A Study of the Mediterranean Oregano Populations. Chemical Composition of Essential Oils of Origanum ehrenbergii Boiss. from Two Populations in Lebanon
Abstract
Seedlings obtained from seeds of three populations of oregano (Origanum ehrenbergii) harvested in Lebanon and sown at a research station in France were planted in three replicates in a Fischer block design. oils produced by water distillation were subjected to analysis by GC and GC/MS. The oils of one of the populations were uniform and carvacrol-rich. The other population displayed a variable chemical composition, including thymol, thymol-carvacrol and p-cymene/y-terpinene chemotypes.
Key Word Index
Origanum ehrenbergii, Lamiaceae, essential oil composition, thymol, carvacrol, γ-terpinene.
Introduction
Origanum ehrenbergii belongs to the Prolaticorolla section of Group C in the classification of letswaart (1). Although many species belonging to the genus Origanum have been studied by Baser and Duman (2-6), Arnold et al. (7), Valentini et al. (8), Skoula et al. (9) and Melegari et al. (10), no work has been done on the chemical composition of O. ehrenbergii. However, O. compactum Benth. and O. laevigatum Boiss. from the same group have been studied by Charai et al. (11), Baser et al. (12) and Tucker and Maciarello (13), respectively. Origanum compactum was found to contain carvacrol (56%), γ-terpinene (15%) andp-cymene (9%), while O. laevigatum was found to be rich in sesquiterpenes, principally bicyclogennacrene (25-38%), germacrene D (20-21%) and β-caryophyllene (17-14%).
Origanum ehrenbergii is endemic in Lebanon where it grows at an altitude of up to 1500 m. It occurs on dry limestonebased stony scrubland and in open pinewood understories in acid soils. It is used as a condiment, although the less strongly flavored O. syriacum L. is preferred.
Origanum ehrenbergii as a crop is similar to the ordinary oregano found in France. It grows rapidly, but its yield of dry matter is moderate. It is a prostrate plant that sends out numerous floral scapes and blooms abundantly.
The work reported here was carried out on batches of seeds from three distinct populations located in Lebanon, grown at the CNPPMAI facility at Milly-La-Forêt in 1999, 2000 and 2001.
Experimental
Plant material: The seeds collected in 1998 from three sample populations designated 98-011, 98-012 and 98-034 located near Aaquoura, Choueir and Bolonia in Lebanon were grown first in 1999 and disposed in separate plots at CNPP-MAI, with three repeats each. Plants were harvested in 1999 and 2000. New plantings were carried out in 2000 for further studies in 2000 and 2001. The 1999 harvest comprised only enough material for a pooled study of each population, but the 2000 and 2001 harvests allowed a plant-by-plant study.
The plant was harvested in full flower, dried away from light and isolated by use of a Clevenger-type water distillation unit for 3 h. Specimens of each were kept at CNPPMAI.
GC: The gas phase chromatography analysis was carried out on a Delsi DI 200 instrument equipped with a flame ionization detector and a DBS column (25 m x 0.25 mm, df: 0.25 µm) with a split flow rate of 60 mL/inin, nitrogen as carrier gas and temperature programming (5 min at 50°C and 3°C/min up to 220°C), injector temperature 220°C and detector temperature 235°C. Quantitative data were obtained from FID area percents without the use of correction factors.
GC/MS: The oils were analyzed on a Hewlett-Packard gas chromatograph Model 6890 coupled to a Hewlett-Packard MS Model 5873 equipped with an HP5 column (30 m x 0.25 mm, df: 0.25 µm) programmed from 50°C (5 min) to 300°C at 5°C/ min, and 5 min hold. The carrier gas was helium (1 mL/min), injection in split mode (1/10); injector and detector temperatures 250°C and 280°C, respectively. The MS ran in electron impact mode at 70 eV, electron multiplier 2200 V, ion source temperature 230°C. Mass spectral data were acquired in the scan mode in the m/z range 33-450.
Identification was carried out by calculating retention indices and comparing mass spectra with those in data banks; personal, Adams (14) and McLafferty and Stauffer (15).
Results and Discussion
The yields of dry matter were similar for all three batches, at around 15 g/plant. The essential oil contents were about 3.6 mL/100g of dry matter, giving an average oil yield of 0.47 mL/plant. GC and GC/MS analysis identified 63 constituents, of which 28 were quantified, the others being present only in trace amounts (
Sample 98-011 was carvacrol-rich (71.5-78.5%) for the three replicates, which were thus fairly uniform. Some γ-terpinene (2.6-6%) and p-cymene (5-7%) were also found. Thymol was present in only trace amounts (0.9-1.3%). The second and third samples (98-012 and 93-034) differed from the first and were highly heterogeneous. Two repeats (98-012) were rich in thymol/carvacrol (35.2-38.3% / 21.2-22.5%) accompanied byyterpinene (14.5-2.1%) and p-cymene (10.6-8.9%), three repeats (one 98-012, two 98-034) were thymol-rich (52.4-59.2%) with very small amounts of carvacrol 1.6-4.9%, and γ-terpinene and p-cymene comprising 11,1-14.7% and 5.5-10.6%. The last 93034 repeat stood out by its high thymol content (28.5%), usual γ-terpinene content (10.6%) and particularly large amounts of p-cymene (33.1%). This population study of O. ehrenbergii revealed four chemotypes illustrated by histograms shown in Figure 1. These results compared with those obtained forother species in the same group and section differed markedly from those obtained for O. laevigatum, which were sesquiterpene-rich. The oil from the carvacrol chemotype in 98-011 was chemically similar to that analyzed by Charai et al. (11).
