Anise Myrtle Research Articles

Phytochemical Composition and Toxicological Screening of Anise Myrtle and Lemon Myrtle Using Zebrafish Larvae.

Plants are an immense source of drugs, and 50% of modern pharmacopeia has a plant origin. With increasing life expectancy in humans, many age-related degenerative diseases converge on oxidative cellular stress pathways. This provides an opportunity to develop broad treatments by targeting the cause of common pathologic cell degeneration. Toxicological effects can be readily assessed in a live animal model system to establish potential fauna for clinical use. Here, we characterized and evaluated the antioxidant potential and toxicological effects of anise myrtle (Syzygium anisatum) and lemon myrtle (Backhousia citriodora) leaves. Using zebrafish larvae, a model for high-throughput pre-clinical in vivo toxicology screening, we identified safe levels of extract exposures for development of future therapeutics. The antioxidant capacity and toxicity were very similar in these two myrtles. The LC50-96h for anise myrtle was 284 mg/L, and for lemon myrtle, it was 270 mg/L. These measurements are comparable to ongoing studies we are performing using the same criteria in zebrafish, which allow for robust testing and prioritization of natural fauna for drug development.

LC-ESI-QTOF-MS/MS Identification and Characterization of Phenolic Compounds from Leaves of Australian Myrtles and Their Antioxidant Activities.

Phenolic compounds, present in plants, provide substantial health advantages, such as antioxidant and anti-inflammatory properties, which enhance cardiovascular and cognitive well-being. Australia is enriched with a wide range of plants with phytopharmacological potential, which needs to be fully elucidated. In this context, we analyzed leaves of aniseed myrtle (Syzygium anisatum), lemon myrtle (Backhousia citriodora), and cinnamon myrtle (Backhousia myrtifolia) for their complex phytochemical profile and antioxidant potential. LC-ESI-QTOF-MS/MS was applied for screening and characterizing these Australian myrtles' phenolic compounds and the structure-function relation of phenolic compounds. This study identified 145 and quantified/semi-quantified 27 phenolic compounds in these Australian myrtles. Furthermore, phenolic contents (total phenolic content (TPC), total condensed tannins (TCT), and total flavonoids (TFC)) and antioxidant potential of phenolic extracts from the leaves of Australian myrtles were quantified. Aniseed myrtle was quantified with the highest TPC (52.49 ± 3.55 mg GAE/g) and total antioxidant potential than other selected myrtles. Catechin, epicatechin, isovitexin, cinnamic acid, and quercetin were quantified as Australian myrtles' most abundant phenolic compounds. Moreover, chemometric analysis further validated the results. This study provides a new insight into the novel potent bioactive phenolic compounds from Australian myrtles that could be potentially useful for functional, nutraceutical, and therapeutic applications.

Formulation, characterization, and stability of food grade oil‐in‐water nanoemulsions of essential oils of Tasmannia lanceolata, Backhousia citriodora and Syzygium anisatum

AbstractOil‐in‐water nanoemulsions were formulated using sunflower oil mixed with each of the essential oils of Tasmannia lanceolata (Tasmanian pepper leaf [TPL]), Backhousia citriodora (lemon myrtle [LM]) and Syzygium anisatum (anise myrtle [AM]) and stabilized with Tween 80 using ultrasonication. An oil‐surfactant ratio of 3:1 was found to produce the lowest emulsion droplet sizes of 96.6 nm for LM, 122.2 nm for AM and 131.8 nm for TPL. Increase in surfactant concentration above 10r resulted in larger droplet sizes, 165.8–2,647.2 nm for LM (radius, r = .82), 153.7–2,573.5 nm for AM (r = .93) and 157.4–2,621.6 nm for TPL (r = .83). Sonication for 3 min produced smaller droplet size; however, sonication for 9 min resulted in increase of droplet size by 1.48, 1.43 and 1.47 times for oils of LM (r = .82), AM (r = .93) and TPL (r = .83), respectively. A positive correlation was found between sonication amplitude (20–50%) and droplet size for nanoemulsions of LM (r = .93), AM (r = .98) and TPL (r = .95). TPL and LM nanoemulsions showed broad‐ spectrum antimicrobial activities against yeasts and bacteria. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) against weak‐acid resistant yeasts were between 0.001–0.003 and 0.002–0.007 mg/ml for nanoemulsion of TPL and between 0.003–0.014 mg/ml and 0.005–0.027 for nanoemulsion of LM, respectively. The stability and antimicrobial activity of TPL and LM essential oil nanoemulsions confirm their potential for application as food preservatives especially in beverage products that are commonly spoiled by weak‐acid resistant yeasts.

