Designing monographs for Rosmarinus officinalis L. and Lavandula angustifolia L.: Two Lebanese species with significant medicinal potentials

The world health organization recommends validating medicinal plants to ensure safety and efficacy. Lavandula is valued for the production of essential oil used in aromatherapy. Rosmarinus is known for its folk use as anti-oxidant and antidiabetic. This study aims to design monographs for Rosmarinus officinalis (Linn.) and Lavandula angustifolia (Linn.) cultivated, in conditions similar to their natural habitat, at Beirut Arab University botanical garden. Also screening the antioxidant activity of both plants, and assessing the antidiabetic effect of R. officinalis (Linn.).  macroscopic and microscopic examination, hydrodistillation to obtain essential oils (EOs), thin layer chromatography, Gas chromatography-Mass spectrometry, and physiochemical properties of the EOs were determined. The antioxidant effect of the EOs is determined by DPPH assay. The antidiabetic effect of Rosemary EO is investigated using alloxan-induced diabetic mice.  the qualities of studied species are in agreement with the standards described in WHO monographs and in the British pharmacopeia. Composition of the EO produced from either Rosemary (REO) or Lavender (LEO) collected at different months showed differences. The EO of both showed significant antioxidant effects in DPPH assay (p<0.05). In vivo investigation of the antidiabetic effect of REO showed a significant antidiabetic effect in diabetic mouse model (p<0.05).  both plant species and their EOs fulfilled most of standards specified by the British Pharmacopeia 2013. Rosemary collected in April (Spring) showed the best antioxidant activity. Lavender collected in September (Early Fall) showed the best antioxidant activity. REO revealed significant lowering in blood glucose level at the dose of 110 mg/kg.

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title: Quality of herbal medicines: challenges and solutions.
abstract: The popularity of herbal medicines has risen worldwide. This increase in usage renders safety issues important. Many adverse events of herbal medicines can be attributed to the poor quality of the raw materials or the finished products. Different types of herbal medicines are associated with different problems. Quality issues of herbal medicines can be classified into two categories: external and internal. In this review, external issues including contamination (e.g. toxic metals, pesticides residues and microbes), adulteration and misidentification are detailed. Complexity and non-uniformity of the ingredients in herbal medicines are the internal issues affecting the quality of herbal medicines. Solutions to the raised problems are discussed. The rigorous implementation of Good Agricultural and Collection Practices (GACP) and Good Manufacturing Practices (GMP) would undoubtedly reduce the risk of external issues. Through the use of modern analytical methods and pharmaceutical techniques, previously unsolved internal issues have become solvable. Standard herbal products can be manufactured from the standard herbal extracts.

title: Availability and needs of herbal medicinal information resources at community pharmacy, Riyadh region, Saudi Arabia.
abstract: A cross-sectional survey of community pharmacists in Riyadh region, Saudi Arabia was conducted over a period of 6 months from July through December 2011. Data collection was carried out using a structured self-administered questionnaire. The survey questionnaire consisted of a brief introduction to the study and eleven questions. The questions consisted of close ended, multiple-choice, and fill-in short answers. A stratified random sample of one thousand and seven hundred registered pharmacy practitioners all over Saudi Arabia were randomly chosen to respond to the survey. The data from each of the returned questionnaire were coded and entered into Statistical Package for the Social Sciences (SPSS) version 19 software (SPSS Inc., Chicago, IL, USA) which was used for statistical analysis. Only one thousand four hundred one pharmacists responded to the survey (response rate is 82.4%) with a completely answered questionnaire. The study results show that 59.7% of the participants sometimes discuss herbal medicine use with their patients, while only 4.25% never discuss it. The study shows 48.5% of participated pharmacists record herbal medicine use sometimes where only 9.4% of them never did so. However, with regard to initiation of the discussion, the study shows that 44.3% of the respondents reported that patients initiate herbal issue discussion while 20.8% reported that pharmacists initiate the discussion. This discussion was reported to be a one time discussion or an ongoing discussion by 14.3% or 9.9% of the respondents respectively. According to the study results, respondents reported that the most common barriers that limit discussing herbal medicines' use with their patients were lack of time due to other obligations assigned to the community pharmacist (46%), lack of reliable resources (30.3%), lack of scientific evidence that support herbal medicine use (15.2%), or lack of knowledge of herbal medicines (13.4%). Yet, a small number of respondents was concerned about interest in herbal medicines (9.1%) and other reasons (2.4%). So it is urgent to ensure that pharmacists are appropriately educated and trained. Extra efforts are needed to increase the awareness of pharmacists to adverse drug reactions reporting system at Saudi Food and Drug Authority. Finally, more consideration to herbal issues should be addressed in both pharmacy colleges' curricula and continuous education program..

title: Future visions for traditional and herbal medicinal products--a global practice for evaluation and regulation?
abstract: Medicinal plants and traditional medicines have been used worldwide since ancient times. Currently, there is neither a globally consented terminology nor a harmonized regulatory approach. Nevertheless, it is common sense that quality, efficacy and safety should be assessed following scientific standards, addressing particulars and considering an adequate level of risk management. A global market for traditional medicines is emerging, if not already existing. Therefore, a constructive communication about regulatory systems for herbal and traditional medicinal products should be enforced. Best practice standards might be developed according to current scientific knowledge in order to improve mutual acceptance of data, sets of monographs and assessments. Overall, a convergence of the diverse regulatory systems might save resources and lead to an adequate availability of herbal and traditional medicinal products to the patients without neglecting public health.

