2.3.10. Calibration Curves for Standard Markers

Standard solutions of kaempferol, rutin, ellagic acid, catechin, and quercetin were applied in triplicate on silica gel 60 F254 plates, using a CAMMAG Linomat-5 Automatic Sample Spotter. The plates were developed in a solvent system, toluene/ethyl acetate/formic acid/methanol (3:3:0.8:0.2 *v*/*v*) in a CAMAG glass twin trough chamber (20 × 10 cm) up to a space of 8 cm. Further, the developed plates were dried and scanned at 254 nm using CAMAG TLC scanner 3 and win CATS 4 software. The peak areas were recorded. The quantification of polyphenols was carried out based on peak area with a linear calibration curve at concentration ranges of 40–200, 100–1600, 20–200, 200–1400, and 10–160 ng/band of kaempferol, rutin, ellagic acid, catechin, and quercetin, respectively. Calibration curves of polyphenols were obtained by plotting peak areas versus applied crude extracts, and the concentrations of kaempferol, rutin, ellagic acid, catechin, and quercetin were calculated, respectively.

### **3. Results**
