Quantitative analysis of penicillins in porcine tissues, milk and animal feed using derivatisation with piperidine and stable isotope dilution liquid chromatography tandem mass spectrometry

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Abstract

Penicillins are used universally in both human and veterinary medicine. The European Union (EU) has established maximum residue levels (MRLs) for most ß-lactam antibiotics in milk and animal tissues and included them in the National Residue Monitoring Programs. In this study, a novel method is described for the determination and confirmation of eight penicillins in porcine tissues, milk and animal feed by liquid chromatography–tandem mass spectrometry (LC–MS/MS). To prevent degradation of penicillin residues during workup, a derivatisation procedure was developed, by which penicillins were converted to stable piperidine derivatives. Deuterated piperidine derivatives were synthesised for all relevant penicillins, enabling the use of isotope dilution for accurate quantification. Penicillin residues were derivatised in the crude extract with piperidine and isolated using solid-phase extraction. The penicillin piperidine derivatives were determined by LC–MS/MS. The method was validated at the current MRLs, which range from 25–300 µg kg-1 in muscle and kidney to 4–30 µg kg-1 in milk as well as at the target value of 100 µg kg-1 chosen for animal feed, according to the EU requirements for a quantitative confirmatory method. Accuracy ranged from 94–113% (muscle), 83–111% (kidney) and 87–103% (milk) to 88–116% (animal feed). Intra-day precision (relative standard deviation (RSD)r) ranged from 5–13% (muscle, n¿=¿18), 4–17% (kidney, n¿=¿7) and 5–18% (milk, n¿=¿7) to 11–32% (animal feed, n¿=¿18). Inter-day precision (RSDRL, n¿=¿18) ranged from 6–23% (muscle) to 11–36% (animal feed). From the results, it was concluded that the method was fit for purpose at the target MRLs in animal tissue and target levels for animal feed
Original languageEnglish
Pages (from-to)3027-3040
JournalAnalytical and Bioanalytical Chemistry
Volume396
Issue number8
DOIs
Publication statusPublished - 2010

Fingerprint

Liquid chromatography
Tandem Mass Spectrometry
Liquid Chromatography
Isotopes
Penicillins
Dilution
Mass spectrometry
Milk
Animals
Swine
Tissue
Chemical analysis
Muscle
Muscles
European Union
Kidney
Derivatives
Veterinary medicine
Lactams
Veterinary Medicine

Keywords

  • beta-lactam antibiotics
  • solid-phase extraction
  • bovine-milk
  • precolumn derivatization
  • antimicrobial residues
  • degradation-products
  • confirmatory assay
  • kidney
  • muscle
  • amoxicillin

Cite this

@article{86074548a8dc40a59b6e3f7998fb413a,
title = "Quantitative analysis of penicillins in porcine tissues, milk and animal feed using derivatisation with piperidine and stable isotope dilution liquid chromatography tandem mass spectrometry",
abstract = "Penicillins are used universally in both human and veterinary medicine. The European Union (EU) has established maximum residue levels (MRLs) for most {\ss}-lactam antibiotics in milk and animal tissues and included them in the National Residue Monitoring Programs. In this study, a novel method is described for the determination and confirmation of eight penicillins in porcine tissues, milk and animal feed by liquid chromatography–tandem mass spectrometry (LC–MS/MS). To prevent degradation of penicillin residues during workup, a derivatisation procedure was developed, by which penicillins were converted to stable piperidine derivatives. Deuterated piperidine derivatives were synthesised for all relevant penicillins, enabling the use of isotope dilution for accurate quantification. Penicillin residues were derivatised in the crude extract with piperidine and isolated using solid-phase extraction. The penicillin piperidine derivatives were determined by LC–MS/MS. The method was validated at the current MRLs, which range from 25–300 µg kg-1 in muscle and kidney to 4–30 µg kg-1 in milk as well as at the target value of 100 µg kg-1 chosen for animal feed, according to the EU requirements for a quantitative confirmatory method. Accuracy ranged from 94–113{\%} (muscle), 83–111{\%} (kidney) and 87–103{\%} (milk) to 88–116{\%} (animal feed). Intra-day precision (relative standard deviation (RSD)r) ranged from 5–13{\%} (muscle, n¿=¿18), 4–17{\%} (kidney, n¿=¿7) and 5–18{\%} (milk, n¿=¿7) to 11–32{\%} (animal feed, n¿=¿18). Inter-day precision (RSDRL, n¿=¿18) ranged from 6–23{\%} (muscle) to 11–36{\%} (animal feed). From the results, it was concluded that the method was fit for purpose at the target MRLs in animal tissue and target levels for animal feed",
keywords = "beta-lactam antibiotics, solid-phase extraction, bovine-milk, precolumn derivatization, antimicrobial residues, degradation-products, confirmatory assay, kidney, muscle, amoxicillin",
author = "{van Holthoon}, F.L. and P.P.J. Mulder and {van Bennekom}, E.O. and H.H. Heskamp and T. Zuidema and {van Rhijn}, J.A.",
year = "2010",
doi = "10.1007/s00216-010-3523-0",
language = "English",
volume = "396",
pages = "3027--3040",
journal = "Analytical and Bioanalytical Chemistry",
issn = "1618-2642",
publisher = "Springer Verlag",
number = "8",

}

TY - JOUR

T1 - Quantitative analysis of penicillins in porcine tissues, milk and animal feed using derivatisation with piperidine and stable isotope dilution liquid chromatography tandem mass spectrometry

AU - van Holthoon, F.L.