title: In vivo assessment of antidiabetic and antioxidant activities of rosemary (Rosmarinus officinalis) in alloxan-diabetic rabbits.
abstract: Rosemary (Rosmarinus officinalis), used in traditional Turkish folk medicine for the treatment of hyperglycaemia, is widely accepted as one of the medicinal herb with the highest antioxidant activity. Accordingly, the present study was designed to investigate the possible actions of ethanolic extract of the leaves of Rosmarinus officinalis on glucose homeostasis and antioxidant defense in rabbits. In the first set of experiments, hypoglycaemic effects of oral administration of various doses (50, 100 and 200 mg/kg) of the extract were examined in normoglycaemic and glucose-hyperglycaemic rabbits. Optimal effect was observed in both of the animal groups with a dose of 200 mg/kg of the extract and this activity was independent from the effects of insulin. In another part of experiments, acute effect of various doses of the Rosmarinus officinalis extract on blood glucose and serum insulin levels was studied in alloxan-induced diabetic rabbits. Of the three doses of extract, the highest dose (200 mg/kg) significantly lowered blood glucose level and increased serum insulin concentration in alloxan-diabetic rabbits. The last set of experiments designed to investigate the subacute effect of the Rosmarinus officinalis extract on repeated administration in alloxan-diabetic rabbits. At the doses of 100 and 200 mg/kg, antihyperglycaemic effect of extract was accompanied by a significant increase in serum insulin levels in diabetic rabbits. Furthermore, during 1 week of treatment of diabetic rabbits with a dose of 200 mg/kg of the extract showed that the extract possessed a capability to inhibit the lipid peroxidation and activate the antioxidant enzymes. It was concluded that probably, due to its potent antioxidant properties, the Rosmarinus officinalis extract exerts remarkable antidiabetogenic effect.

title: Essential oils composition in two Rosmarinus officinalis L. varieties and incidence for antimicrobial and antioxidant activities.
abstract: The essential oil composition of Rosmarinus officinalis var. typicus and var. troglodytorum endemic to Tunisia, and growing wild in different bioclimates, was determined by GC and GC-MS. Oils were assessed for their antimicrobial and antioxidant activity. A variation of the chemical composition attributed to varieties rather than to bioclimates was revealed. 1.8-Cineole (47.2-27.5%) and camphor (12.9-27.9%) were identified as the main constituents of var. typicus and var. troglodytorum, respectively. The principal component analysis performed on oil constituents for all the populations allowed the distinction of two distinct population groups in accordance to the varietal subdivision. Based on the determination of the diameter of inhibition and the determination of the minimum inhibitory concentration, a low to moderate antimicrobial activity according to oils was revealed against eight bacteria tested. However, oils from var. troglodytorum showed higher bactericidal effect than those from var. typicus. The oils' antioxidant activity, determined by 1,1-diphenyl-1-picrylhydrazyl (DPPH) assay, ferric reducing (FRAP) assay and β-carotene bleaching test, was relatively high. The highest activity was found in oils from var. troglodytorum and in one population of var. typicus from the upper semi-arid bioclimate.

title: Plants used as remedies antirheumatic and antineuralgic in the traditional medicine of Lebanon.
abstract: In the present investigation, ethnobotanical and ethnopharmacological research of the traditional use of plants to cure antirheumatic and antineuralgic diseases was carried out in Lebanon.



The information was collected from different cities and villages of the twenty-six districts "aqdya or qadaa" of the six governorates "mohaafazah" of Lebanon. The knowledge on the medicinal use of plants has been recorded directly on the basis of a detailed survey on herbalists "Attarin or dabbous", folk healers, on older experienced people and midwives "daye". This research was carried out for 5 years (2002-2007).



In this survey, 231 species of which 5 are endemic, distributed in 79 botanical families, currently used as antirheumatic herbal remedies, were collected and identified. Among them 53.3% are indigenous, 19.2% are cultivated and 17.5% are imported. Their latin names, local names, local medicinal uses, the used parts of the plant and the preparations and the ways of administration are described. The anti-inflammatory properties due to the plant compounds have been demonstrated and discussed.

title: Antinociceptive effect and GC/MS analysis of Rosmarinus officinalis L. essential oil from its aerial parts.
abstract: The rationale of this investigation was to examine the antinociceptive properties of the essential oil obtained from Rosmarinus officinalis aerial parts, using a rat model of arthritic pain. The essential oil (100, 300 and 600 mg/kg, I. P.) produced a dose-dependent antinociceptive effect, manifested as a significant reduction in the dysfunction in the pain-induced functional impairment model in the rat (PIFIR model), mainly at high doses. Chemical constituents of the essential oil were further analyzed by gas chromatography-mass spectrometry (GC/MS). The major compounds in the essential oil were alpha-pinene (14.10 %), camphene (11.47 %), beta-pinene (12.02 %), myrcene (3.31 %), alpha-phellandrene (7.87 %), eucalyptol (8.58 %), 2-bornanone (3.42 %), camphor (8.75 %), isoborneol (3.48 %), borneol (4.85 %) and borneol acetate (6.49 %). The antinociceptive effects of R. officinalis essential oil were tested in combination with 0.12 mg/kg WAY100635, s. c. (an antagonist of 5-HT(1A) receptors) or 1 mg/kg naloxone, i. p. (an antagonist of endogenous opioids receptors), demonstrating in both cases an inhibition of the antinociceptive response. This study suggests an involvement, at least in part, of the serotonergic system via 5-HT(1A) receptors and endogenous opioids in the antinociceptive effect of R. officinalis essential oil in the PIFIR model.

title: Determination of active components in rosemary by capillary electrophoresis with electrochemical detection.
abstract: Rosemary (Rosmarinus officinalis L.) is a spice and medicinal herb widely used around the world. Among natural antioxidants, rosemary has been widely accepted as one of the spices with the highest antioxidant activity. A capillary electrophoresis method for the determination of its active components using electrochemical detection was developed. Effects of several important factors were investigated to acquire the optimum conditions. The detection electrode was a 300 microm carbon disc electrode at a working potential of +0.90 V (versus SCE). The analytes can be well separated with 25 min in a 75 cm length fused-silica capillary at a separation voltage of 16 kV in an 80 mmol/l borate buffer (pH 9.0). The current response was linear over about three orders of magnitude with detection limits (S/N=3) ranging from 2 x 10(-7) to 1 x 10(-6)g/ml for all the analytes. The method was successfully used in the analysis of rosemary with relatively simple extraction procedures, and the assay results were satisfactory.