Review of the Leaf Essential Oils of the Genus Backhousia Sens. Lat. and a Report on the Leaf Essential Oils of B. gundarara and B. tetraptera.

A review of the leaf oils of the 13 species now recognised in the genus Backhousia is presented. This review carries on from, and incorporates data from, an earlier (1995) review of the then recognised eight species. The leaf oils of two new species of Backhousia, B. gundarara and B. tetraptera are reported for the first time. B. gundarara contains a mixture of mono- and sesquiterpenes, with α-pinene (14%) and spathulenol (11%) being the main members. In B. tetraptera, the principal component of the mainly terpenoid leaf oil is myrtenyl acetate (20–40%). The review also incorporates the two species of the genus Choricarpia, which have been subsumed into Backhousia, viz. B. leptopetala and B. subargentea. Due to its history in Backhousia, Syzygium anisatum, which has been transferred out of Backhousia, is included in the review for historical reasons.

Anti-Yeast Synergistic Effects and Mode of Action of Australian Native Plant Essential Oils

Yeasts are the most common group of microorganisms responsible for spoilage of soft drinks and fruit juices due to their ability to withstand juice acidity and pasteurization temperatures and resist the action of weak-acid preservatives. Food industries are interested in the application of natural antimicrobial compounds as an alternative solution to the spoilage problem. This study attempts to investigate the effectiveness of three Australian native plant essential oils (EOs) Tasmanian pepper leaf (TPL), lemon myrtle (LM) and anise myrtle (AM) against weak-acid resistant yeasts, to identify their major bioactive compounds and to elucidate their anti-yeast mode of action. The minimum inhibitory concentration (MIC), minimum fungicidal concentration (MFC) and minimum bactericidal concentration (MBC) were assessed for EOs against weak-acid resistant yeasts (Candida albicans, Candida krusei, Dekkera anomala, Dekkera bruxellensis, Rhodotorula mucilaginosa, Saccharomyces cerevisiae, Schizosaccharomyces pombe, Zygosaccharomyces bailii and Zygosaccharomyces rouxii) and bacteria (Staphylococcus aureus and Escherichia coli). The EOs showed anti-yeast and antibacterial activity at concentrations ranging from 0.03–0.07 mg/mL and 0.22–0.42 mg/mL for TPL and 0.07–0.31 mg/mL and 0.83–1.67 mg/mL for LM, respectively. The EOs main bioactive compounds were identified as polygodial in TPL, citral (neral and geranial) in LM and anethole in AM. No changes in the MICs of the EOs were observed in the sorbitol osmotic protection assay but were found to be increased in the ergosterol binding assay after the addition of exogenous ergosterol. Damaging of the yeast cell membrane, channel formation, cell organelles and ion leakage could be identified as the mode of action of TPL and LM EOs. The studied Australian native plant EOs showed potential as natural antimicrobials that could be used in the beverage and food industry against the spoilage causing yeasts.

