Dietary Supplementation with an Extract of Aloysia citrodora (Lemon verbena) Improves Sleep Quality in Healthy Subjects: A Randomized Double-Blind Controlled Study.

Seventy-one healthy subjects with sleep disturbances participated in a randomized, double-blind controlled trial in which dietary supplementation with an extract of Aloysia citrodora (lemon verbena) (n = 33) or placebo (n = 38) was administered for 90 days. There were between-group differences in favor of the experimental group in the visual analogue scale (VAS) for sleep quality (6.5 ± 1.6 vs. 5.5 ± 2.1, p = 0.021) as well as in the overall score (5.8 ± 2.4, p = 0.008) and scores for sleep latency (1.6 ± 1.0 vs. 1.9 ± 0.7, p = 0.027) and sleep efficiency (84.5 ± 12.8 vs. 79.8 ± 13.6, p = 0.023) in the Pittsburgh Sleep Quality Index (PSQI). Sleep-related variables (latency, efficiency, wakefulness after sleep onset, awakenings) assessed by actigraphy also showed better scores in the experimental group (p = 0.001). Plasma nocturnal melatonin levels also increased significantly in the experimental group (199.7 ± 135.3 vs. 174.7 ± 115.4 pg/mL, p = 0.048). Changes in anthropometric parameters and physical activity levels were not found. In summary, a dietary supplement of lemon verbena administered for 3 months was associated with a significant improvement in sleep quality as compared with placebo in a population of healthy subjects with sleep problems.

Cistus, Acacia, and Lemon verbena Valorization through Response Surface Methodology: Optimization Studies and Potential Application in the Pharmaceutical and Nutraceutical Industries.

Cistus ladanifer L., Acacia dealbata L., and Aloysia citrodora Paláu were subject to an optimization procedure for two extraction techniques (heat-assisted extraction (HAE) and ultrasound-assisted extraction (UAE)). The extracts were then analyzed by HPLC-DAD-ESI/MS for their phenolic profile (cistus-15 compounds, acacia-21 compounds, and lemon verbena-9 compounds). The response surface methodology was applied, considering four varying factors: ethanol percentage; extraction time; temperature/power; and S/L ratio, generating two responses (the major phenolic compound, or family of compounds, and the extraction yield). For cistus, both techniques optimized the extraction yield of punicalagins, with UAE proving to be the most efficient extraction method (3.22% ethanol, 22 min, 171 W, and 35 g/L). For acacia, HAE maximized the extraction of procyanidin (74% ethanol, 86 min, 24 °C, and 50 g/L), and UAE maximized the content of myricetin (65% ethanol, 8 min, 50 W, and 50 g/L). For lemon verbena, HAE favored the extraction of martynoside (13% ethanol, 96 min, 49 °C and 17 g/L) and forsythiaside UAE (94% ethanol, 25 min, 399 W, and 29 g/L). The optimal conditions for the extraction of compounds with high added value and potential for use in pharmaceuticals and nutraceuticals were defined.

Role of Polyphenols from the Aqueous Extract of Aloysia citrodora in the Inhibition of Aflatoxin B1 Synthesis in Aspergillus flavus.

Aflatoxin B1 (AFB1) is a mycotoxin considered a potent carcinogen for humans that contaminates a wide range of crops. Various strategies have been established to reduce or block the synthesis of AFB1 in food and feed. The use of aqueous extracts derived from plants with high antioxidant activity has been a subject of study in recent years due to their efficacy in inhibiting AFB1. In this study, we assessed the effect of Aloysia citrodora aqueous extract on Aspergillus flavus growth and on AFB1 production. A bio-guided fractionation followed by High Performance Liquid Chromatography (HPLC) and Mass spectrometry analysis of the active fraction were applied to identify the candidate molecules responsible for the dose-effect inhibition of AFB1 synthesis. Our results revealed that polyphenols are the molecules implicated in AFB1 inhibition, achieving almost a total inhibition of the toxin production (99%). We identified luteolin-7-diglucuronide as one of the main constituents in A. citrodora extract, and demonstrated that it is able to inhibit, by itself, AFB1 production by 57%. This is the first study demonstrating the anti-Aflatoxin B1 effect of this molecule, while other polyphenols surely intervene in A. citrodora anti-AFB1 activity.

