Composition and antioxidant and antimicrobial activity of the essential oil and extracts of Stachys inflata Benth from Iran

Composition and antioxidant and antimicrobial activities of essential oil and methanol extract polar and nonpolar subfractions of Stachys inflata were determined. GC and GC/MS analyse of the essential oil showed 45 constituents representing 95.46% of the oil, the major components linalool (28.55%), α-terpineol (9.45%), spathulenol (8.37%) and (2E)-hexenal (4.62%) constituted 50.99% of it. Essential oil and extracts were also tested for their antioxidant activities using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and β-carotene/linoleic acid assays. In the DPPH test, IC₅₀ value for the polar subfraction was 89.50 μg/ml, indicating an antioxidant potency of about 22% of that of butylated hydroxytoluene (IC₅₀ = 19.72 μg/ml) for this extract. In β-carotene/linoleic acid assay, the best inhibition belonged to the nonpolar subfraction (77.08%). Total phenolic content of the polar and nonpolar extract subfractions was 5.4 and 2.8% (w/w), respectively. The plant also showed a week antimicrobial activity against three strains of tested microorganisms. Linalool and α-terpineol were also tested as major components of the oil and showed no antioxidant but considerable antimicrobial activities.     

Antioxidant activity and contents of essential oil and phenolic compounds in flowers and seeds of Alpinia zerumbet (Pers.) B.L. Burtt. & R.M. Sm

Alpinia zerumbet leaves and rhizomes have been extensively studied for their chemical compositions and biological activities. However, less attention has been given to its flowers and seeds. In our study, essential oil, total phenolics and antioxidant capacities assayed by 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging and β-carotene bleaching methods were evaluated in flowers and seeds of A. zerumbet. In addition, their phenolic composition was determined by GC–MS and HPLC. 1,8-Cineol, camphor, methyl cinnamate and borneol were the major constituents in flower oils, whereas the main components in seeds oil were α-cadinol, T-muurolol, α-terpineol, δ-cadinene and terpinene-4-ol. The results showed that the hexane extract of flowers contained a significantly higher quantity of dihydro-5,6-dehydrokawain (DDK) than that of seeds. Total phenolic contents of flower and seed extracts were measured as 56.7 and 13.7 mg gallic acid equivalent per gram extract, respectively. The ethyl acetate extract of flowers and seeds possessed a high antiradical activity and prevented the bleaching of β-carotene. The HPLC analysis indicated that p-hydroxybenzoic acid, ferulic acid and syringic acid were the predominant phenolics in the ethyl acetate extract of flowers, whilst p-hydroxybenzoic acid, syringic acid and vanillin were the major phenolics in seeds.     

Antioxidant activity and chemical constituents of essential oil and extracts of Rhizoma Homalomenae

Antioxidant activity and composition of essential oil and extracts of Rhizoma Homalomenae were determined. The extracts, especially the ethyl acetate extract (QJ4 fraction) of the aqueous residue after oil distillation, had considerable antioxidant potency which was significantly associated with their total phenolic and flavonoid contents, but the essential oil showed only weak or moderate activity. GC–MS analysis of the essential oil (yield: 0.82%, v/w) resulted in the identification of 77 compounds, accounting for 96.5% of the content of the oil. The major components, epi-α-cadinol (14.8%), α-cadinol (14.8%), α-terpineol (13.8%), linalool (11.1%), terpinen-4-ol (4.92%), and δ-cadinene (4.91%) constituted 64.3% of it. LC–MS/MS and HPLC analyses showed seven phenolic compounds (protocatechuic acid, vanillic acid, syringic acid, caffeic acid, p-coumaric acid, ferulic acid and apigenin) with a great amount in the ethyl acetate extract (QJ4 fraction). The strong antioxidant properties of the plant extracts may be attributed to the presence of these phenolics.     