The overall compositions obtained on repeats for each of the three samples were pooled averages, and may have hidden more specific compositions. We therefore isolated oils from the most prolific individual plants. We studied 34 plants in 2000, six of 98-011 and three of 98-012 in 2001, 23 plants of 98-011 and two new individuals planted in 2000. The analytical results are given in Table II.
The 23 individuals of 98-011, planted in 1999, survived two winters and proved hardy. Most of the oils had the same composition, with carvacrol as the main constituent (78.2-87.6%). Five other individual plants differed notably in their linalool contents (7.9-16.4%), and contained less carvacrol (7.9-16.4%). One plant had an unusual thymol content (13.5%). Linalool was present only in traces in the pooled samples, where linalool-rich individuals were presumably few in number (Figure 1).
The few individuals studied in 2000 and planted in 1999 (98-012) or 2000 (98-068) showed that these samples were especially sensitive to harsh climate. There was no 98-034 material. The five individuals studied were all of different composition (Barchart 5); four (98-012) were all thymol-rich to different extents (40.9-72.9%), with correspondingly ranging complementary carvacrol (2.9-24.6%) and p-cymene contents (16.7-15.5%), γ-terpinene content ranging from 1.1-11.5%. The last individual displayed a more marked predominance of carvacrol (55.1%) with thymol at 13.1%, p-cymene at 9.5% and γ-terpinene at 2.8%.
A carvacrol chemotype (Figure 1) was thus clearly defined in both the pooled study of repeats and in individual plants. Thymol-rich, thymol/carvacrol andthymol/carvacrol/p-cymene/ γ-terpinene chemotypes (Figure 1) were represented by only a few individuals. The most abundant (carvacrol) chemotype is thus readily available, but the others would require further selective breeding before they could be exploited.
References
1. J.H.A. letswaart, Taxonomic revision of the genus Origanum (Labiatae). Leiden Botanical Series, Leiden University Press, The Hague, Netherlands, 4, (1980).
2. K.H.C. Baser and H. Duman, Composition of the essential oils of Origanum boissieri letswaart and Origanum bargyli Mouterde. J. Essent. Oil Res., 10, 71-72 (1998).
3. K.H.C. Baser, T. Özek, M. Kürkçüoglu and G. Tümen, Composition of the Essential oil of Origanum sipyleum of Turkish origin. J. Essent. Oil Res., 4, 139-142 (1992).
4. K.H.C. Baser, N Ermin, M. Kürkçüoglu and G. Tumen, Essential oil of Origanum hypericifolium O. Schwarz et P.H. Daw’s. J. Essent. Oil Res., 6, 631-633 (1994).
5. K.H.C. Baser, M. Kürkçüoglu and G. Tümen, Composition of the essential oil of Origanum haussknechtii Boiss. J. Essent. Oil Res., 10, 227-228 (1998).
6. K.H. C. Baser, T. Özek, and G. Tümen, Essentialoil of Origanum rotundifolium Boiss. J. Essent. Oil Res., 7, 95-96 (1995).
7. N. Arnold, B. Bellomaria and G. Valentini, Composition of the essential oil of three different species of Origanum in the Eastern Mediterranean. J. Essent. Oil Res., 12, 192-196 (2000).
8. G.Valentini, N.Arnold, B. Bellomaria and H.J. Arnold, Study of the anatomy and the essential oil of Origanum cordifolium, an endemic of Cyprus. J. Ethnopharm., 35, 115-122 (1991).
9. M. Skoula, R Gotsiou, G. Naxakis and C.B. Johnson, A chemosystematic investigation on the mono and sesquiterpenoi’ds in the genus Origanum (Labiatae). Phytochemistry, 52, 649-657 (1999)
Development of medical countermeasures to chemical terrorism—the NIEHS’s involvement in a government-wide research effort
The attacks of September 11, 2001, using airplanes, followed closely by e biologic attacks using anthrax spores placed in letters, have illustrated how vulnerable our society is to such acts of terrorism. Since these events, the NIH has led the national research effort focused on the development of medical countermeasures to treat civilian mass casualties. The NIH has received more than $1.5 billion annually, starting in FY 2002, to fund research focused primarily on development of products to diagnose, treat, or prevent bacterial and viral infectious diseases and toxemias that could result from bioterrorist attacks. These initial efforts, focused on infectious agents, have been led by the National Institute of Allergy and Infectious Diseases. In FY 2005, the NIH was given an additional $47.1 million to establish a radiation and nuclear medical countermeasures program to identify products to treat civilians injured by a dirty bomb or nuclear attack. Similar plans are under way to initiate research in FY 2006 to develop products to diagnose and treat victims of a chemical attack.
