ABSTRACT
Biomarker rutin was analyzed in methanol extracts of leaves of five different species of genus Ficus [Ficus carica, Ficus nitida, Ficus ingens, Ficus palmata and Ficus vasta] by NP-HPTLC [Method I] and RP- HPTLC methods [Method II]. The development and validation for method I was carried out with silica gel 60F[254] plates using EA: GAA: FA: H[2]O [10:1:1:2.5, v/v/v/v] as developing system. Method II was carried out on silica gel 60F[254] RP-18 plates using mobile phase ACN: H[2]O [4:6 v/v]. Both analyses were scanned at 305 nm and were found to give well resolved peak of rutin at Rf 0.28 +/- 0.01 and 0.68 +/- 0.03 for Method I and Method II, respectively. The percentage of rutin was found to be 0.51% and 0.66% in F. ingens, 0.24% and 0.54% in F. palmata and 0.14% and 0.17% in F. vasta by Method I and Method II, respectively. Method II [RP-HPTLC] was found to be more accurate, precise and sensitive than Method I. Method II can be used as an important tool for standardization and quality control of bulk drugs and in-process formulations of rutin
ABSTRACT
<p><b>OBJECTIVE</b>To develop HPTLC fingerprint profile of anti-inflammatory active extract fractions of Tribulus terrestris (family Zygophyllaceae).</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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 Rf values.</p><p><b>CONCLUSIONS</b>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.</p>