A Rapid, Sensitive, and Selective Method for Quantitation of Lamprey Migratory Pheromones in River Water

Abstract

The methodology of using fish pheromones, or chemical signatures, as a tool to monitor or manage species of fish is rapidly gaining popularity. Unequivocal detection and accurate quantitation of extremely low concentrations of these chemicals in natural waters is paramount to using this technique as a management tool. Various species of lamprey are known to produce a mixture of three important migratory pheromones; petromyzonol sulfate (PS), petromyzonamine disulfate (PADS), and petromyzosterol disulfate (PSDS), but presently there are no established robust methods for quantitation of all three pheromones. In this study, we report a new, highly sensitive and selective method for the rapid identification and quantitation of these pheromones in river water samples. The procedure is based on pre-concentration, followed by liquid chromatography/tandem mass spectrometry (LC/MS/MS) analysis. The method is fast, with unambiguous pheromone determination. Practical quantitation limits of 0.25 ng/l were achieved for PS and PADS and 2.5 ng/l for PSDS in river water, using a 200-fold pre-concentration, However, lower quantitation limits can be achieved with greater pre-concentration. The methodology can be modified easily to include other chemicals of interest. Furthermore, the pre-concentration step can be applied easily in the field, circumventing potential stability issues of these chemicals.

A Sensitive Analytical Method for Quantifying Petromyzonol Sulfate in Water as a Potential Tool for Population Monitoring of the Southern Pouched Lamprey, Geotria Australis, in New Zealand Streams

“…The MS/MS has orders of magnitude sensitivity improvement over the ITMS, which is highlighted by the detection of PADS, even though release rates were much reduced. Furthermore, MS/MS is able to fragment PS, whereas ITMS is not, thus providing selectivity and improved confidence in chemical confirmation (Stewart et al, 2011). As such, MS/MS appears to be more applicable to quantifying pheromones at the extremely low water concentrations observed in the environment.…”

“…Manual integration of peak areas was undertaken and PS concentration calculated from a standard curve. Where higher sensitivity was required (i.e., quantitation of PS from passive samplers and active sampling in the field and quantitation of PS, PADS, and PSDS from lamprey ammocete tankwater), LC/MS/MS was used, as described previously (Stewart et al, 2011). This method afforded instrument practical quantitation limits of 0.05, 0.05, and 0.50 ng/ml for PS, PADS, and PSDS, respectively.…”

“…Final work-up of the sample was as described previously for passive sampler sorbents and discs. The sample was sealed with a lid and septum for LC/MS/MS analysis as described previously (Stewart et al, 2011).…”

“…Detection of such low water concentrations require sensitive analytical techniques that are capable of quantifying pheromone concentrations in natural waters. Liquid chromatography coupled with triple quadrupole mass spectrometry (LC/MS/MS) is suitable for detecting and quantifying environmental concentrations of organic chemicals, and previous work has shown this method -when used in conjunction with pre-concentration -is capable of detecting PS at stream water concentrations as low as 2.5×10 -14 M (Stewart et al, 2011).…”

Determination of the Three Main Components of the Grapevine Moth Pest Pheromone in Grape-Related Samples by Headspace-Gas Chromatography-Mass Spectrometry

“…Pheromone determinations have been employed for different purposes [14][15][16] including the detection of insects [17]. In this sense, our research group has developed some analytical approaches for the detection of the major components of the Tuta absoluta pheromone in tomato, water and air [18,19] and the main component of the Bactrocera oleae pheromone in olives [20].…”