TY - JOUR
T1 - Non-targeted workflow for identification of antimicrobial compounds in animal feed using bioassay-directed screening in combination with liquid chromatography-high resolution mass spectrometry
AU - Wegh, Robin S.
AU - Berendsen, Bjorn J.A.
AU - Driessen-Van Lankveld, Wilma D.M.
AU - Pikkemaat, Mariël G.
AU - Zuidema, Tina
AU - Van Ginkel, Leen A.
PY - 2017/11
Y1 - 2017/11
N2 - A non-targeted workflow is reported for the isolation and identification of antimicrobial active compounds using bioassay-directed screening and LC coupled to high-resolution MS. Suspect samples are extracted using a generic protocol and fractionated using two different LC conditions (A and B). The behaviour of the bioactive compound under these different conditions yields information about the physicochemical properties of the compound and introduces variations in co-eluting compounds in the fractions, which is essential for peak picking and identification. The fractions containing the active compound(s) obtained with conditions A and B are selected using a microbiological effect-based bioassay. The selected bioactive fractions from A and B are analysed using LC combined with high-resolution MS. Selection of relevant signals is automatically carried out by selecting all signals present in both bioactive fractions A and B, yielding tremendous data reduction. The method was assessed using two spiked feed samples and subsequently applied to two feed samples containing an unidentified compound showing microbial growth inhibition. In all cases, the identity of the compound causing microbiological inhibition was successfully confirmed.
AB - A non-targeted workflow is reported for the isolation and identification of antimicrobial active compounds using bioassay-directed screening and LC coupled to high-resolution MS. Suspect samples are extracted using a generic protocol and fractionated using two different LC conditions (A and B). The behaviour of the bioactive compound under these different conditions yields information about the physicochemical properties of the compound and introduces variations in co-eluting compounds in the fractions, which is essential for peak picking and identification. The fractions containing the active compound(s) obtained with conditions A and B are selected using a microbiological effect-based bioassay. The selected bioactive fractions from A and B are analysed using LC combined with high-resolution MS. Selection of relevant signals is automatically carried out by selecting all signals present in both bioactive fractions A and B, yielding tremendous data reduction. The method was assessed using two spiked feed samples and subsequently applied to two feed samples containing an unidentified compound showing microbial growth inhibition. In all cases, the identity of the compound causing microbiological inhibition was successfully confirmed.
KW - animal feed
KW - antibiotics
KW - Antimicrobials
KW - bioassay
KW - LC-MS
UR - https://doi.org/10.6084/m9.figshare.5332189
U2 - 10.1080/19440049.2017.1364431
DO - 10.1080/19440049.2017.1364431
M3 - Article
AN - SCOPUS:85028573949
SN - 1944-0049
VL - 34
SP - 1935
EP - 1947
JO - Food Additives & Contaminants. Pt. A, Chemistry, Analysis, Control, Exposure & Risk Assessment
JF - Food Additives & Contaminants. Pt. A, Chemistry, Analysis, Control, Exposure & Risk Assessment
IS - 11
ER -