Determination of Gastrodin and Ligustrazine Hydrochloride in Plasma and Brain Dialysate by LC–Tandem MS

Abstract

A gradient liquid chromatography-tandem mass spectrometry method has been developed and validated for the determination of gastrodin and ligustrazine hydrochloride in rat plasma and brain dialysates. Zolpidem was used as internal standard. For plasma samples, solid-phase extraction was used and the brain dialysates were collected from freely moving rats using brain microdialysis. Both were followed by HPLC separation and positive electrospray ionization tandem mass spectrometry detection (ESI–MS–MS). Chromatographic separation was achieved on a Symmetry RP-18 column using gradient elution with methanol and water containing 0.5% formic acid and 2 mM ammonium formate. Selected reaction monitoring (SRM) mode was used for quantitation. Good linearities were obtained in the range of 0.05–100 and 0.01–50 μg mL⁻¹ for gastrodin and ligustrazine hydrochloride in rat plasma, and 0.05–1,000 ng mL⁻¹ for both in dialysate. The lower limit of quantitation was 0.01 ng mL⁻¹ for gastrodin and 0.05 ng mL⁻¹ for ligustrazine. The method is precise and reliable and can be applied to pharmacokinetic studies.