Refaat I H
Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy, Assiut University, Assiut, Egypt.
Boll Chim Farm. 2002 Jan-Feb;141(1):57-62.
A simple high-performance liquid chromatographic method was developed and validated for the determination of three alpha-amino acids (glycine, lysine, cysteine) and the amino acid derivative (N-acetylhomocysteine thiolactone) using aqueous mobile phase containing copper (II) ions. These amino acids were extracted from tablets, vials, effervescent salts and syrups and quantitatively diluted by the mobile phase. The separation was carried out on zorbax TMS column. The mobile phase was either 1.0 x 10(-3) M CuSO4 or 1.0 x 10(-3) M CuSO4 with 1.0% trimethylamine, pH 4.0. A linear relationship was obtained between detector response at 232 nm and the concentration of the studied amino acids in the range of 1.5-50 ng/microL. The intraday precision for 10-replicate samples varied from 0.59-2.66 while the interday precision ranged from 0.91-3.55 as CV%. A good recovery percentage was obtained in the analysis of many dosage forms ranging from 96.93 to 100.85 with a CV% from 0.33 to 1.74. The presented chromatographic separation method does not depend on any pretreatment or derivatization. The described chromatographic conditions, for both linearity and sensitivity, are sufficient for the selective determination of the cited amino acids. The interference from other combined pharmaceutical ingredients was resolved.
建立了一种简单的高效液相色谱法，用于测定三种α-氨基酸（甘氨酸、赖氨酸、半胱氨酸）和氨基酸衍生物（N-乙酰高半胱氨酸硫内酯），该方法使用含铜（II）离子的水性流动相。这些氨基酸从片剂、小瓶、泡腾盐和糖浆中提取，并用流动相进行定量稀释。分离在zorbax TMS柱上进行。流动相为1.0×10⁻³ M CuSO₄或含1.0%三甲胺、pH 4.0的1.0×10⁻³ M CuSO₄。在232 nm处检测器响应与所研究氨基酸浓度在1.5 - 50 ng/μL范围内呈线性关系。10次重复样品的日内精密度变异系数（CV%）在0.59 - 2.66之间，日间精密度变异系数范围为0.91 - 3.55。在分析多种剂型时获得了良好的回收率，范围为96.93%至100.85%，CV%为0.33%至1.74%。所提出的色谱分离方法不依赖于任何预处理或衍生化。所描述的色谱条件，无论是线性还是灵敏度，都足以选择性地测定所引用的氨基酸。解决了来自其他复方药物成分的干扰。
