Single-Laboratory Validation for the Determination of Aflatoxin B1, B2, G1, and G2 in Foods Based on Immunoaffinity Column and Liquid Chromatography with Postcolumn Derivatization and Fluorescence Detection

2012 ◽  
Vol 6 (1) ◽  
pp. 36-44 ◽  
Author(s):  
Cemalettin Baltacı ◽  
Huri İlyasoğlu ◽  
Ferhat Yüksel
Keyword(s):  
Liquid Chromatography ◽  
Fluorescence Detection ◽  
Aflatoxin B1 ◽  
Immunoaffinity Column ◽  
Single Laboratory ◽  
Postcolumn Derivatization

scholarly journals Immunoaffinity Column Cleanup with Liquid Chromatography for Determination of Aflatoxin B1 in Corn Samples: Interlaboratory Study

2007 ◽  
Vol 90 (3) ◽  
pp. 765-772 ◽  
Author(s):  
Carlo Brera ◽  
Francesca Debegnach ◽  
Valentina Minardi ◽  
Elena Pannunzi ◽  
Barbara De Santis ◽  
...  
Keyword(s):  
Liquid Chromatography ◽  
Standard Deviation ◽  
Relative Standard Deviation ◽  
Aflatoxin B1 ◽  
Interlaboratory Study ◽  
Precolumn Derivatization ◽  
Immunoaffinity Column ◽  
Relative Standard ◽  
Postcolumn Derivatization

Abstract An interlaboratory study was conducted to evaluate the effectiveness of an immunoaffinity column cleanup liquid chromatography (LC) method for the determination of aflatoxin B1 levels in corn samples, enforced by European Union legislation. A test portion was extracted with methanolwater (80 + 20); the extract was filtered, diluted with phosphate-buffered saline solution, filtered on a microfiber glass filter, and applied to an immunoaffinity column. The column was washed with deionized water to remove interfering compounds, and the purified aflatoxin B1 was eluted with methanol. Aflatoxin B1 was separated and determined by reversed-phase LC with fluorescence detection after either pre- or postcolumn derivatization. Precolumn derivatization was achieved by generating the trifluoroacetic acid derivative, used by 8 laboratories. The postcolumn derivatization was achieved either with pyridinium hydrobromide perbromide, used by 16 laboratories, or with an electrochemical cell by the addition of bromide to the mobile phase, used by 5 laboratories. The derivatization techniques used were not significantly different when compared by the Student's t-test; the method was statistically evaluated for all the laboratories. Five corn sample materials, both spiked and naturally contaminated, were sent to 29 laboratories (22 Italian and 7 European). Test portions were spiked with aflatoxin B1 at levels of 2.00 and 5.00 ng/g. The mean values for recovery were 82% for the low level and 84% for the high contamination level. Based on results for spiked samples (blind pairs at 2 levels) as well as naturally contaminated samples (blind pairs at 3 levels), the values for relative standard deviation for repeatability (RSDr) ranged from 9.9 to 28.7%. The values for relative standard deviation for reproducibility (RSDR) ranged from 18.6 to 36.8%. The method demonstrated acceptable within- and between-laboratory precision for this matrix, as evidenced by the HorRat values.

scholarly journals Immunoaffinity Column Cleanup with Liquid Chromatography Using Post-Column Bromination for Determination of Aflatoxins in Hazelnut Paste: Interlaboratory Study

2005 ◽  
Vol 88 (2) ◽  
pp. 526-535 ◽  
Author(s):  
Hamide Z Senyuva ◽  
John Gilbert
Keyword(s):  
Liquid Chromatography ◽  
Standard Deviation ◽  
Relative Standard Deviation ◽  
Aflatoxin B1 ◽  
The United States ◽  
Interlaboratory Study ◽  
Immunoaffinity Column ◽  
Test Portion ◽  
Relative Standard

