高效液相色谱-串联质谱法测定动物血浆中罗匹尼罗的浓度

Determination of Ropinirole in Animal Plasma by LC-MS/MS

建立高效液相色谱-串联质谱(LC-MS/MS)测定动物血浆中罗匹尼罗浓度的方法。血浆经乙酸乙酯萃取后,以HPLC分离,电喷雾离子化(ESI^+)串联质谱检测。以甲醇-乙腈-0.1‰冰醋酸(36:9:55)为流动相,流速为0.2mL·min^(-1),采用Ultimate XB-C 18柱(150mm×2.1mm,3μm)分离,在三级四极杆串联质谱中经电喷雾电离源(ESI)离子化,以多反应监测(MRM)方式进行检测。盐酸罗匹尼罗、盐酸苯海拉明(内标)的扫描离子对m/Z分别为261→114和m/Z 256→167。LC-MS/MS测定血浆中罗匹尼罗线性范围为0.02—400ng·mL^(-1),范围内线性关系良好(r=0.9998);以3个浓度水平的质量控制样品求得各浓度水平日内、日间精密度(RSD)均小于15.2%。在非临床药代动力学研究中,应用此法测定了受试兔子血浆中罗匹尼罗的浓度。该法灵敏、快速、准确,操作简便,样品处理方便,线性范围宽,该方法检测快速、专一、灵敏,可满足罗匹尼罗临床前药代动力学研究和临床药动学研究的要求。

The plasma was extracted with ethyl acetate,then separated by HPLC,and detected by electrospray ionization （ ESI＋ ） tandem mass spectrometry. The mobile phase of methanol-acetonitrile-0.1‰ acetic acid （36 ： 9 ： 55） with flow rate of 0. 2mL · min-1. The chromatographic system consisted of Ultimate XB-C18 column （150mm× 2. 1 mm, 3um）. A tandem mass spectrometer equipped with electrospray ionization （ESI） source was used as detector and operated in the positive ion mode. Multiple reaction monitoring using the precursor → product ion combinations of m/Z 261 → 114 and m/Z 256→167 were used to quantify ropinirole and the internal standard, respectively. The linear calibration curve showed a good linear relationship（r=0. 9998） in the range of 0. 02--400ng · mL-1. With three concentrations of quality control samples,the obtained concentration levels of all days,the intra-and inter-day precision （RSD） were less than 15.2%. This method was used to detecte ropinirole in rabbit of pre-clinical pharmacokinetic study with sensitive and acceptable preeision, and can be used for the pre-clinical pharmacokinetie study and clinical pharmaeokinetie of ropinirole.