Chemical Composition and Antioxidant Properties of Common and Lemon Verbena.

The nutritional profiles of common and lemon verbena leaves were analyzed (proximate constituents, free sugars, organic acids, tocopherols, and fatty acids) and the leaves were prepared in hydromethanolic and aqueous (decoctions and infusions) extracts. The phenolic compound composition and antioxidant activity (2,2-Diphenyl-1-picrylhydrazyl (DPPH); 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), ferric-reducing antioxidant power (FRAP); and cupric-reducing antioxidant capacity (CUPRAC) assays) of the extracts were characterized. The nutritional composition varied between the studied species, with lemon verbena showing higher amounts of protein, ash, and fat than common verbena, whereas the opposite trend was recorded for the dietary fiber content. The main free sugars detected in both species were fructose, glucose, and sucrose, which were present in higher amounts in the common verbena samples. Succinic acid was the most abundant organic acid in both species while high amounts of oxalic acid were detected in lemon verbena. The main fatty acids in both species were α-linolenic, palmitic, and linoleic acid. Regarding the phenolic compound content, the extracts of lemon verbena presented higher amounts of total phenolic compounds (TPCs), total flavonoids (TFs) and total phenolic acids (TPAs) than the common verbena extracts while the aqueous extracts (infusions and decoctions) were richer in TPCs, TFs, and TPAs than the hydromethanolic ones in both species. Nine phenolic compounds were identified and quantified, including seven phenolic acids and two flavonoids. The lemon verbena samples were characterized by higher antioxidant activity compared to the common verbena samples while the aqueous extracts showed higher antioxidant efficacy than the hydromethanolic ones. In conclusion, both species showed promising results in terms of the nutritional value, chemical composition, and antioxidant activities, which were positively correlated with the phenolic compound contents. Moreover, the extraction protocol may affect the chemical composition and bioactive properties of both species, with aqueous extracts showing better results than hydromethanolic ones.

Protective effect of aqueous and ethanolic extracts of Lippia citriodora Kunth. on acrylamide-induced neurotoxicity.

Objective: Acrylamide (ACR) neurotoxicity is induced by different mechanisms such as oxidative stress and apoptosis. Scientific researchs have indicated the antioxidative properties of Lippia citriodora. The protective effect of L. citriodora aqueous and ethanolic extracts on ACR-induced neurotoxicity was investigated. Materials and methods: Male Wistar rats were randomly divided into 13 groups: (1) control, (2) ACR (50 mg/kg, i.p.), (3-6) ACR+aqueous extract (12.5, 25, 50, and 100 mg/kg, i.p.), (7-10) ACR+ethanolic extract (12.5, 25, 50, and 100 mg/kg, i.p.), (11) aqueous extract (100 mg/kg), (12) ethanolic extract (100 mg/kg), and (13) ACR+Vitamin E (200 mg/kg, every other day, i.p.). After 11 days, gait score, MDA, and GSH levels in brain cortical tissue were measured. In the in vitro test, the viability of PC12 cells (using MTT test), the amount of reactive oxygen species (ROS; using DCFH-DA method), and the protein levels of Bax, Bcl2 and caspase 3 (by western blotting) were measured. Results: In the in vitro study, the IC50 for the treatment of PC 12 cells with ACR after 24 hr was 6 mM. ACR decreased cell viability, but increased ROS level, Bax/Bcl-2 ratio, and caspase-3 protein level. Pre-treatment by L. citriodora extracts (15-120 µg/ml) ameliorated the toxic effects of ACR on PC12 cells. In the in vivo experiment, ACR-induced movement disorders increased MDA but decreased GSH content. The extracts of L. citriodora improved ACR toxic effects. Conclusion: Aqueous and ethanolic extracts of L. citriodora were found to reduce ACR-induced neurotoxicity via inhibiting oxidative stress and apoptosis.