Abstract An interlaboratory study was conducted to evaluate the effectiveness of an immunoaffinity column cleanup liquid chromatography (LC) method for determination of aflatoxin B1 and total aflatoxins in hazelnut paste at European regulatory limits. The test portion was extracted with methanol–water (6 + 4). The extract was filtered, diluted with phosphate-buffered saline (PBS) solution to a specified solvent concentration, and applied to an immunoaffinity column containing antibodies specific to aflatoxins. The aflatoxins were removed from the immunoaffinity column with methanol, and then quantified by reversed-phase LC with post-column derivatization (PCD) involving bromination. The PCD was achieved with electrochemically generated bromine (Kobra Cell®) followed by fluorescence detection (except for one participant who used pyridinum hydrobromide perbromide for bromination). Hazelnut paste, both naturally contaminated with aflatoxins and blank (<0.1 ng/g) for spiking by participants with aflatoxins, was sent to 14 collaborators in Belgium, The Netherlands, Spain, Turkey, the United Kingdom, and the United States. Test portions were spiked at levels of 4.0 and 10.0 ng/g for total aflatoxins by participants using supplied total aflatoxins standards. Recoveries for total aflatoxins and aflatoxin B1 averaged from 86 to 89%. Based on results for naturally contaminated samples (blind duplicates at 3 levels ranging from 4.0 to 11.8 ng/g total aflatoxins), the relative standard deviation for repeatability (RSDr) ranged from 2.3 to 3.4% for total aflatoxins and from 2.2 to 3.2% for aflatoxin B1. The relative standard deviation for reproducibility (RSDR) ranged from 6.1 to 7.0% for total aflatoxins and from 7.3 to 7.8% for aflatoxin B1. The method showed exceptionally good within-laboratory and between-laboratory precision for hazelnut paste, as evidenced by HORRAT values, which in all cases were significantly below target levels, the low levels of determination for both aflatoxin B1 and total aflatoxins.

scholarly journals Determination of Sulfonamides in Edible Salmon Tissue by Liquid Chromatography with Postcolumn Derivatization and Fluorescence Detection

1997 ◽  
Vol 80 (4) ◽  
pp. 751-755 ◽  
Author(s):  
Theresa A Gehring ◽  
Larry G Rushing ◽  
Harold C Thompson
Keyword(s):  
Acetic Acid ◽  
Liquid Chromatography ◽  
Fluorescence Detection ◽  
Coho Salmon ◽  
Gradient Elution ◽  
Reversed Phase ◽  
Aqueous Acetic Acid ◽  
Postcolumn Derivatization

Abstract Fourteen sulfonamides—sulfanilamide, sulfadiazine, sulfathiazole, sulfapyridine, sulfam- erazine, sulfamethazine, sulfamethizole, sulfamethoxypyridazine, sulfachloropyridazine, sulfamonomethoxine, suļfadoxine, sulfamethoxazole, sulfadimethoxine, and sulfaquinoxoline—residues of which could be found in aquacultured species, were separated in <25 min by reversed-phase (C18) liquid chromatography (LC) with gradient elution. Analytes were extracted from edible salmon tissue (muscle and adhering skin) with acetonitrile—2% aqueous acetic acid, isolated with 2 liquid-liquid partitionings, and derivatized with fluorescamine after eluting from the column. The derivatives were detected by fluorescence. Recoveries (n = 4) from coho salmon fortified with sulfonamides at 5,10, and 20 ng/g tissue averaged 79.7± 7.3, 84.6 ± 7.7, and 88.2 ± 7.1%, respectively. Limits of quantitation were 5 ng/g tissue, for sulfanilamide, sulfamethoxypyridazine, and sulfaquinoxoline and 1 ng/g tissue for the remaining sulfonamides.

Method Validation for the Quantitative Analysis of Aflatoxins (B1, B2, G1, and G2) and Ochratoxin A in Processed Cereal-Based Foods by HPLC with Fluorescence Detection

2015 ◽  
Vol 98 (4) ◽  
pp. 939-945 ◽  
Author(s):  
Işil Gazioğlu ◽  
Ufuk Kolak
Keyword(s):  
Quantitative Analysis ◽  
Simultaneous Determination ◽  
Ochratoxin A ◽  
Fluorescence Detection ◽  
Aflatoxin B1 ◽  
Hplc Analysis ◽  
Extraction Procedure ◽  
Immunoaffinity Column ◽  
Rp Hplc

Abstract Modified AOAC 991.31 and AOAC 2000.03 methods for the simultaneous determination of total aflatoxins (AFs), aflatoxin B1, and ochratoxin A (OTA) in processed cereal-based foods by RP-HPLC coupled with fluorescence detection were validated. A KOBRA® Cell derivatization system was used to analyze total AFs. One of the modifications was the extraction procedure of mycotoxins. Both AFs and OTA were extracted with methanol–water (75 + 25, v/v) and purified with an immunoaffinity column before HPLC analysis. The modified methods were validated by measuring the specificity, selectivity, linearity, sensitivity, accuracy, repeatability, reproducibility, recovery, LOD, and LOQ parameters. The validated methods were successfully applied for the simultaneous determination of mycotoxins in 81 processed cereal-based foods purchased in Turkey. These rapid, sensitive, simple, and validated methods are suitable for the simultaneous determination of AFs and OTA in the processed cereal-based foods.

