Immunoaffinity Column Coupled with Liquid Chromatography for Determination of Fumonisin B1 in Canned and Frozen Sweet Corn

1995 ◽  
Vol 78 (3) ◽  
pp. 705-710 ◽  
Author(s):  
Mary W Trucksess ◽  
Michael E Stack ◽  
Shantae Allen ◽  
Noel Barrion
Keyword(s):  
Sweet Corn ◽  
Limit Of Detection ◽  
Fumonisin B1 ◽  
The United States ◽  
Reversed Phase ◽  
Immunoaffinity Column ◽  
Fluorescence Detector ◽  
Corn Sample ◽  
Limit Of Determination ◽  
Canned Corn

Abstract A modified liquid chromatographic (LC) method for determining fumonisin B1 (FB1) in corn was applied to canned and frozen sweet corn. The corn is extracted with methanol–water (8 + 2), and the extract is filtered. The filtrate is diluted with water and passed through an immunoaffinity column. After the column is washed with water, FBi is eluted with methanol–water (8 + 2). The eluate is evaporated to dryness by using a vacuum concentrator, and the residue is dissolved in acetonitrile–water (1 +1). FB1 is derivatized with o-phthaldialdehyde. The derivative is separated on a reversed-phase C18 LC column using acetonitrile–water–acetic acid (50 + 50 + 1) and quantitated with a fluorescence detector. Recoveries of FB1 from canned and frozen corn spiked over the range of 50–200 ng/g were 76–88%. The limit of determination was about 25 ng/g, and the limit of detection was about 4 ng/g. The method was applied to 97 commercial canned and frozen sweet corn samples collected from different areas of the United States. Sixty samples contained no FB1. Low levels (trace–82 ng FB1/g corn) were found in 35 samples; 235 ng FB1/g was found in 1 canned corn sample, and 350 ng FB1/g was found in 1 frozen corn sample.

scholarly journals Determination and Survey of Ochratoxin A in Wheat, Barley, and Coffee—1997

1999 ◽  
Vol 82 (1) ◽  
pp. 85-89 ◽  
Author(s):  
Mary W Trucksess ◽  
John Giler ◽  
Kathryn Young ◽  
Kevin D White ◽  
Samuel W Page
Keyword(s):  
Ochratoxin A ◽  
Limit Of Detection ◽  
Test Sample ◽  
The United States ◽  
Reversed Phase ◽  
Cereal Grains ◽  
Green Coffee ◽  
Roasted Coffee ◽  
Fluorescence Detector ◽  
Penicillium Species

Abstract Ochratoxin A (OA) is a nephrotoxic and nephrocarcinogenic mycotoxin produced by Aspergillus and Penicillium species. It has been found mainly in cereal grains and coffee beans. The purpose of this study was to investigate the occurrence of OA in cereal grains and in coffee imported to the United States. A modified liquid chromatographic (LC) method for determining OA in green coffee was applied to wheat, barley, green coffee, and roasted coffee. The test sample was extracted with methanol–1% NaHCO3 (7 + 3), and the extract was filtered. The filtrate was diluted with phosphate-buffered saline (PBS), filtered, and passed through an immunoaffinity column. After the column was washed with PBS and then with water, OA was eluted with methanol. The eluate was evaporated to dryness, and the residue was dissolved in acetonitrile–water (1 +1). OA was separated on a reversed-phase C18 LC column with acetonitrile-water-acetic acid (55 + 45 + 1) as eluant and quantitated with a fluorescence detector. Recoveries of OA from the 4 commodities spiked over the range 1–4 ng/g were 71–96%. The limit of detection was about 0.03 ng/g. OA contamination at >0.03 ng/g was found in 56 of 383 wheat samples, 11 of 103 barley samples, 9 of 19 green coffee samples, and 9 of 13 roasted coffee samples. None of the coffee samples contained OA at >5 ng/g; only 4 samples of wheat and 1 sample of barley were contaminated above this level.

scholarly journals Preparative Method for Isolating α-Zearalenol and Zearalenone Using Extracting Disk

1999 ◽  
Vol 82 (1) ◽  
pp. 90-94 ◽  
Author(s):  
George M Ware ◽  
Yuhong Zhao ◽  
Shia S Kuan ◽  
Allen S Carman
Keyword(s):  
Limit Of Detection ◽  
Solid Phase ◽  
Control Sample ◽  
Preparative Method ◽  
Reversed Phase ◽  
Fluorescence Detector ◽  
Coefficients Of Variation ◽  
Limit Of Quantitation ◽  
Standard Calibration ◽  
Liquid Chromatographic

