TLC Assay of Thymoquinone in Black Seed Oil (Nigella Sativa Linn) and Identification of Dithymoquinone and Thymol

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Abstract
A simple quantitative TLC method for the determination of thymoquinone in commercially available black seed oil obtained from Nigella Sativa L. (Ranunculaceae) using a scanning densitometer is described. Also the identification of thymol, dithymoquinone in this sample was established. The Rf values for thymoquinone, thymol and dithymoquinone are 0.77, 0.37 and 0.52 respectively. The identification of the thymoquinone spot, obtained from the methanol extract of oil is confirmed by GC/MS which is essentially identical to thymoquinone standard. The solvent system consisted of benzene: isopropyl ether (1:1). All the spots were visualised and quantitated at 254 nm. The method proposed is simple, reproducible with a lower limit of detection of 100 nmoles/ml and can be used in routine analysis of thymoquinone in black seed oil for quality control purposes.