Determination of Ochratoxin A in Feed by Immunoaffinity Cleanup and Liquid Chromatography

2013 ◽  
Vol 96 (3) ◽  
pp. 599-602 ◽  
Author(s):  
Ping Ding ◽  
Ziyou Mi ◽  
Yali Hou ◽  
Yigang He ◽  
Jianhua Xie
Keyword(s):  
Liquid Chromatography ◽  
Detection Limit ◽  
Ochratoxin A ◽  
Cyanogen Bromide ◽  
Polyclonal Antibodies ◽  
Immunoaffinity Column ◽  
Low Levels ◽  
Sepharose 4B ◽  
Feed Samples

Abstract A method using LC was developed for determination of ochratoxin A (OTA) in feeds. The extracted samples were cleaned up by an immunoaffinity column prepared by covalently coupling polyclonal antibodies against OTA to cyanogen bromide-activated Sepharose 4B. The eluates were determined by LC with fluorescence detection. Recoveries of OTA from fortified samples of 1–10 μg/kg levels ranged from 84.3 to 90.0%, with CVs of 3.3–7.8%. The detection limit was 0.045 μg/kg based on an S/N of 3:1. A total of 65 feed samples were screened for OTA with the proposed method. The results showed that only nine samples were contaminated with OTAs at low levels. The presented method was successfully applied to quantify OTAs in real feed samples.

scholarly journals Residue Analysis for Halofuginone in Sturgeon Muscle by Immunoaffinity Cleanup and Liquid Chromatography

2005 ◽  
Vol 88 (6) ◽  
pp. 1644-1648 ◽  
Author(s):  
Shuangyang Ding ◽  
Yali Hou ◽  
Ningpeng Wu ◽  
Jiangzhong Shen
Keyword(s):  
Liquid Chromatography ◽  
High Performance ◽  
Cyanogen Bromide ◽  
Polyclonal Antibodies ◽  
Residue Analysis ◽  
Immunoaffinity Chromatography ◽  
Absorbance Detection ◽  
Coefficients Of Variation ◽  
Sepharose 4B

Abstract A high-performance liquid chromatography (LC) method was developed for the determination of halofuginone (HFG) in sturgeon muscle. The extracted samples were cleaned up by an immunoaffinity chromatography column that was prepared by covalently coupling polyclonal antibodies against HFG to cyanogen bromide (CNBr) activated Sepharose 4B. The eluate was evaporated to dryness, and residues were determined by LC with absorbance detection at 243 nm. Recoveries of HFG from samples fortified at 20–200 μg/kg levels ranged 74.6–81.1%, with coefficients of variation of 0.7–8.6%. The detection limit was estimated to be 10 μg/kg in a 2 g sample.

Comparison of Postcolumn Derivatization-Liquid Chromatography with Thin-Layer Chromatography for Determination of Aflatoxins in Naturally Contaminated Corn

1990 ◽  
Vol 73 (4) ◽  
pp. 579-581 ◽  
Author(s):  
Rodney W Beaver ◽  
David M Wilson ◽  
Mary W Trucksess
Keyword(s):  
Liquid Chromatography ◽  
Detection Limit ◽  
Thin Layer ◽  
Thin Layer Chromatography ◽  
Correlation Coefficient ◽  
Low Levels ◽  
Aflatoxin B2 ◽  
Layer Chromatography ◽  
Postcolumn Derivatization

Abstract Quantitation of aflatoxins by liquid chromatography with postcolumn iodine derlvatization (LC-PCD) and fluorescence detection was compared with quantitation by the AOAC CB method, 968.22. Thirty-seven naturally contaminated corn samples were ground and then divided. One portion was extracted, and the extract was cleaned up and analyzed by thin-layer chromatography according to the CB method. The second portion was extracted and cleaned up In a similar fashion, but quantitation was by the LC-PCD method. For aflatoxin B1, concentrations ranging from 0 to 150 ng/g, results obtained by the 2 methods were fitted to a linear equation with the LC-PCD results as the dependent variable. The correlation coefficient was 0.99, the Intercept was near 0, and the slope was near 1. For aflatoxin B2, the correlation coefficient was 0.97, and the Intercept was near 0. However, the slope of the equation relating LC-PCD concentration to TLC concentration was only 0.5. We believe that this lack of equivalence between the methods for determination of aflatoxin B2 is due to overestlmatlon by the TLC method because the low levels present are near the TLC detection limit for B2.

