Tang P H, Miles M V, Glauser T A, DeGrauw T
Department of Pediatric Neurology, The Children's Hospital Medical Center, Cincinnati, OH 45229-3039, USA.
J Chromatogr B Biomed Sci Appl. 1999 Apr 30;727(1-2):125-9. doi: 10.1016/s0378-4347(99)00077-8.
An automated high-performance liquid chromatographic method for the determination of gabapentin, 1-(amino-methyl)cyclohexaneacetic acid, in serum is described. The procedure involves protein precipitation with methanol followed by using a robotized derivatization with o-phthaldialdehyde reagent and automated high-performance liquid chromatography. The analog of gabapentin, 1-(aminomethyl)cycloheptaneacetic acid, was used as the internal standard. Blank serum was fortified with gabapentin (0.1-10.0 microg/ml) and internal standard. Separation was achieved on a Waters 5-microm reversed-phase column (10 cmx4.6 mm) with mobile phase consisting of 0.02 M phosphate buffer (pH 4.5)-acetonitrile (50:50, v/v). Eluents were monitored by fluorescence spectroscopy with excitation and emission wavelengths of 230 and 420 nm, respectively. The calibration curve for gabapentin in serum was linear (r=0.999) over the concentration range 0.1-10.0 microg/ml. The inter- and intraassay variations for three different gabapentin concentrations were < or =10% throughout. The lower limit of quantitation was found to be 0.1 microg/ml. Chromatography was unaffected by a range of commonly employed antiepileptic drugs or selected amino acids.
本文描述了一种用于测定血清中加巴喷丁（1-（氨基甲基）环己烷乙酸）的自动化高效液相色谱法。该方法包括用甲醇进行蛋白沉淀，随后使用邻苯二甲醛试剂进行自动化衍生化以及自动化高效液相色谱分析。加巴喷丁的类似物1-（氨基甲基）环庚烷乙酸用作内标。向空白血清中加入加巴喷丁（0.1 - 10.0微克/毫升）和内标。在Waters 5微米反相柱（10厘米×4.6毫米）上进行分离，流动相由0.02 M磷酸盐缓冲液（pH 4.5） - 乙腈（50:50，v/v）组成。洗脱液通过荧光光谱法进行监测，激发波长和发射波长分别为230和420纳米。血清中加巴喷丁的校准曲线在0.1 - 10.0微克/毫升的浓度范围内呈线性（r = 0.999）。三种不同加巴喷丁浓度的批间和批内变异始终≤10%。定量下限为0.1微克/毫升。色谱分析不受一系列常用抗癫痫药物或选定氨基酸的影响。
