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Michael S. Filigenzi
California Animal Health and Food Safety Laboratory System, University of California, 620 West Health Sciences Drive, Davis, CA 95616, USA;(M.F.);(R.H.P.)

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Journal article
Published: 15 February 2020 in Toxins
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Globally, mushroom poisonings cause about 100 human deaths each year, with thousands of people requiring medical assistance. Dogs are also susceptible to mushroom poisonings and require medical assistance. Cyclopeptides, and more specifically amanitins (or amatoxins, here), are the mushroom poison that causes the majority of these deaths. Current methods (predominantly chromatographic, as well as antibody-based) of detecting amatoxins are time-consuming and require expensive equipment. In this work, we demonstrate the utility of the lateral flow immunoassay (LFIA) for the rapid detection of amatoxins in urine samples. The LFIA detects as little as 10 ng/mL of α-amanitin (α-AMA) or γ-AMA, and 100 ng/mL of β-AMA in urine matrices. To demonstrate application of this LFIA for urine analysis, this study examined fortified human urine samples and urine collected from exposed dogs. Urine is sampled directly without the need for any pretreatment, detection from urine is completed in 10 min, and the results are read by eye, without the need for specialized equipment. Analysis of both fortified human urine samples and urine samples collected from intoxicated dogs using the LFIA correlated well with liquid chromatography–mass spectrometry (LC-MS) methods.

ACS Style

Candace S. Bever; Kenneth D. Swanson; Elizabeth I. Hamelin; Michael Filigenzi; Robert H. Poppenga; Jennifer Kaae; Luisa W. Cheng; Larry H. Stanker. Rapid, Sensitive, and Accurate Point-of-Care Detection of Lethal Amatoxins in Urine. Toxins 2020, 12, 123 .

AMA Style

Candace S. Bever, Kenneth D. Swanson, Elizabeth I. Hamelin, Michael Filigenzi, Robert H. Poppenga, Jennifer Kaae, Luisa W. Cheng, Larry H. Stanker. Rapid, Sensitive, and Accurate Point-of-Care Detection of Lethal Amatoxins in Urine. Toxins. 2020; 12 (2):123.

Chicago/Turabian Style

Candace S. Bever; Kenneth D. Swanson; Elizabeth I. Hamelin; Michael Filigenzi; Robert H. Poppenga; Jennifer Kaae; Luisa W. Cheng; Larry H. Stanker. 2020. "Rapid, Sensitive, and Accurate Point-of-Care Detection of Lethal Amatoxins in Urine." Toxins 12, no. 2: 123.

Evaluation study
Published: 03 March 2015 in Journal of Agricultural and Food Chemistry
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Bromethalin, a potent neurotoxin, is widely available for use as a rodenticide. As access to other rodenticides is reduced due to regulatory pressure, the use of bromethalin is likely to increase with a concomitant increase in poisonings in nontarget animals. Analytical methods for the detection of bromethalin residues in animals suspected to have been exposed to this rodenticide are needed to support post-mortem diagnosis of toxicosis. This paper describes a novel method for the analysis of desmethylbromethalin (DMB), bromethalin's toxic metabolite, in tissue samples such as liver, brain, and adipose. Samples were extracted with 5% ethanol in ethyl acetate, and an aliquot of the extract was evaporated dry, reconstituted, and analyzed by reverse phase ultrahigh-performance liquid chromatography-mass spectrometry. The mass spectrometer utilized electrospray ionization in negative ion mode with multiple reaction monitoring. This method was qualitatively validated at a level of 1.0 ng/g in liver tissue. The quantitative potential of the method was also evaluated, and a method detection limit of 0.35 ng/g wet weight was determined in fat tissue. DMB was detected in tissue samples from animals suspected to have been poisoned by this compound. To the authors' knowledge, there have been no other methods reported for analysis of DMB in tissue samples using LC-MS/MS.