title: Protective action of ethanolic extract of Rosmarinus officinalis L. in gastric ulcer prevention induced by ethanol in rats.
abstract: The pathology of a gastric ulcer is complex and multifactorial. Gastric ulcers affect many people around the world and its development is a result of the imbalance between aggressive and protective factors in the gastric mucosa. In this study, we evaluated the ethanolic extract of Rosmarinus officinalis L. (eeRo); this plant, more commonly known as rosemary, has attracted the interest of the scientific community due to its numerous pharmacological properties and their potential therapeutic applications. Here, we tested the preventive effects of eeRo against gastric ulcer induced by 70% ethanol in male Wistar rats. In addition, we aimed to clarify the mechanism involved in the preventive action of the eeRo in gastric ulcers. Based on the analysis of markers of oxidative damage and enzymatic antioxidant defense systems, the measurement of nitrite and nitrate levels and the assessment of the inflammatory response, the eeRo exhibited significant antioxidant, vasodilator and antiinflammatory properties.

title: Healing potential of Rosmarinus officinalis L. on full-thickness excision cutaneous wounds in alloxan-induced-diabetic BALB/c mice.
abstract: Rosmarinus officinalis (Rosemary) used in Jordanian folk medicine for wound management and treatment. Therefore, the present study was conducted to assess the healing efficacy of both aqueous extract and essential oil of the aerial parts on alloxan-induced diabetic BALB/c mice.



Two full-thickness round wounds were created in the dorsal area of each mouse. Animals were divided into four groups of twenty mice each: untreated normal, untreated diabetic, aqueous extract- (intraperitoneal injection of 0.2 ml at a dose of 10% for 3 days) and essential oil-treated diabetic mice (topical application of 25 microl/excision wound, twice a day for 3 days). For 15 days, the wounds were visually observed; blood glucose level, body weight, regenerated granulation tissue weight and the percentage of wound contraction were measured. On days 6 and 15 after wounding, the animals were sacrificed and the histology of wound area was examined.



Significant positive differences (p<0.01) between treated and control groups were observed at different aspects of diabetic wound healing process. Reduced inflammation and enhanced wound contraction, re-epithelialization, regeneration of granulation tissue, angiogenesis and collagen deposition were detected in the treated wounds.



Results indicated that the essential oil of Rosmarinus officinalis was the most active in healing diabetic wounds and provide a scientific evidence for the traditional use of this herb in wound treatment. However, further scientific verification is required to confirm and assess the range of wound healing potential of essential oils of Rosemary chemotypes.

title: The antimutagenic activity of Lavandula angustifolia (lavender) essential oil in the bacterial reverse mutation assay.
abstract: Essential oils from Melaleuca alternifolia (tea-tree oil) and Lavandula angustifolia (lavender oil) are commonly used to treat minor health problems. Tea-tree oil possesses broad-spectrum antimicrobial activity, and is increasingly used for skin problems. Lavender oil, traditionally used as an antiseptic agent, is now predominantly used as a relaxant, carminative, and sedative in aromatherapy. Despite their growing use no data are available on their mutagenic potential. In this study, after determining the chemical composition of tea-tree oil and lavender oil, by gas-chromatography and mass spectrometry, we investigated their mutagenic and antimutagenic activities by the bacterial reverse mutation assay in Salmonella typhimurium TA98 and TA100 strains and in Escherichia coli WP2 uvrA strain, with and without an extrinsic metabolic activation system. Neither essential oil had mutagenic activity on the two tested Salmonella strains or on E. coli, with or without the metabolic activation system. Conversely, lavender oil exerted strong antimutagenic activity, reducing mutant colonies in the TA98 strain exposed to the direct mutagen 2-nitrofluorene. Antimutagenicity was concentration-dependent: the maximal concentration (0.80 mg/plate) reduced the number of histidine-independent revertant colonies by 66.4%. Lavender oil (0.80 mg/plate) also showed moderate antimutagenicity against the TA98 strain exposed to the direct mutagen 1-nitropyrene. Its antimutagenic property makes lavender oil a promising candidate for new applications in human healthcare.

title: The effects of habitat type and diurnal harvest on essential oil yield and composition of Lavandula angustifolia Mill.
abstract: The world demand for lavender essential oil is still increasing. It is estimated that over two hundred thousands hectares are being cultivated in Europe and the quality of produced essential oil is important especially for medicinal and pharmaceutical uses as well as for aromatherapy. We tested and evaluated the quantity and the quality of essential oil produced by one lavender genotype, Lavandula angustifolia var etherio, which is well adapted in Greece region, in two different experimental sites (Kato Sholari and Kilkis). The diurnal essential oil yield and oil composition were also tested. The genotype presented an excellent transplantation degree and high survival percentage in the field. Small differences were recorded in production of oil between the experimental sites (for Kato Scholari was 2.67 ±0.12% and for Kilkis region 2.54 ±0.13%). Differences were recorded in essential oil composition between the experimental sites. The main compounds were linalyl acetate (30.62%), linalool (29.56%), 1,8-cineole (5.18%) and camphor (4.03%) for Kato Sholari. The main compounds for Kilkis were linalyl acetate (26.92%), linalool (16.78%), 1,8-cineole (15.55%) and camphor (7.41%). Diurnal differences in oil yield were not observed. In contrast the major compounds percentage showed differences. The high content of linalyl acetate and linalool and low content of 1,8-cineole and camphor for Kato Scholari resulted in a very pleasant and delightful aroma. The selection of a field for lavender cultivation in a big scale should take into consideration the local conditions. Calcareous, well drainage light substrates are suggested as the most suitable for lavender oil production. It is also concluded from this study that the best time for lavender harvesting is after midday and during afternoon where the linalyl acetate is higher.

