State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 210009, China.
Holdwarm Pharmaceutical China Co. Ltd., Nanjing 210008, China.
Chin J Nat Med. 2017 Sep;15(9):710-720. doi: 10.1016/S1875-5364(17)30101-2.
The present study was designed to develop and validate a rapid, sensitive, and reliable ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) method for the simultaneous determination of five major active constituents in the traditional Chinese medicinal preparation Xingxiong injection (XXI) in rat plasma, including quercetin 3-O-rutinoside (QCR), kaempferol 3-O-rutinoside (KFR), isorhamnetin 3-O-rutinoside (ISR), bilobalide (BB), and ligustrazine (LGT). The plasma samples were pretreated by protein precipitation with acetonitrile. The chromatographic separation was achieved on a Waters Symmetry C analytical column (2.1 mm × 100 mm, 3.5 μm) with a mobile phase of 0.1% aqueous formic acid (A)-acetonitrile (B). Quantitation of the five bioactive constituents was achieved. Naringin was used as the internal standard (IS). All the calibration curves showed good linearity (r > 0.996) over the concentration range, with the lowest limit of quantification (LLOQ) between 2-18 ng·mL. The intra- and inter-day accuracy and precision of the analytes were both within acceptable limits. Moreover, satisfactory extraction recoveries (90.92%-104.03%) were obtained by protein precipitation. The validated method was successfully applied to a pharmacokinetic study of XXI in rats after intravenous administration at three doses. The pharmacokinetic parameters of the five compounds varied in a dose-dependent manner within the tested dosage range. The present study was the first report of pharmacokinetic study for XXI.
本研究旨在建立并验证一种灵敏、可靠的超高效液相色谱-串联质谱（UHPLC-MS/MS）方法，用于同时测定传统中药制剂芎香注射液（XXI）在大鼠血浆中的 5 种主要活性成分，包括槲皮素 3-O-鼠李糖苷（QCR）、山柰酚 3-O-鼠李糖苷（KFR）、异鼠李素 3-O-鼠李糖苷（ISR）、银杏内酯 B（BB）和川芎嗪（LGT）。血浆样品经乙腈沉淀蛋白预处理。采用 Waters Symmetry C 分析柱（2.1mm×100mm，3.5μm），以 0.1%甲酸水溶液（A）-乙腈（B）为流动相进行色谱分离。对 5 种生物活性成分进行定量分析。以柚皮苷为内标（IS）。所有校准曲线的线性均良好（r＞0.996），最低定量限（LLOQ）在 2-18ng·mL 之间。日内和日间准确度和精密度均在可接受范围内。此外，通过蛋白沉淀可获得令人满意的提取回收率（90.92%-104.03%）。该方法已成功应用于静脉注射三种剂量后 XXI 在大鼠体内的药代动力学研究。五种化合物的药代动力学参数在测试剂量范围内呈剂量依赖性变化。本研究为首报 XX1 的药代动力学研究。
