Ruian People's Hospital, The Third Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China.
Laboratory Animal Centre, Wenzhou Medical University, Wenzhou 325035, China.
Biomed Res Int. 2019 Jan 6;2019:6262105. doi: 10.1155/2019/6262105. eCollection 2019.
Lappaconitine is extracted from Nakai, which belongs to the Ranunculaceae. Lappaconitine is as a diterpenoid alkaloid used as a nonaddictive analgesic. To assure the rational use of the drug, ultrahigh-pressure liquid chromatography tandem mass spectrometry (UPLC-MS/MS) was conducted to determine lappaconitine in mouse blood and its application to pharmacokinetics. In this study, khasianine was used as internet standard (IS). A UPLC BEH C18 column was used for chromatographic separation and the mobile phase consisted of acetonitrile and 10 mmol/L ammonium acetate (0.1% formic acid). The flow rate of was 0.4 mL/min. Quantitative detection was performed in a multiple reaction monitoring (MRM) mode using an electrospray ionization source in positive mode. Twenty-four mice were randomly divided into four groups, three of which received 2, 4, and 8 mg/kg lappaconitine by intragastric administration, while the other group received 1 mg/kg lappaconitine by intravenous administration. After 0.0833, 0.5, 1, 1.5, 2, 3, 4, and 8 h, blood samples were collected and acetonitrile was used for protein precipitation. A linear calibration relationship (R = 0.9979) in the range of 0.1-500 ng/mL in mouse blood indicated good results. The lower limit of quantitation was 0.1 ng/mL and the limit of detection was 0.04 ng/mL. The intra-day and inter-day precision were below 13% and 14%, respectively. The accuracy was 90.1-107.2%, and the recovery exceeded 81.1%. The matrix effect ranged between 102.1 and 108.8%. The absolute bioavailability of lappaconitine was 2.0%. UPLC-MS/MS achieved high sensitivity, speed, and selectivity. Methodological verification indicated this method as suitable for determination of lappaconitine in mouse blood.
高乌甲素是从乌头属植物中提取的一种二萜类生物碱，用作非成瘾性镇痛药。为了确保药物的合理使用，采用超高效液相色谱串联质谱法（UPLC-MS/MS）测定小鼠血液中的高乌甲素及其药代动力学应用。在本研究中，以khasianine 作为内标（IS）。采用 UPLC BEH C18 色谱柱进行色谱分离，流动相由乙腈和 10 mmol/L 乙酸铵（0.1%甲酸）组成。流速为 0.4 mL/min。采用电喷雾电离源正模式下的多重反应监测（MRM）模式进行定量检测。将 24 只小鼠随机分为四组，其中三组分别灌胃给予 2、4 和 8 mg/kg 高乌甲素，另一组静脉给予 1 mg/kg 高乌甲素。在 0.0833、0.5、1、1.5、2、3、4 和 8 h 后采集血样，用乙腈进行蛋白沉淀。在小鼠血液中 0.1-500 ng/mL 范围内呈良好的线性关系（R = 0.9979）。定量下限为 0.1 ng/mL，检测限为 0.04 ng/mL。日内和日间精密度均低于 13%和 14%。准确度为 90.1-107.2%，回收率超过 81.1%。基质效应在 102.1-108.8%之间。高乌甲素的绝对生物利用度为 2.0%。UPLC-MS/MS 具有高灵敏度、速度和选择性。方法学验证表明，该方法适用于测定小鼠血液中的高乌甲素。
