Qiu Feng, Li Zhaoxia, He Lang, Wang Dan
Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, 100193, China.
Biomed Chromatogr. 2013 Mar;27(3):311-7. doi: 10.1002/bmc.2793. Epub 2012 Aug 2.
Luteoloside is a potential anticarcinogenic component isolated from Lonicera japonica, a traditional Chinese medicine (TCM). This study details the development and validation of a sensitive and accurate HPLC-ESI-MS/MS method for the quantification of luteoloside in dog plasma. Sample pretreatment includes simple protein precipitation using methanol-acetonitrile (1:1, v/v). A Phenomenex Gemini C(18) column (2.0 × 50 mm, i.d., 3.5 µm) was used to separate luteoloside and internal standard by gradient mode with mobile phase consisting of water containing 0.1% formic acid and methanol containing 0.1% formic acid at a flow rate of 0.40 mL/min with a column temperature of 25°C. The detection was performed by positive ion electrospray ionization (ESI) in multiple reaction monitoring mode. The calibration curves were linear (R > 0.995) over the concentration range 1.0-2000 ng/mL and the lower limit of quantification was 1.0 ng/mL. The intra-day and inter-day precisions (RSD) were all <15%, accuracies (RE) were within the range of ±15%, and recoveries were between 85.0 and 115%. The validated HPLC-ESI-MS/MS method was successfully applied to determine plasma concentrations of luteoloside after intravenous administration of luteoloside at a dose level of 20 mg/kg.
木犀草苷是从传统中药金银花中分离出的一种潜在抗癌成分。本研究详细阐述了一种灵敏且准确的HPLC-ESI-MS/MS方法的建立与验证，该方法用于定量犬血浆中的木犀草苷。样品预处理包括使用甲醇-乙腈（1:1，v/v）进行简单的蛋白沉淀。采用Phenomenex Gemini C(18)柱（2.0×50 mm，内径，3.5 µm），以含0.1%甲酸的水和含0.1%甲酸的甲醇为流动相，通过梯度模式分离木犀草苷和内标，流速为0.40 mL/min，柱温为25°C。检测采用正离子电喷雾电离（ESI）在多反应监测模式下进行。校准曲线在1.0 - 2000 ng/mL浓度范围内呈线性（R>0.995），定量下限为1.0 ng/mL。日内和日间精密度（RSD）均<15%，准确度（RE）在±15%范围内，回收率在85.0%至115%之间。经过验证的HPLC-ESI-MS/MS方法成功应用于以20 mg/kg剂量静脉注射木犀草苷后测定其血浆浓度。
