High-Performance Liquid Chromatography—Receptorgram: A Comprehensive Method for Identification of Veterinary Drugs and Their Active Metabolites

Zomer, Eliezer; Quintana, Julio; Scheemaker, J.; Saul, Steven J.; Charm, Stanley E.

doi:10.1021/bk-1996-0636.ch016

Cited by 33 publications

(4 citation statements)

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“…This increase in sensitivity with the ampicillin incurred samples may be explained by a metabolite that is not quantified by the HPLC but that is cumulatively detected along with the parent compound by the screening method. Metabolites in these samples were observed previously by HPLC (24). Similar results have been observed previously with other incurred studies (3,12,13).…”

Section: Ruggednesssupporting

confidence: 92%

“…Samples containing penicillin G, amoxicillin, cephapirin, ampicillin, and ceftiofur were tested and commingled in 2007 and stored at -80°C (12). These concentra tions were verified by HPLC analysis prior to use (2,3,7,9,14,24). In the spring of 2010, CVM collected incurred residue milk samples by treating three cows with flunixin after it was verified that the cows were healthy and had not been treated for the preceding 30 days.…”

Section: Methodsmentioning

confidence: 94%

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Validation of a Rapid Lateral Flow Test for the Simultaneous Determination of β-Lactam Drugs and Flunixin in Raw Milk

Douglas¹,

Banaszewski

Juskelis

et al. 2012

Journal of Food Protection

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β-Lactam antibiotics are the most commonly used drugs on dairy farms. β-Lactam residues in milk are kept out of the human milk supply with good agricultural practices and mandatory truck screening performed by the dairy industry under Appendix N of the Pasteurized Milk Ordinance. Flunixin, a nonsteroidal and anti-inflammatory drug, appears in dairy cattle tissue residues with a frequency similar to the occurrence of penicillin G. This creates concern that flunixin residues could be in milk and would go undetected under current milk screening programs. A single test that combines mandatory β-lactam screening with voluntary flunixin screening is an economical approach for monitoring and controlling for potential flunixin or 5-hydroxyflunixin, the primary flunixin metabolite marker in milk. The objective of this study was to validate a β-lactam and flunixin rapid lateral flow test (LFT) and compare the results obtained with a liquid chromatography-triple quadrupole tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of flunixin and 5-hydroxyflunixin in raw milk with a limit of detection of <1 ppb, equivalent to 1 ng/ml. Using the LFT, three combined manufactured lots of test strips detected penicillin G at 2.0 ppb, ampicillin at 6.8 ppb, amoxicillin at 5.9 ppb, cephapirin at 13.4 ppb, ceftiofur (total metabolites) at 63 ppb, and 5-hydroxyflunixin at 1.9 ppb at least 90% of the time with 95% confidence. The LFT also detected incurred flunixin milk samples that were analyzed with the LC-MS/MS and diluted to tolerance in raw milk. The detection levels for the LFT are lower than the U.S. safe levels or tolerances and qualify the test to be used in compliance with U.S. milk screening programs.

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Section: Ruggednesssupporting

confidence: 92%

Section: Methodsmentioning

confidence: 94%

Validation of a Rapid Lateral Flow Test for the Simultaneous Determination of β-Lactam Drugs and Flunixin in Raw Milk

Douglas¹,

Banaszewski

Juskelis

et al. 2012

Journal of Food Protection

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“…These sulfonamide results indicated a shift in sensitivity of the incurred samples relative to the 90/95% levels calculated from parent drug spiked samples. HPLC-Receptorgram analysis of samples demonstrates some acetyl-sulfonamide metabolites, a liver metabolite ( 26 ). These metabolites were contributing to the test response making incurred samples about 50% more positive than the spiked parent sulfonamide compound.…”

Section: Resultsmentioning

confidence: 99%

TRIO Method for Detection of Beta-Lactams, Sulfonamides, and Tetracyclines in Raw Commingled Cows' Milk

Salter¹,

Markovsky²,

Douglas³

et al. 2020

Journal of AOAC INTERNATIONAL

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A qualitative 3 minute one step assay for detecting beta-lactam, sulfonamide and tetracycline antibiotics was validated following milk screening test guidelines developed by FDA-CVM, AOAC-RI and IDF. The validated 90% detection levels with 95% confidence were penicillin G 2 part per billion (ppb); amoxicillin 4 ppb; ampicillin 9 ppb; ceftiofur plus metabolites 50 ppb; cloxacillin 9 ppb; cephapirin 15 ppb; sulfadimethoxine 8 ppb; sulfamethazine 9 ppb; chlortetracycline 34 ppb; oxytetracycline 53 ppb; and tetracycline 42 ppb. Detection levels were lower than US and Canadian allowable limits for milk and were consistent with most European Maximum Residue Limits. Tests of raw commingled cows’ milk indicated a low positive error rate of < 0.3% with no interferences demonstrated by 1.08 MM/mL somatic cells, Gram – or + bacteria < 300K/mL, freeze/thawing, or non-targeted drugs. Detection of incurred residues were similar to, or more sensitive to, fortified samples. Some cross reactivity across drug families occurred in interference studies and therefore initial positive samples should be confirmed with drug family specific screening methods. The National Conference of Interstate Milk Shipments approval as a bulk tank/tanker screening test was completed in three stages for each drug family, including a tetracycline confirmation procedure to target the USA tolerance levels. Detection and robustness were found to be appropriate for multiple countries’ regulatory requirements for screening tests. The method development, validation and approval was intended to diversify and increase the verification tools for the control of the major antibiotic drug families used in managing cows’ health and welfare.

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“…[1][2][3][4][5][6][7][8][9] Liquid chromatography has been integrated with other assays to determine antibiotic residues in milk. 8,10 To the authors' knowledge, none of the methods for detection and identification of β-lactams in urine uses an integrated approach of LC and a residue screening test designed to detect residues in milk.…”

mentioning

confidence: 99%

Screening method for identification of β-lactams in bovine urine by use of liquid chromatography and a microbial inhibition test

Musser¹,

Anderson²,

Moats³

2001

ajvr

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High-Performance Liquid Chromatography—Receptorgram: A Comprehensive Method for Identification of Veterinary Drugs and Their Active Metabolites

Cited by 33 publications

References 0 publications

Validation of a Rapid Lateral Flow Test for the Simultaneous Determination of β-Lactam Drugs and Flunixin in Raw Milk

Validation of a Rapid Lateral Flow Test for the Simultaneous Determination of β-Lactam Drugs and Flunixin in Raw Milk

TRIO Method for Detection of Beta-Lactams, Sulfonamides, and Tetracyclines in Raw Commingled Cows' Milk

Screening method for identification of β-lactams in bovine urine by use of liquid chromatography and a microbial inhibition test

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