Determination of Deoxynivalenol in Wheat-Based Breakfast Cereals Marketed in Portugal

2001 ◽  
Vol 64 (11) ◽  
pp. 1848-1850 ◽  
Author(s):  
MARIA LÍGIA MARTINS ◽  
H. MARINA MARTINS
Keyword(s):  
Liquid Chromatography ◽  
Detection Limit ◽  
Natural Occurrence ◽  
Fungal Metabolites ◽  
Immunoaffinity Column ◽  
Breakfast Cereals ◽  
Average Recovery ◽  
Genus Fusarium ◽  
Natural Contamination

Deoxynivalenol (DON), also known as vomitoxin, is one of a group of closely related secondary fungal metabolites—the trichothecenes—and is produced predominantly by several species of the genus Fusarium, especially Fusarium graminearum. The present study was carried out to evaluate the natural occurrence of DON in different kinds of wheat-based breakfast cereals widely consumed by the population. A total of 88 commercially available samples of wheat-based breakfast cereals were randomly collected from different supermarkets in Lisbon, Portugal. The samples were analyzed using immunoaffinity column, and DON was quantified by liquid chromatography. Detection limit was 100 μg/kg. Average recovery of DON was 80%. Of 88 analyzed samples, 72.8% contained levels of DON between 103 and 6,040 μg/kg, with mean level of 754 μg/kg, and 24 samples (27.2%) were not contaminated (<100 μg/kg). These results indicate an incidence of this mycotoxin in these products, and the authors suggest a monitoring for the prevention of molds and mycotoxins. This is the first report in Portugal on natural contamination with DON in wheat-based breakfast cereals.

scholarly journals Comparison of two clean up techniques in isolation of ochratoxin A from red wine

2010 ◽  
Vol 28 (No. 3) ◽  
pp. 233-241 ◽  
Author(s):  
E. Belajová ◽  
D. Rauová
Keyword(s):  
Acetic Acid ◽  
Liquid Chromatography ◽  
Detection Limit ◽  
Ochratoxin A ◽  
Red Wine ◽  
Solid Phase ◽  
Phase Extraction ◽  
Immunoaffinity Column ◽  
Experimental Conditions ◽  
Quantification Limit

Two procedures for the extraction of ochratoxin A (OTA) from red wine &ndash; the reference clean up procedure using specific immunoaffinity column (IAC), and solid phase extraction (SPE) in which an active carbon was employed, was compared. In SPE procedure, various mixtures of dichloromethane (D), toluene (T), acetonitrile (AC), methanol (M), and acetic acid (A) were used as OTA desorption agents. Two types of SPE carbonaceous columns were tested &ndash; commercial SPE columns (Supelclean<sup>TM</sup> Envi-Carb) with a nonporous graphitised carbon, and SPE columns prepared in our laboratory (further specified as Lab-Carb) that were filled with a micro particular granular carbon. OTA was extracted from spiked red wine by the use of both carbonaceous columns. The highest OTA mean recovery calculated in relation to the reference IAC procedure was 98.5%, using the Lab-Carb adsorbent and acetonitrile + toluene, 3 + 1 (v + v) as the elution mixture (OTA spike levels of 0.2 &micro;g/l). Using the elution mixture of dichloromethane + methanol, 9 + 1 (v + v), the relative recoveries of 76.4% and 82.9% were reached at the OTA spike levels of 0.2 &micro;g/l and 1.9 &micro;g/l, respectively. The application of Envi-Carb adsorbent generally resulted in a very poor OTA recovery under the experimental conditions used (less than 50%). OTA was detected by liquid chromatography with fluorescence detection (LC-FLD) providing the detection limit of 0.011 &micro;g/l and the quantification limit of 0.033 &micro;g/l. &nbsp;

scholarly journals Simultaneous Determination of Aflatoxins, Ochratoxin A, and Zearalenone in Grains by New Immunoaffinity Column/Liquid Chromatography

