A rapid two-step chromatographic method for the quantitative determination of vitellogenin in fish plasma

Abstract

By combining anion-exchange membrane purification with high-performance size-exclusion chromatography (HPSEC) analysis, a two-step chromatographic method was developed for the determination of vitellogenin (Vtg) in fish plasma. Most plasma protein interferences can be removed during anion-exchange membrane purification process. Vtg is eluted from the size-exclusion chromatography column with a retention time of about 9 min and is characterized based on the native molecular weight, with a limit of quantification of 20 µg Vtg mL -1 plasma. The spiked recovery and interassay variability were better than 80% and 4.8%. This method was successfully applied to analyze the plasma Vtg levels of loach (Misgurnus angaillicaudatus) and sea catfish (Enchelyopus elongatus). In addition to all the female fish, Vtg is detected in 75% of male loaches and 100% of male sea catfish. The result indicates that some chemicals or unknown factors with estrogenic activity have induced male fish to produce Vtg.

Determination of 4-tert-octylphenol, 4-nonylphenol and bisphenol A in surface waters from the Haihe River in Tianjin by gas chromatography–mass spectrometry with selected ion monitoring

“…It was found that many chemical industries including chemical plant of Dagu were near the estuary of the Haihe River. Our study also found that the high level of vitellogenin, a biomarker of estrogen, could be detected in all the male sea catfish collected from the site R1, which indicated the heavy pollution status of this site (Shao et al, 2004). (3) At the south of R1 there was a sewage outlet of the Dagupaiwu River, and the effluents can easily pollute the estuary of the Haihe River during the period of flood tide.…”

Measurement of estrogenic activity in sediments from Haihe and Dagu River, China

“…Relatively high concentrations of OP, NP, BPA and selected organochlorine pesticides were found at the sampling sites (Table 3). In a previous study (Shao et al, 2004) a high level of vitellogenin (VTG), a possible biomarker of estrogen (Sumpter and Jobling, 1995), was detected in all the male sea catfish collected from the Zhah site. Chang et al (2003) compared the effects of BPA, NP and OP on in vitro VTG synthesis in primary cultures of hepatocytes of Chinese minnow Phoxinus oxycephalus.…”

Separation and detection of vitellogenin in fish plasma by capillary zone electrophoresis

“…The main techniques for VTG determination currently are enzyme linked immunosorbent assay (ELISA) [7][8][9], radioimmunoassay (RIA) [10,11], Western blots [12], HPLC [13] and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) [14]. In many related articles, immunoassays were applied in VTG analysis as the commonly used approach.…”