Remmerie B M M, Sips L L A, de Vries R, de Jong J, Schothuis A M, Hooijschuur E W J, van de Merbel N C
Department of Bioanalysis, Johnson and Johnson Pharmaceutical Research and Development, Turnhoutseweg 30, B-2340, Beerse, Belgium.
J Chromatogr B Analyt Technol Biomed Life Sci. 2003 Jan 15;783(2):461-72. doi: 10.1016/s1570-0232(02)00715-8.
Since the first entry of risperidone on to the market in the early 1990s, investigation of the pharmacokinetic behaviour of the compound for which the availability of a bioanalytical method was a condition sine qua non, has received considerable attention. Most of the published methods, however, did not reach the level of sensitivity and selectivity which can be obtained today since the evolution of liquid chromatography-tandem mass spectrometry (LC-MS-MS) towards a routine technique in the bioanalytical laboratory. Therefore, we developed and validated a new LC-MS-MS method for the determination of risperidone and its active metabolite 9-hydroxyrisperidone in human plasma. This paper describes in detail the bioanalytical procedure and summarizes the validation results obtained. In addition, it focuses on the pitfalls one might encounter when developing similar assays. Despite the particular physicochemical characteristics of risperidone and 9-hydroxyrisperidone, the LC-MS-MS method enabled the quantification of both compounds down to 0.1 ng/ml. The method uses a sample preparation step by solid-phase extraction at pH 6 using a mixed-mode phase. In a short chromatographic run, separation of 9-hydroxyrisperidone from the minor metabolite 7-hydroxyrisperidone is achieved. Detection takes place by (turbo)ionspray tandem mass spectrometry in the positive ion mode. The validated concentration range is from 0.100 to 250 ng/ml, using 500 microliter of sample, with accuracy (bias) and precision (coefficient of variation) being below 15%. Although new developments in equipment will allow us to further improve and speed up the method, the assay reported can be used as a routine method to support a wide range of pharmacokinetic studies.
自20世纪90年代初利培酮首次上市以来，在必须具备生物分析方法的条件下，对该化合物药代动力学行为的研究受到了广泛关注。然而，由于液相色谱 - 串联质谱（LC-MS-MS）在生物分析实验室中已发展成为一种常规技术，大多数已发表的方法都未达到如今所能获得的灵敏度和选择性水平。因此，我们开发并验证了一种用于测定人血浆中利培酮及其活性代谢物9-羟基利培酮的新LC-MS-MS方法。本文详细描述了生物分析程序，并总结了获得的验证结果。此外，还重点讨论了在开发类似分析方法时可能遇到的陷阱。尽管利培酮和9-羟基利培酮具有特殊的物理化学特性，但该LC-MS-MS方法能够将两种化合物定量至低至0.1 ng/ml。该方法采用在pH 6下使用混合模式相进行固相萃取的样品制备步骤。在短时间的色谱运行中，可实现9-羟基利培酮与次要代谢物7-羟基利培酮的分离。通过（涡轮）离子喷雾串联质谱在正离子模式下进行检测。使用500微升样品时，验证的浓度范围为0.100至250 ng/ml，准确度（偏差）和精密度（变异系数）均低于15%。尽管设备的新发展将使我们能够进一步改进和加速该方法，但所报道的分析方法可作为一种常规方法，用于支持广泛的药代动力学研究。
