雌性大鼠血清和组织中壬基酚的高效液相色谱测定法

目的建立大鼠血清及组织样品中壬基酚(NP)的液-液萃取和高效液相色谱荧光检测方法。方法清洁级SD雌性大鼠血清采用正己烷-乙醚混合溶剂(V∶V=7∶3)提取,组织采用甲醇-乙酸铵缓冲液(pH值为4.5,V∶V=2∶8)提取,采用C18色谱柱分离,乙腈-乙酸铵缓冲液(pH值为4.5,V∶V=95∶5)为流动相,柱温40℃,在激发波长227nm,发射波长313nm处用荧光检测器检测。结果血清和组织中NP含量与峰面积呈良好的线性关系,线性范围为0.1～30.0μg/ml。检出限为13ng/ml,加标回收率为92.60%～108.00%,日内精密度RSD为0.29%～1.51%,日间精密度RSD为0.74%～1.57%,均小于2%。结论该方法简便易操作、灵敏准确,回收率好,精密度高,适用于动物血清及组织中环境雌激素NP含量的检测。

Method for Determination of Nonylphenol in Serum and Tissues of Female Rats by High Performance Liquid Chromatography with Fluorescence Detector

Objective To establish a simple and highly sensitive method to measure nonylphenoi (NP) in rat serum and tissues by using liquid-liquid extraction and high performance liquid chromatography (HPLC) with fluorescence detector. Methods NP in the serum samples was extracted by a mixed solvent of n-hexane and diethylether (V:V=7:3), the tissue samples were homogenized with methanol ammonium acetate buffer (V:V=2:8),the organic layer was dried with nitrogen flow and dissolved with mobile phase. The operating conditions were optimized as follows: C_(18) column (250 min×4.6 mm, 5 μm), acetonitrile-ammonium acetate bufer (pH= 4.5) as mobile phase at a flow rate of 1 ml/min and fluorescence detector. The excitation and emission wavelength at 227 nm and 313 nm respectively were used for the determination. Results There was a good linear relationship between the concentrations of the NP in rat serum and their peak areas in the range of 0.1 to 30.0 μg/ml. The detection limit was 13 ng/ml. The recovery rates of rat serum samples were 90.60%-108.00%. The RSDs of intra-day precision were 0.29%-1.51%, the RSDs of inter-day precision were 0.74%-1.57%. Conclusion The method is simple, sensitive, reliable and is applicable for the determination of NP in animal serum.