[Determination of 4-nonylphenol and bisphenol A in rat serum by high performance liquid chromatography with fluorescence detection].

ABSTRACT

OBJECTIVE: This study was directed to the development of a simple and highly sensitive method for determination of 4-nonylphenol (4-NP) and bisphenol A (BPA) in rat serum using HPLC with fluorescence detector. METHODS: 4-NP and BPA in serum samples were extracted by a mixed solvent of n-hexane and diethyl ether (7030), the organic layer was dried with nitrogen flow and dissolved with mobile phase. The operating conditions such as C18 column (250 mm x 4.6 mm, 5 microns), acetonitrile-ammonium acetate buffer (pH 4.5) as mobile phase at a flow rate of 1 ml/min and fluorescence detector with the excitation and emission wavelength at 227 nm and 313 nm respectively were used for the determination. RESULTS: There was good linear relationship between the concentrations of the analytes in rat serum and their peak areas in the ranges of 0.013-5.0 micrograms/ml for 4-NP and 0.004-3.0 micrograms/ml for BPA. The detection limit of the method was 13 ng/ml for 4-NP and 4 ng/ml for BPA. Recoveries of rat serum samples were 83.8%-100.0% for 4-NP and 83.3%-100.0% for BPA. The intra-day precision was 1.70%-2.70%, the inter-day precision was 2.06%-9.78%. CONCLUSION: The results showed that the method is suitable for the determination of BPA and 4-NP in serum.