Abstract A liquid chromatographic method is described for the determination of zearalenol and zearalenone in corn. Zearalenol and zearalenone are extracted from corn with methanol–water (1+1) and cleaned up using a solid-phase extraction (SPE) disk, separatedon a reversed-phase analytical column, and detected with a fluorescence detector. The SPE disk concentrated and cleanly separated zearalenol and zearalenone from sample interferences. Standard calibration curves for zearalenol and zearalenone for the concentration range 25–500 ng/mL were linear. The small extract disk had a column capacity equivalent to 1 g extracted corn. Zearalenol and zearalenone were added at levels ranging from 10 to 2000 ng/g to a control sample that contained no detectable levels of zearalenol and zearalenone. Both toxins were recovered from spiked samples at 106.3 and 103.8%, with coefficients of variation of 7.6 and 13.0%, respectively. The method has an estimated reliable limit of detection and limit of quantitation around 10 and 40 ng/g for each toxin, respectively.

scholarly journals Determination of Fumonisins B1 and B2 in Various Maize Products by a Combined SAX + C18 Column and Immunoaffinity Column

2000 ◽  
Vol 83 (1) ◽  
pp. 99-103 ◽  
Author(s):  
Tord E Möller ◽  
Håkan F Gustavsson
Keyword(s):  
Limit Of Detection ◽  
Fumonisin B1 ◽  
Immunoaffinity Column ◽  
Maize Flour ◽  
Main Column ◽  
Flour Sample ◽  
Fumonisin B2 ◽  
Fumonisins B1 And B2

Abstract Fumonisins B1 and B2 were determined in 42 samples of different maize products from the Swedish market by 2 different methods based on cleanup steps using an immunoaffinity column and a combination of SAX + C18 columns, respectively. A simple “precipitation step” was included before the samples were added to the main column(s), giving less column clogging, fewer interfering peaks, and better recoveries for the different sample matrixes. Recovery, repeatability, and results from the survey showed comparable results with the methods. The limit of detection for both methods was 5 μg/kg for fumonisin B1 and 10 μg/kg for fumonisin B2. All 7 maize chips analyzed and 6 of 8 popcorn samples contained fumonisins (B1 + B2) with averages of 180 and 115 μg/kg, respectively. All other samples except a maize flour sample contained little or no fumonisins.

scholarly journals Determination of Zearalenone Content in Cereals and Feedstuffs by Immunoaffinity Column Coupled with Liquid Chromatography

2001 ◽  
Vol 84 (5) ◽  
pp. 1453-1459 ◽  
Author(s):  
Bela Fazekas ◽  
Andrea Tar
Keyword(s):  
Liquid Chromatography ◽  
Recovery Rate ◽  
Limit Of Detection ◽  
Poultry Feed ◽  
Array Detector ◽  
Immunoaffinity Column ◽  
Uv Spectrum ◽  
Fluorescence Detector ◽  
Immunoaffinity Columns

Abstract The zearalenone content of maize, wheat, barley, swine feed, and poultry feed samples was determined by immunoaffinity column cleanup followed by liquid chromatography (IAC–LC). Samples were extracted in methanol–water (8 + 2, v/v) solution. The filtered extract was diluted with distilled water and applied to immunoaffinity columns. Zearalenone was eluted with methanol, dried by evaporation, and dissolved in acetonitrile–water (3 + 7, v/v). Zearalenone was separated by isocratic elution of acetonitrile–water (50 + 50, v/v) on reversed-phase C18 column. The quantitative analysis was performed by fluorescence detector and confirmation was based on the UV spectrum obtained by a diode array detector. The mean recovery rate of zearalenone was 82–97% (RSD, 1.4–4.1%) on the original (single-use) immunoaffinity columns. The limit of detection of zearalenone by fluorescence was 10 ng/g at a signal-to-noise ratio of 10:1 and 30 ng/g by spectral confirmation in UV. A good correlation was found (R2 = 0.89) between the results obtained by IAC–LC and by the official AOAC–LC method. The specificity of the method was increased by using fluorescence detection in parallel with UV detection. This method was applicable to the determination of zearalenone content in cereals and other kinds of feedstuffs. Reusability of immunoaffinity columns was examined by washing with water after sample elution and allowing columns to stand for 24 h at room temperature. The zearalenone recovery rate of the regenerated columns varied between 79 and 95% (RSD, 3.2–6.3%). Columns can be regenerated at least 3 times without altering their performance and without affecting the results of repeated determinations.

scholarly journals Tocopherol Content in Vegetable Oils Using a Rapid HPLC Fluorescence Detection Method

