Indirect resolution of enantiomers of penicillamine by TLC and HPLC using Marfey's reagent and its variants.
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.
Bhushan
R
R
Department of Chemistry, Indian Institute of Technology Roorkee, Roorkee 247 667, India. rbushfcy@iitr.ernet.in
Brückner
H
H
Kumar
Virender
V
D000409:Alanine / Q000031:analogs & derivatives* / Q000737:chemistry; D018501:Antirheumatic Agents / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002855:Chromatography, Thin Layer / Q000379:methods*; D004136:Dinitrobenzenes / Q000737:chemistry*; D015394:Molecular Structure; D010396:Penicillamine / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D013237:Stereoisomerism
D018501:Antirheumatic Agents; D004136:Dinitrobenzenes; C047134:Marfey's reagent; D010396:Penicillamine; D000409:Alanine