ACS Style

Michael S. Filigenzi; Adrienne C. Bautista; Linda S. Aston; Robert H. Poppenga. Method for the Detection of Desmethylbromethalin in Animal Tissue Samples for the Determination of Bromethalin Exposure. Journal of Agricultural and Food Chemistry 2015, 63, 5146 -5151.

AMA Style

Michael S. Filigenzi, Adrienne C. Bautista, Linda S. Aston, Robert H. Poppenga. Method for the Detection of Desmethylbromethalin in Animal Tissue Samples for the Determination of Bromethalin Exposure. Journal of Agricultural and Food Chemistry. 2015; 63 (21):5146-5151.

Chicago/Turabian Style

Michael S. Filigenzi; Adrienne C. Bautista; Linda S. Aston; Robert H. Poppenga. 2015. "Method for the Detection of Desmethylbromethalin in Animal Tissue Samples for the Determination of Bromethalin Exposure." Journal of Agricultural and Food Chemistry 63, no. 21: 5146-5151.

Original articles
Published: 14 July 2011 in Food Additives & Contaminants: Part A
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A method combining QuEChERS extraction, ultra-high pressure liquid chromatography and full scan high resolution mass spectrometry was evaluated for its use in screening for chemical residues and contaminants in animal-related food matrices. The method was evaluated by analysis of multiple replicates of whole milk, muscle tissue, liver tissue and corn silage. Analytes tested included plant alkaloids, carbamate and organophosphate pesticides, and several types of veterinary drugs. A database containing the chemical formula for each analyte was used to calculate accurate mass-to-charge ratios for expected pseudo-molecular ions. This information, as well as retention times, was used to identify analytes. Of 118 compounds chosen for analysis, 86 were detectable in all fortified replicates of at least one matrix at levels ranging from 1.0 to 5000 ng/g. Variability of response, as measured in % relative standard deviation of peak areas over seven replicate fortified sample analyses, was found to differ among the classes of analytes, ranging from 100%. Retention times were stable and analytes were routinely detected at measured mass-to-charge ratios within 2 ppm of their theoretical mass-to-charge ratios. These results indicate that the combination of generic extraction and chromatographic procedures with full scan high resolution mass spectrometry can serve as a useful method for screening complex matrices.

ACS Style

Michael S. Filigenzi; Nanette Ehrke; Linda S. Aston; Robert H. Poppenga. Evaluation of a rapid screening method for chemical contaminants of concern in four food-related matrices using QuEChERS extraction, UHPLC and high resolution mass spectrometry. Food Additives & Contaminants: Part A 2011, 28, 1324 -1339.

AMA Style

Michael S. Filigenzi, Nanette Ehrke, Linda S. Aston, Robert H. Poppenga. Evaluation of a rapid screening method for chemical contaminants of concern in four food-related matrices using QuEChERS extraction, UHPLC and high resolution mass spectrometry. Food Additives & Contaminants: Part A. 2011; 28 (10):1324-1339.

Chicago/Turabian Style

Michael S. Filigenzi; Nanette Ehrke; Linda S. Aston; Robert H. Poppenga. 2011. "Evaluation of a rapid screening method for chemical contaminants of concern in four food-related matrices using QuEChERS extraction, UHPLC and high resolution mass spectrometry." Food Additives & Contaminants: Part A 28, no. 10: 1324-1339.

Journal article
Published: 10 September 2008 in Journal of Agricultural and Food Chemistry
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In 2007, it was determined that melamine, ammeline, ammelide, and cyanuric acid (abbreviated as MARC for melamine and related contaminants) had been added to wheat gluten and rice protein that were subsequently incorporated into pet food. The consumption of food tainted by MARC compounds was implicated in numerous instances of renal failure in cats and dogs. A method for the analysis of MARC compounds in kidney tissue using high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) has been developed. MARC analytes were extracted by homogenization of kidney tissue in 50/40/10 acetonitrile/water/diethylamine. The homogenate was centrifuged, and an aliquot of supernatant was diluted with acetonitrile, concentrated, and fortified with a stable isotope-labeled analogue of melamine. Analytes were detected using atmospheric pressure chemical ionization and multiple reaction monitoring. Quantitation of positive samples was performed using the internal standard method and five-point calibration curves ranging between 50 and 1000 ng/mL of each analyte. The method was validated by analysis of replicate kidney tissue samples fortified with the individual analytes and by analysis of kidney samples fortified with melamine cyanurate powder at two different concentrations. This method was successfully used for routine postmortem diagnosis of melamine toxicosis in animals. Melamine was also detected by this method in paraffin-embedded tissue from animals suspected to have died of melamine toxicosis.