title: Protective effects of Origanum majorana L. against Neurodegeneration: Fingerprinting, Isolation and In vivo Glycine Receptors Behavioral Model
abstract: Extracts from Origanum majorana L. are used in Asia and Africa as sedatives and neurotonics. Few studies done to explore the active chemical constituents responsible for this apparent contrast. The inhibitory glycine receptors (GlyRs) are key mediators of synaptic signaling in spinal cord, brain stem, and higher central nervous system regions. Neurodegeneration may cause alteration of the GlyRs causing strychnine-like convulsions and stiffness. Here, modulation of GlyRs in-vivo was studied in a mouse model of strychnine toxicity. Total phenolics in Egyptian marjoram (Mj-eg) and Lebanese marjoram(Mj-lb) were calculated and fingerprinted. The Mj-eg and Mj-lb ethanolic extracts revealed to be potent modulators to GlyR; with potential anticonvulsant properties in low doses. Chlorogenic acid (CGA), p-coumaric acid (pCA) and p-hydroxy benzoic acid (pHBA) were detected in the active fractions via column chromatography and RP-HPLC fractionation. The active fraction phenolics at harmless low doses, showed anticonvulsant activity by reversing strychnine toxicity in mice. By applying Berenbaum isoblographic method, we confirmed that at low concentrations the protective effects of CGA and pCA on strychnine toxicity are synergistic. It could be concluded that both marjoram and active phenolics could be used as sedatives in low doses and as neurotonics in high doses. In order to fight against neurodegenerative diseases is to improve body antioxidant, marjoram provided to be good sources for antioxidant potential. In brief, the marjoram phenolics, especially CGA, pCA and pHBA, suggested to be novel GlyR modulators, good phytotherapy, pharmacological tool and a dose sensitive drug to treat convulsions, stiffness and neurodegenerative disorders.

title: Chemotypes investigation of Lavandula essential oils growing at different North African soils
abstract: The chemical composition of the hydrodistilled essential oil of Lavandula augustifolia Mill., collected from Batna (Eastern Algerian), was analyzed by GC and GC/MS. 32 components representing 89.6% of the essential oil were detected with 1,8cineole (29.4%) and camphor (24.6%) as the major components. A comparative study on the compositions of essential oils of Lavandula growing at different North African soils is investigated.

title: Alloxan-induced diabetic thermal hyperalgesia, prophylaxis and phytotherapeutic effects of Rheum ribes L. in mouse model
abstract: Rheum ribes L., known as Syrian rhubarb, is used in traditional Lebanese folk medicine for the treatment of diabetes. The present study aims to investigate the activities of R. ribes aqueous extract for glucose homeostasis, in vivo antioxidant and diabetic neuropathy protection in mice. The acute and the subacute effects of various doses of R. ribes on blood glucose and in vivo antioxidant activity utilizing serum catalase level (CAT) were studied in alloxan-diabetic mice. The high doses significantly lowered glucose level and increased serum CAT in alloxan-diabetic mice. Pretreatment with the extract prior to alloxination, protected the mice from acquiring diabetes and diabetic neuropathy. Treatment with the extract for 8 weeks alleviated hyperalgesia in diabetic mice. Our findings provide clinicians with promising drugs intended for the management of the symptoms of diabetic complications. The protective activity of R. ribes against acquiring diabetes and diabetic neuropathy might pave the way for preparing a prophylactic treatment for diabetes risk groups.

title: Evolution of rosmarinic acid biosynthesis.
abstract: Rosmarinic acid and chlorogenic acid are caffeic acid esters widely found in the plant kingdom and presumably accumulated as defense compounds. In a survey, more than 240 plant species have been screened for the presence of rosmarinic and chlorogenic acids. Several rosmarinic acid-containing species have been detected. The rosmarinic acid accumulation in species of the Marantaceae has not been known before. Rosmarinic acid is found in hornworts, in the fern family Blechnaceae and in species of several orders of mono- and dicotyledonous angiosperms. The biosyntheses of caffeoylshikimate, chlorogenic acid and rosmarinic acid use 4-coumaroyl-CoA from the general phenylpropanoid pathway as hydroxycinnamoyl donor. The hydroxycinnamoyl acceptor substrate comes from the shikimate pathway: shikimic acid, quinic acid and hydroxyphenyllactic acid derived from l-tyrosine. Similar steps are involved in the biosyntheses of rosmarinic, chlorogenic and caffeoylshikimic acids: the transfer of the 4-coumaroyl moiety to an acceptor molecule by a hydroxycinnamoyltransferase from the BAHD acyltransferase family and the meta-hydroxylation of the 4-coumaroyl moiety in the ester by a cytochrome P450 monooxygenase from the CYP98A family. The hydroxycinnamoyltransferases as well as the meta-hydroxylases show high sequence similarities and thus seem to be closely related. The hydroxycinnamoyltransferase and CYP98A14 from Coleus blumei (Lamiaceae) are nevertheless specific for substrates involved in RA biosynthesis showing an evolutionary diversification in phenolic ester metabolism. Our current view is that only a few enzymes had to be "invented" for rosmarinic acid biosynthesis probably on the basis of genes needed for the formation of chlorogenic and caffeoylshikimic acid while further biosynthetic steps might have been recruited from phenylpropanoid metabolism, tocopherol/plastoquinone biosynthesis and photorespiration.

title: Physiological investigations on the effect of olive and rosemary leaves extracts in male rats exposed to thioacetamide.
abstract: Physiologically, it is known that thioacetamide (TAA) toxicity is generally associated with hepatic fibrosis induction, complicated metabolic disorders and health problems. The capability of extracts of olive and rosemary leaves to attenuate the severe physiological disturbances induced by thioacetamide (TAA) intoxication in male rats has been evaluated. Healthy male Wistar rats were used in the present study and were divided randomly into eight groups. Rats of the first group were served as normal control. Rats of the second group were administrated with TAA. Rats of the third, fourth and fifth groups were exposed to TAA plus olive leaves extract, TAA plus rosemary leaves extract and TAA plus olive and rosemary leaves extracts respectively. The sixth, seventh and eighth groups were supplemented with olive leaves extract, rosemary leaves extract, and olive and rosemary leaves extracts respectively. After 12 weeks of experimental treatments, the levels of serum glucose, total protein, albumin and high density lipoprotein cholesterol were significantly decreased, while the levels of triglycerides, cholesterol, low density lipoprotein cholesterol, very low density lipoprotein cholesterol, creatine kinase and lactate dehydrogenase were statistically increased in rats exposed to TAA. Administration of the studied extracts inhibited the hematobiochemical parameters and improved the physiological disturbances induced by TAA intoxication. Additionally, most improvements were noted in rats administrated with rosemary leaves extract followed by olive and rosemary leaves extracts and olive leaves extract. These results suggested that the effect of these extracts might be due to their antioxidant activities against TAA toxicity.