Antimicrobial Activity of Nanoencapsulated Essential Oils of Tasmannia lanceolata, Backhousia citriodora and Syzygium anisatum against Weak-Acid Resistant Zygosaccharomyces bailii in Clear Apple Juice

The anti-yeast activity of oil-in-water encapsulated nanoemulsion containing individual or a combination of the three essential oils of Tasmanian pepper leaf (Tasmannia lanceolata), lemon myrtle (Backhousia citriodora), and anise myrtle (Syzygium anisatum) against weak-acid resistant Zygosaccharomyces bailii in clear apple juice was investigated. The effectiveness of the shelf-life extension of Z. bailii-spiked (1 × 103 CFU/mL) clear apple juice was evaluated and compared between natural (essential oils) and synthetic (sodium benzoate) antimicrobial agents. Essential oils showed an immediate reduction in the Z. bailii cell population at day-0 and exerted a fungicidal activity at day-4 of storage, with no further noticeable growth at the end of the experiment (day-28). At lower concentrations, Tasmanian pepper leaf oil of 0.0025% had >6 log CFU/mL at day-12 of storage. For lemon myrtle essential oils, the yeast population reached >6 log CFU/mL at day-24 and day-20 for concentrations of 0.02% and 0.01%, respectively. The fungicidal activity of Tasmanian pepper leaf oil reduced from 0.005% to 0.0025% v/v when mixed at a ratio of 1:1 with anise myrtle oil. The results of the present study suggest that these three native Australian herbs have the potential to be used in the beverage industry by controlling Zygosaccharomyces bailii in clear apple juice products.

Exploring Common Culinary Herbs and Spices as Potential Anti-Quorum Sensing Agents.

Quorum sensing controls bacterial pathogenesis and virulence; hence, interrupting this system renders pathogenic bacteria non-virulent, and presents a novel treatment for various bacterial infections. In the search for novel anti-quorum sensing (AQS) compounds, 14 common culinary herbs and spices were screened for potential antipathogenicity activity against Chromobacterium violaceum ATCC 12472. Extracts of Glycyrrhiza glabra (liquorice), Apium graveolens (celery), Capsicum annuum (cayenne pepper) and Syzygium anisatum (aniseed) demonstrated good AQS potential, yielding opaque halo zones ranging from 12–19 mm diameter at sub-minimum inhibitory concentrations (0.350–4.00 mg/mL). For the same species, the percentage reduction in violacein production ranged from 56.4 to 97.3%. Zones with violacein inhibitory effects were evident in a celery extract analysed using high performance thin layer chromatography-bio-autography. The major active compound was isolated from celery using preparative-high performance liquid chromatography-mass spectrometry and identified using gas chromatography-mass spectrometry (GC-MS) as 3-n-butyl-4,5-dihydrophthalide (sedanenolide). Potent opaque zones of inhibition observed on the HPTLC-bio-autography plate seeded with C. violaceum confirmed that sedanenolide was probably largely responsible for the AQS activity of celery. The bacteriocidal properties of many herbs and spices are reported. This study, however, was focussed on AQS activity, and may serve as initial scientific validation for the anti-infective properties ascribed to several culinary herbs and spices.

Mechanism of Action against Food Spoilage Yeasts and Bioactivity of Tasmannia lanceolata, Backhousia citriodora and Syzygium anisatum Plant Solvent Extracts.

Bioactive properties of solvent extracts of Tasmannia lanceolata, Backhousia citriodora and Syzygium anisatum investigated. The antimicrobial activities evaluated using agar disc diffusion method against two bacteria (Escherichia coli and Staphylococcus aureus) and six weak-acid resistant yeasts (Candida albicans, Candida krusei, Dekkera anomala, Rhodotorula mucilaginosa, Saccharomyces cerevisiae and Schizosaccharomyces pombe). The antioxidant activities determined using DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging and reducing power assays. Quantification of major active compounds using ultra-high performance liquid chromatography. Extracts showed broad-spectrum antifungal activity against weak-acid resistant yeasts in comparison to the standard antifungal agents, fluconazole and amphotericin B. Dekkera anomala being the most sensitive and strongly inhibited by all extracts, while Escherichia coli the least sensitive. Polygodial, citral and anethole are the major bioactive compounds identified in Tasmannia lanceolata, Backhousia citriodora and Syzygium anisatum, respectively. Hexane extracts contain the highest amount of bioactive compounds and demonstrate the strongest antimicrobial activities. Methanol and ethanol extracts reveal the highest phenolic content and antioxidant properties. Fluorescence microscopic results indicate the mechanism of action of Backhousia citriodora against yeast is due to damage of the yeast cell membrane through penetration causing swelling and lysis leading to cell death.

