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Fluorimetric determination of the enantiomers of vigabatrin, an antiepileptic drug, by reversed‐phase HPLC with a novel diastereomer derivatization reagent
Author(s) -
Uekusa Shusuke,
Onozato Mayu,
Sakamoto Tatsuya,
Umino Maho,
Ichiba Hideaki,
Fukushima Takeshi
Publication year - 2021
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.5060
Subject(s) - chemistry , chromatography , derivatization , enantiomer , diastereomer , reagent , high performance liquid chromatography , vigabatrin , solid phase extraction , stereochemistry , anticonvulsant , organic chemistry , neuroscience , epilepsy , biology
Herein, determination of an antiepileptic drug, (±)‐vigabatrin (VB), was performed by reversed‐phase HPLC with fluorimetric detection using a newly designed and synthesized fluorescence derivatization reagent, 2,5‐dioxopyrrolidin‐1‐yl (4‐{[(2‐nitrophenyl)sulfonyl]oxy}‐6‐(3‐oxomorpholino)quinoline‐2‐carbonyl)prolinate [Ns‐MOK‐( R )‐ or ( S )‐Pro‐OSu]. During the derivatization of VB with Ns‐MOK‐( R )‐Pro‐OSu at 60°C, the nosyl (Ns) group, which was introduced to protect a phenolic hydroxy group, was released within 30 min to produce MOK‐( R )‐Pro‐VB, which was detected fluorimetrically at 448 nm with an excitation wavelength of 333 nm. The VB enantiomers were separated on an octadecylsilica (ODS) column with a resolution value of 5.57, because Ns‐MOK‐( R )‐Pro‐OSu bears an optically active D‐proline structure. A complete separation of MOK‐( R )‐Pro‐( R )‐ and ‐( S )‐VB enantiomers was achieved on the ODS column within 40 min using stepwise gradient elution, and the detection limits were ~0.80 and 0.37 pmol on the column, respectively. The proposed HPLC with fluorimetric detection method was successfully used for determining VB enantiomers in VB‐spiked human serum following solid‐phase extraction with an anion‐exchange cartridge.