2 research outputs found
Determination of antibacterials in animal feed by pressurized liquid extraction followed by online purification and liquid chromatography-electrospray tandem mass spectrometry
A fully automated method has been developed for
analysis of eighteen antibacterial compounds, including
penicillins, cephalosporins and sulfonamides, in animal feed
with limits of quantification in the range 0.25–5.79 μgkg−1.
The method is based on pressurized liquid extraction of 3 g
homogenized feed with water and online clean-up of 500 μL
of the extract with C18HD cartridges. The purified sample
was directly analysed by liquid chromatography–electrospray
tandem mass spectrometry (SPE–LC–ESI-MS–MS). Chromatographic
separation was achieved within 10 min by use of
a C12 Phenomenex Hydro-RP reversed-phase analytical
column and a mobile phase gradient (water + 0.1% formic
acid–methanol + 0.1% formic acid). The method was
validated, revealing capability for detection of concentrations
as low as 0.09 μgkg−1, decision limits (CCα) and detection
capabilities (CCβ) in the range 10–174 μgkg−1 and 22–
182 μgkg−1, respectively, and inter-day precision ranging
from 0.7 to 8.3%. Recovery, with internal standard correction,
was in the range 93–134% for all analytes. The method was
then applied to analysis of fifteen feed samples, nine of which
contained at least one antimicrobial at concentrations between
0.006 and 1.526 mgkg−1.The performance data and results
from the method were compared with those from a previous
method developed by our group, using offline SPE, by
analyzing the same set of samples by both methods. The
online SPE approach resulted in slightly improved sensitivity,
with LODs of 0.09–2.12 μgkg−1 compared with 0.12–
3.94 μgkg−1 by the offline approach. In general, better
recovery was achieved by use of online purification (for 72%
of the analytes) and the correlation between the two methods
was good. The main advantages of the new online method
are rapid and automated sample pre-treatment, and reduction
of sample manipulation, enabling high-throughput analysis
and highly accurate results. Because of all these characteristics,
the proposed method is applicable and could be
deemed necessary within the field of food control and safety.This work was supported by the Spanish
Ministry of Science and Innovation, through project CEMAGUA
(CGL2007-64551/HID). Waters is acknowledged for providing the
SPE cartridges. Lina Kantiani acknowledges the Generalitat de
Catalunya (2009FI_B 00660) and the Alexander S. Onassis
foundation (F-ZD 029/2007-2008).Peer reviewe