As part of the planning for an expanded effort related to chemical agents, the NIEHS and the National Institute on Neurological Diseases and Stroke co-sponsored, in FY 2005, an administrative supplement program to existing grants. The intent of these supplements is to develop improved detection, diagnosis, and treatment strategies for likely chemical threat agents. As a result of this program, NIEHS has funded six supplements for a total of $450,000. The NIEHS supplements went to:
* Paul Bishop and Joseph Caruso, University of Cincinnati, to work on increasing the sensitivity of methods they have developed for detecting hydrolysis products of nerve agents and to extend their analysis to spiked food and water samples
* Brian Day, National Jewish Medical and Research Center, to test the ability of lipoic acid, dihydrolipoic acid, and thioredoxin to reverse or prevent pulmonary injury caused by sulfur mustard
* Richard DiGiulio and Ted Slotkin, Duke University, to study the developmental neurotoxicity of nerve agents and to assess some possible protective treatment strategies
* Clem Furlong, University of Washington, to attempt to increase the activity of human PON1 enzyme to levels sufficient to protect against paraoxon exposure in a mouse model. If successful, he will test the modified enzyme’s ability to protect against exposure to sarin, soman, and VX agents in the same mouse model.
* Bruce Hammock and Ian Kennedy, University of California, Davis, to work on the development of miniaturized sensors for use in detecting botulinum toxin, ricin, and abrin
* Cary Pope, Oklahoma State University, to study the differential toxicity of the organophosphorus pesticides chlorpyrifos and parathion, as well as the threat agents sarin and soman, and their inhibition of acetylcholinesterase and the accumulation of acetylcholine in a cannulated rat brain model
This administrative supplement program for selected grant mechanisms funded by NIEHS has just been reannounced for FY 2006 in the NIH Guide (http://grants.nih.gov/grants/guide/notice-files/NOT-ES-06-001.html).
Eiken Chemical Develops LAMP-Based Reagent for Detecting Norovirus
Tokyo, Japan, Nov 2, 2005 - (JCNN) - Eiken Chemical has developed a reagent for detecting norovirus using its proprietary nucleic acid amplification method, Loop-mediated isothermal amplification (LAMP).
The new reagent can specifically identify norovirus RNA in one hour.
Eiken Chemical plans to commercialize the new reagent within the year. Norovirus, which is orally transmitted, causes acute gastroenteritis; at worst, it leads to death.
Source: JCN http://www.japancorp.net
Variation on the Chemical Composition of the Oil from Damaged Branches of Guarea guidonia (L.) Sleumer (Meliaceae)
Abstract
The damaged branches of one specimen of Guarea guidonia, which had been blown down after a high wind, were submitted to steam distillation. The oil was analyzed by GC and GC/MS. The chromatographic profile indicated a difference in the composition of this oil in comparison to an oil obtained from healthy-branch oil analyzed previously. The crude oil was thus submitted to chromatographic separation, and the main components identified by NMR and GC/MS.
Key Word Index
Guarea guidonia, Meliaceae, essential oil composition, germacrene D.
Plant Name
Guarea guidonia (L.) Sleumer (Meliaceae).
Source
The branches of one specimen of Guarea guidonia, which had been blown down after a high wind, were collected in Araraquara, São Paulo State, Brazil. These branches, taken on the same day, came from the same specimen previously analyzed (1). The plant material was identified by Gilberto L. Pozetti from the Institute de QuÃmica, UNESP, Araraquara, and was compared with avoucher specimen (LAG 17311) kept at Institute Agronômico de Campinas, Secretaria de Agricultura do Estado de São Paulo.
Previous Work
G. guidonia L. Sleumer (Meliaceae) is a widespread tree, which grows in Brazil from Rio Grande do Sul to Amazonas States (2). The bark of this plant has been used in folk medicine as an abortive and febrifugal agent; the leaves and fruits are reported to be quite toxic to cattle (3). Previous chemical studies on species of G. guidonia describe the occurrence of meliacines, triterpenes, steroids, diterpenes, sesquiterpenes and coumarins (4,5). The first work as G. guidonia volatiles describes the identification of 11 sesquiterpenes from stem bark oil, including β-caryophyllene (25.0%) and germacrene D (24.0%) as the major components (1). The leaf oil from G. guidonia was also analyzed; seven sesquiterpenes were identified, with the oxygenated derivatives eudesm-6-en-4β-ol (21.0%) and guai-6-en-10β-ol (21.0%) being the major metabolites (6,7).
Experimental
Plant material: The fresh branches of G. guidonia were separated in wood and stem bark. The stem bark portion (720 g) was submitted to hydrodistillation in a Clevenger-type apparatus for 4 h yielding 300 mg of the crude oil (0.04%).
Separation and identification methodology: The crude oil was immediately analyzed by GC and GC/MS. The retention time associated to mass-spectrum for each component indicated the presence of sesquiterpene hydrocarbons and oxygenated sesquiterpene derivatives (7). Prior to GC analysis, the oil was submitted to column chromatography on an Si-gel coated with AgNO^sub 3^ and eluted with CH^sub 2^Cl^sub ^2 and mixtures of CH^sub 2^Cl^sub 2^: Me^sub 2^CO (97:3, 93:7 and 9:1). The chromatographic separation yielded 60 fractions (10 mL each), which were pooled together into 13 groups after GC analysis of each fraction. This procedure afforded pure compounds as well as mixtures. After analysis of the ^sup 1^H- and ^sup 13^C-NMR spectra beside GC/MS of each one of the groups, following a known methodology (6,7), 14 sesquiterpenes were identified (Table I).
