Second Clinical Medical College of Guangzhou University of Chinese Medicine, Guangdong Provincial Hospital of Chinese Medicine Affiliated to Guangzhou University of Chinese Medicine, 111 Dade Road, Guangzhou 510120, China.
Second Clinical Medical College of Guangzhou University of Chinese Medicine, Guangdong Provincial Hospital of Chinese Medicine Affiliated to Guangzhou University of Chinese Medicine, 111 Dade Road, Guangzhou 510120, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2019 Aug 1;1122-1123:18-28. doi: 10.1016/j.jchromb.2019.05.019. Epub 2019 May 17.
A simple, accurate, and reliable liquid chromatography-mass spectrometry (LC-MS/MS) method was developed and validated for the determination of mangiferin in rat plasma and tissue homogenates using rutin as an internal standard (IS). Chromatographic separation was achieved on a Shiseido CAPCELL PAK C column (150 × 2.0 mm, 5 μm) using a gradient elution of 1% acetic acid in water and methanol at a flow rate of 300 μL·min. Quantification was performed on an API 4000 triple-quadrupole mass spectrometer equipped with a turbo electrospray ionization (ESI) source in positive ion multiple reaction monitoring (MRM) mode. Selected ion monitoring transitions of 423.1 → 273.1 and 611.4 → 303.3 were chosen to quantify mangiferin and IS. Biological samples were pretreated via protein precipitation with acetonitrile-acetic acid. The standard calibration curves were above the ranges of 2 to 500 ng·mL and 5 to 2000 ng·mL for tissues, and 1 to 600 ng·mL for plasma. All calibration curves for tissue and plasma samples showed good linearity (r ≥ 0.9974) over the concentration ranges. Intra- and inter-day precisions were <14.0%, and accuracy ranged from 97.2% to 111.7%. The established method was successfully applied on mangiferin tissue distribution following the intragastric administration of mangiferin, Rhizoma Anemarrhenae (A. Rhizoma) decoction, or Rhizoma Anemarrhenae-Phellodendron (herb pair, HP) decoction under healthy or diabetic conditions. Mangiferin was detected from all the tested tissues (except for brain) after monomer administration, and the concentrations were lower compared with the decoction groups. Distributions in the HP group were lower than those in the A. Rhizoma group, but mangiferin content in pancreas was obviously higher than in other tissues and in the A. Rhizoma group. Compared with healthy rats, mangiferin distributions in pancreas and intestine were lower in diabetic rats administered with the same dose of the herb pair, but still higher than those in other tissues. In addition, distributions in liver, spleen, lung, kidney, stomach, and plasma were higher than those in the normal group.
建立并验证了一种简单、准确、可靠的液相色谱-串联质谱（LC-MS/MS）方法，用于大鼠血浆和组织匀浆中芒果苷的测定，以芦丁为内标（IS）。色谱分离在 Shiseido CAPCELL PAK C 柱（150×2.0mm，5μm）上进行，采用梯度洗脱，水相为 1%乙酸，甲醇为流动相，流速为 300μL·min。定量分析采用 API 4000 三重四极杆质谱仪，配有涡轮电喷雾电离（ESI）源，正离子多反应监测（MRM）模式。选择离子监测转换为 423.1→273.1 和 611.4→303.3，用于定量测定芒果苷和 IS。生物样品经乙腈-乙酸沉淀预处理。组织的标准校准曲线范围为 2 至 500ng·mL和 5 至 2000ng·mL，血浆的标准校准曲线范围为 1 至 600ng·mL。组织和血浆样品的所有校准曲线在浓度范围内均具有良好的线性（r≥0.9974）。日内和日间精密度均<14.0%，准确度在 97.2%至 111.7%之间。该方法成功应用于芒果苷经灌胃给药、知母（A. Rhizoma）汤或知母-黄柏（药对，HP）汤后在健康或糖尿病条件下的组织分布。单体给药后，所有检测组织（脑除外）均检测到芒果苷，其浓度低于汤剂组。HP 组的分布低于 A. Rhizoma 组，但胰腺中的芒果苷含量明显高于其他组织和 A. Rhizoma 组。与健康大鼠相比，糖尿病大鼠给予相同剂量的药对后，胰腺和肠道中芒果苷的分布较低，但仍高于其他组织。此外，肝脏、脾脏、肺、肾脏、胃和血浆中的分布高于正常组。
