High-performance liquid chromatographic separation of penicillamine enantiomers labelled with N-[4-(6-dimethylamino-2-benzofuranyl)phenyl] maleimide on a chiral stationary phase.
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 D- or L-penicillamine to give each two kinds of strong fluorescent derivatives (D1-, D2-, L1- and L2-DBPM), which could be separated on a Pirkle-type chiral stationary phase using an eluent of 75% aqueous methanol solution containing 0.15 M CH3COONH4 and 0.05 M tetra-n-butylammonium bromide. Two of the peaks (D1- or L1-DBPM), having a shorter retention time than the others, had almost the same retention times (25 min for D1-DBPM and 25.7 min for L1-DBPM). The retention times of the peaks eluted later were 28 min and 31.6 min for D2- and L2-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-to-noise ratio of 3. Using the proposed method, the absence of contamination of L-penicillamine in a commercially available D-penicillamine preparation (capsule) was confirmed.
Nakashima
K
K
School of Pharmaceutical Sciences, Nagasaki University, Japan.
Ishimaru
T
T
Kuroda
N
N
Akiyama
S
S
D001572:Benzofurans / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D008301:Maleimides / Q000737:chemistry; D010396:Penicillamine / Q000737:chemistry / Q000302:isolation & purification*; D013050:Spectrometry, Fluorescence; D013237:Stereoisomerism; D013696:Temperature
D001572:Benzofurans; D005456:Fluorescent Dyes; D008301:Maleimides; C049623:N-(4-(2-(6-dimethylamino)benzofuranyl)phenyl)maleimide; D010396:Penicillamine