Gas chromatography: GC analysis were performed in a Hewlett-Packard 5890 Series II (using helium as carrier gas) equipped with a capillary column HP-5, crosslinked5% phenyl in methyl silicone (30 m x 0.32 mm, film thickness 0.25 µm) and automatic injector (HP 7673) and electronic integrator (HP 3396A). Injector and detector temperatures were set at 180°C and 260°C, respectively; the oven temperature was programmed from 100°C isothermal for 2 min, 100°-240°C at 5°C/min then isothermal at 240°C for 5 min (7).
Gas chromatography/mass spectrometry: All the samples were analyzed by GC/MS, using quadrupole EI-MS (70 eV) system Hewlett-Packard HP-5973 coupled with a Hewlett-Packard HP-6890 with a HP-5 column (30 m x 0.35 mm, film thickness 0.25 µm), using the same temperature programming conditions described above.
NMR: NMR spectra were recorded at 50 M Hz for ^sup 13^C and 200 MHz to ^sup 1^H in a Bruker AC-200 spectrometer, using CDCI^sub 3^ as solvent and TMS as internal standard.
Results and Discussion
The stem bark oil from damaged branches of G. guidonia in was analyzed by GC/MS and submitted to chromatographic separation. The use of the following spectrometric techniques: GC, GC/MS, ^sup 1^H-and ^sup 13^C-N M R, allowed the identification of fourteen sesquiterpenes, representing 75.7% of the total oil (Table I). We could observe that it was composed only of sesquiterpenes, with germacrene D (20.9%), isocaryophyllene oxide and caryophyllene oxide (12.1%), γ-cadinene (7.2%), T-muurolol (8.6%) and α-muurolol (9.6%) being the major components. As has been mentioned before, the absence of monoterpenes chemotaxonomically characterizes the oils of Guarea species (8).
In the present study, the oil obtained from the damaged branches of the same plant previously investigated, was analyzed (Table I). These data indicated a variation in the composition of G. guidonia stem bark oil because in the previously investigated oil (1), the main characterized components were β-caryophyllene (25.0%) and gennacrene D (24.0%). A comparison of the oils indicates a decrease in the sesquiterpene hydrocarbon content (70.7% [arrow right] 40.1%), while the oxygenated sesquiterpenoids content increases (7.3% [arrow right] 35.6%). We also observed that no previously detected caclinane derivative sesquiterpenoids were identified in this oil (T-muurolol and α-inuurolol), which should have been formed by direct oxidation of the γ-muurolene. A similar oxidation pathway can be suggested to explain the formation of isocaryophyllene oxide and caryophyllene oxide from β-caryophyllene, this last one described as major metabolite in the oil previously investigated and detected in low concentration in the damaged branches. As this analysis has been done immediately after the plant material collection, the possibility of oxidation of the β-caryophyllene, α-humulene and cadinene derivatives may be discarded, thereby suggesting that due to the branches being recently blown down, the enzymatic system could have directed the biosynthesis of the sesquiterpenes to produce oxidative derivatives. However, to prove this suggestion, more studies need to be done.
Acknowledgements
The authors are grateful to Massuo Jorge Kato (IQ-USP) for the plant material collection. This article was supported to FAPESP and CNPq.
References
1. C.V. Nuñez and N.F. Roque, Sesquiterpenes from the stem bark of Guarea guidonia (L.) Sleumer (Meliaceae). J. Essent. Oil Res., 11, 439-440 (1999).
2. M.P. Correa, Dicionário das Plantas Úteis do Brasil e das Exóticas Cultivadas. volume 2, Imprensa Nacional, Rio de Janeiro (1984).
3. A.P. Lins, M.M. Bragiio, J.D. FelÃcio. A.M. Giuriatti and J.C. FelÃcio, Chemical and pharmacological aspects ol Guarea guidona. Rev. Latinoamer. QuÃm., 23, 30-33 (1992).
4. F.R. Garcez, C.V. Nuñez, W.S. Garcez, R.M. Almeida and N.F. Roque, Sesquiterpenes, limonoid and coumarin from the wood bark of Guarea guidonia. Planta Med., 64, 79-80 (1998).
5. M. Furlan, N. F. Roque and W. W. Filho, Cycloartane derivatives from Guarea trichilioides. Phytochemistry, 32, 1519-1522 (1993).
6. J.H.G. Lago, C.B. Brochini and N.F. Roque, Terpenoids from Guarea guidonia. Phytochemistry, 60, 333-338 (2002).
7. C.B. Brochini, C.V. Nuñez, I.C. Moreira, N.F. Roque, M.H. Chavesand D. Martins, Identificação de componentes de óleos voláteis: Análise espectroscópica de misturas de sesquiterpenos. QuÃmica Nova, 22, 37-40 (1999).