title: Variation of free phenolic acids in medicinal plants belonging to the Lamiaceae family.
abstract: Ten species belonging to the family Lamiaceae and representing the most popular medicinal plants used in Polish phytotherapy were examined for the content of free phenolic acids (PhAs). Two depsides, rosmarinic and chlorogenic acids, as well as eight simple PhAs, protocatechuic, gentisic, p-hydroxybenzoic, caffeic, vanillic, syringic, p-coumaric and ferulic acids, in different qualitative and quantitative proportions depending on the plant examined were determined by the rapid, selective and accurate method combining solid-phase extraction and high-performance liquid chromatography.

title: Authenticity of essential oils
abstract: Essential oils are natural materials widely used in many fields all over the world and have become an integral part of everyday life. There is increasing demand for essential oils, which has resulted in cases of adulteration. Authentication is thus a matter of critical importance for both consumers and chemical companies. This comprehensive overview covers known adulterations in essential oils, and some analytical methodologies adopted for their detection. We first list recommended tests, and then we explain and discuss common analytical techniques, such as chiral gas chromatography, isotope-ratio mass spectrometry, and nuclear magnetic resonance spectroscopy. We also present (high-performance) thin-layer chromatography, vibrational spectroscopy, coupled and multidimensional chromatography, high-performance liquid chromatography, and combination with chemometrics-metabolomics. This review provides a critical overview of existing techniques.

title: Comparison of two isolation methods for essential oil from rosemary leaves: Hydrodistillation and microwave hydrodiffusion and gravity
abstract: Traditional hydrodistillation (HD) and innovative Microwave Hydrodiffusion and Gravity (MHG) methods have been compared and evaluated for their effectiveness in the isolation of essential oil from fresh Rosmarinus officinalis leaves. The microwave method offers important advantages over traditional alternatives, namely: shorter isolation times (15 min against 3 h for hydrodistillation), environmental impact (energy cost is fairly higher to perform HD than that required for rapid MHG isolation), cleaner features (as no residue generation and no water or solvent used), increases antimicrobial activities, increases antioxidant activity and provides a more valuable essential oil (with high amount of oxygenated compounds). It offers also the possibility for a better reproduction of natural aroma of the essential oil from rosemary leaves than the HD essential oil. Moreover, microwave procedure yielded essential oils that could be analysed or used directly without any clean-up, solvent exchange or centrifugation steps. Scanning electron microscopy shows important structural changes for MHG extraction in contrast to those obtained by HD. Electron micrographs show clearly that the cells are broken and damaged during microwave treatment. Finally, the mechanism of Microwave Hydrodiffusion and Gravity is proposed and discussed.

title: Hydrodistillation-headspace solvent microextraction, a new method for analysis of the essential oil components of Lavandula angustifolia Mill.
abstract: A new method involving concurrent headspace solvent microextraction combined with continuous hydrodistillation (HD-HSME) for the extraction and pre-concentration of the essential oil of Lavandula angustifolia Mill. into a microdrop is developed. A microdrop of n-hexadecane containing n-heptadecane (as internal standard) extruded from the needle tip of a gas chromatographic syringe was inserted into the headspace above the plant sample. After extraction for an optimized time, the microdrop was retracted into the syringe and injected directly into a GC injection port. The effects of the type of extracting solvent, sample mass, microdrop volume and extraction time on HD-HSME efficiency were investigated and optimized. Using this method, thirty-six compounds were extracted and identified. Linalool (32.8%), linalyl acetate (17.6%), lavandulyl acetate (15.9%), alpha-terpineol (6.7%) and geranyl acetate (5.0%) were found to be the major constituents. To the best of our knowledge this is the first report on the use of continuous headspace solvent microextraction coupled with hydrodistillation for investigation of essential oil components.

title: Variation in chemical composition of essential oil of populations of Lavandula × intermedia collected from Western Iran
abstract: Knowledge of population diversity is a main prerequisite and the first step in plant breeding and domestication. Essential oils from different species of Lavandula L. (Lamiaceae) are extensively used in cosmetics, hygiene products, and alternative medicines. The essential oils of Lavandula × intermedia leaves collected from ten regions from the Zagros regions, Western Iran were investigated for their chemical components and oil yield diversity. The hydrodistillated essential oil analyzed by GC-FID and GC/MS. Results indicated differences ( p  ≤ 0.05) among the various populations of L. × intermedia for essential oil yield. The major components of the essential oils from various populations of L. × intermedia were 1,8-cineole (31.64–47.94%), borneol (17.11–26.14%), and camphor (8.41–14.4%). In conclusion, the main source of variability in chemical composition and oil yield seemed to be a difference in environmental conditions.

title: Environmental and developmental factors affect essential oil production and quality of Lavandula angustifolia during flowering period
abstract: The world demand for lavender essential oil is still increasing. Despite of growing interest and the commercial importance of lavender oil it is not clear how environmental and developmental factors influence the optimum period of harvesting lavender flowers for essential oil production. In this report, it was evaluated how the quality and quantity of Lavandula angustifolia cv. Etherio essential oil (EO) is influenced during blooming by environmental factors. It was examined if lavender EO is correlated with gene expression during blooming, targeting to manage the optimum EO harvesting time. The EO content was positively regulated by temperature and flowering stage development while it was negatively affected by rainfall during flowering period. The quality of EO and specifically linalool content was influenced by temperature, flower development, LaLINS expression and rainfall. Although rainfalls remarkably decrease linalool production, the quality of lavender oil was recovered after ten days, suggesting that linalool biosynthesis is regulated during flowering period by both environmental and developmental cues. It is suggested, that the optimum harvesting period to obtain valuable lavender essential oil is when flowering reaches at 60%, ambient temperature is over 26 °C and no precipitations were occurred ten days before harvesting.

