Bhushan R, Brückner H, Kumar Virender
Department of Chemistry, Indian Institute of Technology Roorkee, Roorkee 247 667, India.
Biomed Chromatogr. 2007 Oct;21(10):1064-8. doi: 10.1002/bmc.854.
TLC and HPLC methods were developed for indirect chiral separation of penicillamine (3,3-dimethylcysteine) enantiomers after derivatization with Marfey's reagent (FDNP-Ala-NH(2)) and two of its structural variants, FDNP-Phe-NH(2) and FDNP-Val-NH(2). The binary mobile phase of phenol-water (3:1 v/v) and solvent combinations of acetonitrile and triethylamine phosphate buffer were found to give the best separation in normal and reversed-phase TLC, respectively. The diastereomers were also resolved on a reversed-phase C18 HPLC column with gradient elution of acetonitrile and 0.01 m trifluoroacetic acid. The results due to these three reagents were compared. The method was successful for checking the enantiomeric impurity of l-penicillamine in d-penicillamine and to check the enantiomeric purity of pharmaceutical formulations of d-penicillamine. The method was validated for linearity, repeatability, limit of detection and limit of quantification.
采用薄层色谱法（TLC）和高效液相色谱法（HPLC），在青霉胺（3,3-二甲基半胱氨酸）对映体与马费试剂（FDNP-Ala-NH₂）及其两种结构变体FDNP-Phe-NH₂和FDNP-Val-NH₂衍生化后，对其进行间接手性分离。发现苯酚-水二元流动相（3:1 v/v）以及乙腈和磷酸三乙胺缓冲液的溶剂组合，分别在正相和反相TLC中实现了最佳分离。非对映异构体也在反相C18 HPLC柱上通过乙腈和0.01 m三氟乙酸的梯度洗脱得以分离。比较了这三种试剂的实验结果。该方法成功用于检测d-青霉胺中l-青霉胺的对映体杂质，并用于检测d-青霉胺药物制剂的对映体纯度。该方法针对线性、重复性、检测限和定量限进行了验证。
