Szabo G K, Pylilo R J, Perchalski R J, Browne T R
Department of Neurology, Boston University School of Medicine, MA 02118.
J Chromatogr. 1990 Dec 28;535(1-2):271-7. doi: 10.1016/s0021-9673(01)88952-4.
The use of stable isotope-labeled tracer compounds is the safest and most effective method to perform many steady state pharmacokinetic and drug interaction studies. We describe a method by which the heavily deuterated 2H10 analogues of carbamazepine (2H10 CBZ) and phenytoin (2H10 PHT) can be chromatographically separated by high-performance liquid chromatography from unlabeled CBZ and PHT. All compounds are quantitated against an internal standard (IS) (10,11-dihydrocarbamazepine) and measured using conventional UV detection rather than mass spectrometry. Baseline resolution of extracted serum containing 2H10 CBZ, CBZ, 2H10 PHT, PHT and IS is achieved on a heated (55 degrees C) 25 cm x 4.6 mm BioAnalytical Systems Phase II 5 microns ODS column with an isocratic mobile phase consisting of water-acetonitrile-tetrahydrofuran (80:16:4, v/v/v) at 1.2 ml/min. Eluting compounds were monitored at a UV wavelength of 214 nm. Calculated resolution of 2H10 CBZ from CBZ and of 2H10 PHT from PHT were 1.3. Serum standard curves were linear (R greater than or equal to 0.999) over a range of 0.5-14 micrograms/ml for 2H10 CBZ, 0.5-20 micrograms/ml for CBZ, 0.5-20 micrograms/ml for 2H10 PHT, and 0.5-30 micrograms/ml for PHT. Within-day percent relative standard deviations (precision) were less than 6% in all cases.
使用稳定同位素标记的示踪化合物是进行许多稳态药代动力学和药物相互作用研究的最安全、最有效的方法。我们描述了一种方法，通过该方法，卡马西平（2H10 CBZ）和苯妥英（2H10 PHT）的重氢代2H10类似物可以通过高效液相色谱从未标记的CBZ和PHT中进行色谱分离。所有化合物均以内标（IS）（10,11-二氢卡马西平）进行定量，并使用传统的紫外检测而非质谱进行测量。在加热（55摄氏度）的25厘米×4.6毫米BioAnalytical Systems II相5微米ODS柱上，使用由水-乙腈-四氢呋喃（80:16:4，v/v/v）组成的等度流动相，流速为1.2毫升/分钟，可实现含有2H10 CBZ、CBZ、2H10 PHT、PHT和IS的提取血清的基线分离。在214纳米的紫外波长下监测洗脱化合物。2H10 CBZ与CBZ以及2H10 PHT与PHT的计算分离度为1.3。血清标准曲线在2H10 CBZ为0.5 - 14微克/毫升、CBZ为0.5 - 20微克/毫升、2H10 PHT为0.5 - 20微克/毫升以及PHT为0.5 - 30微克/毫升的范围内呈线性（R大于或等于0.999）。在所有情况下，日内相对标准偏差（精密度）均小于6%。