8. J.H.G. Lago, A.A. Reis and N.F. Roque, Chemical composition of the volatile oil from stem bark of Guarea macrophylla (Meliaceae). Flav. Fragr. J., 17, 255-257 (2002).
CecÃlia V. Nuñez
INFA - Instituto National de Pesquisas da Amazônia, CPPN - Coordenação de Pesquisas em Produtos Naturals, CEP 69083-000, Manaus, AM, Brazil
João Henrique G. Lago*
Faculdade de Ciências Biológicas, Exatas e Experimentais, Universidade Presbiteriana Mackenzie, CEP 01302-907, São Paulo - SP, Brazil
NÃdia F. Roque
Instituto de QuÃmica, Universidade Federal da Bahia, Campus Universitário de Ondina, CEP 40170-290, Salvador, BA, Brazil
* Address for correspondence
Received: February 2003
Revised: March 2003
Accepted: April 2003
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Copyright Allured Publishing Corporation Nov/Dec 2005
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Chemical Composition and Microbial Activity of the Essential Oil of Calamintha nepeta (L.) Savi ssp. nepeta var. subisodonda (Borb.) Hayek from Serbia
Abstract
An oil of Calainintha nepeta (L.) Savi ssp. nepeta van suhisodonda (Borb.) Hayek obtained by hydrodistillation was analyzed by GC and GC/MS. Eleven constituents were identified (96.5%). The main constituents in the oil were pulegone (75.5%), piperitenone oxide (6.0%), menthone (5.3%) and menthol (4.3%). The microbial activity of the oil was screened against Aspergillus niger, Escherichia coli, Staphylococcus aureus, Salmonella enteritidis, Bacillus subtilis and Pseudomonas aeruginosa. It was found that all mentioned microorganisms were sensitive to the oil.
Key Word Index
Calamintha nepeta ssp. nepeta var. suhisodonda, Lamiaceae, essential oil composition, pulegone, menthone, piperitenone oxide, menthol, antimicrobial activity.
Plant Name
Calamintha nepeta (L.) Savi ssp. nepeta var. stibisodonda (Borb.) Hayek [syii.: Calamintha tardarensvs Pavl. non Silic, Calamintha subnuda N. Randj. non (Wald. & Kit.) Host.]
Previous Work
Calamintha species has been the subject of intensive studies over the last few decades (4-45), particularly the oil of C. nepeta due to a great diversity in a chemical composition, microbial and fungicidal activities (4-31). It can be seen that the oil of C. nepeta has been found to be rich in monoterpenes. In all studies on oils obtained from wild growing species of the Calamintha genus, the compounds of monoterpenoids were dominant, especially the oxygenated p-menthane-type monoterpenoids. The monoterpenoids of the pinane, thujane and camphanetypes were present only as minor components or they were not detected. Two atypical oil samples have been characterized; one with isopinocamphone as the major component (38), and the other with carvone and 1,8-cineole as main components (22). The contents of sesquiterpenoids were present up to 5% while the other compound were very low (less than 2%).
Source
The aerial parts of flowering plant were collected in August 2000 on Vratarnica mountain (Knjazevac, South Serbia). A voucher specimen has been deposited in the Herbarium Moesicum Doljevac (HMD; No 395) (1).
Plant Part
Dried and pulverized aerial parts of the plant (100 g) were hydrodistilled for 2.5 h using a Clevenger-type apparatus (2). The oil was extracted from the distillate with Et^sub 2^O and then dried with anhydrous Na^sub 2^SO^sub 4^. After filtration, the solvent was removed by distillation under the atmospheric pressure and pure yellow oil, 1.0 m L, was kept at 4°C until analysis.
Present Work
The oil analysis was performed using GC and GC/MS. Constituents of the oil were identified by comparison of their mass spectra to those from the MS library (Wiley275.1) using a computer search and literature. For the purpose of the quantitative analysis the area percentage obtained by FID was used as the base without the use of correction factors.
Experimental
GC: The GC analysis of the oils was carried out on a GC HP-5890 II apparatus, equipped with the split-splitless injector, SPB 5 capillary column (30 m x 0.25 mm, 0.25 µm film thickness) with helium as the carrier gas (1 mL/min) and fitted to FID. Operating conditions: injector temperature 250°C, temperature program isothermal 50°C for 3 min, then 50°-250°C at 5°C/min and finally isothennal at 250°C for 15 min.
GC/MS: GC/MS analyses were performed on a Hewlett Packard apparatus, Model 5890, Series II at 70 eV coupled with a mass selective detector 8MSD 5971A, under the same gas-chromatograph conditions.
Identification procedure: The constituents were identified by comparison of their mass spectra to those from MS library Wiley 275.1. Obtained results were correlated with retention index(3). The areapercentage was obtained electronically from the GC-FID response without the use of internal standard or correction factors.
Antimicrobial activity: The oil was tested in vitro using the diffusion discs method with the following microorganisms: Aspergillus niger ATCC 16404, Escherichia coli 95 TORLAK, Staphylococcus aureus ATCC 6538, Salmonella enteritidis ATCC 13076, Bacillus subtilis 201 and Pseudomonas aeniginosa ATCC 9027.
