固相萃取前处理HPLC-MS法测定口服给药后家兔血浆中的灯盏乙素

摘要

目的建立灵敏、专属的HPLC-ESI-MS测定法, 用于测定家兔口服灯盏花素缓释片(2片 60 mg/片)后的血药浓度.方法该定量方法采用芦丁作内标,用固相萃取(SPE)进行样品前处理.以甲醇10 mmol·L-1醋酸铵水溶液(pH 8.0)梯度洗脱,流动相流速0.4 mL·min-1,柱温35 ℃,经过Zorbax Extend-C18 (150 mm×2.1 mm ID, 5 μm)色谱柱分离,电喷雾质谱检测器在负离子模式下检测灯盏乙素和芦丁.结果在2～200 ng·mL-1范围内灯盏乙素与内标峰面积与浓度线性关系良好,且准确度、精密度满足生物样品分析的要求.结论本实验建立的兔血浆中灯盏乙素的HPLC-MS分析方法灵敏度高,可用于口服给药后灯盏乙素的血药浓度测定.%Aim To establish a sensitive and specific liquid chromatography-mass spectrometry method for determination of scutellarin in rabbit plasma after oral administration. Methods For the quantitative analysis, rutin was used as an internal standard and solid-phase extraction (SPE) was performed by using a Phenomenex C8 cartridge. HPLC was carried out using a Zorbax Extend-C18column (150 mm×2.1 mm ID, 5 μm) with a guard cartridge (Phenomenex). Gradient elution was selected with the mobile phase of methanol 10 mmol·L-1 ammonium acetate solution (pH adjusted to 8.0 with ammonia solution). The flow rate of mobile phase was 0.4 mL·min-1 and the column temperature was 35 ℃. Both scutellarin and the internal standard rutin in rabbit plasma extracts were detected by mass spectrometry using an ESI interface in the negative ion mode. Results The linear range was from 2 to 200 ng·mL-1, with acceptable accuracy and precision (RSD). Conclusion A sensitive, reliable and accurate method for the quantitation of scutellarin in rabbit plasma has been established.