Chemometric Profiling and Bioactivity of Verbena (Aloysia citrodora) Methanolic Extract from Four Localities in Tunisia.

This research aimed to study the chemical composition of Aloysia citrodora methanolic extract and its biological activities as an antioxidant, and its antibacterial, antifungal and anti-inflammatory activities based on four bioclimatic collection stages. The contents of total phenols, total flavonoids and total tannins were determined. Nine phenolic compounds were identified by LC-DAD-ESI-MS/MS. The major compound was acteoside, a phenylpropanoid which represented about 80% of the methanolic fraction in the various regions. The antioxidant activities of different locations were measured by different analytical assays, such as DPPH, ABTS and iron reducing power. The results showed that phenolic compounds and antioxidant activities varied with climatic and environmental factors. Moreover, there was a significant dependency between regions and biological activities. The use of a principal component analysis showed that there was a close relationship among phenylpropanoids, phenolic compounds and the studied biological activities.

Andean Plants Essential Oils: A Scented Alternative to Synthetic Insecticides for the Control of Blowflies.

Blowflies are vectors of microorganisms and human pathogens, and their maggots cause myiasis in vertebrates and infest and spoil meat and fish products. Essential oils (EOs) from spices were proven to be a safer and more sustainable alternative to synthetic insecticides for the control of blowflies and are suitable for protecting food from such pests. However, some EOs are not acceptable for environmental or topical applications due to their strong, unpleasant odour. In this study, we measured the acute toxicity and the repellence of two EOs extracted from the Andean plants Aloysia citrodora Palau and Bursera graveolens (Kunth) Triana and Planch., both known for their pleasant odour, against the blue blowfly Calliphora vomitoria (L.) (Diptera: Calliphoridae). We also compared their bioactivity with that of the Allium sativum L. EO, which is very effective but bad-smelling. The A. citrodora EO was mainly rich in oxygenated monoterpenes, the most abundant of which were geranial (26.8%) and neral (21.0%). The B. graveolens EO was chiefly composed of monoterpene hydrocarbons, mostly represented by limonene (46.2%). According to the sensory description, the best odour profile was associated with the A. citrodora EO, while the olfactory expression of the EO from B. graveolens was negatively affected by a strong odour of "mouldy lemon". The behavioural test showed that the A. citrodora EO was more repellent than that of A. sativum and, on the contrary, that the B. graveolens EO was attractive. The toxicity tests showed that the EOs of both Andean plants have a clear toxic effect on the C. vomitoria eggs and adults. In terms of ovicidal activity, there were no significant differences among the effects of the three tested EOs. On the adult flies, the toxicity of the two EOs of the Andean plants was much lower than that of A. sativum (LC50 fumigation = 1.86 µL EO L-1 air; LC50 ingestion = 8.10 µL EO mL-1 gel) both by fumigation (LC50 = 23.66 and 25.30 µL EO L-1 air for A. citrodora and B. graveolens, respectively) and ingestion (LC50 = 36.65 and 44.97 µL EO mL-1 gel for A. citrodora and B. graveolens, respectively), while, by contact, the A. citrodora EO (LD50 = 0.27 µL EO insect-1) was more toxic than the A. sativum EO (LD50 = 0.46 µL EO insect-1).

Aloysia Citrodora Essential Oil Inhibits Melanoma Cell Growth and Migration by Targeting HB-EGF-EGFR Signaling.

