In vitro pro-inflammatory enzyme inhibition and anti-oxidant potential of selected Sri Lankan medicinal plants.

BACKGROUND: The extracts of the ten selected Sri Lankan medicinal plants have been traditionally used in the treatment of inflammatory mediated diseases. The extracts were investigated for anti-inflammatory and anti-oxidant potential in vitro to identify bio-active extracts for further chemical characterization. METHODS: In vitro anti-inflammatory activities of total ethanol extracts were investigated measuring the inhibitory activities of four pro-inflammatory enzymes, arachidonate-5- lipoxygenase (A5-LOX), hyaluronidase (HYL), xanthine oxidase (XO) and inducible nitric oxide (iNO) synthase. Cytotoxicity of extracts were determined by MTT assay. Oxidative burst inhibition (OBI) on human whole blood (WB) and isolated polymorphoneutrophils (PMNs) was carried out for a selected bio-active extract. Anti- oxidant activities of the extracts were determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging, ferric reducing antioxidant power (FRAP), ferrous ion chelation (FIC) and oxygen radical absorbance capacity (ORAC) assays. Total polyphenol and total Flavonoid contents of the extracts were also determined. The most active plant extract was analysed using Gas chromatography-Mass spectrometry (GC-MS) and High Performance Liquid Chromatography (HPLC). RESULTS: The ethanol bark extract of Flacourtia indica showed the highest A5-LOX (IC50: 22.75 ± 1.94 g/mL), XO (70.46 ± 0.18%; 250 µg/mL) and iNOs inhibitory activities on LPS- activated raw 264.7 macrophage cells (38.07 ± 0.93%; 500 µg/mL) with promising OBI both on WB (IC50: 47.64 2.32 µg/mL) and PMNs (IC50: 5.02 0.38 µg/mL). The highest HYL inhibitory activity was showed by the leaf extracts of Barathranthus nodiflorus (42.31 ± 2.00%; 500 µg/mL) and Diospyros ebenum (41.60 ± 1.18%; 500 µg/mL). The bark and leaf extracts of Callophyllum innophyllum (IC50: 6.99 ± 0.02 µg/mL) and Symplocus cochinchinesis (IC50: 9.85 ± 0.28 µg/mL) showed promising DPPH free radical scavenging activities. The GC-MS analysis of ethanol bark extract of F. indica showed the presence of two major bio-active compounds linoleic acid ethyl ester and hexadecanoic acid, ethyl ester (> 2% peak area). The HPLC analysis showed the presence polyphenolic compounds. CONCLUSION: The ethanol bark extract of F. indica can be identified as a potential candidate for the development of anti-inflammatory agents, which deserves further investigations. The bio-active plant extracts may be effectively used in the applications of cosmetic and health care industry.

Comparative Phytochemical Analysis and Antioxidant Activities of Tamalakyadi Decoction with Its Modified Dosage Forms.

BACKGROUND AND OBJECTIVE: Tamalakyadi decoction (TD) is a classical formulation mentioned in authentic traditional medicine text Sarasankshepaya under nasal diseases and used as a remedy for allergic rhinitis. It consists of 12 plant ingredients. Decoction preparations are widely used in Sri Lankan traditional system and considered effective and safe for treating many disorders. However, decoctions have to be used only in fresh state due to shorter shelf life and loss of stability. This decoction preparation method leads to decreasing the patient compliance and is also time consuming. Hence, the objective of the present study was to convert TD to consumer friendly novel dosage form, namely, freeze dried, spray dried, and traditional ganasara forms. METHODOLOGY: Therefore, we compared the phytochemical constituents and antioxidant activities of TD with its modified dosage forms. The chemical comparison of four dosage forms comprises phytochemical screening, TLC and HPTLC fingerprint profiles and the antioxidant activities by DPPH free radical scavenging activity, Ferric reducing antioxidant power (FRAP), total polyphenol content (TPC), and total flavonoid content (TFC). RESULTS: Phytochemical screening revealed the presence of alkaloids, saponins, tannins, steroids, flavonoids, phenols, and terpenoids in all dosage forms. However, the saponins, alkaloids, flavonoids, terpenoids, and steroids were more prominent in TD and freeze dried preparation than the other two preparations. HPTLC fingerprint pattern of freeze dried dosage was more similar with HPTLC fingerprint pattern of TD in terms of number of peaks and their intensity compared to that of spray dried and ganasara dosage forms. Antioxidant activities such as DPPH, FRAP, TPC, and TFC were higher in decoction and freeze dried preparation than in spray dried and ganasara preparation. CONCLUSION: Freeze dried TD is the most suitable ready to use preparation having similar chemical properties and antioxidant activities to TD.