高效液相色谱法测定貉子肉中褪黑素残留量的研究

Study on determination of the residual quantity of melatonin in nyctereutes procyonoides tissues using high performance liquid chromatography

本研究建立一种检测貉子肉中褪黑素药物残留的高效液相色谱——荧光检测法。貉子肉经乙酸乙酯提取，旋转蒸发至干，溶解残留物，加入水饱和的正己烷去脂肪，Waters Oasis HLB柱净化。以20mmol／LNaAc—HAc溶液和甲醇溶液为流动相，等度洗脱，检测波长：Ex=285nm，Em=345nm。结果表明，褪黑素在0．1～100g／L呈良好的线性关系，r均大于0．9999。方法的最低检出限为0．25g／kg，定量限为1g／kg。在1、5和10g／kg的添加水平上，其回收率范围在72．0％-90．4%之间，相对标准偏差（RSD）为1．2％～2．4％。

A high performance liquid chromatography method has been developed for the simultaneous determination melatonin in nyctereutes procyonoides tissues muscle by fluorescence detector. Sample was extracted by ethyl acetate, the solution was purified with Waters Oasis HLB. Mobile phase of 20 mmol/L balanced solution （sodium acetate trihydrate-glacial acetic acid） -methanol was used with isocratic elution. The detection wavelength was Ex=285 nm, Em=345 nm. The results showed that the calibration curves of melatonin had good linearity relationship in the range from 0. i to 100 ~ g/L, and the correlation coefficient was greater than 0.9999. The average recovery rates of melatonin in muscle tissue were from 72.0% to 90.4% at three adding levels of i, 5 and 10 μ g/kgfor melatonin, and the values of relative standard deviation（RSD） for the average recovery rates of melatonin were from 1.2% to 2.4%,the limit of detection and limit of quantification were 0.25 μg/kg and 1 μg/kg, respectively.