Chemical composition, in vitro cytotoxic and antioxidant activities of the essential oil and major constituents of Cymbopogon jawarancusa (Kashmir)

The chemical composition of the hydrodistillate of aerial parts of Cymbopogon jawarancusa, a natural grass considered as major forage for animal nutrition, used in food because of the presence of sufficient concentration of minerals like calcium and potassium was analysed by capillary GC–FID, GC–MS and ¹³C NMR. Seventeen constituents representing 97.8% of the total oil with piperitone (58.6%) and elemol (18.6%) as major constituents were identified. In vitro cytotoxicity of the oil and its constituents on human cancer cell lines THP-1 (leukemia), A-549 (lung), HEP-2 (liver) and IGR-OV-1 (ovary) was evaluated by Sulphorhodamine-B assay. The oil was found to be more potent than its components against cancer cell lines tested with IC₅₀ of 6.5 μg/ml (THP-1), 6.3 μg/ml (A-549), 7.2 μg/ml (HEP-2) and 34.4 μg/ml (IGR-OV-1). Antioxidant activity of oil and its constituents was evaluated by DPPH assay. In conclusion, the results demonstrate potent cytotoxic and antioxidant effects of oil, and its components like piperitone, α-pinene, β-caryophyllene and β-elemene.     

Antimicrobial and antioxidant properties of the essential oils and methanol extract from Mentha longifolia L. ssp. longifolia

This study was designed to evaluate antimicrobial and antioxidant activities of the essential oil and methanol extract from Mentha longifolia ssp. longifolia. The essential oil showed strong antimicrobial activity against all 30 microorganisms tested whereas the methanol extract almost remained inactive. In contrast, the extract showed much better activity than the essential oil in antioxidant activity assays employed, e.g. in the inhibition of free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) and β-carotene/linoleic acid systems. In the former, the extract was able to reduce the stable free radical DPPH with an IC₅₀ of 57.4 μg/ml while that of the oils was 10 700 μg/ml. When compared to BHT, a synthetic antioxidant, both showed weaker antioxidative potential. Similarly, in β-carotene/linoleic acid assay, these samples were not effectively able to inhibit the linoleic acid oxidation; exhibiting only 24% and 36% inhibitions at 2 mg/ml, respectively; both were far below than that of BHT. Total phenolic constituent of the extract was 4.5 g/100 g as gallic acid equivalent. GC–MS analysis of the oil resulted in the identification of 45 constituents, cis-piperitone epoxide, pulegone and piperitenone oxide being the main components.     

Study on chemical composition and biological activities of essential oil and extract from Salvia pisidica

In this study, total contents of phenolic, flavanol and flavonol, antioxidant activities and antimicrobial activities of the Turkish endemic Salvia pisidica Boiss. & Heldr. ex Bentham (Lamiaceae) extract and essential oil were assessed in vitro. Total phenolic, flavanol and flavonol contents in the extract were 54.57 mg gallic acid equivalents (GAE)/g, 16.70 mg catechine equivalents (CE)/g and 18.19 mg rutin equivalents (RE)/g, respectively. Antioxidant activities (IC₅₀ value) of the extract and essential oil were determined as 4.88 and 6.41 mg/mL by DPPH assay, respectively. 31 compounds were determined in the essential oil using GC-MS and the major compounds (%) were camphor (23.76), sabinol (19.2), α-thujone (14.2) and eucalyptol (1.8-cineole) (5.8).The antimicrobial activity of the methanolic extract and the essential oil against 13 bacterial and two yeast strains was determined. The extract (concentration 5 g/100 ml or 10 g/100 ml) was effective against most of the strains tested, yet not against Bacillus cereus, Staphylococcus aureus, Aeromonas hydrophila and the two yeast strains tested. The essential oil (2 g/100 ml) showed an antimicrobial effect against all the gram (+) bacteria tested, against Saccharomyces cerevisiae, but was not effective against all gram (−) bacteria and Candida albicans. These results show that S.piscidica essential oil and extract could be considered as a natural alternative to traditional food preservatives and be used to enhance food safety and shelf life.     

Chemical composition and antimicrobial activity of essential oil of Tarchonanthus camphoratus

The essential oil of Tarchonanthus camphoratus (Asteraceae), obtained by hydro-distillation, was analysed by gas chromatography–mass spectrometry (GC–MS) and also evaluated for antimicrobial activity. Out of 45 peaks representing 99.8% of the oil, 38 components which constitute 95.8% of the total oil were identified. The oil was dominated by monoterpenes, which accounted for 80.9% of the oil. This study indicates the presence of a high percentage of oxygenated monoterpenes (62.3%), of which the main constituents were fenchol (15.9%), 1,8-cineole (14.3%) and α-terpineol (13.2%). Other monoterpenes present in fairly good amounts were α-pinene (6.87%), trans-pinene hydrate (6.51%), terpinen-4-ol (4.74%) and camphene (3.76%). The oil was screened for antimicrobial activity against both Gram positive (Staphylococcus aureus, Bacillus ssp.) and Gram negative (Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi, Klebsiella pneumoniae, Proteus mirabilis) bacteria and a pathogenic fungus Candida albicans. Except for P. aeruginosa, which showed resistance, the oil had pronounced antibacterial and antifungal activities.     

