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«Determination of tetracycline residues in soil by pressurized liquid extraction and liquid chromatography tandem mass spectrometry Vicente Andreu · ...»

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Ecotoxicological studies, especially on soil microorganisms, should be performed to estimate the risk for the soil flora and the spread of antibiotic resistance.

Acknowledgements The authors thank the Spanish Ministry of Science and Innovation together with the European Regional Development Funds (ERDF) (projects GCL2007-66687-C02-01/BOS, CGL2007-66687C02-02 and CGL2008-01693/BTE) and the Conselleria de Sanitat of the Generalitat Valenciana (EVES2008-011) for financial support. P.V.R thanks the Spanish Ministry of Science and Innovation for the FPU grant.


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Table 1. LC-MS/MS conditions for confirming and quantifying the selected tetracyclines in soil samplesa

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a The criteria for residue identification were (i) four identification points through the measurement of two product ions plus the precursor ion; (ii) retention time of suspected analyte and reference standard within the tolerance interval of ±2.5%, and (iii) the ion ratio for each analyte in samples matches that of the standards within the maximum permitted tolerances (± 20 % for TC, e-TC, CTC. e-CTC and e-OTC; ± 25 for OTC, and ± 50 for DC).

b CV = cone voltage c CE = collision energy d Ion ratios=the ratio of the intensities of the two most abundant transitions of each tetracycline determined from the analysis of standards prepared in methanol-water (10:90 v/v) (n = 15).

Table 2. Linear regression parameters of TCs from soil calibration curvesa ranging from 10 to 500 µg kg-1 (6 points, triplicate analyses), LODs and LOQs

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a The linear regression analysis was carried out by plotting the peak area ratio of analyte and IS versus the analyte concentration Table 3. Recoveries and the relative standard deviations (RSDs) of the studied TCs in soil spiked at different concentrations.

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Table 4. Results of the investigation of 100 samples collected of the Ap horizon (0-20 cm) from 50 fields located in the South surrounding of Valencia city in February 2007.

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Fig. 1 Efficiency of the PLE system using different extractants. Soil samples were spiked at 50 µg kg-1 and the extract were analyzed by LC-ESI-MS/MS without further cleanup step.

Fig 2. Efficiency on the SPE clean-up using different SPE platforms at two different concentrations 1.25 and 12.5 μg l−1.

Fig. 3 LC-MS/MS chromatograms of soil spiked at 10 µg kg-1 of each tetracycline including the internal standard (A) TIC of the ten precursor → product ion transitions reported in Table 1, (B) mass chromatogram for DC (445 → 428 transition), (C) mass chromatogram for TC (445 → 427 transition), (D) mass chromatogram for OTC (461 → 443 transition) and (E) mass chromatogram for CTC (479 → 444 transition).

Fig. 4 LC-MS/MS chromatograms of soil spiked at 10 µg kg-1 of each tetracycline epimer and the the internal standard (A) TIC of the ten precursor → product ion transitions reported in Table 1, (B) mass chromatogram for e-TC (445 → 427 transition), (C) mass chromatogram for e-OTC (461 → 443 transition) and (D) mass chromatogram for e-CTC (479 → 444 transition).

Fig. 5 LC-MS/MS chromatogram corresponding to sample 1b.

Figure 1 Figure 2 Figure 3 Soil sample spiked with 10 µg kg-1 of each TC and of the IS

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% % 7.52

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2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00

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