High-performance liquid chromatographic separation of penicillamine enantiomers labelled with N-[4-(6-dimethylamino-2-benzofuranyl)phenyl]maleimide on a chiral stationary phase

✍ Scribed by Kenichiro Nakashima; Tomoko Ishimaru; Naotaka Kuroda; Shuzo Akiyama

• Publisher: John Wiley and Sons
• Year: 1995
• Tongue: English
• Weight: 312 KB
• Volume: 9
• Category: Article
• ISSN: 0269-3879
• DOI: 10.1002/bmc.1130090207

✦ Synopsis

Penicillamine enantiomers derivatized with N-[4-(6-dimethylamino-2-benzofuranyl)phenyl]maleimide (DBPM) were separated and determined by high-performance liquid chromatography. A fluorogenic reagent, DBPM easily reacted with 1)-or L-penicillamine to give each two kinds of strong fluorescent derivatives (D,-, D2-, L,-and L,-DBPM), which could be separated on a Pirkle-type chiral stationary phase using an eluent of 75% aqueous methanol solution containing 0.15 M CH,COONH, and 0.05 M tetra-n-butylammonium bromide. Two of the peaks (D,-or L,-DBPM), having a shorter retention time than the others, bad almost the same retention times (25 min for D,-DBPM and 25.7 min for L,-DBPM). The retention times of the peaks eluted later were 28 min and 31.6 min for D2-and L,-DBPM respectively. Linear calibration curves over the range of 2-50 pmol per injection were obtained for D-and L-penicillamines with a detection limit of 290 and 350 fmol at respectively at a signal-tonoise ratio of 3. Using the proposed method, the absence of contamination of i,-penicillamine in a commercially available wpenicillamine preparation (capsule) was confirmed.