Department of Pharmaceutical Sciences, University of Ferrara, via Fossato di Mortara 19, 44121 Ferrara, Italy.
J Chromatogr B Analyt Technol Biomed Life Sci. 2012 Jul 15;901:72-8. doi: 10.1016/j.jchromb.2012.06.001. Epub 2012 Jun 12.
A fast HPLC-ESI-MS/MS method has been developed and validated for the quantification of the potent and selective antimigraine zolmitriptan in rat blood and cerebrospinal fluid (CSF). The assay has been then applied for in vivo preclinical studies. The analytical determination has been used to obtain pharmacokinetics of zolmitriptan in the two biological matrices after its intravenous or nasal administration. Liquid-liquid extraction of zolmitriptan was performed from 100 μL rat blood samples in the presence of N(6)-cyclopentyladenosine (internal standard) with the employment of ethyl acetate. Calibration standards were prepared by using blood matrix and following the same liquid-liquid extraction procedure. CSF samples were analyzed without any pre-treatment steps and by using an external calibration method in pure water matrix. Chromatographic separation was performed under reversed phase and a gradient elution condition on a C18 packed column (100 × 2.0 mm, 2.5 μm particles diameter). The mobile phase was a mixture between acetonitrile, water and formic acid (0.1% v/v). The applied HPLC-MS/MS method allowed low limits of detection, as calculated from calibration curves, of 6.6 and 24.4 ng/mL for water matrix and rat blood extracts, respectively. Linearity of the calibration curves was established up to 5 μM (1.44 μg/mL), as well as good assay accuracy. The intravenous infusion of 20 μg zolmitriptan to male Sprague-Dawley rats produced blood concentrations ranging from 9.4±0.7 to 1.24±0.07 μg/mL within 10 h, with a terminal half-life of 3.4±0.2h. The nasal administration of a water suspension of 20 μg zolmitriptan produced blood concentrations ranging from 2.92±0.21 to 0.85±0.07 μg/mL within 6h. One hour after zolmitriptan intravenous infusion or nasal administration, its CSF concentrations were 0.0539±0.0016 and 0.0453±0.0012 μg/mL, respectively. This study determined the suitability of the herein proposed method to investigate the pharmacokinetics of zolmitriptan after its administration by means of novel formulations and, hence, to evaluate the efficacy of innovative nose-to-brain drug delivery in preclinical studies.
一种快速高效液相色谱-电喷雾串联质谱（HPLC-ESI-MS/MS）法已被开发并验证用于定量检测强效且选择性的偏头痛治疗药物佐米曲普坦（zolmitriptan）在大鼠血液和脑脊液（CSF）中的浓度。该分析方法随后被应用于体内临床前研究。该分析方法用于在静脉或鼻内给予佐米曲普坦后，从大鼠血液和脑脊液样本中获得佐米曲普坦的药代动力学数据。使用乙腈从 100 μL 大鼠血样中提取佐米曲普坦，同时加入 N(6)-环戊基腺苷（内标），并用乙酸乙酯萃取。使用血液基质并遵循相同的液液萃取程序制备校准标准。CSF 样品无需预处理，在纯水基质中采用外部校准方法进行分析。在反相条件下，通过梯度洗脱在 C18 填充柱（100×2.0mm，2.5μm 粒径）上进行色谱分离。流动相为乙腈、水和甲酸（0.1%v/v）的混合物。应用的 HPLC-MS/MS 方法允许从校准曲线计算出低检测限，水基质和大鼠血液提取物的检测限分别为 6.6 和 24.4ng/mL。校准曲线的线性范围为 5μM（1.44μg/mL），并且具有良好的分析准确性。雄性 Sprague-Dawley 大鼠静脉输注 20μg 佐米曲普坦后，10 小时内血液浓度范围为 9.4±0.7 至 1.24±0.07μg/mL，半衰期为 3.4±0.2h。鼻内给予 20μg 佐米曲普坦水混悬液后，6 小时内血液浓度范围为 2.92±0.21 至 0.85±0.07μg/mL。静脉输注佐米曲普坦或鼻内给予佐米曲普坦 1 小时后，CSF 浓度分别为 0.0539±0.0016 和 0.0453±0.0012μg/mL。本研究确定了所提出方法用于通过新型制剂研究佐米曲普坦给药后的药代动力学的适用性，从而评估创新的鼻内递药在临床前研究中的疗效。
