A reversed phase high performance liquid chromatographic(RP-HPLC) method was developed for the determination of nodularin in hepatopancreas,kidney,muscle and plasma tissues of Carassius auratus(C-auratus) by optimizing conditions of pretreament and chromatography.Samples were extracted with 90% methanol solution by ultrasonic,followed by purification with a CNW Bond C18 solid phase extration column,and then determined by RP HPLC equipped with ultraviolet(UV) detector.The steps of purification included activating columns,sample absorbing onto columns,flushing the columns and eluting the columns.The eluent was 90% methanol containing 0.1% trifluoroacetic acid(TFA).The detection of target compound was performed with RP-HPLC using 0.1% trifluoroacetic acid(TFA)-methanol(42∶58) as mobile phase at a constant flow of 1.0 mL/min.The column temperature was set at 30 ℃ and the wavelength of UV detector was set at 238 nm while the injection volume was 20 μL.The experimental results showed that the concentration of nodularin in four crucian tissues showed good linearity(r2>0.99) with peak area in the range of 0.05-5.00 mg/L.The recoveries of nodularin from crucian tissues at three spiked concentration levels of 0.08,0.40,2.50 mg/L ranged from 79.5% to 106.4%,with RSDs of 0.54%-5.3%.The method detection limit was 11.30 μg/kg for muscle and 24.00 μg/L for plasma sample.The method was accurate and reliable,and could be applied in the dynamic distribution and residues determination of nodularin in C-auratus,providing an effective tool for the detection of nodularin in other aquatic products and for the safety control of aquatic products quality.
关键词
反相高效液相色谱肝胰脏肾脏血浆肌肉节球藻毒素
Keywords
reversed phase high performance liquid chromatography(RP HPLC)hepatopancreaskidneyplasmamusclenodularin