Simultaneous high-performance liquid chromatographic analysis of pregabalin, gabapentin and vigabatrin in human serum by precolumn derivatization with o-phtaldialdehyde and fluorescence detection.
A rapid, simple and robust method is presented for the simultaneous determination of the gamma-amino-n-butyric acid (GABA) derivatives pregabalin (PGB), gabapentin (GBP) and vigabatrin (VGB) in human serum by high-performance liquid chromatography (HPLC). Serum is deproteinized with trichloroacetic acid and aliquots of the supernatant are precolumn derivatized with o-phtaldialdehyde (OPA) and 3-mercaptopropionic acid. Separation is achieved on a Alltima 3C18 column using isocratic elution; the drugs are monitored using fluorescence detection. Norvaline is used as an internal standard. Within-day precision (COV; n = 10) is 1.2% for PGB (serum concentration 10.0 mg/l), 1.1% for GBP (serum concentration 15.8 mg/l) and 0.3% for VGB (serum concentration 15.5 mg/l). The method is linear up to at least 63 mg/l for PGB, 40 mg/l for GBP and 62 mg/l for VGB. Lower limits of quantitation (LOQ) are 0.13 mg/l for PGB, 0.53 mg/l for GBP and 0.06 mg/l for VGB. No interferences were found from commonly coadministered antiepileptic drugs (AEDs) and from endogenous amino acids. Experimental design in combination with statistical evaluation (ANOVA) was used to study the robustness of chromatography and sample preparation. The method is very suitable for routine therapeutic drug monitoring and for pharmacokinetic studies.
Vermeij
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Laboratory of Clinical Chemistry and Clinical Pharmacology, Epilepsy Institute of the Netherlands, SEIN, P.O. Box 540, 2130 AM Hoofddorp, Heemstede, The Netherlands.
Edelbroek
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D000588:Amines / Q000097:blood*; D002851:Chromatography, High Pressure Liquid; D003509:Cyclohexanecarboxylic Acids / Q000097:blood*; D018691:Excitatory Amino Acid Antagonists / Q000097:blood*; D005456:Fluorescent Dyes; D000077206:Gabapentin; D006801:Humans; D007202:Indicators and Reagents; D000069583:Pregabalin; D012015:Reference Standards; D015203:Reproducibility of Results; D012996:Solutions; D013048:Specimen Handling; D013050:Spectrometry, Fluorescence; D020888:Vigabatrin / Q000097:blood*; D005680:gamma-Aminobutyric Acid / Q000031:analogs & derivatives* / Q000097:blood*; D009764:o-Phthalaldehyde
D000588:Amines; D003509:Cyclohexanecarboxylic Acids; D018691:Excitatory Amino Acid Antagonists; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D012996:Solutions; D000069583:Pregabalin; D005680:gamma-Aminobutyric Acid; D009764:o-Phthalaldehyde; D000077206:Gabapentin; D020888:Vigabatrin