Direct Determination of Propranolol in Serum or Plasma by Fluoroimmunoassay

Al-Hakiem, Mohammad H.H.; White, G. W.; Smith, David S.; Landon, J.

doi:10.1097/00007691-198102000-00007

Cited by 19 publications

(9 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…21,22 With regard to the ability to detect low concentrations of (S)-propranolol, the improvements made here place the aqueous and organic solvent based assays using MIPs on the same level as immunoassays using biological antibodies. [34][35][36][37][38] Further work to improve the sensitivity of the aqueous assay may be based on the observation that the ratio of specific to nonspecific binding increases with increasing concentrations of ethanol and buffer (Figures 4 and 5). Hence, despite the fact that the total response is reduced, the portion that is sensitive to analyte concentration increases, which probably lead to an improvement of the overall assay performance.…”

Section: Discussionmentioning

confidence: 99%

Application of Molecular Imprinting to the Development of Aqueous Buffer and Organic Solvent Based Radioligand Binding Assays for (S)-Propranolol

1996

View full text Add to dashboard Cite

Antibody mimics have been prepared by molecular imprinting of (S)-propranolol and applied in the development of radioligand binding assays for the imprint species. In the assays, polymer particles, radioligand, and analyte were incubated either in an organic solvent or an aqueous buffer, with similarly high sensitivity under both set of conditions. Optimization of the assay conditions led to 100-1000-fold improvements in the limit of determination and 100-1000-fold reductions of the amount of imprinted polymer used, compared with previous studies of this novel type of assay. The present assay uses only 10-50 μg of polymer, and the limit of determination is about 6 nM. The toluene-based assay showed excellent enantioselectivity, the cross-reactivity of the R enantiomer being only 1%, which is better than that demonstrated by biological antibodies. The aqueous buffer based assay showed high substrate selectivity for propranolol in the presence of structurally similar β-blockers. The different selectivity profiles obtained are due to a different balance between hydrophobic and polar interactions in toluene and water, since polar interactions, such as hydrogen bonds, are strong in apolar solvents and hydrophobic interactions are strong in water.

show abstract

Section: Discussionmentioning

confidence: 99%

Application of Molecular Imprinting to the Development of Aqueous Buffer and Organic Solvent Based Radioligand Binding Assays for (S)-Propranolol

1996

View full text Add to dashboard Cite

show abstract

“…Clinically relevant plasma concentrations are in the range of approximately 3-300 nm (1-100 ng ml 21 ). [8][9][10] The inter-individual variation in response for blank plasma and samples spiked with known amounts of S-propranolol at three different concentrations was investigated. Samples were prepared using plasma drawn from six individuals (5 males and 1 female) and the inter-individual variation was found to be in the range of 3-8% (RSD).…”

Section: Resultsmentioning

confidence: 99%

Molecular Imprint Based Radioassay for Direct Determination of S-Propranolol in Human Plasma

Bengtsson¹,

Roos²,

Andersson³

1997

Anal. Commun.

View full text Add to dashboard Cite

Molecular imprints of S-propranolol have been applied to the development of a radiolabelled assay, which is conceptually identical to radioimmunoassay, for direct determination of the concentration of S-propranolol in biofluids. Determination of S-propranolol in human plasma and urine was used as a model system. The imprint based assay measured plasma and urine concentrations in the range of 20 to 1000 nM with accuracies of 89-107% and 91-125%, and precisions of 3-13% and 1-7%, respectively. The results demonstrate that it is possible to carry out molecular imprint based assays of biological samples without prior sample clean up.

show abstract

“…3). If an assay to detect lower concentrations of cotinine were re~uired, a separation fluorimmunoassay" L. 19 could be developed using anticotinine serum coupled to solid-phase particles and the same fluorescein-labelled cotinine. The separation step would enable the removal of sample components that interfere with the fluorescence end-point measurement, and therefore allow larger samp.le (urine or serum) volumes to be employed.…”

