LC-MS Applications for Environmental Analysis

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Application Note 63552
The calculations, of limit of detection (LOD) and limit
of quantification (LOQ) were made in accordance with
the UNICHIM Manual N. 179/0 where the calculation
of the limit of detection is made through the calibration
curve of the instrument used for analysis.5
To estimate the LOD and LOQ of the method (Table 4),
ten samples were prepared by adding standard solution
to 500 mg of homogenized mussel flesh and repeating
the extraction procedure according to the method in
“EU-Harmonised Standard Operating Procedure for
determination of Lipophilic marine biotoxins in
molluscs by LC-MS/MS – Version 3”. LOD and LOQ
are expressed in µg/Kg. Recoveries are shown in Table 5.
Conclusion
This method proved to be selective, sensitive, accurate,
and reproducible. It can be successfully applied for the
quantitative determination of several classes of lipophilic
marine biotoxins in bivalve mollusc samples.
References
1. Regulation (EC) No 853/2004 of the European Parliament and of the
Council of 29 April 2004. Official Journal of the European
Communities L139, 55-255.
2. Commission Regulation (EU) No 15/2011 of 10 January 2011.
Official Journal of the European Union . L6, 3-9.
3. Commission Regulation (EC) No 2074/2005 of 5 December 2005.
Official Journal of the European Communities, L388, 27-59.
4. European Union Reference Laboratory for Marine Biotoxins
(EU-RL-MB), Agencia Española de Seguridad Alimentaria y
Nutrición (AESAN). EU-Harmonised Standard Operating Procedure
for determination of Lipophilic marine biotoxins in molluscs by
LC-MS/MS – Version 3. 2011.
5. UNICHIM, Manual N. 179/0. Linee guida per la convalida di
metodi analitici nei laboratori chimici - criteri generali. Milano. 1999.
Hypersil GOLD
Analyte LOD (µg/Kg) LOQ (µg/Kg) Outliers (Huber Test)
AZA-1 0.56 ± 0.18 1.11 NO
AZA-2 0.93 ± 0.31 1.86 NO
AZA-3 1.28 ± 0.43 2.57 NO
DTX-1 5.66 ± 1.02 10.45 NO
DTX-2 0.71 ± 0.23 1.42 NO
YTX 1.67 ± 0.56 3.33 NO
PTX-2 1.40 ± 0.46 2.79 NO
OA 3.95 ± 1.32 7.91 NO
Table 4: LOD and LOQ of the method
Analyte Spiked Purified Extract Spiked Extract
AZA-1 96 ± 11 97 ± 11
AZA-2 101 ± 9 94 ± 14
AZA-3 104 ± 10 99 ± 6
DTX-1 101 ± 6 101 ± 7
DTX-2 101 ± 6 108 ± 42
YTX 99 ± 15 102 ± 17
PTX-2 103 ± 13 102 ± 20
OA 95 ± 7 93 ± 18
Table 5: Recovery values, where
R% = {[(μg/Kg)CALCULATED/(μg/Kg) THEORETICAL]*100}
Figure 1: Chromatogram of sample containing 40 ppb of toxins
(Retention Time: 7.44 min – AZA-1; 7.77 min – AZA-2; 6.88 min – AZA-3;
5.69 min – OA; 7.O5 min – DTX-1; 6.06 min – DTX-2; 5.54 min – YTX;
6.28 min – PTX-2
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