Substrate for the bacteria was Antibiotic Medium 1 (Difco Laboratories Detroit Michigan USA), for the fungus: Tripton Soja Agar (Torlak, Beograd). The ethanolic solution of the oil (60 µL) in the dilutions of 1:101:201:30 and 1:50 (oil:abc. ethanol) was put on discs Antibiotica Test Blattchen (Schleicher and Schuell, Dassel, Germany; diameter 12.7 mm). The discs then were put on the antibiotic medium sowed with microorganisms and kept at 37°C. After 18 h the activities were determined on a Fisher-Lilly Antibiotic Zone Reader (Ficher Scientific Co. USA) by measuring the inhibition diameter around the discs.
Results and Discussion
Compared with other Calamintha species (3-45), and following the grouping of Lamiacea species, proposed by Kokldni et al., in oil-rich (> 2%), oil-intermediate (0.5-2%) and oil-poor (
The results of the oil analysis of C. nepeta ssp. nepeta var. subisodonda can be seen in Table I. About 96.3% (11 components) of the oil has been identified. Pulegone (75.5%) was the main compound of the oil; the other compounds with noticeable percentage in the oil were piperitenone oxide (6.0%), menthone (5.3%) and menthol (4.3%). According to the literature, the chemical composition of C. nepeta oil is independent of the nature of the subspecies nepeta or glandulosa (28). Generally, there are two main C. nepeta oil types. The first oil type is characterized by the dominance of pulegone along with menthone and menthol and/or their isomers, piperitenone and piperitone and their oxides (4,8,12,13,15,17,20-24,29,30,31), and second type is distinguished by dominance of piperitenone oxide and/orpiperitone oxide (11,16,18,26,27). By comparison with those types the oil of C. nepeta ssp. nepeta var. subisodonda belongs to the pulegone-rich group, which is the most common one.
The results of the microbial activity of the oil of C. nepeta ssp. nepeta var. subisodonda determinated by measuring the inhibition zone diameter of examined microorganisms (given in mm) are summarized in Table II. It was found that all the microorganisms were susceptible to the oil, at all oil dilutions, however, the oil activity declined with dilution. Microbial investigation of the oil of C. nepeta ssp. glandulosa gave the similar results, under the same conditions (31). The oils show the different activities only against Aspergillus niger.
Acknowledgment
We thank the Ministry of Science if Technology of Republic of Serbia for the financial support of this study.
References
1. N. Randjelovic and V. Randjelovic, Taxonomical and horological problems in the frame of the aggregate Calamintha nepeta s. lato on the territory of Serbia. Proceeding of the 7th Symposium of Flora of Southeastern Serbia and Neighbouring Regions, Dimitrograd, SCG, 25-28 (2002).
2. Pharmacopeia. Jugoslavia, 4,128 (1984).
3. R.P. Adams, Identification of Essential oils by Ion Trap Mass Spectroscopy. Academic Press, New York (1989).
4. L. Panizzi, G. Flamini, P.L. Cioni and I. Morelli, Composition and antimicrobial properties of essential oils of four Mediterranean Lamiaceae. J. Ethnopharm., 39, 167-170 (1993).
5. A.M. Pagni, S. Catalano, P.L. Cioni, C. Coppi and I. Morelli, Morphological, anatomical and phytohemical studies of Calamintha nepeta (L) Savi (Labiatae). Plant. Med. Phytother., 24, 203-213 (1990).
6. S. Perrucci, F. Mancianti, P.L.Cioni, G. Flamini, I. Morelli and G. Macchioni, In vitro antifugal actyvity of essential oils againnst some isolated of Microsporum canis andMicrosporumgypseum. Planta Med., 60, 184-187 (1994).
7. G. Flamini, P.L. Cioni, R. Puelio, I. Morelli and L. Panizzi, Antimicrobial activity of the essential oils of Calamintha nepeta and its constituent pulegone against bacteria and fungi. Phytother. Res., 13, 349-351 (1999).
8. P.Bandini and M.Pacchiani, Constituents, properties and use of Calamintha nepeta. Essenze Deriv. Agrum., 51, 325-330 (1981).
9. S. Pavlovic, R. Ivanic, K. Savin, R Zivanovic, R. Jancic, D. Miljkovic and S. Vujcic, Essential oil, tannins and flavonoids from wild species of Micromeria, Satureja, Calamintha, Teucrium and Acinos genera occuring in Yugoslavia. Arh. Farm., 33, 287-291 (1983).
10. B.Bellomaria and G. Valentini, Composition of the essential oil of Calamintha nepeta subsp. glandulosa. G. Bot. liai., 119, 237-245 (1985).
11. H.L. De Pooler, L.F. De Buyck and N.M. Schamp, The volatiles of Calamintha nepeta (L) Savi subsp. glandulosa. Phytochemistry, 25, 691-694 (1986).
12. A. Velasco Negueruela, MJ. Perez-Alonso, J. Maria and M. Marta Rico, Essential oil of Iberian Lamiaceae with pulegone as basic component. An. Bromatol., 39, 357-372 (1987).