title: Flavour compounds of Lavandula angustifolia L. to use in food manufacturing: Comparison of three different extraction methods
abstract: Sixty compounds of Lavandula angustifolia L . cultivated in Friuli Venezia Giulia (North-East Italy) were identified and quantified by GC-MS and GC-FID from essential oils obtained by means of hydrodistillation, and from extracts obtained by supercritical CO 2 extraction (SFE) and ultrasound-assisted extraction (US). Using absolute calibration, a true quantification of 1-8 cineol, camphor, linalool, linalyl acetate and β-caryophyllene was carried out. The best extracts, in terms of amount of isolated compounds, flavour quality and stability were those obtained with SFE. Sonication performed at low amplitude for 5 min offered respect to high amplitude a promising alternative to hydrodistillation as a source of lavender flavouring ready to use for alcoholic beverages or/and confectionery products.

title: The Biological Activities of Cinnamon, Geranium and Lavender Essential Oils
abstract: Acinetobacter sp. represent an important cause of nosocomial infections. Their resistance to some antibiotics, their ability to survive on inanimate surfaces in the hospital environment and their ability to produce biofilms contributes to their virulence. The aim of the study was to determine the antibacterial properties of cinnamon, lavender and geranium essential oils against bacteria of the genus Acinetobacter isolated from several clinical materials and from the hospital environment. A comprehensive evaluation of the susceptibility of Acinetobacter sp. clinical strains to recommended antibiotics was performed. The constituents of cinnamon, lavender and geranium essential oils were identified by GC-FID-MS analysis, and their Minimal Inhibitory Concentrations (MICs) against tested clinical strains were determined by the micro-dilution broth method. In addition, the effects of essential oils on the viability of human microvascular endothelial cells (HMEC-1) and glioblastoma cell line (T98G) were evaluated. Cinnamon bark oil was the most active against clinical and environmental strains of Acinetobacter baumannii with MIC values ranging from 0.5 to 2.5 µL/mL. The MIC values for geranium oil were between 7.5 and 9.5 µL/mL, and between 10.5 and 13.0 µL/mL for lavender oil. These essential oils can be best employed in the fight against infections caused by bacteria from Acinetobacter genus as components of formulations for hygiene and disinfection of hospital environment.

title: Lavender essential oil inhalation suppresses allergic airway inflammation and mucous cell hyperplasia in a murine model of asthma.
abstract: Lavender essential oil (Lvn) has been reported to have anti-inflammatory effects. Bronchial asthma is characterized by bronchial allergic inflammation with airway remodeling. Therefore, we evaluated the anti-inflammatory effect of Lvn on experimentally induced bronchial asthma in a murine model.



BALB/c mice were sensitized by an intraperitoneal injection of ovalbumin (OVA) at days 0 and 14, and subsequently challenged with nebulized OVA on days 28-30 (Control-Asthma group). Mice in the treatment group inhaled Lvn on days 14-31 (Lvn-Asthma group). The allergic inflammatory response was determined on days 32 and 33.



An increase in airway resistance was inhibited in the Lvn-Asthma group than in the Control-Asthma group. The Lvn-Asthma group showed lower total cell numbers and eosinophils in bronchoalveolar lavage (BAL) fluids and peribronchial and perivascular tissues when compared with the Control-Asthma group. The Lvn-Asthma group also had less mucin hyperplasia than the Control-Asthma group. Furthermore, the Lvn-Asthma group showed lower interleukin (IL)-5 and IL-13 cytokine levels in BAL fluids, as well as reduced IL-4 and IL-5 mRNA expression in lung tissue, compared with the Control-Asthma group and determined by FlowCytomix and reverse transcriptase-polymerase chain reaction (RT-PCR), respectively. In addition, Lvn inhalation reduced Muc5b mRNA expression in the lungs without significantly changing the expression of Muc5ac mRNA.



Lvn inhibits allergic inflammation and mucous cell hyperplasia with suppression of T-helper-2 cell cytokines and Muc5b expression in a murine model of asthma. Consequently, Lvn may be useful as an alternative medicine for bronchial asthma.

title: Distillation time effect on lavender essential oil yield and composition.
abstract: Lavender (Lavandula angustifolia Mill.) is one of the most widely grown essential oil crops in the world. Commercial extraction of lavender oil is done using steam distillation. The objective of this study was to evaluate the effect of the length of the distillation time (DT) on lavender essential oil yield and composition when extracted from dried flowers. Therefore, the following distillation times (DT) were tested in this experiment: 1.5 min, 3 min, 3.75 min, 7.5 min, 15 min, 30 min, 60 min, 90 min, 120 min, 150 min, 180 min, and 240 min. The essential oil yield (range 0.5-6.8%) reached a maximum at 60 min DT. The concentrations of cineole (range 6.4-35%) and fenchol (range 1.7-2.9%) were highest at the 1.5 min DT and decreased with increasing length of the DT. The concentration of camphor (range 6.6-9.2%) reached a maximum at 7.5-15 min DT, while the concentration of linalool acetate (range 15-38%) reached a maximum at 30 min DT. Results suggest that lavender essential oil yield may not increase after 60 min DT. The change in essential oil yield, and the concentrations of cineole, fenchol and linalool acetate as DT changes were modeled very well by the asymptotic nonlinear regression model. DT may be used to modify the chemical profile of lavender oil and to obtain oils with differential chemical profiles from the same lavender flowers. DT must be taken into consideration when citing or comparing reports on lavender essential oil yield and composition.

title: Etherio, a new variety of Lavandula angustifolia with improved essential oil production and composition from natural selected genotypes growing in Greece
abstract: Lavender oil is a popular essential oil which unfortunately is not produced in any Greek region due to problems reported before from insufficient lavender cultivars or varieties used. The aim of the present study was to create a synthetic Lavandula angustifolia variety from native Greek plants which however present very well adaptation in local fields. Thirty native plants of L. angustifolia from 10 different habitats were quantified and qualified for essential oil. The best two plants of each population were cross pollinated and the seeds came out from this pollination cultivated until new plants test again for oil quantity and quality. This cycle process was being repeated for 6 years until it resulted in a final plant genotype with high amount of essential oil. The essential oil yield reached under laboratory extraction 2.6% (w/fw) while under field steam distillation was 2.3%. The name of this synthetic variety is L. angustifolia var. etherio and the major essential oil compounds are linalool 26.9% and linalyl acetate with 22.8%. L. angustifolia var. etherio was reproduced by tissue culture and 2 ha of this variety are cultivated for 6 years now. The variety showed high transplantation degree, high adaptation, plant viability, rich flower production and high essential oil yield.

