In Situ Sample Cleanup during Immunoassay: A Simple Method for Rapid Detection of Aflatoxin B1 in Food Samples
IR@IICB: CSIR-Indian Institute of Chemical Biology, Kolkata
View Archive Info| Field | Value | |
| Title |
In Situ Sample Cleanup during Immunoassay: A Simple Method
for Rapid Detection of Aflatoxin B1 in Food Samples
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| Creator |
Pal, Arindam
Acharya, Debopam Saha, Debjani Roy, Dipika Dhar, Tarun K |
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| Subject |
Drug Development/Diagnostics & Biotechnology
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| Description |
A strategy for rapid in situ elimination of interfering substances that are present in extracts of food samples during assay is described in this article. The novel feature of this method is that the sample purification is carried out as a part of the assay, and a separate sample cleanup step is not required. The assay procedure involves the sequential addition of standard or sample,
cleaning solutions, and aflatoxin B1–horseradish peroxidase conjugate (AFB1-HRP) over antibody-spotted zones of a membrane, and 3,39-diaminobenzidine was used as the substrate for visualization. We have determined that trifluoroacetic acid and propionic acids at concentrations of 100 mM are highly effective for cleaning groundnut, wheat, corn, and poultry feed samples and that NaHCO3 (100 mM) is successful in cleaning processed soybean. In all cases, subsequent washing was performed with phosphate-buffered saline solution to facilitate the removal of traces of adhering interfering substances. A batch of 12
samples can be analyzed within 8 min either by visual comparison of the color intensity (inversely related to the analyte concentration) of a sample spot with those of reference standards or, more precisely, by densitometry. The method was tested for the analysis of AFB1 in groundnut, wheat, corn, processed soybean, chili, and poultry feed. The detection limit obtained was 5 mg/kg, except for chili, where it was 10 mg/kg. The average recoveries from different noninfected food samples spiked
with AFB1 at concentrations of 5 to 100 mg/kg were between 99 and 105%. The values obtained for infected corn and
groundnut samples correlated well with the estimates obtained by high-pressure liquid chromatography. The absence of a sample extraction step reduces the cost and labor involved in the assay. The method may be potentially applicable to the assay of other mycotoxins and environmental pollutants.
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| Date |
2005-05-17
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| Type |
Article
PeerReviewed |
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| Format |
application/pdf
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| Identifier |
http://www.eprints.iicb.res.in/2674/1/Journal_of_Food_Protection%2C_Vol._68%2C_No._10%2C_2005.pdf
Pal, Arindam and Acharya, Debopam and Saha, Debjani and Roy, Dipika and Dhar, Tarun K (2005) In Situ Sample Cleanup during Immunoassay: A Simple Method for Rapid Detection of Aflatoxin B1 in Food Samples. Journal of Food Protection, 68 (10). 2169-2177. |
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| Relation |
http://www.eprints.iicb.res.in/2674/
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