2013 ◽  
Vol 41 (1) ◽  
pp. 93 ◽  
Author(s):  
Constantin BELE ◽  
Cristian T. MATEA ◽  
Camelia RADUCU ◽  
Vioara MIRESAN ◽  
Octavian NEGREA
Keyword(s):  
High Performance ◽  
Detection Method ◽  
Limit Of Detection ◽  
Direct Method ◽  
Tocopherol Content ◽  
Reversed Phase ◽  
Excitation Wavelength ◽  
Plant Oils ◽  
Fluorescence Detector ◽  
Edible Plant

A quick and direct method based on reversed phase high performance liquid chromatography with fluorescence detector for measuring tocopherols (α , β + γ and δ) has been developed. Oils are diluted in methanol: hexane: tetrahydrofuran (neither previous extraction of tocopherols nor saponification procedure are required) and after being vortexed and centrifuged, an aliquot of the overlay was injected directly into an Alltima C 18 column. Acetonitrile and methanol (50: 50) mixture was used as a mobile phase with a flow rate of 1 mL min-1. Quantification of tocopherols was performed by fluorescence detector at 290 nm excitation wavelength and 325 nm emission wavelength. Tocopherols were separated at 25°C in less than 10 min after injection. The method has good limit of detection (9 ng g-1 for α-tocopherol and 8 ng g-1 for β-, γ- and δ- tocopherols) and reproducibility (CV< 2.9 %). This method can be used to assess the influence of genetic modification of oil seeds on the distribution of tocopherols or the effect of tocopherols on the oxidative stability of edible plant oils.

scholarly journals Determination of Emamectin Benzoate in Freshwater and Seawater at Picogram-Per-Milliliter Levels by Liquid Chromatography with Fluorescence Detection

1997 ◽  
Vol 80 (5) ◽  
pp. 1098-1103 ◽  
Author(s):  
Michael B Hicks ◽  
Lori D Payne ◽  
Sunil V Prabhu ◽  
Teresa A Wehner
Keyword(s):  
Ethyl Acetate ◽  
Fluorescence Detection ◽  
Ammonium Acetate ◽  
Limit Of Detection ◽  
Solid Phase ◽  
Tap Water ◽  
Reversed Phase ◽  
Emamectin Benzoate ◽  
Fluorescence Detector ◽  
Solid Phase Extraction Cartridge

Abstract Emamectin benzoate is a very effective agricultural insecticide applied to various crops at very low rates. A method to analyze emamectin benzoate in seawater and freshwater was developed. The method yields good recoveries with limits of quantitation of 20 pg/mL for freshwater and 24 pg/mL for seawater (signal-to-noise ratio [S/N] ≥ 10). The limit of detection is 10 pg/mL (S/N ≥ 3). Emamectin benzoateis concentrated from a 50 mL sample on a C8 solid-phase extraction cartridge and eluted with 1% ammonium acetate in methanol. The eluate is evaporated to 1 mL, diluted with 1 % ammonium acetate in water, and partitioned into ethyl acetate. The ethyl acetate extract is concentrated to about 0.5 mL and diluted to about 1 mL with acetonitrile. 1-Methylimidazole is added to the diluted sample, and then trifluoroacetic anhydride is added to form a fluorescent derivative. The derivatized sample is injected onto a reversed-phase liquid chromatographic system equipped with a fluorescence detector. Overall method recoveries for concentrations ranging from 20 to 1010 pg/mL were 84 ± 9.5% for tap water and 89 ± 11% for seawater. These results are equivalent to recoveries previously obtained with a more elaborate method (90 ± 8.8% for freshwater). Controls were free of interferences or coeluting impurities. Fluorescence detection provides high sensitivity and selectivity, enabling trace-level analysis.

scholarly journals Rapid Immunoaffinity-Based Method for Determination of Zearalenone in Corn by Fluorometry and Liquid Chromatography

1999 ◽  
Vol 82 (6) ◽  
pp. 1364-1368 ◽  
Author(s):  
Scott C Kruger ◽  
Barb Kohn ◽  
Catherine S Ramsey ◽  
Reginaldo Prioli
Keyword(s):  
Limit Of Detection ◽  
Official Method ◽  
Immunoaffinity Column ◽  
Fluorescence Detector ◽  
Percentage Recovery ◽  
Relative Standard ◽  
Chloride Hexahydrate ◽  
Aoac Official ◽  
Sensitive Method ◽  
Aoac Official Method

