Chromatographic finger print analysis of anti-inflammatory active extract fractions of aerial parts of Tribulus terrestris by HPTLC technique

Objective:To develop HPTLC fingerprint profile of anti-inflammatory active extract fractions of Tribulus terrestris(family Zygophyllaceae).Methods:The anti-inflammatory activity was tested for the methanol and its fractions(chloroform,ethyl acetate,n-butanol and aqueous)and chloroform extract of Tribulus terrestris(aerial parts)by injecting different groups of rats(6 each)with carrageenan in hind paw and measuring the edema volume before and 1,2 and 3 h after carrageenan injection.Control group received saline i.p.The extracts treatment was injected i.p.in doses of 200 mg/kg 1 h before carrageenan administration.Indomethacin(30 mg/kg)was used as standard.HPTLC studies were carried out using CAMAG HPTLC system equipped with Linomat IV applicator.TLC scanner 3.Reprostar 3,CAMAG ADC 2 and WIN CATS-4 software for the active fractions of chloroform fraction of methanol extract.Results:The methanol extract showed good antiedematous effect with percentage of inhibition more than 72%,indicating its ability to inhibit the inflammatory mediators.The methanol extract was re-dissolved in 100 mL of distilled water and fractionated with chloroform,ethyl acetate and n-butanol.The four fractions(chloroform,ethyl acetate,n-butanol and aqueous)were subjected to anti-inflammatory activity.Chloroform fraction showed good anti-inflammatory activity at dose of 200 mg/kg.Chloroform fraction was then subjected to normal phase silica gel column chromatography and eluted with petroleum ether-chloroform,chloroform-ethyl acetate mixtures of increasing polarity which produced 15 fractions(F1-F15).Only fractions F1,F2,F4,F5,F7,F9,F11 and F14 were found to be active,hence these were analyzed with HPTLC to develop their finger print profile.These fractions showed different spots with different R_f values.Conclusions:The different chloroform fractions F1,F2,F4,F5,F7,F9,F11 and F14 revealed 4,7,7,8,9,7,7 and 6 major spots,respectively.The results obtained in this experiment strongly support and validate the traditional uses of this Sudanese medicinal plant.