Biochemical and functional properties of indigenous Australian herbal infusions

The phytochemical profile, organic acid content, minerals, various antioxidant assays and consumers acceptability of indigenous Australian herbal infusions namely gulban (Melaleuca citrolens), anise myrtle (Syzygium anisatum), and lemon myrtle (Backhousia citriodora) were compared with a commercial green tea (Camellia sinensis). Total phenolic content and catechin derivatives were higher in green tea as compared to indigenous herbal infusions (P < 0.05). Phytochemical profiles showed high levels of caffeine in green tea, but, it was not found in herbal infusions (P < 0.05). Australian indigenous herbal infusions were a good source of calcium and magnesium compared to green tea (P < 0.05). Oxalic acid was higher in green tea, whereas gulban and anise myrtle infusions were rich in citric acid (P < 0.05). Antioxidant activities of green tea and gulban herbal infusions were comparable (P ≥ 0.05). Overall liking scores were higher for herbal infusions compared to green tea (P < 0.05). Indigenous Australian herbal infusions particularly gulban has a potential to become a successful commercial herbal beverage.

Formulation, characterisation and antibacterial activity of lemon myrtle and anise myrtle essential oil in water nanoemulsion

This study focussed on the formulation, characterisation of lemon myrtle (LM) and anise myrtle (AM) essential oil (EO) in water nanoemulsion and their antibacterial activity. The required hydrophilic lipophilic balance (rHLB) value of LM EO and AM EO was 14 and 12, respectively. The Central Composite Rotatable Design (CCRD) model produces the smallest droplet size and polydispersity index (PDI) for LMEO (d ≈ 16.07 nm; PDI ≈ 0.209) and AMEO (d ≈ 30.23 nm; PDI ≈ 0.216) at 1% EO and 10% surfactant mixture (Smix) ratio using ultrasonication for 5 min. Whereas, increased in EO, decrease in Smix concentrations and ultrasonication time produces higher droplet size of nanoemulsions. LMEO (LM-15, LM-17) nanoemulsions was clear and transparent compared to AMEO (AM-15, AM-17). All the selected nanoemulsions showed good stability at 4, 25 and 40 °C during storage, except LM-15 at 40 °C. LMEO nanoemulsion showed enhanced antibacterial activity compared to LMEO alone (P < 0.05).

Growth inhibitory properties of Backhousia myrtifolia Hook. &amp; Harv. and Syzygium anisatum (Vickery) Craven &amp; Biffen extracts against a panel of pathogenic bacteria

Growth inhibitory properties of Backhousia myrtifolia Hook. &amp; Harv. and Syzygium anisatum (Vickery) Craven &amp; Biffen extracts against a panel of pathogenic bacteria

Anti-inflammatory potential of native Australian herbs polyphenols

The anti-inflammatory potential of hydrophilic polyphenolic-rich extracts obtained from native Australian herbs: anise myrtle, lemon myrtle and Tasmannia pepper leaf, and a reference sample bay leaf, was evaluated using the lipopolysaccharide (LPS)-activated murine macrophage RAW 264.7 model. Pretreatment with all herbal extracts at non-cytotoxic concentrations reduced the LPS-induced protein levels of pro-inflammatory enzymes, cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS). Concomitant decrease in accumulation of their products, prostaglandin E2 (PGE2) and nitric oxide (NO), respectively, was observed. A suppression of LPS-induced expression of COX-2 and iNOS and decrease of NO and PGE2 levels suggests potential anti-inflammatory properties of the extracts.Anise myrtle, lemon myrtle and bay leaf selectively inhibited COX-2 and iNOS enzymes, while Tasmannia pepper leaf extract exhibited a pronounced inhibitory activity toward COX-1 and was the least effective inhibitor of iNOS. Anise myrtle and lemon myrtle are potentially more efficient anti-inflammatory agents than Tasmannia pepper leaf.