Patients diagnosed with melanoma have a poor prognosis due to regional invasion and metastases. The receptor tyrosine kinase epidermal growth factor receptor (EGFR) is found in a subtype of melanoma with a poor prognosis and contributes to drug resistance. Aloysia citrodora essential oil (ALOC-EO) possesses an antitumor effect. Understanding signaling pathways that contribute to the antitumor of ALOC-EO is important to identify novel tumor types that can be targeted by ALOC-EO. Here, we investigated the effects of ALOC-EO on melanoma growth and tumor cell migration. ALOC-EO blocked melanoma growth in vitro and impaired primary tumor cell growth in vivo. Mechanistically, ALOC-EO blocked heparin-binding-epidermal growth factor (HB-EGF)-induced EGFR signaling and suppressed ERK1/2 phosphorylation. Myelosuppressive drugs upregulated HB-EGF and EGFR expression in melanoma cells. Cotreatment of myelosuppressive drugs with ALOC-EO improved the antitumor activity and inhibited the expression of matrix metalloproteinase-7 and -9 and a disintegrin and metalloproteinase domain-containing protein9. In summary, our study demonstrates that ALOC-EO blocks EGFR and ERK1/2 signaling, with preclinical efficacy as a monotherapy or in combination with myelosuppressive drugs in melanoma.

Effect of Aloysia citrodora Essential Oil on Biochemicals, Antioxidant Characteristics, and Shelf Life of Strawberry Fruit during Storage.

Strawberry fruits are highly susceptible to cold burning, resulting in low storage periods at low temperatures. Plant extracts or essential oils (EOs) can potentially be used as preservatives in fruits throughout the refrigerated period. In the present study, the biochemicals, antioxidant characteristics, and shelf life of treated strawberries with Aloysia citrodora essential oil (ACEOs) were evaluated during keeping time. The treatments were produced as follows: T1, control; T2, 250 ppm ACEOs; T3, 500 ppm ACEOs; and T4, 750 ppm ACEOs. Total soluble solids (TSS), weight loss, titratable acidity (TA), antioxidant activity (DPPH assay), total phenolic (TPC), flavonoid and anthocyanin contents (TFC), and enzymes activity (peroxidase and ascorbate peroxidase) were evaluated during the refrigerated period (5 °C with relative humidity of 85-90% for 20 days). The results revealed that weight loss and TA were reduced in all treatments during storage, being that the rates were lower in samples treated with ACEOs. TPC, TFC, TSS, antioxidant, and enzymes activity were higher in treated fruits than control.

Dinámica de polifenoles de “Cedrón” (Aloysia citrodora Palau -Verbenaceae-) en relación al desarrollo foliar / Polyphenol dinamics on “cedrón” (Aloysia citrodora Palau -Verbenaceae-) according to leave development

Leaf infusions from Aloysia citrodora Palau –Verbenaceae-, known as “cedrón” were analyzed in order to establish the possible influence of their development on polyphenols profile. Chromatographic fingerprints of infusions from young and full expanded leaves were performed and total phenols, total tannins, total flavonoids and total hydroxycinnamic acids were quantified. Young leaves had significative higher total phenols, total flavonoids and total hydroxycinnamic acids concentrations than full expanded leaves. This research suggest that the analyzed parameter should be considered when elaborating products from “cedrón” leaves, as the variation in compound concentration may translate in different biological activities.

Se analizaron las infusiones realizadas a partir de las hojas de Aloysia citrodora Palau –Verbenaceae-, conocida vulgarmente como “cedrón”, con el objeto de establecer la influencia del grado de desarrollo foliar en el perfil de polifenoles. Se obtuvieron los perfiles cromatográficos y se cuantificaron los fenoles totales, taninos totales, flavonoides totales y ácidos hidroxicinámicos totales provenientes de los extractos acuosos de las hojas jóvenes y adultas. Se observaron diferencias cuali-cuantitativas en el perfil de polifenoles cuando se comparan las hojas jóvenes con las hojas adultas. En las hojas jóvenes, las concentraciones de fenoles totales, flavonoides totales y ácidos hidroxicinámicos totales resultaron ser significativamente superiores a las determinadas en las hojas adultas. Los estudios realizados sugieren que la variable aquí analizada debe ser considerada al momento de elaborar productos en base a hojas de cedrón, dado que las diferentes concentraciones detectadas podrían traducirse en diferentes actividades biológicas.