ACS Style

Michael S. Filigenzi; Birgit Puschner; Linda S. Aston; Robert H. Poppenga. Diagnostic Determination of Melamine and Related Compounds in Kidney Tissue by Liquid Chromatography/Tandem Mass Spectrometry. Journal of Agricultural and Food Chemistry 2008, 56, 7593 -7599.

AMA Style

Michael S. Filigenzi, Birgit Puschner, Linda S. Aston, Robert H. Poppenga. Diagnostic Determination of Melamine and Related Compounds in Kidney Tissue by Liquid Chromatography/Tandem Mass Spectrometry. Journal of Agricultural and Food Chemistry. 2008; 56 (17):7593-7599.

Chicago/Turabian Style

Michael S. Filigenzi; Birgit Puschner; Linda S. Aston; Robert H. Poppenga. 2008. "Diagnostic Determination of Melamine and Related Compounds in Kidney Tissue by Liquid Chromatography/Tandem Mass Spectrometry." Journal of Agricultural and Food Chemistry 56, no. 17: 7593-7599.

Validation study
Published: 13 November 2007 in Rapid Communications in Mass Spectrometry
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In early 2007 it was determined that the compound melamine, suspected of having been involved in the deaths of numerous pets, had been fed to hogs intended for human consumption. This report describes a method for the analysis of melamine in porcine muscle tissue using solid‐phase extraction (SPE) and high‐performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS). Melamine was extracted in 50% acetonitrile in water. Homogenates were centrifuged and supernatants were acidified and washed with methylene chloride. The aqueous extracts were cleaned up using mixed‐mode C8/strong cation exchange SPE and then concentrated, fortified with a stable isotope‐labeled analog of melamine, and analyzed by HPLC/MS/MS. Gradient HPLC separation was performed using an ether‐linked phenyl column with ammonium acetate/acetic acid and acetonitrile as the mobile phase. Multiple reaction monitoring (MRM) mode of two precursor‐product ion transitions for melamine and one for the internal standard was used. A five point calibration curve ranging from 50 to 2000 ng/mL of melamine in solvent was used to establish instrument response. The method was validated by analysis of seven replicate porcine muscle tissue samples fortified with 10 ng/g of melamine. The mean recovery for the seven replicates was 83% with 6.5% relative standard deviation and the calculated method detection limit was 1.7 ng/g. Copyright © 2007 John Wiley & Sons, Ltd.

ACS Style

Michael S. Filigenzi; Elizabeth R. Tor; Robert H. Poppenga; Linda A. Aston; Birgit Puschner. The determination of melamine in muscle tissue by liquid chromatography/tandem mass spectrometry. Rapid Communications in Mass Spectrometry 2007, 21, 4027 -4032.

AMA Style

Michael S. Filigenzi, Elizabeth R. Tor, Robert H. Poppenga, Linda A. Aston, Birgit Puschner. The determination of melamine in muscle tissue by liquid chromatography/tandem mass spectrometry. Rapid Communications in Mass Spectrometry. 2007; 21 (24):4027-4032.

Chicago/Turabian Style

Michael S. Filigenzi; Elizabeth R. Tor; Robert H. Poppenga; Linda A. Aston; Birgit Puschner. 2007. "The determination of melamine in muscle tissue by liquid chromatography/tandem mass spectrometry." Rapid Communications in Mass Spectrometry 21, no. 24: 4027-4032.