Effect of Packaging Materials and Storage on Major Volatile Compounds in Three Australian Native Herbs

Lemon myrtle, anise myrtle, and Tasmanian pepper leaf are commercial Australian native herbs with a high volatile or essential oil content. Packaging of the herbs in high- or low-density polyethylene (HDPE and LDPE) has proven to be ineffective in preventing a significant loss of volatile components on storage. This study investigates and compares the effectiveness of alternate high-barrier property packaging materials, namely, polyvinylidene chloride coated polyethylene terephthalate/casted polypropylene (PVDC coated PET/CPP) and polyethylene terephthalate/polyethylene terephthalate/aluminum foil/linear low-density polyethylene (PET/PET/Foil/LLDPE), in prevention of volatile compound loss from the three native herbs stored at ambient temperature for 6 months. Concentrations of major volatiles were monitored using gas chromatography-mass spectrometry (GC-MS) techniques. After 6 months of storage, the greatest loss of volatiles from lemon myrtle was observed in traditional LDPE packaging (87% loss) followed by storage in PVDC coated PET/CPP (58% loss) and PET/PET/Foil/LLDPE (loss of 23%). The volatile loss from anise myrtle and Tasmanian pepper leaf stored in PVDC coated PET/CPP and PET/PET/Foil/LLDPE packaging was <30%. This study clearly indicates the importance of selecting the correct packaging material to retain the quality of herbs with high volatile content.

Cytoprotective and pro-apoptotic activities of native Australian herbs polyphenolic-rich extracts

Three commercially grown native herbs unique to Australia, Tasmannia pepper leaf (Tasmannia lanceolata R. Br., Winteracea; TPL), anise myrtle (Syzygium anisatum Vickery, Craven & Biffen, Myrtaceae; AM) and lemon myrtle (Backhousia citriodora F. Muell, Myrtaceae; LM) as well as a reference sample bay leaf (Laurus nobilis L., Lauraceae; BL) were examined for potential cytoprotective properties. All native herbs exhibited greater cellular antioxidant activity as measured by the cellular antioxidant activity (CAA) assay than bay leaf and reduced the hydrogen peroxide (H2O2) induced death of hepatocellular carcinoma (HepG2) cells by 25–50%. All herb extracts reduced the proliferation of colon (HT-29; IC50=0.75–1.39mg/ml), stomach (AGS; IC50=0.59–1.88mg/ml), bladder (BL13; IC50=0.56–1.12mg/ml) and liver (HepG2; IC50=0.38–1.36mg/ml) cancer cells. No significant reduction of cell viability of non-transformed colon (CCD-18Co; IC50>2.0mg/ml) and mixed stomach and intestine (Hs 738.St/Int; IC50>2.0mg/ml) cells was observed. Flow cytometry analysis and the results of the cytokinesis block micronucleus cytome (CBMNCyt) assay conducted with respectively, promyelocytic leukaemia (HL-60) and colon adenocarcinoma (HT-29) cells suggest an increase in apoptosis following treatment with the herb extracts. The occurrence of apoptotic cells coincided with an increase in caspase-3 enzyme activity. The results of the CBMNCyt assay suggested no direct DNA damage in colon adenocarcinoma (HT-29) cells as a result of treatment with all extracts, applied at final concentrations of 0.5 and 1.0mg/ml.

Composition of native Australian herbs polyphenolic-rich fractions and in vitro inhibitory activities against key enzymes relevant to metabolic syndrome

Polyphenolic-rich fractions obtained from three native Australian herbs: Tasmannia pepper leaf, anise myrtle and lemon myrtle were characterised with regards to their composition, antioxidant capacities and inhibitory activities against α-glucosidase, pancreatic lipase and angiotensin I-converting enzyme, using in vitro models. Ellagic acid and derivatives were the dominant compounds of anise myrtle and lemon myrtle fractions, accompanied by flavonoids (catechin, myricetin, hesperetin, and quercetin). Tasmannia pepper leaf fraction comprised chlorogenic acid and quercetin derivatives, exhibited the highest oxygen radical absorbance capacity and effectively inhibited α-glucosidase (IC(50): 0.83 mg/ml) and pancreatic lipase (IC(50): 0.60 mg/ml). Anise myrtle and lemon myrtle fractions had pronounced α-glucosidase-inhibitory activities (IC(50): 0.30 and 0.13 mg/ml, respectively) and were less effective against lipase. Enzyme-inhibitory activities showed various levels of correlation with the levels of total phenolics and antioxidant capacities, indicating a specificity of individual phenolic compounds present in the isolated fractions to complex with proteins.

