口服大蓟提取物后大鼠血浆中七种黄酮类化合物的LC-MS/MS测定及药代动力学研究。

LC-MS/MS determination and pharmacokinetic study of seven flavonoids in rat plasma after oral administration of Cirsium japonicum DC. extract.

作者信息

Zhang Zhiyong, Jia Peipei, Zhang Xiaoxu, Zhang Qiaoyue, Yang Haotian, Shi He, Zhang Lantong

机构信息

Department of Pharmaceutical Analysis, School of Pharmacy, Hebei Medical University, Shijiazhuang, 050017, PR China.

出版信息

J Ethnopharmacol. 2014 Dec 2;158 Pt A:66-75. doi: 10.1016/j.jep.2014.10.022. Epub 2014 Oct 23.

DOI:10.1016/j.jep.2014.10.022

PMID:25456423

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE

Cirsium japonicum DC., a Traditional Chinese Medicine, has the curative effect of antihemorrhagic and antitumor. Pharmacological studies prove that the curative effect may relate to the flavonoids. A simple and rapid resolution liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was first developed and validated for the quantification of seven flavonoids including pectolinarin, linarin, pectolinarigenin, hispidulin, diosmetin, acacetin and apigenin in rat plasma after oral administration of Cirsium japonicum DC. extract.

MATERIALS AND METHODS

The chromatographic separation was achieved on a C18 column with gradient elution by using a mixture of 0.1% formic acid aqueous solution and methanol as the mobile phase at a flow rate of 0.8mL/min. A tandem mass spectrometric detection was conducted using multiple-reaction monitoring (MRM) via an electrospray ionization (ESI) source in positive and negative ionization mode simultaneously. Samples were pre-treated by a single-step protein precipitation with methanol, and sulfamethoxazole was used as internal standard (IS).

RESULTS

The optimized mass transition ion-pairs (m/z) for quantization were 623.4/315.2 for pectolinarin, 593.3/285.1 for linarin, 315.3/300.2 for pectolinarigenin, 301.2/286.2 for hispidulin, 301.2/258.2 for diosmetin, 283.0/267.9 for acacetin, 269.0/117.0 for apigenin and 252.2/155.8 for IS. After oral administration of 6mL/kg Cirsium japonicum DC. extract in rats, the maximum plasma concentration (Cmax) of pectolinarin, linarin, pectolinarigenin, hispidulin, diosmetin, acacetin and apigenin were 876.77±97.34ng/mL, 86.79±1.70ng/mL, 6.13±0.12ng/mL, 32.85±2.50ng/mL, 37.2±2.04ng/mL, 19.02±1.29ng/mL and 148.26±20.63ng/mL, respectively. The time to reach the maximum plasma concentration (Tmax) was 5min for pectolinarin, linarin, pectolinarigenin, hispidulin, diosmetin, acacetin and 360min for apigenin. The intra-day and inter-day precisions (RSD%) for seven compounds were less than 13.16% and 7.77% and the accuracy (RE%) range from -7.92% to 14.77%.

CONCLUSIONS

This is the first research on the pharmacokinetic study of bioactive components in rat plasma after oral administration of Cirsium japonicum DC. extract. The results provided a meaningful basis for better understanding the absorption mechanism of Cirsium japonicum DC. and evaluating the clinical application of this herb medicine.

摘要

民族药理学相关性

中药大蓟具有止血和抗肿瘤的疗效。药理学研究证明，其疗效可能与黄酮类化合物有关。首次建立并验证了一种简单快速的液相色谱 - 串联质谱（LC-MS/MS）方法，用于定量大鼠口服大蓟提取物后血浆中的七种黄酮类化合物，包括果胶菌素、木犀草苷、果胶菌素元、圣草酚、香叶木素、刺槐素和芹菜素。

材料与方法

采用C18柱，以0.1%甲酸水溶液和甲醇的混合物为流动相进行梯度洗脱，流速为0.8mL/min，实现色谱分离。通过电喷雾电离（ESI）源，在正离子和负离子模式下同时采用多反应监测（MRM）进行串联质谱检测。样品采用甲醇一步蛋白沉淀法进行预处理，磺胺甲恶唑用作内标（IS）。

结果

用于定量的优化质量转移离子对（m/z）分别为：果胶菌素623.4/315.2，木犀草苷593.3/285.1，果胶菌素元315.3/300.2，圣草酚301.2/286.2，香叶木素301.2/258.2，刺槐素283.0/267.9，芹菜素269.0/117.0，内标252.2/155.8。大鼠口服6mL/kg大蓟提取物后，果胶菌素、木犀草苷、果胶菌素元、圣草酚、香叶木素、刺槐素和芹菜素的最大血浆浓度（Cmax）分别为876.77±97.34ng/mL、86.79±1.70ng/mL、6.13±0.12ng/mL、32.85±2.50ng/mL、37.2±2.04ng/mL、19.02±1.29ng/mL和148.26±20.63ng/mL。达到最大血浆浓度的时间（Tmax）：果胶菌素、木犀草苷、果胶菌素元、圣草酚、香叶木素、刺槐素为5min，芹菜素为360min。七种化合物的日内和日间精密度（RSD%）均小于13.16%和7.77%，准确度（RE%）范围为-7.92%至14.77%。