Laboratory of Clinical and Analytical Chemistry, College of Pharmaceutical Sciences, Ritsumeikan University, 1-1-1 Nojihigashi, Kusatsu, Shiga, 525-8577, Japan.
Department of Pharmacy, Japanese Red Cross Kyoto Daiichi Hospital, 15-749 Honmachi, Higashiyama-Ku, Kyoto, Kyoto, 605-0981, Japan.
Anal Sci. 2023 Apr;39(4):447-454. doi: 10.1007/s44211-023-00266-z. Epub 2023 Jan 13.
A single reference high-performance liquid chromatographic (SR-HPLC) method was developed and validated for the therapeutic drug monitoring (TDM) of phenytoin (PHT) and carbamazepine (CBZ) in plasma from patients. The analytical parameters evaluated were linearity, limit of quantification (LOQ), selectivity, accuracy, and stability according to the US Food and Drug Administration (FDA) guideline. The developed method shows good linearity (r2 > 0.999; LOQ-50 µg/mL), and LOQ values were 1.56 µg/mL for PHT and 0.40 µg/mL for CBZ at 254 nm. For the development of SR-HPLC method, we evaluated to improve the detection wavelength, stirred retention time, and stability for SR, and selected 5-(p-methylphenyl)-5-phenylhydantoin for PHT (relative molar sensitivity, RMS = 0.848) and 10-methoxyiminostilbene for CBZ (RMS = 0.263). The established differential definite quantities of PHT and CBZ in plasma samples were similar using the RMS and absolute calibration methods based on RSD < 5.10%. A preliminary application was performed using chemiluminescent immunoassay and SR-HPLC method, in which the detectable values of the correlation coefficient and the slope of the intercept were PHT: 0.964 and 0.992647, and CBZ: 0.969 and 1.072089, respectively. Based on these results, we propose that the SR-HPLC method with RMS would be offered to the useful and accurate TDM of various medicines in plasma/serum samples.
建立并验证了一种用于患者血浆中苯妥英（PHT）和卡马西平（CBZ）治疗药物监测（TDM）的单参考高效液相色谱法（SR-HPLC）。根据美国食品和药物管理局（FDA）的指南，评估了分析参数的线性、定量下限（LOQ）、选择性、准确性和稳定性。所开发的方法具有良好的线性（r2>0.999；LOQ-50μg/mL），在 254nm 处 PHT 的 LOQ 值为 1.56μg/mL，CBZ 的 LOQ 值为 0.40μg/mL。为了开发 SR-HPLC 方法，我们评估了改进检测波长、搅拌保留时间和 SR 的稳定性，并选择 5-（对甲基苯基）-5-苯基海因（相对摩尔灵敏度，RMS=0.848）作为 PHT 的检测物质，10-甲氧基亚氨基苯乙烯（RMS=0.263）作为 CBZ 的检测物质。基于 RSD<5.10%，使用 RMS 和绝对校准方法建立的血浆样品中 PHT 和 CBZ 的定量差异相似。初步应用采用化学发光免疫分析法和 SR-HPLC 法，相关系数和截距斜率的可检测值分别为 PHT：0.964 和 0.992647，CBZ：0.969 和 1.072089。基于这些结果，我们提出 RMS 的 SR-HPLC 方法将为各种血浆/血清样品中的药物提供有用和准确的 TDM。
