Korst A E, Vermorken J B, van der Vijgh W J
Department of Medical Oncology, University Hospital Vrije Universiteit, Amsterdam, Netherlands.
J Chromatogr B Biomed Sci Appl. 1997 May 23;693(1):167-74. doi: 10.1016/s0378-4347(97)00016-9.
A high-performance liquid chromatographic (HPLC) method was developed for the combined analysis of the chemoprotective agent, amifostine, its active metabolite, WR 1065, and the (symmetrical and mixed) disulfides of WR 1065 in plasma. These three compounds were quantified by measuring WR 1065 after three different sample pretreatment procedures. During these procedures, amifostine and the disulfides were quantitatively converted into WR 1065, by incubating the sample either at a low pH or in the presence of dithiothreitol, respectively. The resulting amounts of WR 1065 were determined by HPLC with electrochemical detection (Au electrode, + 1.00 V). The lower limit of quantitation of WR 1065 was 0.15 microM. The within-day and between-day precision were < or =4.4 and < or =8.2% for WR 1065, < or =4.9 and < or =13.1% for amifostine and < or =8.5 and < or =5.5% for the disulfides, respectively. The within-day and between-day accuracy ranged from 97.2 to 109.8% and from 97.6 to 101.5% for WR 1065, from 88.3 to 110.7% and from 99.4 to 101.5% for amifostine and from 99.2 to 110.2% and from 103.3 to 104.9% for the disulfides, respectively. This method is superior to other described methods due to its simple and relatively rapid analysis of all three compounds in one system. Furthermore, it is at least as sensitive as earlier reported methods for one of the compounds and the application of the gold electrode requires only minor maintenance. Therefore, this method is very suitable for pharmacokinetic studies of amifostine and its metabolites. As an example, the plasma concentrations of amifostine, WR 1065 and the disulfides are shown in a patient after receiving an i.v. dose of 740 mg/m2 amifostine.
建立了一种高效液相色谱（HPLC）方法，用于联合分析血浆中的化学保护剂氨磷汀、其活性代谢物WR 1065以及WR 1065的（对称和混合）二硫化物。通过三种不同的样品预处理程序测定WR 1065的量来对这三种化合物进行定量。在这些程序中，分别通过在低pH下孵育样品或将样品置于二硫苏糖醇存在的条件下，将氨磷汀和二硫化物定量转化为WR 1065。通过带有电化学检测（金电极，+1.00 V）的HPLC测定所得的WR 1065的量。WR 1065的定量下限为0.15 microM。WR 1065的日内和日间精密度分别≤4.4%和≤8.2%，氨磷汀分别≤4.9%和≤13.1%，二硫化物分别≤8.5%和≤5.5%。WR 1065的日内和日间准确度分别为97.2%至109.8%和97.6%至101.5%，氨磷汀分别为88.3%至110.7%和99.4%至101.5%，二硫化物分别为99.2%至110.2%和103.3%至104.9%。该方法优于其他已描述的方法，因为它能在一个系统中对所有三种化合物进行简单且相对快速的分析。此外，它对其中一种化合物的灵敏度至少与早期报道的方法相同，并且金电极的应用只需少量维护。因此，该方法非常适合氨磷汀及其代谢物的药代动力学研究。例如，给出了一名患者静脉注射740 mg/m²氨磷汀后血浆中氨磷汀、WR 1065和二硫化物的浓度。
