HPLC chromatogram of C. sativum root extract. Reverse phase separation was performed at 40°C using a Merck Purospher STAR RP-18 endcapped column (5 μm). The mobile phase consisted of trifluoroacetic acid in water at pH 2.6 (solvent A) and acetonitrile (solvent B). The gradient program consisted of: 0% to 12.5% B for 2.5 min, 12.5% to 100% B for 17.5 min and 100% B for 10 min. The flow rate was kept at 1 ml/min and injection volume was 10 μl. The eluted peaks were monitored at 254 nm. 1: ascorbic acid; 2: p-coumaric acid; 3: butylated hydroxytoluene; S1: unidentified peaks collected for GC-MS analysis. (BSTFA = N,O-bis(trimethylsilyl)trifluoroacetamide).