Antonian L, DeMontigny P, Wislocki P G
Department of Drug Metabolism, Merck Research Laboratories, Rahway, NJ 07065, USA.
J Pharm Biomed Anal. 1998 Apr;16(8):1363-71. doi: 10.1016/s0731-7085(97)00183-0.
Eprinomectin is a potent anthelmintic compound that kills certain parasitic nematodes and arthropods of cattle. A sensitive and automated bioanalytical assay was developed for quantitation of eprinomectin in bovine plasma in support of clinical development of eprinomectin for use in all classes of cattle. This assay determined the concentration of eprinomectin in plasma by reversed-phase high performance liquid chromatography (HPLC) with fluorometric detection. Plasma sample preparation included liquid extraction performed by the Packard MultiPROBE robotics workstation, followed by solid phase extraction performed by the Gilson ASPEC XL automated workstation. The HPLC assay included automated pre-column derivatization with a fluorogenic reagent system which included trifluoroacetic anhydride and N-methylimidazole as the catalyst. This reversed-phase chromatographic analysis was based on the fluorescence detection of derivatized eprinomectin and an internal standard, L-648 548, which was similarly derivatized by the fluorogenic reagents. The assay was automated and validated for two concentration ranges of 0.05-10 and 0.5-200 ng ml-1. The lower limit of quantitation of eprinomectin in plasma was 0.05 ng ml-1. The %RSD of the assay was 10% or better at all concentrations. This automated analysis of eprinomectin was used for high-throughput clinical assays with acceptable accuracy and precision.
依普菌素是一种强效驱虫化合物，可杀死牛的某些寄生线虫和节肢动物。为了支持依普菌素在所有牛类中的临床开发，开发了一种灵敏且自动化的生物分析方法，用于定量牛血浆中的依普菌素。该方法通过反相高效液相色谱法（HPLC）和荧光检测来测定血浆中依普菌素的浓度。血浆样品制备包括由帕卡德MultiPROBE机器人工作站进行的液液萃取，随后由吉尔森ASPEC XL自动工作站进行固相萃取。HPLC分析包括用荧光试剂系统进行自动柱前衍生化，该荧光试剂系统包括三氟乙酸酐和作为催化剂的N-甲基咪唑。这种反相色谱分析基于衍生化依普菌素和内标L-648 548的荧光检测，内标也由荧光试剂进行类似的衍生化。该方法经过自动化处理，并在0.05 - 10和0.5 - 200 ng ml-1两个浓度范围内进行了验证。血浆中依普菌素的定量下限为0.05 ng ml-1。该方法在所有浓度下的相对标准偏差（%RSD）均为10%或更低。这种依普菌素的自动化分析用于高通量临床检测，具有可接受的准确度和精密度。
