Independent Laboratory of Chemistry of Natural Products, Medical University of Lublin, 1 Chodzki St., 20-093 Lublin, Poland.
Department of Pharmacognosy, Medical University of Lublin, 1 Chodzki St., 20-093 Lublin, Poland.
Molecules. 2021 Mar 15;26(6):1626. doi: 10.3390/molecules26061626.
Alkaloids of the Lycopodiaceae family are of great interest to researchers due to their numerous properties and wide applications in medicine. They play a very important role mainly due to their potent antioxidant, antidepressant effects and a reversible ability to inhibit acetylcholinesterase (AChE) enzyme activity. This property is of immense importance due to the growing problem of an increasing number of patients with neurodegenerative diseases in developed countries and a lack of effective and efficient treatment for them. Numerous studies have shown that Lycopodiaceae alkaloids are a rich source of AChE inhibitors. In the obtaining of new therapeutic phytochemicals from plant material, the extraction process and its efficiency is crucial. Therefore, the aim of this work was to optimize the conditions of modern PLE to obtain bioactive alkaloids from two species: L. and L. Five different solvents of different polarity were used for prepared plant extracts in order to compare the alkaloid content in and thereby effectiveness of the entire extraction. PLE parameters were used based on multiple studies conducted that gave the highest alkaloids recovery. Crude extracts were purified using solid-phase extraction (SPE) on Oasis HLB cartridge and examined by HPLC/ESI-QTOF-MS of the highly abundant alkaloids. To the best of our knowledge, this is the first time such high recoveries have been obtained for known Lycopodiaceae alkaloids. The best extraction results of alkaloid-lycopodine were detected in the dichloromethane extract from , where the yield exceeded 45%. The high recovery of annotinine above 40% presented in was noticed in dichloromethane and ethyl acetate extracts. Moreover, chromatograms were obtained with all isolated alkaloids and the best separation and quality of the bands in methanolic extracts. Interestingly, no alkaloid amounts were detected in cyclohexane extracts belonging to the non-polar solvent. These results could be helpful for understanding and optimizing the best conditions for isolating potent AChE inhibitors.
石松科生物碱因其众多的特性和在医学上的广泛应用而引起了研究人员的极大兴趣。它们主要由于其强大的抗氧化、抗抑郁作用和可逆抑制乙酰胆碱酯酶（AChE）活性的能力而发挥着非常重要的作用。由于发达国家越来越多的神经退行性疾病患者以及缺乏有效和高效的治疗方法，这种特性变得非常重要。许多研究表明，石松科生物碱是 AChE 抑制剂的丰富来源。在从植物材料中获得新的治疗性植物化学物质时，提取过程及其效率至关重要。因此，这项工作的目的是优化现代 PLE 的条件，从两个物种中获得生物活性生物碱：石松和卷柏。为了比较生物碱含量，从而比较整个提取过程的效率，使用了五种不同极性的不同溶剂来制备植物提取物。PLE 参数是基于多项研究得出的，这些研究给出了最高的生物碱回收率。粗提取物通过固相萃取（SPE）在 Oasis HLB 卡上进行纯化，并通过 HPLC/ESI-QTOF-MS 分析高度丰富的生物碱。据我们所知，这是首次获得如此高的已知石松科生物碱回收率。在从 中获得的二氯甲烷提取物中检测到生物碱石松宁的最佳提取结果，其收率超过 45%。在二氯甲烷和乙酸乙酯提取物中观察到Annotinine 的回收率超过 40%。此外，还获得了所有分离的生物碱的色谱图，并且在甲醇提取物中获得了最佳的分离效果和质量。有趣的是，属于非极性溶剂的环己烷提取物中未检测到任何生物碱。这些结果可能有助于理解和优化分离强效 AChE 抑制剂的最佳条件。