13. C. Souleles, N. Argyriadou and S. Philianos, Constituents of Calamintha nepeta. J. Nat. Prod., 50, 510-522 (1987).
Multiple chemical sensitivity sufferer requests help
My daughter, 24, is dying and no one is able to help us. I had never heard of MCS until a few weeks ago, when my sister in Florida emailed me to say she had read something about MCS, that it sounded like Kim, my daughter, and I should check it out.
My daughter’s problems started over a dozen years ago, and have steadily evolved. Her first “episode” is of unknown origin. My husband and I came home after an evening out, and checked on Kim in bed. She looked like she had been beaten up. Her face was swollen and bruised and almost unrecognizable. We still have no idea what caused this. Seemed to be allergies. Over the next few years she would have repeated swellings in her face for seemingly no reason. The swelling would be so extreme that small blood vessels burst, resulting in bruising. She began to see an allergist for testing and subsequently shots. Mold, dust, etc, etc. High school was especially rough. One day the swelling started in school, and a counselor thought we were abusing Kim. Despite Kim’s protests that she was fine before she got to school, the counselor called the authorities. They wouldn’t believe that we had not harmed Kim, and sent someone over to visit us at home. The man was skeptical, despite our explanations and Kim’s. I finally ran my finger over the top of the hutch, rubbed the dust under Kim’s nose, and told him to watch. He said he’d never seen anything like that in his life–in front of his eyes Kim’s face began to swell and bruise. Her chin was always the most affected, and her lips. The rest of high school passed similarly; there was remodeling being done and the smell of the paint bothered her. This was also during the time that the local dump fumes were being carried into the building and the town was in an uproar. The dump was later capped and closed, but not before we had to remove Kim from school on a permanent basis and home school her for her senior year.
Over the next few years, after completing her allergy shots, the swelling and bruising ceased. Kim developed asthma instead. We could not figure out why, nor was there any rhyme or reason to her attacks. Exercise doesn’t affect her as one would expect, and she can just be sitting somewhere and suddenly have an attack. Doctors have tried to blame it on our pets, but we point out that she’s best at home. The animals are not affecting her. During one period of disability from work she spent a couple of months at home, exclusively surrounded by animals. She was fine until the day she left the house to go to the doctor for an appointment.
The past three years have been a nightmare, and getting progressively worse. I’m afraid she is going to die because no one understands or can help her.
She works for a supermarket chain, mid-management. She went months with no problems, then was moved to another store. Episode after episode followed–mostly just asthma attacks. It was decided that the ventilation system in the store might be harboring something that bothered her, so they cleaned it. She was fine after that. Kim was transferred to another store, and had more problems. The asthma attacks became worse, and Kim was taken out of the store a couple of dozen times via ambulance. She started to notice that the attacks were brought on by smells. Perfumes are the worst. Paint and just about anything else can trigger this as well.
Kim has been on so many medications for so long I don’t see how her system can take much more. This past December, during another hospitalization, she was on massive doses of IV steroids. After two weeks she was unable to walk. The doctors wanted to put her into a nursing home for rehab. I refused, and drove her to the Mayo Clinic in Florida, hoping to get her accepted as a patient. She was, but has yet to return. I knew it was the steroids that caused the muscle weakness; I was right. Getting her off the steroids saved her mobility.
She has had many close calls. She’ll leave work ok, and start to feel “funny” on the way home. She’s been stopped numerous times by law enforcement for erratic driving and ended up in the hospital. Her one goal during an attack is to get home. That’s where she feels safe. Luckily she’s never had an accident so far, but I’m sure the day will come. When she’s at the height of an attack she’s disoriented and confused.
Kim had a baby in July. During the pregnancy she had a few episodes, but stayed home for the most part. Now she’s back to work, and getting worse. Three weeks ago we went out to dinner, and a man had chest pains. The ambulance came and took him away, but not before he had vomited. The cleaning solution that the staff used to clean up caused Kim to have an attack. I could see she wasn’t going to last long, so we finished up dinner. We tried to get Kim outside to a car to take her to the hospital, and she collapsed. My husband and sister-in-law stabilized her while I called for an ambulance. (My husband and I are EMT’s, my sister-in-law is a Paramedic). Vital signs were not good, and she wasn’t moving any air. She was confused and disoriented. She got to the hospital, was kept for a few hours while she was observed, and released. She did have one more incident while at the hospital, which was triggered by the cleaning cart outside the door. Oxygen, breathing treatments, steroids. The usual.
Two weeks ago she called me from her car. She had been to an appointment with the baby, and the lady had perfume on. Kim was trying to get home. She assured me she could make it; she was two miles from the house at the most. I called home to alert my son (14) to look for her, and headed home. It took me 15 minutes to get home–no Kim. We managed to get her on her cell phone. She was lost. We told her to pull over and park. It was another 15 minutes before we found her. My husband called the police to help find her. We found her a few miles from home on a road that is not on the way home by any stretch of the imagination, parked as she had been instructed. She had no idea where she was or how she got there. I called the police to update them and order an ambulance. Kim was incoherent for the most part. Totally out of it. One of the officers had oxygen, so we hooked her up to that while we waited for the ambulance. When the ambulance arrived there was no time to wait for a paramedic intercept, they basically loaded her and went straight to the hospital. At the hospital Kim failed all the questions. Wrong birthdate, wrong day of the week, and so on. She kept insisting she was late for school. After some breathing treatments, IV steroids, and oxygen she was ok to return home.
