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A rapid imethod™ test for analysis of carbamate pesticides in beverages
A Rapid iMethod™ Test for the Analysis of Tetracyclines in Meat using Manual SPE
iMethod™ Test for Tetracycline Antibiotics Version 1.0 for Cliquid® Software
Meat and other animal products need to be routinely monitored
for veterinary drug residues that are used to fight disease and
infection in animals, but are harmful to humans if present upon
ingestion. However, with wide availability, variable legislation and
variable animal husbandry practices from country-to-country,
Strata™-X 33 µm polymeric SPE cartridge. This user-contributed
high drug residue levels in produce continue to be an issue.
iMethod™ test for Tetracyclines includes method documentation,
Over-use and inappropriate use of antimicrobial drugs can lead
MS and LC method files, processing method files and report
to increased antimicrobial resistance, reducing the ability to fight
templates, all deployed as a new iMethod test in your Cliquid®
human infection. On top of this, exposure to such drugs can
software. More in-depth sample preparation and instrument
affect human reproduction, cause cancer and have a toxic effect.
parameter information is included as part of the standard
The following description outlines the instrument requirements
operating procedure provided with the method, as are the
and expected results obtainable from the iMethod™ test for the
required analytical columns; solvents, standards and any
quantification of five tetracycline antibiotics and metabolites
supplies required for sample preparation are not included.
when using an AB SCIEX 3200 QTRAP® LC/MS/MS instrument.
The mobile phase consists of water with formic acid and
Sample preparation is based on extraction with acidified
acetonitrile with separation on Phenomenex Gemini C18 5µm
methanol, followed by filtration, centrifugation, dilution with
4.6 x 50 mm HPLC column. An example chromatogram of the
acidified water, and manual SPE clean-up on a Phenomenex
separation achieved is shown below in figure 1.
Figure 1. Example chromatogram for an injection of a 10 ppb standard, showing all the tetracyclines overlaid in one window (between 2.5 and
Figure 2 illustrates the performance of the method for two
Please note that the results presented were obtained using a
transitions for all five analytes. In all cases the S/N is greater
single instrument and single set of standards and samples, and
than 7:1 for a 1 ppb injection on column.
the results here may not be typical for all instruments. Prior to
production use, the method should be fully validated with real
samples. Variations in LC column properties, chemicals,
environment, instrument performance and sample preparation
procedures will impact performance, thus these results should be
considered as informative rather than representative.
Figure 2. Example Chromatograms for 1 ppb standards (signal-to-noise measured using PEAK:PEAK script).
The following two calibration curves are representative of the
performance obtained on the instrument using the method
described here. No internal standards were used in the analysis
Figure 3. Calibration curve for chlortetracycline 1, standards 1 to 500 ppb (no internal standard used).
Figure 4. Calibration curve for methacycline 1, standards 1 to 500 ppb (no internal standard used).
In order to run this method as outlined above, the following
The purchase and use of certain of the chemicals listed above
may require the end user to possess any necessary licenses,
permits or approvals, if such are required in accordance with
• An AB SCIEX 3200 QTRAP® LC/MS/MS System
local laws and regulations. It is the responsibility of the end
• Shimadzu XR HPLC system, or equivalent
user to purchase these chemicals from a licensed supplier, if
required in accordance with local laws and regulations. The
suppliers and part numbers listed below are for illustrative
purposes only and may or may not meet the aforementioned
• A Phenomenex Gemini C18 5µm 4.6 x 50 mm HPLC
local requirements. AB SCIEX is not responsible for user’s
compliance with any statute or regulation, or for any permit or
• Phenomenex Strata™-X 33 µm, Polymeric Sorbent
approval required for user to implement any iMethod™
• Pipettes and standard laboratory glassware
The iMethod™ test described above has been designed by
Please note that the Phenomenex HPLC column is required
AB SCIEX to provide the sample prep and instrument
but not included with this iMethod™ test. This method can also
parameters required to accelerate the adoption of this method
be run on other HPLC systems, given that they are supported
for routine testing. This method is provided for information
for use by Cliquid® Software and the retention times are
purposes only. The performance of this method is not
updated to reflect the configuration used.
guaranteed due to many different potential variations, including
instrument performance, tuning, and maintenance, chemical
variability and procedures used, technical experience, sample
iMethod™ Test for Tetracyclines for Cliquid®
matrices, and environmental conditions. It us up to the end
user to make adjustments to this method to account for slight
differences in equipment and/or materials from lab-to-lab as
well as to determine and validate the performance of this
method for a given instrument and sample type. Please note
that a working knowledge of Analyst® Software may be
For Research Use Only. Not for use in diagnostic procedures.
2010 AB SCIEX. The trademarks mentioned herein are the property of AB Sciex Pte. Ltd. or their respective owners. AB SCIEX™ is being used under license.
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