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ANALYTICAL METHOD FOR THE DETERMINATION OF PHENYLGLUCURONIDE IN RAINBOW TROUT GILL WATER
Citation:
Kuehl, D W. AND C. T. Jenson. ANALYTICAL METHOD FOR THE DETERMINATION OF PHENYLGLUCURONIDE IN RAINBOW TROUT GILL WATER. Presented at SETAC 8th Annual Meeting, Bloomington, MN, April 13-14, 2000.
Description:
Phenylglucuronide (PG), a primary phase II metabolite of phenol, can be excreted by fish through urine and feces, similar to mammals. In addition, it may also be possible to eliminate it through a fish's gills. In order to assess the significance of gill water elimination, analytical methods were needed to identify and quantify PG at concentrations in water to 1 ug/L. These analyses are difficult because fish are constantly excreting a water soluble high density mixture of materials referred to as the mucous coat, or mucin, from their gills. In preliminary experiments using distilled or Lake Superior water, very good recoveries of PG (>90%) using either liquid/liquid partitioning or extraction with carbon, XAD, C18, or SAX sorbents were observed; however, poor recoveries (<10%) were obtained when gill water (water passed over gills in a special trout bioassay chamber) was used. Good recoveries of PG from gill water were finally obtained using Diazem C8-SAX (MetaChrom), a sorbent with an antifouling coated surface and C8 and SAX in the interior of the bead. It was found that a maximum of 50 mL of acidified (0.1% acetic acid) gill water containing methanol (1%) could be extracted per 500 mg of resin, before fouling of the resin resulted in decreased recoveries. Quantification of PG was done using a Beckman Nouveau UV/HPLC system equipped with a Beckman Ultrasphere C18 column. PG was monitored for at 265 nm. Gill water samples were spiked with an internal standard, p-nitrophenyl-glucuronide (10 ug/L) for quantification of PG. Confirmation of the presence of PG in the gill water extract was done using a Waters 2690 HPLC/MicroMass QTOF mass spectrometer.