| Résumé | Quantitation of trace levels of domoic acid (DA) in seawater samples usually requires labour-intensive protocols involving chemical derivatization with 9-fluorenylmethylchloroformate and liquid chromatography with fluorescence detection (FMOC–LC–FLD). Procedures based on LC–MS have been published, but time-consuming and costly solid-phase extraction pre-concentration steps are required to achieve suitable detection limits. This paper describes an alternative, simple and inexpensive LC method with ultraviolet detection (LC–UVD) for the routine analysis of trace levels of DA in seawater without the use of sample pre-concentration or derivatization steps. Qualitative confirmation of DA identity in dubious samples can be achieved by mass spectrometry (LC–MS) using the same chromatographic conditions. Addition of an ion-pairing/acidifying agent (0.15% trifluoroacetic acid) to sample extracts and the use of a gradient elution permitted the direct analysis of large sample volumes (100 μl), resulting in both high selectivity and sensitivity (limit of detection = 42 pg ml−1 by LC–UVD and 15 pg ml−1 by LC–MS). Same-day precision varied between 0.4 and 5%, depending on the detection method and DA concentration. Mean recoveries of spiked DA in seawater by LC–UVD were 98.8% at 0.1–10 ng ml−1 and 99.8% at 50–1000 ng ml−1. LC–UVD exhibited strong correlation with FMOC–LC–FLD during inter-laboratory analysis of Pseudo-nitzschia multiseries cultures containing 60–2000 ng DA ml−1 (r2 > 0.99), but more variable results were obtained by LC–MS (r2 = 0.85). This new technique was used to confirm the presence of trace DA levels in low-toxicity Pseudo-nitzschia spp. isolates (0.2–1.6 ng ml−1) and in whole-water field samples (0.3–5.8 ng ml−1), even in the absence of detectable Pseudo-nitzschia spp. cells in the water column. |
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