Gao C X, Krull I S
Department of Chemistry, Northeastern University, Boston, MA 02115.
J Pharm Biomed Anal. 1989;7(10):1183-98. doi: 10.1016/0731-7085(89)80054-8.
A new analytical approach has been developed for the determination of d,l-amphetamine in urine using on-line solid phase derivatizations in HPLC-UV/FL. Several other enantiomeric drugs were also investigated using the same method. The method was validated by several experiments, including: (1) kinetic studies for the reaction of each enantiomeric drug with the solid phase chiral reagent; (2) "single blind spiking" experiments; and (3) polarimetry for confirmation of the enantiomeric composition determined by the solid phase diastereomer formation-HPLC-UV/FL method. The resulting diastereomers were well resolved (Rs = 1.05-1.40) under typical reversed-phase HPLC conditions. Enantiomeric contamination at the 1.1% level could be detected. The lowest amount of d,l-amphetamine that could be simultaneously derivatized and detected was about 50 ng ml-1. The linearity of the overall measurement was 3-4 orders of magnitude. d,l-Amphetamine spiked into urine at different concentration levels and different d,l-ratios, only followed by filtration, were directly injected into the on-line solid phase derivatization-HPLC-UV/FL system for quantitation with relative standard deviations 1.8-6.4% and relative errors 0.6-9.8%.
已开发出一种新的分析方法，用于在高效液相色谱 - 紫外/荧光检测中采用在线固相衍生化法测定尿液中的d，l - 苯丙胺。还使用相同方法研究了其他几种对映体药物。该方法通过多项实验进行了验证，包括：（1）每种对映体药物与固相手性试剂反应的动力学研究；（2）“单盲加样”实验；（3）旋光测定法，以确认通过固相非对映体形成 - 高效液相色谱 - 紫外/荧光法测定的对映体组成。在典型的反相高效液相色谱条件下，生成的非对映体得到了很好的分离（分辨率Rs = 1.05 - 1.40）。可以检测到1.1%水平的对映体污染。能够同时进行衍生化和检测的d，l - 苯丙胺的最低量约为50 ng/ml。整体测量的线性范围为3 - 4个数量级。以不同浓度水平和不同d，l - 比例添加到尿液中的d，l - 苯丙胺，仅经过过滤后，直接注入在线固相衍生化 - 高效液相色谱 - 紫外/荧光系统进行定量，相对标准偏差为1.8 - 6.4%，相对误差为0.6 - 9.8%。