Integrated process for co-production of alkaline lipase and R-(+)-α-terpineol by Fusarium oxysporum

This article complements a series of studies on the strain Fusarium oxysporum 152b describing an extracellular alkaline lipase production and limonene biotransformation. In this study, it was shown that the enzyme responsible for the latter process is enantioselective and enantiospecific for the biotransformation of R-(+)-limonene into R-(+)-α-terpineol, has an intracellular nature and acts in anaerobic conditions. Also, it was demonstrated that the biomass stored in frozen or lyophilised forms could be used for α-terpineol production, where the reaction rate was significantly increased in the first case. Finally, an integrated process for the co-production of lipase and α-terpineol was proposed and, although the maximal α-terpineol concentration (∼2 g l⁻¹) was ∼50% lower than the conventional process (∼4 g l⁻¹), this is an interesting methodology from an industrial point of view and encourages future research in this field.     

Supercritical fluid extraction and hydrodistillation for the recovery of bioactive compounds from Lavandula viridis L’Hér

The chemical profiles of bioactive essential oil and extracts obtained by hydrodistillation (HD) and supercritical fluid extraction (SFE), respectively, from Lavandula viridis were compared. The SFE was performed at 40 °C and at extraction pressures of 12 or 18 MPa in two different separators. Evaluation of the essential oil and SFE extracts by GC–FID and GC–IT–MS revealed that oxygen-containing monoterpenes were the major constituents in both cases, but there were important differences between the chemical profiles produced by the different extraction techniques. More compounds were isolated by HD but higher yields were achieved by SFE. Camphor was the main component identified in the essential oil (31.59 ± 1.32%), and in extracts from the first (1.61 ± 0.34%) and second SFE separators (22.48 ± 1.49%) at 12 MPa. In contrast, the first separator SFE extract at 18 MPa (heavy compounds) was dominated by myrtenol (5.38 ± 2.04%) and camphor (4.81 ± 1.93%), whereas the second separator SFE extract (volatiles) was dominated by verbenone (13.97 ± 5.27%). The essential oil and heavy compound extracts from the first separator possessed antioxidant and anti-cholinesterase activities. Our data show that phytochemicals from the aerial parts of L. viridis could be developed as natural antioxidant and anti-cholinesterase drugs, with particular applications in the symptomatic treatment of Alzheimer’s disease.     

Antibacterial activity of the essential oils of Salvia officinalis L. and Salvia triloba L. cultivated in South Brazil

The essential oils of Salvia officinalis and Salvia triloba cultivated in South Brazil were analyzed by GC–MS. The major constituents of the oil of S. officinalis were α-thujone, 1,8-cineole, camphor, borneol and β-pinene, whereas those of S. triloba were α-thujone, 1,8-cineole, camphor, and β-caryophyllene. The essential oils of both species exhibited remarkable bacteriostatic and bactericidal activities against Bacillus cereus, Bacillus megatherium, Bacillus subtilis, Aeromonas hydrophila, Aeromonas sobria, and Klebsiella oxytoca. Moreover, the essential oil of S. triloba efficiently inhibited the growth of Staphylococcus aureus. S. aureus and A. hydrophila growth were drastically reduced even in the presence of 0.05 mg/ml of the essential oil of S. triloba.     

Fingerprint of volatile flavour constituents and antioxidant activities of teas from Thailand

The volatile flavour components of different teas growing in Thailand were extracted using the simultaneous distillation and extraction (SDE) technique. These volatiles were investigated by GC–MS. At least 54 components representing 76.51–83.32% of all samples were identified. Hotrienol, geraniol and linalool were found to be the major components in Green Oolong tea. Green Assam tea contained linalool, geraniol and α-terpineol as the key flavour constituents. Chin Shin Oolong tea was dominated by linalool, indole and cis-jasmone whilst the major flavour volatiles of Chin Hsuan Oolong tea were trans-nerolidol, cis-jasmone and geraniol. Indole, geraniol and cis-jasmone were detected as the main constituents in Four Season tea. Change of quality and quantity of volatile flavour components was related to fermentation methods that increased volatiles were illustrated by the semi-fermented tea processing method. Green Assam tea infusion extract was evaluated to have the strongest antioxidant activities with the highest amount of phenol content followed by Four Season tea, Chin Shin Oolong tea, Chin Hsuan Oolong tea and Green Oolong tea, respectively.     

