Helleborus purpurascens—Amino Acid and Peptide Analysis Linked to the Chemical and Antiproliferative Properties of the Extracted Compounds

Abstract

There is a strong drive worldwide to discover and exploit the therapeutic potential of a large variety of plants. In this work, an alcoholic extract of Helleborus purpurascens (family Ranunculaceae) was investigated for the identification of amino acids and peptides with putative antiproliferative effects. In our work, a separation strategy was developed using solvents of different polarity in order to obtain active compounds. Biochemical components were characterized through spectroscopic (mass spectroscopy) and chromatographic techniques (RP-HPLC and GC-MS). The biological activity of the obtained fractions was investigated in terms of their antiproliferative effects on HeLa cells. Through this study, we report an efficient separation of bioactive compounds (amino acids and peptides) from a plant extract dependent on solvent polarity, affording fractions with unaffected antiproliferative activities. Moreover, the two biologically tested fractions exerted a major antiproliferative effect, thereby suggesting potential anticancer therapeutic activity.

Bioactive Molecules Profile from Natural Compounds

“…Advanced extraction processes (solid-phase extraction, ultra-sound-assisted extraction, microwave-assisted extraction, supercritical fluid extraction, pulsed electric field extraction, pressurized liquid extraction, enzyme-assisted extraction, surfactant-mediated extraction) have minimized many of these shortcomings. Usually, the separation of a particular group of bioactive compounds from a complex natural product required a selective separation strategy based on phytochemicals partition in several different polarity solvents [43]. Nevertheless, natural product chemistry research concerns the development of new and highly efficient extraction techniques.…”

“…The cells will be analyzed by fluorescence microscopy and will be quantified to establish the degree of apoptosis and implicitly the cell viability. Also, the time-lapse video microscopy can be used to evaluate the bioactive phytochemicals [43]. The in vivo biotests are applied on animals (mice, rats, pigs, etc.…”

“…In an effort to detect the amino acids and thionins from Viscum album a selective partition strategy based on solvents with different polarities (methanol, hexane and carbon tetrachloride) was developed [43]. The plant material (Viscum album leaves and young leaves from Quercus robur) was obtained from a collection taken in December 2015 in Timis, Romania.…”

“…Then the solution was filtered through a 0.30 μm pore size filter and subsequently extracted with the following organic solvents: n-hexane (V 1 ) and carbon tetrachloride and (V 2 ). The separation of thionins was carried on the next experiment: 2 g of sample was extracted successively with petroleum ether (30 mL) and acetone (30 mL) [43]. Identity of the compounds from the obtained viscum fractions: V 1 (hexane), V 2 (CCl 4 ), V 3 (petroleum ether) and respectively, fraction V 4 (acetone) was performed using GC-MS and TOF MS methods.…”

“…The results of design isolation strategy based on different solvent polarity were analyzed through GC-MS [43]. The identified compounds are presented in Table 5; after a careful comparison with spectral database, NIST/NBS was used to compare the results of analysis [43].…”