Antioxidant capacity and phenolic compounds in commercially grown native Australian herbs and spices

Abstract The antioxidant capacities and phenolic composition in six native, commercially grown, Australian herbs and spices were investigated. Tasmannia pepper leaf, followed by anise myrtle and lemon myrtle contained the highest levels of total phenolics (TP; 102.1; 55.9 and 31.4 mg gallic acid equivalents (GAE)/g dry weight (DW), respectively). Tasmannia pepper leaf exhibited the highest oxygen radical absorbance capacity (ORAC assay) followed by lemon myrtle and anise myrtle. Anise myrtle exhibited the highest total reducing capacity [TRC; Ferric Reducing Antioxidant Power (FRAP) assay], followed by Tasmannia pepper leaf and lemon myrtle. Australian bush tomato, with TP content of 12.4 ± 0.9 mg GAE/gDW and TRC of 206.2 μMol Fe+2/gDW, resembled the Chinese Barbary Wolfberry fruit. The TP content of Tasmannia pepper berry (16.86 mg GAE/gDW) was similar to that of black pepper, but it’s TRC was 25% lower. Cinnamic acids and flavonoids, tentatively identified by mass spectrometry, were identified as the main sources of antioxidant activities.

Evaluation of the Phytochemical Anethole as an Anti-Tumor Agent in MCF-7 Cells.

Abstract Background: Chinese herbal medicines have gained considerable interest from the cancer community as potential anti-tumor agents. Both clinical as well as pre-clinical studies have demonstrated that certain Chinese herbal medicines can improve survival, increase tumor response, improve quality of life, and reduce chemotherapy toxicity. The specific mechanisms by which these compounds mediate their effects remain largely unknown, inhibiting their development as clinical agents. One such compound is anethole. Anethole is an aromatic compound that occurs widely in nature as a component of anise, fennel anise myrtle, licorice, and star anise and is used in traditional Chinese medicine as an anti-inflammatory agent. Anethole has been identified as one of the potential phytochemicals with anti-cancer activities that could be developed into clinical treatments for breast cancers.Objective: In this current study, we evaluated the effect of anethole on physiological responses and specific survival pathways in human breast cancer MCF-7 cells.Results: Cell proliferation and cell survival were suppressed in a dose-dependent manner in response to anethole treatment, but these dosages had differential effect on specific pro-survival pathways. Anethole increased the cleavage of Poly (ADP-ribose) polymerase (PARP), suggesting an increase in apoptotic activity, but concurrently increased the expression of pro-survival proteins Bcl-2 and p21 while decreasing the expression of tumor suppressor protein p53. Phosphorylation of Akt kinase and S6 pro-survival proteins were modulated in an alternating pattern with increasing concentrations of anethole. Studies were also conducted to investigate the effect of anethole on the sensitivity of MCF-7 cells to tamoxifen treatments. The effect of tamoxifen on cell proliferation was not affected significantly by co-treatment with anethole but importantly the suppression of cell survival by tamoxifen was enhanced when anethole is present at a concentration of 10-5 M. Preliminary data suggest that one mechanism by which anethole is mediating some of its suppressive effects is through modulation of ER activity, as measured by luciferase assays. Studies are on-going to evaluate the relevance of this modulation to the inhibitory effects of anethole on the MCF-7 cells, as well as the modulation of the signal transduction pathways indicated on survival.Conclusion: These data suggest that the anethole is a potential anti-tumor agent, and that co-treatment of tamoxifen and anethole is a potential approach to sensitize human breast MCF-7 cells to tamoxifen treatments. Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3100.