2004 ◽  
Vol 87 (2) ◽  
pp. 411-416 ◽  
Author(s):  
Roswitha Göbel ◽  
Klaus Lusky
Keyword(s):  
Column Liquid Chromatography ◽  
Liquid Chromatography ◽  
Simultaneous Determination ◽  
Ochratoxin A ◽  
Fluorescence Detection ◽  
Trifluoroacetic Acid ◽  
Detection Limits ◽  
Immunoaffinity Column ◽  
Recovery Rates

Abstract The simultaneous determination of mycotoxins was performed in 3 steps: extraction, cleanup, and detection. For extraction, a mixture of acetonitrile–water (60 + 40, v/v) was proved appropriate. For cleanup, a new Afla-Ochra-Zea immunoaffinity column was used. After derivatization with trifluoroacetic acid, the mycotoxins aflatoxins, ochratoxin A (OTA), and zearalenone (ZEA) were determined simultaneously by liquid chromatography with fluorescence detection. The detection limits in different matrixes after cleanup with the new immunoaffinity column were very low: aflatoxins, 0.002–0.7 μg/kg; OTA, 0.07–0.25 μg/kg; ZEA, 1–3 μg/kg. The limits of determination were: aflatoxins, 0.25 μg/kg; OTA, 0.5 μg/kg; ZEA, 5 μg/kg. The recovery rates for aflatoxins, OTA, and ZEA for rye and rice were between 86 and 93% when a 0.5 g sample matter per immunoaffinity column was used.

scholarly journals Determination of Aflatoxins and Ochratoxin a in Animal Liver Using HPLC-FD Method with Immunoaffinity Column Clean-Up

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Biljana Stojanovska-Dimzoska ◽  
Elizabeta Dimitrieska-Stojkovic ◽  
Zehra Hajrulai-Musliu ◽  
Risto Uzunov ◽  
Aleksandra Angeleska ◽  
...  
Keyword(s):  
Liquid Chromatography ◽  
Ochratoxin A ◽  
Immunoaffinity Column ◽  
Acceptable Range ◽  
Animal Liver ◽  
Relative Standard ◽  
Validation Parameters ◽  
The Mean ◽  
Concentration Levels

Abstract Analytical methods based on immunoaffinity column clean-up and quantitative determination with liquid chromatography-fluorescence detection were used to determine aflatoxins and ochratoxin A in liver samples. The validation of the procedures was performed. The linearity of the methods was checked, and a good coefficient of correlation was found for all aflatoxins and OTA as well. The LOD and LOQ were acceptable: 0.003 µg/kg and 0.009 µg/kg for AFB1; 0.001 µg/kg and 0.005 µg/kg for AFB2; 0.006 µg/kg and 0.020 µg/kg for AFG1; 0.007 µg/kg and 0.022 µg/kg for AFG2; 0.08 µg/kg and 0.27 µg/kg for OTA. The results for the repeatability estimated by the relative standard deviation (RSDr) were satisfactory and the obtained values were in the acceptable range (1.97–14.41% for all aflatoxins and 3.76-8.31% for OTA) at three proposed concentration levels. RSDR values showed acceptable correlation between two analysts for all four aflatoxins and OTA. The RSDR values were as followed: 2.37% and 5.60% for AFB1, 6.71% and 8.78% for AFB2, 4.40% and 7.00% for AFG1 and 10.30% and 13.91% for AFG2 (for the first and second analyst, respectively). The RSDR values for OTA were 4.91% and 3.15% (1 µg/kg); 3.76% and 4.12% (5 µg/kg) and 8.31% and 8.21% (10 µg/kg). The mean recovery for total aflatoxins and OTA were 78.10% and 93.34%, respectively. All validation parameters were in accordance to European legislation. They indicate that the proposed analytical procedures are suitable and they could be methods of choice for the determination of aflatoxins and OTA in liver samples.

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