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为建立一种合理高效液相色谱法,测定丁香酚药浴麻醉鲤鱼后,丁香酚在鲤鱼血中的消除规律,为丁香酚作为鲤鱼麻醉剂使用的安全性评价支撑。本研究采取鲤鱼血样经甲醇+乙腈提取,过滤的方法。流动相比例甲醇-乙腈-水(31:31:38),流速为1 m L/min,紫外吸收为280 nm,柱温为35℃,进样量为20μL。丁香酚在鲤鱼血浆中回归方程:Y=19.146X+0.130 26(r~2=0.999 01),检测限是0.828 ng/m L,定量限为1.656 ng/m L。平均添加回收率为86.74%。日内精密度RSD为4.135%,日间精密度RSD为4.558%。稳定性实验和重现性实验结果良好。说明本方法是测定药浴鲤鱼血中丁香酚较理想的方法。
In order to establish a reasonable high performance liquid chromatography (HPLC), the eradication rule of eugenol in carp blood was determined after the eugenol medicated bath was anesthetized with carp, which is the safety evaluation support of eugenol used as carp anesthetic. In this study, carp blood samples were extracted by methanol + acetonitrile and filtered. The mobile phase was methanol-acetonitrile-water (31:31:38), the flow rate was 1 m L / min, the UV absorption was 280 nm, the column temperature was 35 ℃ and the injection volume was 20 μL. The regression equation of eugenol in carp plasma was Y = 19.146X + 0.130 26 (r ~ 2 = 0.999 01), the limit of detection was 0.828 ng / mL, and the limit of quantification was 1.656 ng / mL. The average recovery rate was 86.74%. The intraday RSD was 4.135% and the intraday RSD was 4.558%. The stability experiment and reproducibility experiment result is good. This method is to determine the medicine bath carp blood eugenol a more ideal method.