenzidine (TMB) and luminol as its colorimetric and its chemiluminescence substrate, respectively. A fentanyl polyclonal antibody was used as a capture antibody for fentanyl and fentanyl-bovine serum albumin (BSA) conjugate. The latter was first biotinylated and then bound by streptavidin labeled with HRP, resulting in the development of two novel competitive immunoassays. The detection limits were 0.045 and 0.0048 ng ml−1 for spectrophotometric and chemiluminescence detection, respectively, and were much lower than existing HRP-fentanyl based kits. Intra- and inter-assay CVs were 2.6-4.5 and 5.4-11.2%, respectively, at concentrations of 0.050-5.000 ng ml−1 for the colorimetric assay, whilst for the chemiluminescent assay intra- and inter-assay CVs were 3.7-6.2 and 6.2-12.3%, respectively, for the linear range of the assay at concentrations of 0.008-0.800 ng ml−1. The methods in this study were developed in order to measure maternal and neonatal fentanyl plasma samples during cesarean section, after the application of a novel subarachnoid analgesia technique. The 28 maternal and neonatal plasma samples were measured by both assays, providing data for subarachnoid administration of fentanyl that had never been presented before. |
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Keywords: | Fentanyl Immunoassay TMB Luminol Subarachnoid administration Umbilical cord samples |
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