Abstract An immunoaffinity-based method was developed to determine zearalenone in corn. Corn samples were extracted in acetonitrile-water (90 + 10, v/v), applied to an immunoaffinity column, and eluted with methanol. The isolated toxin was quantitated either by reaction with aluminum chloride hexahydrate (AlCl33.6H2O) prior to measurement with a fluorometer or injection into a liquid chromatographic (LC) system with a fluorescence detector. Performance was evaluated in terms of antibody specificity, limit of detection, percentage recovery, precision, column capacity, assay linearity, and comparison with AOAC Official Method 985.18. With the immunoaffinity column cleanup procedure, only zearalenone and its metabolites were recognized by the antibody (≥75% recovery). Limits of detection were 0.10 μg/g for the fluorometer and 0.10 or 0.0025 μg/g (sensitive method) for the LC method. Percentage recovery averaged 105% (fluorometer) and 93% (LC method), with average relative standard deviations (RSDs) of 15.7 and 9 .3%. Naturally contaminated samples gave comparable RSDs of 8.3 and 9.9% for the fluorometer and LC methods, respectively. Column capacity was 4.0 μg with 89% recovery. Assay linearity was comparable for both methods (r2 = 0.998). Optimum assay ranges were 0.10-5.0 μg/g for the fluorometer and 0.10-50 or 0.0025-5.0 μg/g (sensitive method) for the LC method. Comparative analysis of 17 naturally contaminated corn samples using ZearalaTest LC and the official AOAC LC method for detection of zearalenone showed that ZearalaTest is statistically comparable to the AOAC Official Method 985.18 (r2 = 0.747).

scholarly journals Chromatographic Analysis of Aflatoxigenic Aspergillus flavus Isolated from Malaysian Sweet Corn

Separations ◽  
2021 ◽  
Vol 8 (7) ◽  
pp. 98
Author(s):  
Rahim Khan ◽  
Farinazleen Mohamad Ghazali ◽  
Nor Ainy Mahyudin ◽  
Nik Iskandar Putra Samsudin
Keyword(s):  
Aspergillus Flavus ◽  
High Performance ◽  
Sweet Corn ◽  
Limit Of Detection ◽  
High Sensitivity ◽  
Field Trials ◽  
Fluorescence Detector ◽  
Efficient Tool ◽  
Limit Of Quantification ◽  
Testing Medium

High-performance liquid chromatography (HPLC) provides a quick and efficient tool for accurately characterizing aflatoxigenic and non-aflatoxigenic isolates of Aspergillus flavus. This method also provides a quantitative analysis of AFs in Aspergillus flavus. The method’s recovery was assessed by spiking a mixture of AF at different concentrations to the testing medium. The validity of the method was confirmed using aflatoxigenic and non-aflatoxigenic strains of A. flavus. The HPLC system, coupled with a fluorescence detector and post-column photochemical reactor, showed high sensitivity in detecting spiked AFs or AFs produced by A. flavus isolates. Recovery from medium spiked with 10, 20, 60, and 80 ppb of AFs was found to be 73–86% using this approach. For AFB1 and AFB2, the limit of detection was 0.072 and 0.062 ppb, while the limit of quantification was 0.220 and 0.189 ppb, respectively. The AFB1 concentrations ranged from 0.09 to 50.68 ppb, while the AFB2 concentrations ranged between 0.33 and 9.23 ppb. The findings showed that six isolates produced more AFB1 and AFB2 than the acceptable limit of 5 ppb. The incidence of aflatoxigenic isolates of A. flavus in sweet corn and higher concentrations of AFB1 and AFB2 emphasize the need for field trials to explore their real potential for AF production in corn.

Development and Validation of A Reversed Phase-High Performance Liquid Chromatography Method for the Quantitative Estimation of Ramipril Drug Content from Formulated Dosage Form

2016 ◽  
Vol 6 (3) ◽  
Author(s):  
Raju Chandra ◽  
Manisha Pant ◽  
Harchan Singh ◽  
Deepak Kumar ◽  
Ashwani Sanghi
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Active Ingredient ◽  
High Performance ◽  
Limit Of Detection ◽  
Quantitative Estimation ◽  
Reversed Phase ◽  
Uv Detector ◽  
Chromatography Method ◽  
Drug Content

A reliable and reproducible reversed-phase high performance liquid chromatography (RP-HPLC) was developed for the quantitative determination of Remipril drug content from marketed bulk tablets. The active ingredient of Remipril separation achieved with C18 column using the methanol water mobile phase in the ratio of 40:60 (v/v). The active ingredient of the drug content quantify with UV detector at 215 nm. The retention time of Remipril is 5.63 min. A good linearity relation (R2=0.999) was obtained between drug concentration and average peak areas. The limit of detection and limit of quantification of the instrument were calculated 0.03 and 0.09 µg/mL, respectively. The accuracy of the method validation was determined 102.72% by recoveries method.

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