Calamus Rhizome Research Articles

Isolation and characterization of insecticidal activity of (Z)‐asarone from Acorus calamus L.

AbstractThe insecticidal activity of Acorus calamus L. rhizome‐derived material against adults of Sitophilus zeamais Motschulsky was examined by using repellency method and contact toxicity. The biologically active constituent of the A. calamus rhizome was separated and identified. The results showed that the ethanol extract of A. calamus had strong repellency and contact effect to S. zeamais and the active constituent of the A. calamus was characterized as (Z)‐asarone by spectroscopic analysis. Responses from the tests varied with exposure times and doses. In the repellency test, ethanol extract of A. calamus had 93.92% repellency at 629.08 μg/cm2 but only 71.38% at 157.27 μg/cm2 12 h after treatment. As a contrast, (Z)‐asarone showed 84.50% repellency at 314.54 μg/cm2 and 77.02% at 78.63 μg/cm2 12 h after treatment. In the filter paper diffusion test, ethanol extract of A. calamus caused 95.56% and 17.78% mortality to S. zeamais at 314.54 μg/cm2 and 78.63 μg/ cm2 4 days after treatment, while (Z)‐asarone brought about 100.00% and 15.56% mortality at 40.89 μg/cm2 and 15.73 μg/cm2 respectively. These results indicate that the insecticidal activity of the A. calamus extract may be due to (Z)‐asarone.

6β,10β-Cadinene-6,10-diol: a sesquiterpenoid from the rhizome ofAcorus calamusL.

The title compound [systematic name: (1S,6R,7S,10S)-7-isopropyl-10-methyl-4-methyl­enedeca­hydro­naphthalene-6,10-diol], C15H26O2, was isolated from the rhizome of Acorus calamus L. The mol­ecule contains two fused six-membered rings, each in a chair conformation. The mol­ecules are linked together by inter­molecular O—H⋯O hydrogen bonding.

In vitro Acetylcholinesterase Inhibitory Activity of the Essential Oil from Acorus calamus and its Main Constituents

The in vitro acetylcholinesterase (AChE) inhibitory potential of the hydroalcoholic extract and of the essential oil from Acorus calamus (AC) rhizomes and that of its major constituents were evaluated based on the Ellman's method. GC/MS analysis of the oil revealed that the major constituents were beta-asarone (79.54%) and alpha-asarone (8.47%). The IC50 values were obtained for the hydroalcoholic extract, the essential oil, beta-asarone and alpha-asarone and were 182.31+/-16.78 microg/mL, 10.67+/-0.81 microg/mL, 3.33+/-0.02 microM and 46.38+/-2.69 microM, respectively. Physostigmine was used as standard inhibitor with an IC50 value of 0.28+/-0.015 microM. The experimental observations revealed that the AC essential oil and its constituents have significant AChE inhibitory potential. beta-Asarone, the major phytoconstituent present in the essential oil, showed the maximum inhibitory potential.

Acetylcholinesterase inhibition of oil from Acorus calamus rhizome

Acorus calamus L. has been used in traditional Indian prescriptions and its beneficial effects on memory disorder, learning performance and other aspects of ageing effect have been reported [1]. The hydro-alcoholic extract and essential oil of Indian Acorus calamus rhizomes were tested for in vitro AChE inhibitory activity based on Ellman's method in 96-well micro plates [2] using acetylcholinesterase (AChE) from bovine erythrocytes. The hydro-alcoholic extract gave an inhibition of AChE (IC50 value 182.31±16.78µg/mL) but the oil was stronger (IC50 value 106.75±8.08µg/mL). GC analysis of the oil was performed on a Varian 3400 programmable capillary GC, using D.B.5 'Wax' capillary column [30m x 0.32mm i.d., film thickness 0.25µ]. Oven temperature was programmed at 140–1800C, at 30C/min and held isothermal at 1800C for 7.67 minutes. Injector temperature was 2100C; Detector temperature was 2500C [FID]; carrier gas Helium. 2µL samples were injected with the Frit-splitter at ratio 12:1. GC analysis of the oil revealed that β-asarone 1 was the major constituent (52.33% w/w with respect to dried rhizomes) while α-asarone content was 1.026% w/w. 1 and α-asarone (the trans isomer of 1) were tested for AChE inhibition and found to have IC50 values of 3.33±0.02µM and 46.38±2.69µM respectively. Physostigmine was used as standard and showed inhibition of AChE with an IC50 value of 0.28±0.015µM.