title: Oregano and lavender essential oils as antioxidant and antimicrobial additives of biogenic gelatin films
abstract: The chemical composition of the essential oils obtained by hydrodistillation from fully-formed, dried oregano leaves ( Origanum vulgareOriganum vulgare ) and lavender leaves and flowers ( Lavandula officinalis ) were analyzed by GC/MS. The effectiveness of oregano (OEO) and lavender (LEO) essential oils and a mixture LEO:OEO (50:50) in inhibiting Escherichia coli and Staphylococcus aureus growth were determined. Both essential oils inhibited the growth of the microorganisms tested, being more sensitive to gram-positive bacteria. OEO yielded the lowest values of minimum inhibitory concentration (MIC OEO  = 1600–1800 ppm vs. MIC LEO  = 2000 ppm against E. coli ; MIC OEO  = 800–900 ppm vs. MIC LEO  = 1000–1200 ppm against S. aureus ), due to the higher content of phenolic compound, which also provides antioxidant capacity (IC50 OEO  = 297 ± 89 ppm vs. IC50 LEO  >> 6000 ppm). Mixture results indicated an antagonist antimicrobial effect between OEO and LEO. Gelatin-based films added with OEO or LEO, were prepared by casting (2000–6000 ppm). Mechanical, optical and water vapor barrier properties were determined to observe film functionality. OEO effect on the functional properties of gelatin films was not significant. LEO, in the highest concentration analyzed, promotes a slight change in water vapor permeability of Ge-based films (1.46 × 10 −13 to 6.8 × 10 −14  Kg.m/Pa.s.m 2 ), due to its high hydrophobic nature. Oregano containing gelatin films exhibited the highest antimicrobial and antioxidant properties.

title: Protective effects of citrus and rosemary extracts on UV-induced damage in skin cell model and human volunteers.
abstract: Ultraviolet radiation absorbed by the epidermis is the major cause of various cutaneous disorders, including photoaging and skin cancers. Although topical sunscreens may offer proper skin protection, dietary plant compounds may significantly contribute to lifelong protection of skin health, especially when unconsciously sun UV exposed. A combination of rosemary and citrus bioflavonoids extracts was used to inhibit UV harmful effects on human HaCaT keratinocytes and in human volunteers after oral intake. Survival of HaCaT cells after UVB radiation was higher in treatments using the combination of extracts than in those performed with individual extracts, indicating potential synergic effects. The combination of extracts also decreased UVB-induced intracellular radical oxygen species (ROS) and prevented DNA damage in HaCaT cells by comet assay and decreased chromosomal aberrations in X-irradiated human lymphocytes. The oral daily consumption of 250 mg of the combination by human volunteers revealed a significant minimal erythema dose (MED) increase after eight weeks (34%, p<0.05). Stronger protection was achieved after 12 weeks (56%, p<0.01). The combination of citrus flavonoids and rosemary polyphenols and diterpenes may be considered as an ingredient for oral photoprotection. Their mechanism of action may deserve further attention.

title: Protective effect of linalool, myrcene and eucalyptol against t-butyl hydroperoxide induced genotoxicity in bacteria and cultured human cells.
abstract: We studied the protective effect of monoterpenes myrcene, eucalyptol and linalool against t-butyl hydroperoxide (t-BOOH) induced genotoxicity in reverse mutation assay with Escherichia coli WP2 IC185 strain and its oxyR mutant IC202, and with the comet assay in human hepatoma HepG2 and human B lymphoid NC-NC cells. The monoterpenes were tested in concentration ranges 0.05-1.5 mg/plate and 0.01-1.0 microg/ml in bacteria and mammalian cells, respectively. Suppression of t-BOOH induced mutagenesis was detected only in IC202 strain, and correlated with the observed inhibition of lipid peroxidation by the three monoterpenes. Linalool and myrcene strongly suppressed t-BOOH induced mutagenesis. Eucalyptol, in addition to moderate suppression of t-BOOH induced mutagenesis, suppressed also spontaneous mutagenesis. In NC-NC cells linalool and myrcene reduced t-BOOH induced DNA damage by about 50% at 0.01 microg/ml, while eucalyptol was less efficient (about 50% reduction at 1.0 microg/ml). In HepG2 cells linalool and eucalyptol reduced DNA damage by 30% and 40%, respectively, while myrcene was ineffective. The repair of t-BOOH induced DNA damage, studied in HepG2 cells, was not affected by monoterpenes. The results indicate that linalool, eucalyptol and myrcene have substantial protective effect against oxidant induced genotoxicity, which is predominately mediated by their radical scavenging activity.

title: Lavandula stoechas essential oil from Spain: Aromatic profile determined by gas chromatography–mass spectrometry, antioxidant and lipoxygenase inhibitory bioactivities.
abstract: Lavandula stoechas essential oils (EOs), obtained from plants grown in the South East of Spain, were analyzed by gas chromatography coupled with mass spectrometry (GC–MS) detection to determine their composition, in both relative (peak area) and absolute (using standard curves) concentrations. Fenchone (33–37%), camphor (16–24%) and eucalyptol (17–18%) were determined as the most abundant molecules among the principal compounds. This characterization was completed with the enantioselective gas chromatography, depicting limonene, fenchone or camphor as the main dextrorotatory components and camphene, linalool or E-β-caryophyllene as the main levorotatory components. These enantiomers provide proof of the natural origin of the EOs. Antioxidant activity was evaluated positively by several methods: activity against free radicals (ORAC, DPPH, ABTS), chelating and reducing power, probably due to linalool and thymol. Mild inhibitory activity on lipoxygenase (LOX) was observed indicating a possible anti-inflammatory activity, mainly due to thymol, fenchone and camphor. These properties support the potential use of L. stoechas EOs as natural cosmetic and natural pharmaceutical ingredients for several skin diseases.