Section: Discussionmentioning

confidence: 99%

“…The product (IV) (50 mg) was diazotised, coupled to KLH (64 mg), and the conjugate isolated using previously described methodsY Anti-cotinine serum Three mature female sheep were immunised with the cotinine immunogen by a previously described protocol. 19 …”

Section: Preparation Of Cotinine-4' -Amidoglycyl-paminophenylalaninementioning

confidence: 99%

Single-Reagent Polarisation Fluoroimmunoassay for Cotinine (a Nicotine Metabolite) in Urine

et al. 1986

View full text Add to dashboard Cite

SUMMARY. A rapid, single-reagent, non-separation, non-isotopic immunoassay was developed for determining levels of the nicotine metabolite, cotinine, in urine.The single reagent was prepared by pre-mixing an appropriate dilution of sheep anti-cotinine serum with a fluorescein-labelled cotinine tracer.All normal reliability criteria were satisfied. The assay was found to be specific for cotinine and there was no cross reactivity with other available nicotine metabolites and structurally-related compounds. The results obtained correlated closely with those of an established radioimmunoassay. The assay was well-suited to application in the discrimination of active smokers from non-smokers (and passive smokers).A dose-response relationship exists between the number of cigarettes smoked and the risk of developing smoke-related diseases. 1 This explains the interest in ways of distinguishing between smokers and non-smokers (passive smokers) shown by insurance companies, surgeons and forensic scientists. Suitable methods include measurement of serum thiocyanate and carboxyhaemoglobin concentrations/: 3 and the concentrations of nicotine and cotinine in biological fluids.v" The measurement of urinary cotinine is the preferred approach for several reasons. Firstly, cotinine is only derived from the metabolism of nicotine," whereas carboxyhaemoglobin and thiocyanate levels can be raised due to environmental influences. 2 Secondly, the methods for measuring cotinine are less complex than those for thiocyanate and carboxyhaemoglobin. Thirdly, urine is more convenient to assay than blood. Finally, cotinine has a half-life of between 7 and 40 h,? whereas the half-life of nicotine is less than 30 rnin.l'' There are several methods available for the detection of cotinine concentrations in urine, including chromatographic procedures. However, despite recent developments in highperformance liquid chromatography and gasCorrespondence: M C Hansel, Department of Chemical Pathology, St Bartholomew's Hospital, 48--50Bartholomew Close, London ECIA 7HL, UK. 596 liquid chromatography, these methods are still complex and time consuming, and require sample preparation and large sample volumes." Radioimmunoassar (RIA) are the most widely used technique.P' . 11, 12 being both sensitive and specific, but their use may involve administrative and regulatory complications, and the methods are technically complex, due to the need for a separation step. Also, assays involving tritiated tracer suffer from the inconvenience and cost of liquid scintillation counting, while radioiodinated tracers have a limited shelf-life. This paper describes the development and validation of a single reagent polarisation fluoroimmunoassay (PFIA) for measuring urinary cotinine, based on the increase in signal associated with antibody binding of a fluorescent-labelled antigen. 13 Its use permits a rapid, non-separation assay, with reagents that have a virtually indefinite shelf-life and avoid any biohazard and administrative problems. The assay covers the range of cot...

show abstract

Direct Determination of Propranolol in Serum or Plasma by Fluoroimmunoassay

Cited by 19 publications

References 0 publications

Application of Molecular Imprinting to the Development of Aqueous Buffer and Organic Solvent Based Radioligand Binding Assays for (S)-Propranolol

Application of Molecular Imprinting to the Development of Aqueous Buffer and Organic Solvent Based Radioligand Binding Assays for (S)-Propranolol

Molecular Imprint Based Radioassay for Direct Determination of S-Propranolol in Human Plasma

Single-Reagent Polarisation Fluoroimmunoassay for Cotinine (a Nicotine Metabolite) in Urine

Contact Info

Product

Resources

About