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This work reports on a sensitive colorimetric immunoassay for aflatoxin B-1 (AFB(1)) in foodstuff. The reagent L-ascorbic acid 2-phosphate (AAP) is added to the system, and alkaline phosphatase (ALP) hydrolyzes AAP under formation of ascorbic acid and phosphate. The ascorbic acid produced reduces chloroauric acid to form zero-valent gold (in the form of nanoparticles). Hence, Au(III) is no longer available to oxidize 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonate) (ABTS) to form a green product. Rather, the solution remains colorless. By using ALP-labelled monoclonal anti-AFB(1) antibody, a competitive enzyme-label immunoassay was developed for AFB(1) in a microplate coated with the AFB(1)-BSA conjugate. Under optimal conditions, the absorbance of the solution at 415 nm increases linearly with increasing AFB(1) concentration in range from 10 pg center dot mL(-1) to 100 ng center dot mL(-1) (while the color gradually turns to green), and the detection limit is 7.8 pg center dot mL(-1). The precision of the method (expressed as RSD) is +/- 9.7%. The accuracy was validated by analyzing both naturally contaminated and spiked food samples, and the results matched the results obtained by ELISA very well.
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MICROCHIMICA ACTA
ISSN: 0026-3672
Year: 2017
Issue: 7
Volume: 184
Page: 2387-2394
5 . 7 0 5
JCR@2017
5 . 4 0 0
JCR@2023
ESI Discipline: CHEMISTRY;
ESI HC Threshold:226
JCR Journal Grade:1
CAS Journal Grade:2
Cited Count:
WoS CC Cited Count: 23
SCOPUS Cited Count: 22
ESI Highly Cited Papers on the List: 0 Unfold All
WanFang Cited Count:
Chinese Cited Count:
30 Days PV: 1
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