Yesterday we received a phone call from her manager at work. He used her cell phone to contact us. He had found Kim wandering around, dazed and confused and wheezing. He admitted it seemed as though she was high on drugs, but he knows her condition and knew that wasn’t the case. He had called an ambulance. He told me when she was lying down, before the ambulance came, that her eyes rolled back in her head and she passed out for a few moments. According to the paramedic, her pulse was up, respirations were up, audible wheezing, blood sugar 64, and the scariest–pulse ox of 74. That’s darn close to dead. Kim was again very confused and incoherent during most of the long ambulance ride, but somewhat more “normal” by the time she got to the hospital. While at the hospital the lady in the next cubicle sprayed perfume. Kim was later admitted, and remains there. She’s on oxygen, breathing treatments, and IV steroids again. They’re saying she’s anemic. No surprise. And a bunch of other things. But I think they’re missing the whole concept here. “MCS” is a “figment of our imagination.” The pulmonologist who has been treating her for two years doesn’t seem to take this seriously. He thinks steroids are the answer. Steroids and drugs. Those will kill her themselves. To keep her breathing she requires massive doses of too many things.
She’s getting progressively worse. The memory goes. No rational thoughts during an attack. “I’m late for work. I’m late for school. I need to call my Mommy.” And so on. She has a baby to support, yet allowing her to work or drive is dangerous. She doesn’t recognize her own mother, me. (She still lives with us) The incidents are coming more often and are more severe each time. She’s going to die if we can’t find someone to help us!
What can I do, where can I go? HELP!
Heidi Evans
Evans Group at Keller Williams
275 Greenwood Avenue
Bethel, Connecticut 06801 USA
203-744-7355
www.ctrealtor.net
Master mind; the rise and fall of Fritz Haber, a Nobel laureate who launched the age of chemical warfare
([c] 2005 Book News, Inc., Portland, OR)
0060562722
Master mind; the rise and fall of Fritz Haber, the Nobel laureate who launched the age of chemical warfare.
Charles, Daniel.
Ecco Press
2005
$24.95
Hardcover
QD22
Haber’s name has not so much been forgotten as driven from public view, says Charles, a popular writer of science and technology, but he thinks people should know about a man whose nameless ghost appeared in every headline, on every kitchen table. He was hero and villain, a Jew who was also a German patriot, a victim of the Nazis who was accused of war crimes himself, the founder of the military-industrial complex, and the inventor of the chemistry by which the world now feeds itself.
([c] 2005 Book News, Inc., Portland, OR)
CRC handbook of chemistry and physics; a ready-reference book of chemical and physical data, 86th ed., 2005-2006
CRC handbook of chemistry and physics; a ready-reference book of chemical and physical data, 86th ed., 2005-2006.
Ed. by David R. Lide.
CRC Press
2005
2544 pages
$139.95
Hardcover
QD65
At three and one-quarter inches thick, the 86th edition of this essential reference has been printed in slightly larger format for ease of use. With the same goal, each of the 16 sections, the appendices, and index are marked with thumbtabs, and the layout and font have been changed. Much new material has been added to this edition, including entirely new topics, such as electron inelastic mean free paths, proton affinities, vapor pressures (solvent activities) for binary polymer solutions, selected properties of semiconductor solid solutions, and electrical conductivity of aqueous solutions. Several of the tables have been expanded and updated, including: bond dissociation energies, NIST atomic transition probability tables, properties of semiconductors, atomic masses and abundances, threshold limits for airborne contaminants, and the standard transfomed Gibbs energy of formation for important biochemical species.
([c] 2005 Book News, Inc., Portland, OR)
Chemical composition of everyday products
Chemical composition of everyday products.
Toedt, John et al.
Greenwood Pr.
2005
205 pages
$49.95
Hardcover
QD75
Not just tables of arcane symbols and numbers, but essays for general readers explain the components and their action in such products as soaps and laundry, cosmetics and bathroom, health and medical, baby, cleaning, lighting, common household and lawn, automotive and general repair, and office supplies. Toedt (physical sciences), Darrell Koza (chemistry, both, Eastern Connecticut State U.) and Kathleen van Cleef-Toedt (physiology and neurobiology, U. of Connecticut) also cover such common materials as paper, plastic, cement, and spray paint. The appendix sets out protocols for laboratory exercises.
([c] 2005 Book News, Inc., Portland, OR)
Chemical calculations at a glance
Chemical calculations at a glance.
Yates, Paul.
Blackwell Publishing
2005
194 pages
$29.95
Paperback
QD39
Written for undergraduate chemistry majors, this study aid reviews the algebra, functions, and calculus operations commonly used to solve chemistry problems. The British author illustrates how to calculate the rate of a chemical reaction, the potential energy of a three- dimensional harmonic oscillator, macroscopic thermodynamic quantities, heat capacity, variation of enthalpy with temperature, and an expectation value.
([c] 2005 Book News, Inc., Portland, OR)