Antioxidative activity of Rosmarinus officinalis L. essential oil compared to its main components

This study was designed to examine the in vitro antioxidant activities of Rosmarinus officinalis L. essential oil compared to three of its main components (1,8-cineole, α-pinene, β-pinene). GC–MS analysis of the essential oil resulted in the identification of 19 compounds, representing 97.97% of the oil, the major constituents of the oil were described as 1,8-cineole (27.23%), α-pinene (19.43%), camphor (14.26%), camphene (11.52%) and β-pinene (6.71%). The oil and the components were subjected to screening for their possible antioxidant activity by means of 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay and β-carotene bleaching test. In the DPPH test system, free radical-scavenging activity of R. officinalis L. essential oil, 1,8-cineole, α-pinene and β-pinene were determined to be 62.45% ± 3.42%, 42.7% ± 2.5%, 45.61% ± 4.23% and 46.21% ± 2.24% (v/v), respectively. In the β-carotene bleaching test system, we tested series concentration of samples to show the antioxidant activities of the oil and its main components, whereas the concentrations providing 50% inhibition (IC₅₀) values of R. officinalis L. essential oil, 1,8-cineole, α-pinene and β-pinene were 2.04% ± 0.42%, 4.05% ± 0.65%, 2.28% ± 0.23% and 2.56% ± 0.16% (v/v), respectively. In general, R. officinalis L. essential oil showed greater activity than its components in both systems, and the antioxidant activities of all the tested samples were mostly related to their concentrations. Antioxidant activities of the synthetic antioxidant, ascorbic acid and BHT, were also determined in parallel experiments as positive control.     

Chemical fingerprinting and bioactivity of Amazonian Ecuador Croton lechleri Müll. Arg. (Euphorbiaceae) stem bark essential oil: A new functional food ingredient?

Croton lechleri essential oil has been obtained by steam distillation of fresh stem bark from Amazonian Ecuador adult plants (yield: 0.61 ml/kg [0.061%]; density: 1.01 g/ml), and then chemically characterised by GC (Gas Chromatography) and GC–MS (gas chromatography–mass spectrometry). Seventy-four chemicals were detected and identified; the most abundant in descending order, were the sesquiterpenes sesquicineole (17.29%), α-calacorene (11.29%), 1,10-di-epi-cubenol (4.75%), β-calacorene (4.34%) and epi-cedrol (4.09%). Monoterpenes checked with a relative peak area higher than 2.0% were α-pinene (2.01%), p-cymene (2.61%), limonene (4.20%) and borneol (2.67%). The structure of the main chemicals were confirmed by GC–MS and ¹H NMR analyses. Spectrophotometric 1,1-diphenyl-2-picrylhydrazyl (DPPH) and DPPH-(high performance) thin layer chromatography (DPPH-(HP)TLC) bioautographic assays showed a lower radical scavenging capacity (IC₅₀) with respect to commercial thyme essential oil and BHA (butylated hydroxyl anisole), pointing out, however, that the C. lechleri essential oil fraction, characterised by α-calacorene, β-calacorene and δ-cadalene, was the most involved in the bioactivity. Similar results were obtained with β-carotene bleaching assay, where the IC₅₀ values were 0.291 ± 0.024 mg/ml for C. lechleri essential oil, 0.164 ± 0.013 and 1.34 × 10⁻⁴ ± 10⁻⁵ mg/ml for thyme essential oil and BHA, respectively. (HP)TLC-bioautographic assay performed with Gram positive and Gram negative bacteria revealed a minimum inhibitory concentration (MIC) values comprised between 0.10 mg/ml (Escherichia coli) and 10.10 mg/ml (for e.g. Pseudomonas aeruginosa), and the fraction mainly characterised by sesquicineole (97.38%) as the most involved in antibacterial capacity. Ames test employing Salmonella typhimurium TA98 and TA100 with and without a metabolic activation mixture (S9 mix) demonstrated the absence of mutagenicity of the C. lechleri essential oil between a concentration range of 10⁻² and 100 mg/plate. The same results were achieved by Saccharomyces cerevisiae D7 strain assay. An interesting mutagen-protective efficacy was evidenced by a 30% and 33% revertants reduction of TA98 strain treated with 2-aminoanthracene and nitrofluorene (2 μg/plate), suggesting, above all, the possibility to employ C. lechleri essential oil as a new flavouring protective ingredient for foods or dietary supplements against potential mutagens formed during cooking and/or processing in general.     

