Chintiroglou Panagiota-Iro, Krigas Nikos, Chatzopoulou Paschalina, Karioti Anastasia
Laboratory of Pharmacognosy, School of Pharmacy, Aristotle University of Thessaloniki, University Campus, Thessaloniki, Greece.
Hellenic Agricultural Organization - DEMETER, Institute of Breeding and Plant Genetic Resources, Thermi, Thessaloniki, Greece.
Planta Med. 2021 Nov;87(14):1219-1230. doi: 10.1055/a-1578-8731. Epub 2021 Sep 2.
An HPLC-PDA method was developed for the determination of the flavonoids in the flowers of from Epirus, Greece. The aim was to investigate the chemical content of the over-harvested populations of Epirus and to develop and optimize an extraction protocol to allow fast, exhaustive, and repeatable extraction. Qualitative analysis revealed that the flowers from Epirus were particularly rich in flavonoids, especially flavonol triglycosides including derivatives of quercetin, isorhamnetin, and kaempferol. A phytochemical investigation of a 70% hydromethanolic extract from the flowers afforded a new flavonoid, namely, isorhamnetin-3---glucopyranosyl-(1 → 2)--glucopyranosyl-(1 → 6)--glucopyranoside, which is also the main constituent of the flower extracts. Its structure elucidation was carried out by means of 1D and 2D NMR and mass spectrometry analyses. The HPLC-PDA method was developed and validated according to the International Council for Harmonisation guidelines. Since the main flavonol glycoside of the plant is not commercially available, rutin was used as a secondary standard and the response correction factor was determined. Finally, the overall method was validated for precision (% relative standard deviation ranging between 1.58 and 4.85) and accuracy at three concentration levels. The recovery ranged between 93.5 and 102.1% with relative standard deviation values < 5%, within the acceptable limits. The developed assay is fast and simple and will allow for the quality control of the herbal drug.
建立了一种HPLC-PDA方法用于测定希腊伊庇鲁斯地区花朵中的黄酮类化合物。目的是研究伊庇鲁斯过度采摘种群的化学组成，并开发和优化一种提取方案，以实现快速、彻底和可重复的提取。定性分析表明，伊庇鲁斯的花朵富含黄酮类化合物，尤其是黄酮醇三糖苷，包括槲皮素、异鼠李素和山奈酚的衍生物。对花朵70%的甲醇提取物进行植物化学研究，得到一种新的黄酮类化合物，即异鼠李素-3- -吡喃葡萄糖基-(1→2)- -吡喃葡萄糖基-(1→6)- -葡萄糖苷，它也是花朵提取物的主要成分。通过一维和二维核磁共振及质谱分析对其结构进行了鉴定。根据国际协调理事会指南开发并验证了HPLC-PDA方法。由于该植物的主要黄酮醇苷没有商业供应，因此使用芦丁作为二级标准品并测定了响应校正因子。最后，在三个浓度水平上对整个方法的精密度（相对标准偏差在1.58%至4.85%之间）和准确度进行了验证。回收率在93.5%至102.1%之间，相对标准偏差值<5%，在可接受范围内。所开发的分析方法快速简便，可用于草药的质量控制。
