Aqueous and hydroalcoholic extracts from Verbena officinalis L were obtained and characterized. The analysis by HPLC-DAD and LC-MS allowed the detection and identification of three iridoids, fifteen flavonoids and four phenolic acid derivatives. Four flavonoids, scutellarein 7-diglucuronide (9), scutellarein 7-glucuronide (13), pedalitin 6-galactoside (15) and scutellarein 7-glucoside (19) are reported for the first time from this plant. In addition, three new flavonoids have been isolated: scutellarein 7-O-(2-O-feruloyl)-diglucuronide (5), pedalitin 6-O-diglucuronide (6) and pedalitin 6-O-(2-O-feruloyl)-diglucuronide (13). To our knowledge, these flavonoids have not been reported as natural products. Both extracts showed significant antioxidant activity using three in vitro model systems and the results have been correlated with total phenolic and total flavonoid contents. The results have allowed establishing an important relation structure-activity and significant correlations have also been found between the mineral content and the flavonoids present in both extracts.

Melissa officinalis L. (Lamiaceae) is consumed as a traditional herbal tea in the Mediterranean region. The cytotoxic effect of the 50% ethanolic and aqueous extract, determined by the MTT and NR assays, was evaluated in vitro on Human Colon Cancer Cell Line (HCT-116), using Triton 10% as positive control. The 50% ethanolic extract showed significant differences after 72 h of treatment, reducing cell proliferation to values close to 40%, even the lowest dose tested (5 μg/ml). In the MTT assay, the same extract caused the lowest cell viability with 13% at a concentration of 1,000 μg/ml after 72 h of treatment, being a value lower than Triton 10%. The antioxidant activity was also confirmed evaluating the capacity of the extracts to scavenge ABTS and DPPH radicals, and IC(50) values were highly correlated with the total phenolic and flavonoid content. Bioassay guided fractionation led to the isolation of an anti-proliferative compound, rosmarinic acid. Its structural elucidation was performed by HPLC/DAD/ESI/MS analysis. High dose of rosmarinic acid (1,000 μg/ml) was clearly cytotoxic against HCT-116 cells, with a significant decrease in cell number since the earliest time point (24 h).

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A new formulation of Bologna-type sausage enriched in ω-3 polyunsaturated fatty acids (PUFA) (8.75% linseed oil) was developed, using a lyophilized aqueous-ethanolic extract of Melissa officinalis. A comparison with the effectiveness of butylhydroxy anisole (BHA) synthetic antioxidant to decrease the oxidation of PUFAs was performed.The formulation increased the ω-3 PUFAs content, especially α-linolenic acid, decreasing significantly the ω-6/ω-3 ratio from 17.3 to 1.9, and also the Atherogenic Index and Thrombogenic Index (0.38-0.31 and 1.03-0.54, respectively).Modified sausages with BHA and Melissa extract showed significantly lower peroxides value (2.62 and 6.11meqO 2 /kg) and thiobarbituric acid value (0.26 and 0.27mg malondialdehyde/kg) and higher antioxidant capacity (hydrophilic fraction ABTS: 0.45 and 0.74meq Trolox/g product; lipofilic fraction ABTS: 0.44 and 0.37meq Trolox/g product) than those without these ingredients (16.49meq O 2 /kg, 2.08 mg malondialdehyde /kg, 0.26 and 0.27meq Trolox/g product, respectively). Sensorial tests showed that acceptability of the new formulations was similar to control products.

A new formulation of dry fermented sausage, including ingredients that contribute to improve the nutritional and health benefits of this type of products is presented. Se yeast (2 g/kg), iodized salt (26 g/kg), linseed:algae (3:2) emulsion (62.5 g/kg), and lyophilized water extract of Melissa officinalis L. as source of natural antioxidants (686 mg/kg), allowed to obtain dry fermented sausages with technological and sensorial properties similar to traditional ones.From the nutritional standpoint, a 50 g portion of this product would cover a 100 % of the recommended intake value for Se, a 70 % of DRI for iodine, and a 40 % and 100 % of the labeling reference intake for α-linolenic and EPA+DHA, respectively. The ω-6/ω-3 ratio decreased from 15.7 in the control product to 1.96 in the modified one.Despite the high PUFA content, no oxidation signs were detected by TBARS (<0.15 mg MDA/kg) and volatile compounds, pointing at the effectiveness of the proposed natural antioxidant from Melissa officinalis.The modified formulation presented good acceptability for panelists and similar appearance, odor, taste and juiciness with control products.