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摘要
An analytical method was developed for the quantification in plasma of the R and S enantiomers of vigabatrin (VGB), a drug used for the treatment of some refractory pediatric epileptic syndromes. After adding 50 mu L of the internal standard, which consisted of a 15 mg/L solution of deuterated racemic VGB, and 100 mu L of water to 100 mu L of plasma samples, a protein precipitation was performed by adding 600 mu L of methanol. The supernatant was evaporated to dryness under a stream of nitrogen and the dry residue was reconstituted with 500 mu L of water. Then, 100 mu L of 0.01 M o-phthaldialdehyde and 0.01 M N-acetyl-L-cysteine in borate buffer (0.1 M, pH = 9.5) were added for pre-column derivatization of the enantiomers as diastereomeric isoindoles. One microliter of the resulting mixture was injected in the chromatographic system. The chromatographic separation was performed in gradient elution mode at a flow rate of 400 mu L/min using a phenomenex EVO C-18 column with a mobile phase composed of 5 mM ammonium acetate and a methanol:acetonitrile (63:37 v/v) mixture. Detection was performed by mass spectrometry In selected reaction monitoring mode using heated electrospray ionization in positive mode as the ion source. Intra- and inter-day precision and accuracy were lower than 15% over the calibration range (0.2-50 mg/L for each enantiomer) and the method was successfully used to assess plasma concentrations of VGB in epileptic children.
- 出版日期2017-12-1
- 单位中国地震局
