Gao Mingjie, Yang Jing, Wang Zhibin, Yang Bingyou, Kuang Haixue, Liu Lu, Wang Liqian, Yang Chunjuan
Department of Pharmaceutical Analysis and Analytical Chemistry, College of Pharmacy, Harbin Medical University, 157 Baojian Road, Nangang District, Harbin 150081, Heilongjang, China.
Analytical Department, Johnson & Johnson, 199 Grandview Road, Skillman, NJ 08558, USA.
Molecules. 2016 Jun 1;21(6):717. doi: 10.3390/molecules21060717.
A specific, simple, sensitive Ultra High Performance Liquid Chromatography-tandem Mass Spectrometry (UHPLC-MS/MS) method has been developed and validated for the simultaneous determination and pharmacokinetic study of purpurin, munjistin, and mollugin in rat plasma. Chromatographic separation was carried out using a C18 column (ACQUITY UPLC(®) HSS T3, 1.8 μm, 2.1 × 100 mm) with gradient elution. The compounds were detected on a 6430 triple-quadrupole tandem MS with an electrospray ionization (ESI) interface using multiple reaction monitoring (MRM) in positive ionization mode. The samples were prepared by a liquid-liquid extraction (LLE) method with ethyl acetate after being spiked with an internal standard (bifendate). The current UHPLC-MS/MS assay was validated for its linearity, intra-day and inter-day precisions, accuracy, extraction recovery, matrix effect and stability in different conditions. The method was linear for all analytes over the investigated range with all determined correlation coefficients exceeding 0.9900. The intra-day and inter-day precisions were in the range of 4.21% to 14.84%, and the relative errors of accuracies were in the range of -14.05% to 14.75%. The mean recoveries and matrix effects of purpurin, munjistin, and mollugin were higher than 78.87% and 92.56%, repectively. After oral administration of 0.82 g/kg of Rubia cordifolia extract, the maximum plasma concentrations (Cmax) were 70.10 ± 11.78 ng/mL for purpurin, 26.09 ± 6.6 ng/mL for munjistin, and 52.10 ± 6.71 ng/mL for mollugin. The time for maximal concentration (Tmax) was 1.61 ± 0.24 h for purpurin, 2.58 ± 0.19 h for munjistin, and 1.99 ± 0.21 h for mollugin. The established method was further applied to a pharmacokinetic study of purpurin, munjistin, and mollugin in rat plasma. It was concluded from the pharmacokinetic parameters that the three analytes showed a process of slow absorption and metabolism after oral administration of R. cordifolia extract to rats.
已开发并验证了一种特定、简单、灵敏的超高效液相色谱-串联质谱法（UHPLC-MS/MS），用于同时测定大鼠血浆中紫红素、蒙花苷和摩罗皂苷元，并进行药代动力学研究。使用C18柱（ACQUITY UPLC® HSS T3，1.8μm，2.1×100mm）进行梯度洗脱，实现色谱分离。在配备电喷雾电离（ESI）接口的6430三重四极杆串联质谱仪上，采用正离子模式下的多反应监测（MRM）对化合物进行检测。样品在加入内标（联苯双酯）后，通过乙酸乙酯液-液萃取（LLE）法制备。对当前的UHPLC-MS/MS测定法进行了线性、日内和日间精密度、准确度、萃取回收率、基质效应以及不同条件下稳定性的验证。在所研究的范围内，所有分析物的方法均呈线性，所有测定的相关系数均超过0.9900。日内和日间精密度在4.21%至14.84%范围内，准确度的相对误差在-14.05%至14.75%范围内。紫红素、蒙花苷和摩罗皂苷元的平均回收率和基质效应分别高于78.87%和92.56%。口服给予0.82 g/kg茜草提取物后，紫红素的最大血浆浓度（Cmax）为70.10±11.78 ng/mL，蒙花苷为26.09±6.6 ng/mL，摩罗皂苷元为52.10±6.71 ng/mL。紫红素的达峰时间（Tmax）为1.61±0.24 h，蒙花苷为2.58±0.19 h，摩罗皂苷元为1.99±0.21 h。所建立的方法进一步应用于大鼠血浆中紫红素、蒙花苷和摩罗皂苷元的药代动力学研究。从药代动力学参数得出结论，大鼠口服茜草提取物后，这三种分析物呈现出缓慢吸收和代谢的过程。
