HPLC-SPE-NMR Characterization of Major Metabolites in Salvia fruticosa Mill. Extract with Antifungal Potential: Relevance of Carnosic Acid, Carnosol, and Hispidulin

Abstract

Plant pathogenic fungi are considered of significant economic importance for adversely affecting both quantitatively and qualitatively fresh and processed produce. Extracts of Salvia fruticosa were initially screened for their antifungal activity, and the ethyl acetate fraction, being the most active, was further analyzed using HPLC-SPE-NMR hyphenation. The methoxylated flavones hispidulin, salvigenin, and cirsimaritin and the diterpenes carnosic acid, carnosol, and 12-methoxycarnosic acid were identified as the major components of the extract. In addition, the concentration levels of all identified components were determined using q-NMR. The antifungal activity of the crude extract and selected phytochemicals was estimated against the fungal species Aspergillus tubingensis, Botrytis cinerea, and Penicillium digitatum. The estimated MIC and MFC values of the ethyl acetate extract of S. fruticosa, as well as three of its major constituents, carnosic acid, carnosol, and hispidulin, support their antifungal activity, especially against B. cinerea and P. digitatum, suggesting their potential use in food and agricultural systems.

Edible coating composed of chitosan andSalvia fruticosaMill. extract for the control of grey mould of table grapes

“…In particular, these phytochemicals were attributed to carnosol, carnosic acid and 12‐methoxy carnosic acid. The identification of six phytochemicals was described in detail in our previous work . NMR data for all identified compounds are provided in the Supporting Information.…”

“…In addition, quantitative NMR (q‐NMR) was used to quantify the major phytochemicals (Supporting Information 1). HPLC‐SPE‐NMR and q‐NMR methods were described in a previous work …”

“…Free radical scavenging activity was determined as follows: 2 mL of diluted extract were mixed with 1 mL of 0.3 mmol L −1 solution of 2,2‐diphenyl‐2‐picrylhydrazyl (DPPH) in methanol, incubated in the dark for 30 min and the absorbance of the mixture was monitored at 517 nm. A standard curve of (±)‐6‐hydroxy‐2,5,7,8‐tetramethylchromane‐2‐carboxylic acid (Trolox) was prepared …”

“…Salvia fruticosa Mill., commonly known as Greek sage, has received attention due to its health‐promoting properties and economic importance. Research so far has emphasised the antimicrobial activity of the EOs of the species, while the in vitro activity of its polar extracts was recently screened …”

Protective effects of carnosic acid against mitochondria‑mediated injury in H9c2 cardiomyocytes induced by hypoxia/reoxygenation

“…1) that has been identified as the major bioactive phytochemical in numerous medicinal plants, including Rosmarinus officinalis (8), Salvia fruticosa (9) and Ocimum sanctum (10). Previous pharmacological studies have demonstrated that carnosic acid affords various biological activities, such as neuroprotection (11), prevention of advanced glycation end-product formation (12), attenuation of Alzheimer's disease (13), anticancer activity (14), anti-inflammation (15) and renoprotection (16).…”