Fluorescence polarization immunoassay for rapid determination of dehydroepiandrosterone in human urine

In this paper, five fluorescein-labeled dehydroepiandrosterone (DHEA) derivatives (tracers) with different chain lengths between the fluorescein and hapten were synthesized and featured so as to establish a fluorescence polarization immunoassay (FPIA) for DHEA detection in human urine samples with p...

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Bibliographic Details
Published in:Analytical and bioanalytical chemistry 2021-07, Vol.413 (17), p.4459-4469
Main Authors: Yang, Huiyi, He, Qiyi, Eremin, Sergei A., Pan, Junkang, Zou, Yikui, Cui, Xiping, Zhao, Suqing
Format: Article
Language:English
Subjects:
Analysis
Analytical Chemistry
Antibodies
Biochemistry
Characterization and Evaluation of Materials
Chemistry
Chemistry and Materials Science
Correlation coefficient
Correlation coefficients
Dehydroepiandrosterone
Enzyme-linked immunosorbent assay
Fluorescein
Fluorescence
Fluorescence polarization
Fluorescent indicators
Food Science
Human wastes
Immunoassay
Laboratory Medicine
Methods
Monitoring/Environmental Analysis
Polarization
Polarization (Light)
Polyclonal antibodies
Research Paper
Sensitivity
Urine
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Summary:In this paper, five fluorescein-labeled dehydroepiandrosterone (DHEA) derivatives (tracers) with different chain lengths between the fluorescein and hapten were synthesized and featured so as to establish a fluorescence polarization immunoassay (FPIA) for DHEA detection in human urine samples with previously prepared polyclonal antibody against DHEA. The outcomes of the structure of tracer on FPIA sensitivity were investigated. Under the optimal condition, the fluorescence polarization value (FP) decreases linearly in DHEA concentration, ranging from 1.6 to 243.3 ng mL −1 , with the limit of detection of 1.1 ng mL −1 and IC 50 value of 25.1 ng mL −1 . Moreover, the developed FPIA was time-saving as it could complete the detection within 3 min. FPIA and commercial enzyme-linked immunosorbent assay kit were both applied to analyze the spiked human urine samples with DHEA. Excellent recoveries (92.1–108.0%) and satisfactory correlation coefficient ( R 2  = 0.98) were acquired with the two methods, indicating that the developed FPIA was a fast and efficient screening immunoassay with accuracy and sensitivity for DHEA detection in human urine samples. Graphical abstract
ISSN:1618-2642
1618-2650
DOI:10.1007/s00216-021-03403-7