Hassan Mohamed G, Emara Kamla M, Mohamed Horria A, Abdel-Wadood Hanaa M, Ikeda Rie, Wada Mitsuhiro, Kuroda Naotaka, Nakashima Kenichiro
Department of Pharmaceutical Analytical Chemistry, Faculty of Pharmacy, Al-Azhar University, Assiut Branch, Assiut 71524, Egypt.
Biomed Chromatogr. 2012 Feb;26(2):214-9. doi: 10.1002/bmc.1648. Epub 2011 May 19.
A sensitive high-performance liquid chromatographic method with fluorescence detection was developed to determine memantine (MT) in rat plasma. The method consists of pre-column labeling of MT with 4-(4,5-diphenyl-1H-imidazol-2-yl)benzoyl chloride (DIB-Cl) and a clean-up step with solid-phase extraction. A good separation of DIB-MT was achieved within 12 min on an octadecylsilica (ODS) column (150 × 4.6 mm i.d.; 5 µm) with a mobile phase of acetonitrile-water (70:30, v/v). The calibration curve prepared with fluoxetine as an internal standard showed good linearity in the range of 10-400 ng/mL (r = .999). The limits of detection and quantitation at signal-to-noise ratios of 3 and 10 were 2.0 and 6.6 ng/mL, respectively. The method was shown to be reliable with precisions of <5% for intra-day and <9% for inter-day as relative standard deviation. The fluorescence property and reaction yield of authentic DIB-MT were also examined. The proposed method was successfully applied to study the pharmacokinetic interaction between MT and methazolamide.
建立了一种采用荧光检测的灵敏高效液相色谱法测定大鼠血浆中的美金刚（MT）。该方法包括用4-（4,5-二苯基-1H-咪唑-2-基）苯甲酰氯（DIB-Cl）对MT进行柱前标记以及固相萃取净化步骤。在十八烷基硅胶（ODS）柱（150×4.6 mm内径；5 µm）上，以乙腈-水（70:30，v/v）为流动相，12分钟内实现了DIB-MT的良好分离。以氟西汀为内标制备的校准曲线在10-400 ng/mL范围内显示出良好的线性（r = 0.999）。信噪比为3和10时的检测限和定量限分别为2.0和6.6 ng/mL。该方法可靠，日内精密度<5%，日间精密度<9%（相对标准偏差）。还考察了真实DIB-MT的荧光性质和反应产率。所提出的方法成功应用于研究MT与甲醋唑胺之间的药代动力学相互作用。
