Antioxidant activity and rosmarinic acid changes in salicylic acid-treated Thymus membranaceus shoots

In order to characterise the antioxidant activity of Thymus membranaceus Boiss. subsp. membranaceus, an endemic species in Southeast Spain, five different analytical methods were used. Water, methanol and hexane extracts obtained from 60-day-old in vitro-grown shoots were assayed for their antioxidative properties using DPPH (1,1-diphenyl-2-picrylhydrazyl radical), ABTS (2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonate), FRAP (ferric reducing antioxidant power), reducing power and conjugated diene assays. Total soluble phenol content, as well as rosmarinic acid, in the different extracts were also determined. Methanolic extracts exhibited the best antioxidant activity and the highest amounts of total soluble phenolics. A single application of salicylic acid (10 μM) on culture media resulted in an increase in rosmarinic acid and phenolic levels, which in turn improved the extracts’ antioxidant properties. These current findings open new opportunities for obtaining valuable natural antioxidants for commercial exploitation by using tissue culture systems.