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通过 UPLC-MS/MS 检测口服马兜铃提取物后大鼠组织中 8 种生物活性成分的分布。

Distributions of eight bioactive components in rat tissues administered Marsdenia tenacissima extract orally detected through UPLC-MS/MS.

机构信息

College of Pharmacy, Liaoning University of Traditional Chinese Medicine, Dalian, P. R. China.

College of Traditional Chinese Medicine, Macau University of Science and Technology, Macau, P. R. China.

出版信息

Biomed Chromatogr. 2021 Apr;35(4):e5034. doi: 10.1002/bmc.5034. Epub 2021 Jan 14.

Abstract

Marsdenia tenacissima (Roxb.) Wight et Arn. (M. tenacissima) is considered an anticancer medicine in traditional Chinese medicine, which is extensively used in clinical application since it has great therapeutic effects. Currently, although a number of articles have examined M. tenacissima in terms of its pharmacology and quality control, few have investigated the in vivo mechanism of M. tenacissima active ingredients. Previously, we have studied the pharmacokinetics of eight active ingredients after oral administration of M. tenacissima extracts in rat plasma. This study constructed a new scientific ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) approach to simultaneously quantify the contents of tenacissosides B, G, H and I, cryptochlorogenic acid, chlorogenic acid, neochlorogenic acid and caffeic acid in rats orally administered M. tenacissima extract. The proposed approach was successfully used for investigating the distributions of those eight analytes in rat tissues, with digoxin being used as an internal control. The Eclipse Plus C RRHD column was used for determination at a column temperature of 30°C. The mobile phase system consisted of acetonitrile and water (supplemented with 0.1% formic acid) under optimal gradient elution conditions. Afterwards, this approach was validated according to the requirements for the analysis of biological samples developed by the US Food and Drug Administration, including precision, accuracy, stability and matrix effects. Based on tissue distribution analysis, those eight analytes showed rapid distribution within all the tested tissues. With regard to organic acid distribution, it followed the order stomach > liver > kidney > small intestine > lung > spleen > heart, whereas the four steroids followed the order stomach > lung > spleen > small intestine > liver > kidney > heart. The present study lays the theoretical foundation for the use and development of M. tenacissima in clinical practice.

摘要

密花藤(Marsdenia tenacissima (Roxb.) Wight et Arn.)在传统中药中被认为是一种抗癌药物,由于其疗效显著,在临床应用中被广泛使用。目前,虽然有许多文章研究了密花藤的药理学和质量控制,但很少有研究调查密花藤活性成分的体内机制。之前,我们研究了密花藤提取物灌胃给药后大鼠血浆中八种活性成分的药代动力学。本研究构建了一种新的科学超高效液相色谱-串联质谱(UPLC-MS/MS)方法,用于同时定量测定大鼠口服密花藤提取物中十种有效成分的含量:纤细薯蓣皂苷 B、G、H 和 I、隐绿原酸、绿原酸、新绿原酸和咖啡酸。该方法成功地用于研究这八种分析物在大鼠组织中的分布,以地高辛为内标。采用 Eclipse Plus C RRHD 柱,柱温为 30°C。在最佳梯度洗脱条件下,流动相体系由乙腈和水(添加 0.1%甲酸)组成。之后,根据美国食品和药物管理局(US Food and Drug Administration)制定的生物样品分析要求,对该方法进行了验证,包括精密度、准确度、稳定性和基质效应。基于组织分布分析,这八种分析物在所有测试组织中均迅速分布。就有机酸分布而言,其顺序为胃>肝>肾>小肠>肺>脾>心,而四种甾体则为胃>肺>脾>小肠>肝>肾>心。本研究为密花藤在临床实践中的应用和开发奠定了理论基础。

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