AU - Mulder, P.P.J.

AU - van Bennekom, E.O.

AU - Heskamp, H.H.

AU - Zuidema, T.

AU - van Rhijn, J.A.

PY - 2010

Y1 - 2010

N2 - Penicillins are used universally in both human and veterinary medicine. The European Union (EU) has established maximum residue levels (MRLs) for most ß-lactam antibiotics in milk and animal tissues and included them in the National Residue Monitoring Programs. In this study, a novel method is described for the determination and confirmation of eight penicillins in porcine tissues, milk and animal feed by liquid chromatography–tandem mass spectrometry (LC–MS/MS). To prevent degradation of penicillin residues during workup, a derivatisation procedure was developed, by which penicillins were converted to stable piperidine derivatives. Deuterated piperidine derivatives were synthesised for all relevant penicillins, enabling the use of isotope dilution for accurate quantification. Penicillin residues were derivatised in the crude extract with piperidine and isolated using solid-phase extraction. The penicillin piperidine derivatives were determined by LC–MS/MS. The method was validated at the current MRLs, which range from 25–300 µg kg-1 in muscle and kidney to 4–30 µg kg-1 in milk as well as at the target value of 100 µg kg-1 chosen for animal feed, according to the EU requirements for a quantitative confirmatory method. Accuracy ranged from 94–113% (muscle), 83–111% (kidney) and 87–103% (milk) to 88–116% (animal feed). Intra-day precision (relative standard deviation (RSD)r) ranged from 5–13% (muscle, n¿=¿18), 4–17% (kidney, n¿=¿7) and 5–18% (milk, n¿=¿7) to 11–32% (animal feed, n¿=¿18). Inter-day precision (RSDRL, n¿=¿18) ranged from 6–23% (muscle) to 11–36% (animal feed). From the results, it was concluded that the method was fit for purpose at the target MRLs in animal tissue and target levels for animal feed

AB - Penicillins are used universally in both human and veterinary medicine. The European Union (EU) has established maximum residue levels (MRLs) for most ß-lactam antibiotics in milk and animal tissues and included them in the National Residue Monitoring Programs. In this study, a novel method is described for the determination and confirmation of eight penicillins in porcine tissues, milk and animal feed by liquid chromatography–tandem mass spectrometry (LC–MS/MS). To prevent degradation of penicillin residues during workup, a derivatisation procedure was developed, by which penicillins were converted to stable piperidine derivatives. Deuterated piperidine derivatives were synthesised for all relevant penicillins, enabling the use of isotope dilution for accurate quantification. Penicillin residues were derivatised in the crude extract with piperidine and isolated using solid-phase extraction. The penicillin piperidine derivatives were determined by LC–MS/MS. The method was validated at the current MRLs, which range from 25–300 µg kg-1 in muscle and kidney to 4–30 µg kg-1 in milk as well as at the target value of 100 µg kg-1 chosen for animal feed, according to the EU requirements for a quantitative confirmatory method. Accuracy ranged from 94–113% (muscle), 83–111% (kidney) and 87–103% (milk) to 88–116% (animal feed). Intra-day precision (relative standard deviation (RSD)r) ranged from 5–13% (muscle, n¿=¿18), 4–17% (kidney, n¿=¿7) and 5–18% (milk, n¿=¿7) to 11–32% (animal feed, n¿=¿18). Inter-day precision (RSDRL, n¿=¿18) ranged from 6–23% (muscle) to 11–36% (animal feed). From the results, it was concluded that the method was fit for purpose at the target MRLs in animal tissue and target levels for animal feed

KW - beta-lactam antibiotics

KW - solid-phase extraction

KW - bovine-milk

KW - precolumn derivatization

KW - antimicrobial residues

KW - degradation-products

KW - confirmatory assay

KW - kidney

KW - muscle

KW - amoxicillin

U2 - 10.1007/s00216-010-3523-0

DO - 10.1007/s00216-010-3523-0

M3 - Article

VL - 396

SP - 3027

EP - 3040

JO - Analytical and Bioanalytical Chemistry

JF - Analytical and Bioanalytical Chemistry

SN - 1618-2642

IS - 8

ER -