HPLC快速测定胺碘酮及去乙基胺碘酮血药浓度的方法建立

Establishment of an HPLC method for the therapeutic drug monitoring of amiodarone and desethylamiodarone

目的:建立高效液相色谱方法,监测血清中胺碘酮及去乙基胺碘酮的血药浓度。方法:以决奈达隆为内标,血清经蛋白沉淀后采用Alltima C18色谱柱(4.6 mm×250 mm,5μm)进行分析,以乙腈-100 mmol·L^(-1)磷酸二氢钠(磷酸调pH至3.0)(55∶45)为流动相,流速1.0 mL·min^(-1),柱温30℃,检测波长241 nm。并将测定结果和薄层扫描法的测定结果进行统计分析和比较。结果:方法具有良好的专属性,血清中的内源性物质和临床常见合用药物不干扰测定;胺碘酮质量浓度在0.048~3.842μg·mL^(-1)的范围内线性良好(r=0.999 6),去乙基胺碘酮质量浓度在0.051~4.084μg·mL^(-1)的范围内线性良好(r=0.999 8);胺碘酮和去乙基胺碘酮的最低检测限均为0.02μg·mL^(-1),定量限均为0.05μg·mL^(-1);准确度和精密度良好,相对回收率和绝对回收率均在97.5%~104.2%之间,日内精密度和日间精密度均小于8.0%;稳定性良好,在室温、冻融和冷藏(2~4℃)条件下胺碘酮和去乙基胺碘酮浓度的相对误差(RE)均小于±9%。方法学比较显示:新建的HPLC法和薄层扫描法的测定结果具有良好的相关性,且无显著性差异(P>0.05),但在<0.02μg·mL^(-1)的低浓度区,2种方法有显著性差异(P<0.05),HPLC法测定结果高于薄层扫描法。结论:该方法快速、简便,易推广,可作为血清中胺碘酮及去乙基胺碘酮的血药浓度监测方法。

Objective：To establish an HPLC method for the therapeutic drug monitoring of amiodarone and desethylamiodarone.Methods：The serum was deproteinized and analyzed on a column of Alltima C18 column（4.6 mm×250 mm,5 μm）with dronedarone as the internal standard.The mobile phase of acetonitrile-100 mmol·L-1 sodium dihydrogen phosphate（adjusted to pH 3.0 with phosphoric acid）55∶45 was used with a flow rate of 1.0 mL·min-1.The column temperature was 30 ℃.The detection wavelength was 241 nm.The determination results were compared with those of a reported TLC method with statistical analysis.Results：The method showed good specificity,no interference from the endogenous substances and potential co-administration drugs was observed;amiodarone showed good linearity in the range of 0.048-3.842 μg·mL-1 with a correlation coeffcient（r）of 0.999 6,while desethylamiodarone was with a correlation coeffcient（r）of 0.999 8 in the range of 0.051-4.084 μg·mL-1;the LODs for both amiodarone and desethylamiodarone were 0.02 μg·mL-1,while the LOQs were 0.05 μg·mL-1;the method indicated good accuracy and precision with relative and absolute recoveries in the range of 97.5%-104.2% and all intra-day and inter-day precisions 8.0%;the stability investigation showed 9% relative errors in the conditions of room temperature,freeze thawing and refrigeration（2-4 ℃）.The method comparison indicated no significant difference（P〈0.05）and good correlations between the present HPLC method and the TLC method,however significant differences between two methods（P〈0.05）were found in the low concentration samples（〈0.02 μg·mL-1）with higher results of HPLC method than TLC method.Conclusion：This method is fast,convenient and propagable for the therapeutic drug monitoring of amiodarone and desethylamiodarone.