HPLC快速测定胺碘酮及去乙基胺碘酮血药浓度的方法建立

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&#215;250 mm，5 μm）进行分析，以乙腈-100mmol&#183;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&#183;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0.048~3.842 μg&#183;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0.051~4.084 μg&#183;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0.02 μg&#183;mL^-1，定量限均为0.05 μg&#183;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&#177;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<0.02 μg&#183;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&#215;250 mm,5 μm)with dronedarone as the internal standard.The mobile phase of acetonitrile-100 mmol&#183;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&#183;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&#183;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&#183;mL^-1;the LODs for both amiodarone and desethylamiodarone were 0.02 μg&#183;mL^-1,while the LOQs were 0.05 μg&#183;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&#183;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.