Journal article
Published: 01 April 2007 in Journal of Agricultural and Food Chemistry
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This paper describes a rapid LC-MS/MS/MS method for the analysis of alpha-amanitin in serum and liver. Serum was initially prepared by precipitation of proteins with acetonitrile and subsequent removal of acetonitrile with methylene chloride. Liver was prepared by homogenization with aqueous acetonitrile and subsequent removal of acetonitrile using methylene chloride. For both matrices, the aqueous phase was then extracted using mixed-mode C18/cation exchange SPE cartridges and analyzed on a linear ion trap LC-MS system. Standards were prepared in extracts of control matrix. Seven replicate fortifications of serum at 0.001 mug/g (1 ng/g) of alpha-amanitin gave a mean recovery of 95% with 8.8% CV (relative standard deviation) and a calculated method detection limit of 0.26 ng/g. Seven replicates of control liver fortified at 1 ng/g gave a mean recovery of 98% with 17% CV and a calculated method detection limit of 0.50 ng/g. This is the first report of a positive mass spectrometric identification and quantitation of alpha-amanitin in serum and liver from suspect human and animal intoxications.

ACS Style

Michael S. Filigenzi; Robert H. Poppenga; Asheesh K. Tiwary; Birgit Puschner. Determination of α-Amanitin in Serum and Liver by Multistage Linear Ion Trap Mass Spectrometry. Journal of Agricultural and Food Chemistry 2007, 55, 2784 -2790.

AMA Style

Michael S. Filigenzi, Robert H. Poppenga, Asheesh K. Tiwary, Birgit Puschner. Determination of α-Amanitin in Serum and Liver by Multistage Linear Ion Trap Mass Spectrometry. Journal of Agricultural and Food Chemistry. 2007; 55 (8):2784-2790.

Chicago/Turabian Style

Michael S. Filigenzi; Robert H. Poppenga; Asheesh K. Tiwary; Birgit Puschner. 2007. "Determination of α-Amanitin in Serum and Liver by Multistage Linear Ion Trap Mass Spectrometry." Journal of Agricultural and Food Chemistry 55, no. 8: 2784-2790.

Journal article
Published: 01 April 2004 in Journal of Agricultural and Food Chemistry
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An LC-MS/MS method was developed for the semiquantitative determination of strophanthidin glycosides in ingesta from animals. Strophanthidin glycosides were simultaneously extracted and hydrolyzed to the strophanthidin aglycone using aqueous methanolic hydrochloric acid and the extracts cleaned up using solid-phase extraction. Extracts were analyzed using reverse-phase HPLC coupled with positive ion electrospray mass spectrometry. Characteristic product ion spectra were produced by fragmentation of the [M + H](+) precursor ion for each analyte. Quantitation was performed using the internal standard method with digitoxigenin serving as the internal standard. The method detection limit was calculated to be 0.075 microg/g, and the limit of quantitation was calculated to be 0.24 microg/g for strophanthidin in control rumen samples. This method was used in diagnostic investigations to confirm fatal strophanthidin glycoside poisonings in horses.

ACS Style

Michael S. Filigenzi; Leslie W. Woods; Marcia C. Booth; Elizabeth R. Tor; Birgit Puschner. Determination of Strophanthidin in Ingesta and Plant Material by LC-MS/MS. Journal of Agricultural and Food Chemistry 2004, 52, 2174 -2178.

AMA Style

Michael S. Filigenzi, Leslie W. Woods, Marcia C. Booth, Elizabeth R. Tor, Birgit Puschner. Determination of Strophanthidin in Ingesta and Plant Material by LC-MS/MS. Journal of Agricultural and Food Chemistry. 2004; 52 (8):2174-2178.

Chicago/Turabian Style

Michael S. Filigenzi; Leslie W. Woods; Marcia C. Booth; Elizabeth R. Tor; Birgit Puschner. 2004. "Determination of Strophanthidin in Ingesta and Plant Material by LC-MS/MS." Journal of Agricultural and Food Chemistry 52, no. 8: 2174-2178.