Pure Isolates and Preparative HPLC Fractions or Crude Extract of Inula viscosa: Effect on β-hematin Inhibition in vitro

Abstract

The antimalarial activity of wild Inula viscosa (I. viscosa) plant leaves was investigated. The efficacy of the whole crude extract versus preparative HPLC fractions versus pure isolates were compared by measuring their effect on β-hematin inhibition in-vitro. The preparative HPLC experiments consisted of reversed phase preparative column (22.2mm x 250mm, 10μm) and linear gradient of water, acetonitrile as the mobile phase. Injection volume was 1000μl and the wavelengths range were from 200-450 nm using photodiode array detector (PDA). While fractions (IV, V and VI) showed antimalarial potential in comparison to chloroquine positive control, the rest of the fractions did not show any significant inhibition to the β-hematin formation. The antimalarial results showed that whole crude exact of the plant works better than the preparative fractions or the pure isolates presumably due to synergistic effect. The chemical identity of some of the pure isolates was explored using UHPLC-ESi-MS. Moreover, I. viscosa extract powder stayed stable over several years, while many other products such as Artemisia annua extract or Artemisin Combined Therapy (ACT) drugs rapidly lost their efficiency under tropical storage conditions. Keywords:

In Vitro Analysis of Antimalarial Activity of Achillea fragrantissima (Forssk.) Sch.Bip Extracts Based on Beta-Hematin Formation

“…In Figure 6, the chromatogram clearly demonstrates the separation and detection of 17 distinct compounds. These compounds include gallic acid (1), 3,4-dihydroxybenzoic acid (2), 3,4-dihydroxyphenylacetic acid (3), chlorogenic acid (4), 4-hydroxyphenylacetic acid (5), vanillic acid (6), caffeic acid (7), syringic acid (8), isovanillic acid (9), p-coumaric acid (10), ferulic acid (11), sinapic acid (12), rutin (13), verbascoside (14), quercetin (15), trans-cinnamic acid (16), and kaempferol (17). Each compound is assigned a specific number to facilitate their identification.…”

“…This process serves as a survival mechanism against the harm posed by the heme to the parasite. 10,13,14 A synthetic crystalline counterpart of hemozoin, known as β-hematin, is utilized as a tool in the search for potential new antimalarial drugs. The need for such drugs is crucial due to the emergence of drug-resistant strains of Plasmodium , including resistance to widely used drugs like chloroquine.…”