高效液相色谱与共振散射光联用方法测定家兔血浆中阿米卡星的含量

Determination of amikacin in rabbit plasma by high performance liquid chromatography coupled with resonance scattering light technology

目的建立高效液相色谱—共振散射光法联用技术测定家兔血浆中阿米卡星含量的方法。方法采用Agilent ZORBAX SB C18(4.6×250 mm,5μm)为色谱柱,流动相为水:乙腈=97:3(V/V),流速0.5 mL/min,荧光检测器λex=λem=488 nm;探针达旦黄为20μg/mL的BR溶液,单泵流速0.21 mL/min。结果家兔血浆中阿米卡星的浓度在10～600μg/mL范围线性关系良好,最低检测限为5μg/mL,低、中、高三种浓度的提取回收率均大于90%,日内日间精密度RSD值均小于10%。结论该方法灵敏度高,选择性好,专属性强,可应用于家兔血浆中阿米卡星药代动力学研究,同时为氨基糖苷类抗生素测定提供一种新的方法。

Objective To establish the method to determine the plasma level of amikacin in rabbit by high performance liquid chromatography coupled with resonance scattering light technology. Methods The chromatographic separation was performed on a Agilent ZORBAX SB C18column （4.6 × 250 mm, 5μm）. The mobile phase consisted of water- acetonitrile （97：3, V/V） at flow rate of 0.5 mL/min. The fluorescence detertor was set at λex =λem = 488nm. Titan Yellow containing 20 ug/mL BR solution was used as the molecular recognition probe and the flow rate of probe solution was 0.21mL/min at post - column. Results The linearity was obtained within tile range of amikacin concentrations from 10 to 600 μg/mL. The minimal inspect level was 5 μg/mL. The recoveries of low, medium and high concentra- tions of extraction were more than 90%. The inter - day and intra - day RSD were less than 10%. Con- clusion The method has high sensitivity, good selectivity and specificity and can be applied to do the amikacin pharmacokinetic study in rabbit plasma and further provide an alternative method for the deter- mination of aminoglycoside antibiotics.