scholarly journals Occurrence of aflatoxins in peanuts and peanut products determined by liquid chromatography with fluorescence detection

2013 ◽  
pp. 27-35 ◽  
Author(s):  
Biljana Stojanovska-Dimzoska ◽  
Zehra Hajrulai-Musliu ◽  
Elizabeta Dimitrieska-Stojkovic ◽  
Risto Uzunov ◽  
Pavle Sekulovski
Keyword(s):  
Liquid Chromatography ◽  
Fluorescence Detection ◽  
Limit Of Detection ◽  
Total Content ◽  
Immunoaffinity Column ◽  
Limit Of Quantification ◽  
Three Samples ◽  
The Mean ◽  
Positive Results

Liquid chromatography with fluorescence detection using immunoaffinity column clean-up was a method described for determination of aflatoxins (AFB1, AFB2, AFG1 and AFG2) in peanuts and peanut based products. The validation of the procedure was performed. Good coefficient of correlation was found for all aflatoxins in the range of 0.9993-0.9999. Limit of detection (LOD) and limit of quantification (LOQ) ranged from 0.003-0.005 mg/kg and 0.009-0.023 mg/kg, respectively, which was acceptable. The mean recovery for total aflatoxins was 88.21%. The method also showed acceptable precision values in the range of 0.171-2.626% at proposed concentration levels for all four aflatoxins. RSDR values (within laboratory reproducibility) calculated from the results showed good correlation between two analysts for all aflatoxins and they ranged from 4.93-11.87%. The developed method was applied for the determination of aflatoxins in 27 samples of peanuts and peanut based products. The results showed that 21 peanut samples (77.7%) were below LOD of the method. Three samples had positive results over the MRL. There was one extreme value recorded for the total aflatoxins in peanut (289.2 mg/kg) and two peanut based products, peanut snack and peanut, with total content of aflatoxins being 16.3 mg/kg and 8.0 mg/kg, respectively. The obtained results demonstrated that the procedure was suitable for the de?termination of aflatoxins in peanuts and peanut based products and it could be implemented for the routine analysis.

scholarly journals Determination of Aflatoxin B1 in Baby Food (Infant Formula) by Immunoaffinity Column Cleanup Liquid Chromatography with Postcolumn Bromination: Collaborative Study

2001 ◽  
Vol 84 (4) ◽  
pp. 1116-1124 ◽  
Author(s):  
Joerg Stroka ◽  
Elke Anklam ◽  
Urban Joerissen ◽  
John Gilbert ◽  
A Barmark ◽  
...  
Keyword(s):  
Liquid Chromatography ◽  
Standard Deviation ◽  
Relative Standard Deviation ◽  
Infant Formula ◽  
Aflatoxin B1 ◽  
Collaborative Study ◽  
Baby Food ◽  
Immunoaffinity Column ◽  
Relative Standard

Abstract A collaborative study was conducted to evaluate the effectiveness of an immunoaffinity column cleanup liquid chromatography (LC) method for determination of aflatoxin B1 in a milk powder based infant formula at a possible future European regulatory limit (0.1 ng/g). The test portion was extracted with methanol–water (8 + 2 [v + v]), filtered, diluted with water, and applied to an immunoaffinity column. The column was washed with water to remove interfering compounds, and the purified aflatoxin B1 was eluted with methanol. The separation and determination of the aflatoxin B1 was performed by reversed-phase LC and detected by fluorescence after postcolumn derivatization (PCD) involving bromination. PCD was achieved with either pyridinum hydrobromide perbromide (PBPB) or an electrochemical (Kobra) cell by addition of bromide to the mobile phase. The baby food (infant formula) test samples, both spiked and naturally contaminated with aflatoxin B1, were sent to 14 laboratories in 13 different European countries. Test portions were spiked at levels of 0.1 and 0.2 ng/g for aflatoxin B1. Recoveries ranged from 101 to 92%. Based on results for spiked test samples (blind pairs at 2 levels) and naturally contaminated test samples (blind pairs at 3 levels), the relative standard deviation for repeatability (RSDr) ranged from 3.5 to 14%. The relative standard deviation for reproducibility (RSDR) ranged from 9 to 23%. Nine participants used PBPB derivatization, and 5 particpants used the Kobra cell. There was no evidence of method performance depending on the derivatization method used. The method showed acceptable within- and between-laboratory precision for baby food matrix, as evidenced by HORRAT values, at the target levels of determination for aflatoxin B1.

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