HPLC and Anti-Inflammatory Studies of the Flavonoid Rich Chloroform Extract Fraction of Orthosiphon Stamineus Leaves

Abstract

The aim of the present study was to verify the anti-inflammatory activity of Orthosiphon stamineus leaf extracts and to identify the active compound(s) contributing to its anti-inflammatory activity using a developed HPLC method. Active chloroform extract of O. stamineus was fractionated into three fractions using a dry flash column chromatography method. These three fractions were investigated for anti-peritoneal capillary permeability, in vitro nitric oxide scavenging activity, anti-inflammatory and nitric oxide (NO) inhibition using carrageenan-induced hind paw edema method. The flavonoid rich chloroform extract fraction (CF2) [containing sinensetin (2.86% w/w), eupatorin (5.05% w/w) and 3’-hydroxy-5,6,7,4’-tetramethoxyflavone (1.101% w/w)], significantly reduced rat hind paw edema, NO and decreased dye leakage to peritoneal cavity at p < 0.05. IC50 of in vitro NO scavenging of CF2 was 0.3 mg/mL. These results suggest that the anti-inflammatory properties of these CF2 may possibly be due to the presence of flavonoid compounds capable of affecting the NO pathway.

Antiproliferative and antiangiogenic effects of flavone eupatorin, an active constituent of chloroform extract of Orthosiphon stamineus leaves

“…Tablets or capsules containing dried leaves are also available as a dietary supplement. For its demonstrated efficacy the extracts from O. stamineus have been more closely examined and the results confirmed its diuretic, uricosuric [3], antioxidant, hepatoprotective [4], antiinflammatory [5] and other properties. Extracts of O. stamineus leaves consist of a wide range of compounds, including flavonoids, terpenoids or organic acids such as rosmarinic or caffeic acid [6].…”

“…The extract reduced cell viability of cancer cells tested as shown in Table 1. It was reported previously that eupatorin is one of the most biologically active constituents of O. stamineus leaves [5]. To evaluate its contribution to the antiproliferative activity of the extract found in this study, the concentration of eupatorin in the extract was determined by using UPLC-MS/MS (0.53 ± 0.08 mg/g DW; Suppl.…”

“…Extracts of O. stamineus leaves consist of a wide range of compounds, including flavonoids, terpenoids or organic acids such as rosmarinic or caffeic acid [6]. The polymethoxylated flavonoids sinensetin, eupatorin and 3′-hydroxy-5,6,7,4′-tetramethoxyflavone, found as dominant compounds in the chloroform fraction of such extracts, are among the most therapeutically important constituents [5].…”

“…This observation led us to study the antiproliferative and antiangiogenic effects of the compound in detail and in an effort to explain the mechanism of its action. We also tried to evaluate the contribution of eupatorin to the overall activity of the chloroform extract of O. stamineus leaves where eupatorin is among the main constituents [5].…”

Topical piroxicam in vitro release and in vivo anti-inflammatory and analgesic effects from palm oil esters-based nanocream

“…Edema volume, in terms of thickness, was measured in all four groups at hours 2, 4, and 6 after carrageenan injection using a micrometer (Ozaki Ltd, Tokyo, Japan). 25 The induced thickness was measured by placing the foot of the rat between the anvil and spindle of the micrometer.…”