Comparative evaluation of the antibacterial activities of the essential oils of Rosmarinus officinalis L. obtained by hydrodistillation and solvent free microwave extraction methods

Rosmarinus officinalis L. is a perennial herb that belongs to the Lamiaceae family. It is used as a food flavouring agent, and well known medicinally for its powerful antimutagenic, antibacterial and chemopreventive properties. Essential oils were obtained from this plant by hydrodistillation (HD) and solvent free microwave extraction (SFME). GC–MS analyses of the oils revealed the presence of 24 and 21 compounds in the essential oils obtained through HD and SFME, respectively. The total yield of the volatile fractions obtained through HD and SFME was 0.31% and 0.39%, respectively. Higher amounts of oxygenated monoterpenes such as borneol, camphor, terpene-4-ol, linalool, α-terpeneol (28.6%) were present in the oil of SFME in comparison with HD (26.98%). However, HD oil contained more monoterpene hydrocarbons such as α-pinene, camphene, β-pinene, myrcene, α-phellanderene, 1,8-cineole, trans β-ocimene, γ-terpenene, and cis sabinene hydrate (32.95%) than SFME extracted oil (25.77%). The essential oils obtained using the two methods of extraction were active against all the bacteria tested at a concentration of 10 mg ml⁻¹. Minimum inhibitory concentration (MIC) values for all the susceptible bacteria ranged between 0.23 mg ml⁻¹ and 7.5 mg ml^−1.     

Effect of lyophilized water extracts of Melissa officinalis on the stability of algae and linseed oil-in-water emulsion to be used as a functional ingredient in meat products

Previous work pointed out the possibility to enhance the nutritional value of meat products using long chain ω − 3 PUFA enriched emulsions. Oil-in-water emulsions elaborated with a mixture of algae and linseed oils (15:10) in order to be used as functional ingredient were stabilized with BHA (butylhydroxyanisol) or with a lyophilized water extract of Melissa officinalis L. (Lemon balm). The lipid profile of the oil mixture showed a high amount of DHA (31.7%), oleic (25.4%) and α-linolenic acid (12.7%) resulting in a very low ω − 6/ω − 3 ratio (0.12). The lyophilized extract of M. officinalis showed a high antioxidant activity (being 62 ppm of the lyophilized water extract of Melissa equivalent to 200 ppm of BHA, using the DPPH assay as reference), and high total phenolic content. Studying the oxidation process in the emulsions during 15 days at room temperature, it could be concluded that this extract was as efficient as BHA in order to control the thiobarbituric acid reactive substances (TBARS) formation.     

Neuroprotective potential of some terebinth coffee brands and the unprocessed fruits of Pistacia terebinthus L. and their fatty and essential oil analyses

In the current study, neuroprotective effects of the ethyl acetate and methanol extracts of four terebinth coffee brands and the fruits of Pistacia terebinthus L. were investigated through enzyme inhibition tests against acetylcholinesterase, butyrylcholinesterase, and tyrosinase as well as antioxidant test systems. Antioxidant activity was measured using radical scavenging activity tests and metal-related tests including metal-chelation capacity, ferric-reducing antioxidant power (FRAP), and phosphomolybdenum reducing power (PRAP). The fatty oils of the coffee brands and the fruits and the fruit essential oil were examined by GC–MS. Total phenol and flavonoid contents were calculated spectrophotometrically. The extracts had moderate inhibition against butyrylcholinesterase (9.78–45.74% at 200 μg mL⁻¹) and potent scavenging activity against DPPH. They exerted strong activity in FRAP and metal-chelation tests and modest activity in PRAP test. Oleic acid was identified as the major fatty acid in the fatty oils, while α-pinene (26.31%) was dominant in the essential oil.     

