Anti-inflammatory, Hypoglycemic, Hepatoprotective, Antihyperlipidemic, Anti-Ulcerative, Cardioprotective Stimulant, Sedative, Hypnotic, Anticonvulsant Activity, Memory Retention, Stroke Preventive Activity, Antimicrobial Activity, Antimycobacterial, Antiviral, Larvicidal, Antiparasitic, Chemopreventative, Chemo-modulatory, Anticancer, Cytoprotective activities were found in a variety of herbal plants, including Ocimum basilicum. The purpose of this study was to design and develop a new HPTLC method that was accurate, precise and cost-effective for simultaneous measurement of rutin and quercetin in a hydroalcoholic extract of Ocimum basilicum seeds. The mobile phase was Toluene: Ethyl Acetate: Methanol: formic acid (6:4:3:1, v/v/v/v/v) and densitometric scanning was performed at 254 nm. Merck TLC aluminium sheets of silica gel 60 F254, (10 x 10 cm) with a thickness of 250 mm was used as stationary phase. At a wavelength of 254 nm, rutin and quercetin were detected. The constituents were resolved satisfactorily, with Rf values of 0.25 ± 2.01 for rutin and 0.80 ± 0.64 for quercetin, respectively. Linearity (300-1300 ng/spot for rutin and quercetin) was used to test the method’s accuracy and reproducibility. For both analytes, intra- and inter-day precision, as evaluated by coefficient of variation, was less than 3%. For rutin, the detection and quantification limits were 46.52 and 140.96 ng/spot, respectively, and for quercetin, they were 81.79 and 247.84 ng/spot. The proposed approach was found to be precise, accurate, repeatable, and specific, and it could be used to measure quercetin and rutin in samples at the same time.
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