title: Does antioxidant properties of the main component of essential oil reflect its antioxidant properties? The comparison of antioxidant properties of essential oils and their main components
abstract: This study discusses the similarities and differences between the antioxidant activities of some essential oils: thyme (Thymus vulgaris), basil (Ocimum basilicum), peppermint (Mentha piperita), clove (Caryophyllus aromaticus), summer savory (Satureja hortensis), sage (Salvia hispanica) and lemon (Citrus limon (L.) Burm.) and of their main components (thymol or estragole or menthol or eugenol or carvacrol or camphor or limonene) estimated by using 2,2′-Diphenyl-1-picrylhydrazyl, 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt and β-carotene bleaching assays. The obtained data show that the antioxidant properties of essential oil do not always depend on the antioxidant activity of its main component, and that they can be modulated by their other components. The conclusions concerning the interaction of essential oil components depend on the type of method applied for assessing the antioxidant activity. When comparing the antioxidant properties of essential oils and their main components, the concepts of synergism, antagonism and additivity are very relevant.

title: Chemical composition, antioxidant and antimicrobial activities of basil (Ocimum basilicum) essential oils depends on seasonal variations.
abstract: Chemical composition, antioxidant and antimicrobial activities of the essential oils from aerial parts of basil (Ocimum basilicum L.) as affected by four seasonal, namely summer, autumn, winter and spring growing variation were investigated. The hydro-distilled essential oils content ranged from 0.5% to 0.8%, the maximum amounts were observed in winter while minimum in summer. The essential oils consisted of linalool as the most abundant component (56.7-60.6%), followed by epi-α-cadinol (8.6-11.4%), α-bergamotene (7.4-9.2%) and γ-cadinene (3.2-5.4%). Samples collected in winter were found to be richer in oxygenated monoterpenes (68.9%), while those of summer were higher in sesquiterpene hydrocarbons (24.3%). The contents of most of the chemical constituents varied significantly (p<0.05) with different seasons. The essential oils investigated, exhibited good antioxidant activity as measurements by DPPH free radical-scavenging ability, bleaching β-carotene in linoleic acid system and inhibition of linoleic acid oxidation. Evaluation of antimicrobial activity of the essential oils and linalool, the most abundant component, against bacterial strains: Staphylococcus aureus, Escherichia coli, Bacillus subtilis, Pasteurella multocida and pathogenic fungi Aspergillus niger, Mucor mucedo, Fusarium solani, Botryodiplodia theobromae, Rhizopus solani was assessed by disc diffusion method and measurement of determination of minimum inhibitory concentration. The results of antimicrobial assays indicated that all the tested microorganisms were affected. Both the antioxidant and antimicrobial activities of the oils varied significantly (p<0.05), as seasons changed.

title: Antioxidant and antibacterial effects of Lavandula and Mentha essential oils in minced beef inoculated with E. coli O157:H7 and S. aureus during storage at abuse refrigeration temperature.
abstract: The essential oils (EOs) of Lavandula angustifolia L. and Mentha piperita L. were analyzed by gas chromatography mass spectrometry (GC/MS). The major constituents were linalool (22.35%), linalyl acetate (21.80%), trans-ocimene (6.16%) and 4-terpineol (5.19%) for L. angustifolia and menthol (33.28%), menthone (22.03%), and menthyl acetate (6.40%) for M. piperita. In vitro antibacterial activity of both EOs against Escherichia coli O157:H7 and Staphylococcus aureus CECT 4459 showed high inhibition against S. aureus. The lowest minimal inhibitory concentrations (MIC) were obtained with L. angustifolia (0.25 μL/mL) against S. aureus; M. piperita exhibited a MIC of 0.50 μL/mL against both microorganisms. Both EOs caused a significant decrease of bacterial growth in minced beef (p<0.05) stored at 9±1 °C. Minced beef treated with EOs showed the lowest TBARS values (lipid oxidation). Moreover, the results showed that the addition of EOs significantly extended fresh meat odor even at abuse temperature.

title: Chemical Composition and Antioxidant Properties of Juniper Berry (Juniperus communis L.) Essential Oil. Action of the Essential Oil on the Antioxidant Protection of Saccharomyces cerevisiae Model Organism
abstract: The essential oil of juniper berries (Juniperus communis L., Cupressaceae) is traditionally used for medicinal and flavoring purposes. As elucidated by gas chromatography/flame ionization detector (GC/FID) and gas chromatography/mass spectrometry (GC/MS methods), the juniper berry oil from Bulgaria is largely comprised of monoterpene hydrocarbons such as α-pinene (51.4%), myrcene (8.3%), sabinene (5.8%), limonene (5.1%) and β-pinene (5.0%). The antioxidant capacity of the essential oil was evaluated in vitro by 2,2-Diphenyl-1-picrylhydrazyl (DPPH) scavenging, 2,2-azino-bis-3-ethylbenzothiazoline-6 sulfonic acid (ABTS) radical cation scavenging, hydroxyl radical (ОН•) scavenging and chelating capacity, superoxide radical (•O2−) scavenging and xanthine oxidase inhibitory effects, hydrogen peroxide scavenging. The antioxidant activity of the oil attributable to electron transfer made juniper berry essential oil a strong antioxidant, whereas the antioxidant activity attributable to hydrogen atom transfer was lower. Lipid peroxidation inhibition by the essential oil in both stages, i.e., hydroperoxide formation and malondialdehyde formation, was less efficient than the inhibition by butylated hydroxytoluene (BHT). In vivo studies confirmed these effects of the oil which created the possibility of blocking the oxidation processes in yeast cells by increasing activity of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx).

title: Antidiabetic potential of Butea monosperma in rats.
abstract: The antihyperglycemic activity of the ethanolic extract of Butea monosperma (BMEE) was studied in glucose-loaded and alloxan-induced diabetic rats. Single dose treatment of BMEE (200 mg/kg, p.o.) significantly improved glucose tolerance and caused reduction in blood glucose level in alloxan-induced diabetic rats. Repeated oral treatment with BMEE (200 mg/kg/day) for 2 weeks significantly reduced blood glucose, serum cholesterol and improved HDL-cholesterol and albumin as compared to diabetic control group.