Chemical composition,antimicrobial, cytotoxic and antioxidant activities of the essential oil of Artemisia indica Willd

Essential oil from the aerial parts of Artemisia indica was analysed by GC-FID and GC–MS. A total of 43 compounds representing 96.8% of the oil were identified and the major components were found to be artemisia ketone (42.1%), germacrene B (8.6%), borneol (6.1%) and cis-chrysanthenyl acetate (4.8%). Antimicrobial activity of the oil was evaluated against seven clinically significant bacterial and two fungal strains. The essential oil and its major constituents exhibited moderate to potent, broad-spectrum antibacterial and antifungal activities targeting both Gram-positive and Gram-negative bacteria. In vitro cytotoxicity evaluation against four human cancer cell lines THP-1 (leukemia), A-549 (lung), HEP-2 (liver) and Caco-2 (colon) showed that the essential oil exhibited concentration dependant growth inhibition in the 10–100 μg/ml dilution range, with IC₅₀ values of 10 μg/ml (THP-1), 25 μg/ml (A-549), 15.5 μg/ml (HEP-2) and 19.5 μg/ml (Caco-2). It was interesting to note that the essential oil also exhibited potent antioxidant activity.     

Antioxidant and antiacetylcholinesterase activities of essential oils from Cymbopogon schoenanthus L. Spreng. Determination of chemical composition by GC–mass spectrometry and C NMR

Cymbopogon schoenanthus L. Spreng, is an aromatic herb consumed in salads and used to prepare traditional meat recipes in Tunisia. The chemical composition, antioxidant activities and acetylcholinesterase inhibitory properties of the essential oils from fresh leaves, dried leaves and roots collected from three different locations in southern Tunisia, were evaluated. Essential oils were analysed by GC–mass spectrometry and ¹³C NMR. The major components were limonene (10.5–27.3%), β-phellandrene (8.2–16.3%), δ-terpinene (4.3–21.2%) and α-terpineol (6.8–11.0%). Antioxidant activity was measured by DPPH assay. The results ranged from 36.0% to 73.8% (2 μl of essential oil per mL of test solution).     

Chemical and biological characteristics of Cuminum cyminum and Rosmarinus officinalis essential oils

Essential oils extracted by hydrodistillation from Cuminum cyminum and Rosmarinus officinalis were characterized by means of GC and GC–MS. C. cyminum and R. officinalis contained α-pinene (29.1%, 14.9%), 1,8-cineole (17.9%, 7.43%) and linalool (10.4%, 14.9%), respectively, as the major compounds. C. cyminum oil exhibited stronger antimicrobial activity than did R. officinalis oil against E. coli, S. aureus and L. monocytogenes. Complete death time on exposure to Cuminum cyminum L. and Rosmarinus officinalis L. oils were 20 and 25 min 180 and 240 min and 90 and 120 min for E. coli, S. aureus and L. monocytogenes, respectively. Radical-scavenging and antioxidant properties were tested by means of 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay and the β-carotene bleaching test. These properties were compared to those of Thymus x-porlock essential oil, used as a reference ingredient. The radical scavenging performance of the rosemary oil was better than that of C. cyminum. Results from the antioxidant test were better than those provided by the radical-scavenging activity. C. cyminum and R. officinalis essential oils may be considered as potent agents in food preservation.     

Inhibitory effect of Turkish Rosmarinus officinalis L. on acetylcholinesterase and butyrylcholinesterase enzymes

In the current study, we have tested acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory activity of the petroleum ether, ethyl acetate, chloroform, and methanol extracts, rosmarinic acid as well as the essential oil obtained from Rosmarinus officinalis L. growing in Turkey by a spectrophotometric method of Ellman using ELISA microplate-reader at 0.2, 0.5, and 1.0 mg/mL concentrations. In addition, quantification of rosmarinic acid, a common phenolic acid found in rosemary, was carried out by reversed-phase HPLC in the methanolic extract of the plant, which was found to have 12.21 ± 0.95% (122.1 ± 9.5 mg/g extract) of rosmarinic acid. Rosmarinic acid was also tested for its AChE and BChE inhibitory effect and found to cause 85.8% of inhibition against AChE at only 1.0 mg/mL. Besides, the essential oil was analyzed by GC–MS technique, which was shown to be dominated by 1,8-cineol (44.42%) and followed by α-pinene (12.57%).