Abstract
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 Lycopodium species: L. clavatum L. and L. annotinum 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 L. clavatum, where the yield exceeded 45%. The high recovery of annotinine above 40% presented in L. annotinum 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.
Highlights
Species belonging to the Lycopodiaceae family, such as Huperzia or Lycopodium, are widely known around the world and have become the subject of numerous research
The extracts from dried aerial parts of L. clavatum and L. annotinum prepared by using Pressurized liquid extraction (PLE) with different solvents were compared in terms of total alkaloids content
It gave highly reproducible results, and it could be combined with other techniques such as high performance liquid chromatography/electrospray-ionizationtime-of-flight-mass spectrometry (HPLC/ESI-quadrupole time-of-flight mass (QTOF)–MS)
Summary
Species belonging to the Lycopodiaceae family, such as Huperzia or Lycopodium, are widely known around the world and have become the subject of numerous research. Most in vivo and in vitro studies have confirmed that Lycopodium alkaloids have beneficial effects in the treatment of brain diseases. Many scientific studies describe results proving alkaloid huperzine A (HupA) to be a great acetylcholinesterase (AChE) inhibitor. This alkaloid was obtained for the first time from Chinese medicinal herb Huperzia serrata belonging to the Lycopodiaceae family [4,5,6,7]. It has the ability to reversibly and selectively inhibit the activity of AChE This property gave hope in the treatment of different types of dementia such as Alzheimer’s disease, which is the fourth most common cause of death in developed countries. The therapy consists only in increasing the concentration of acetylcholine (ACh) at the synaptic site in the brain, stopping the symptoms of the disease, improving memory and alleviating behavioral disturbances [4,10]
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