Possibility of Acorus calamus L. rhizome oil as repellent against Culex quinquefasciatus L. mosquito

Culex quinquefasciatus L. is a vector mosquito for bancroftian filariasis in an urban area. One way to prevent the mosquito bites is the use of repellents. The study aims to reveal the potency of Acorus calamus rhizome oil as a repellent against Cx. quinquefasciatus. Experimental 90 mosquitoes were divided into three groups and two groups were treated with different repellents. One group was treated with sari puspa and another group with Calamus rhizome oil. The fluid group was the control group. Each group was treated four times. ANOVA test gave F test > F table, and t test (α = 5%) showed that t test > t table. This statistic test supported a conclusion that Calamus rhizome oil (Acorus calamus L.) was a potential repellent against Cx. quinquefasciatus.

Rapid assessment of beta-asarone content of Acorus calamus by micellar electrokinetic capillary chromatography.

This report outlines a rapid, reproducible method for the determination of beta-asarone, a known carcinogen, using micellar electrokinetic capillary chromatography (MEKC)-UV-vis absorbance and a simple alcohol extraction. The MEKC method is based on a running buffer comprised of 100 mM sodium dodecyl sulfate (SDS), pH 10. The method is reproducible and provides baseline separation of alpha-asarone and beta-asarone. This protocol was used to determine the beta-asarone content of Acorus calamus rhizome of a diploid variety harvested from the wetlands of the United States and the triploid variety from India obtained commercially. The results indicate raw product that originated from India contained 4.4% w/w beta-asarone, while that from the United States contained 0.2% w/w beta-asarone. Neither sample contained detectable concentrations of alpha-asarone. This is the first report of the use of MEKC to determine asarone in a natural source.

Demonstration of Biological Standardization of Selected Herbal Extracts

Objective : To demonstrate biological standardization of selected herbal extracts using rapid and convenient bioassays. Methods Boswellia serrata alcoholic extract (resin), Emblica officinalis water extract (fruit) and Salacia oblonga 80%alcoholic extract (root) were tested with reference to lipoxygenase inhibition, anti-oxidant and α-glucosidase inhibitory activity respectively. Brine Shrimp lethality assay (BSLA) was used for standardization of Bacopa monnieri alcoholic extract (leaves). Piper longum extracts with varying concentration of piperine were also tested in BSLA. To evaluate the use of BSLA in detecting adulteration, samples of Alpinia galanga deliberately adulterated with Acorus calamus rhizomes were tested. Results : IC 50 limits were established for extracts of B.serrata , E.officinalis and S. oblanga . Limits of acceptance were also achieved for B. monnieri in terms of LC 50 values. BSLA was also capable of detecting the difference in Piper longum extracts, which were varying in Piperine content, the active constituent of Piper longum . BSLA was also sensitive to detect change in bioactivity of Alpinia galanga (rhizome) samples, which were deliberately adulterated with Acorus calamus rhizomes. Conclusion : The use of bioassays is suggested for effective quality control of herbal products.

Anticellular and immunosuppressive properties of ethanolic extract of Acorus calamus rhizome

Modulation of immune response to alleviate disease has been of interest since long. Plant extracts have been widely investigated for possible immunomodulatory properties. We have evaluated the anticellular and immunomodulatory properties of ethanolic extract of Acorus calamus rhizome. This extract inhibited proliferation of mitogen (phytohaemagglutinin; PHA) and antigen (purified protein derivative; PPD)-stimulated human peripheral blood mononuclear cells (PBMCs). In addition, A. calamus extract inhibited growth of several cell lines of mouse and human origin. It also inhibited production of nitric oxide (NO), interleukin-2 (IL-2) and tumor necrosis factor-α (TNF-α). Intracytoplasmic interferon-γ (IFN-γ) and expression of cell surface markers, CD16 and HLA-DR, on human PBMC, were not affected on treatment with A. calamus extract but CD25 expression was down regulated. Our study demonstrates the antiproliferative and immunosuppressive potential of ethanolic extract of A. calamus rhizome in vitro.

Protective effect of acorus calamus against acrylamide induced neurotoxicity.

Exposure of rats to acrylamide (ACR) caused hind limb paralysis in 58% of the animals on day 10 and decreased behavioural parameters, namely distance travelled, ambulatory time, stereotypic time and basal stereotypic movements compared with the control group. These rats also had a decrease in the reduced glutathione (GSH) content and glutathione-S-transferase (GST) activity in the corpus striatum and an increase in striatal dopamine receptors, as evident by an increase in the binding of 3H-spiperone to striatal membranes. Treatment with the ethanol:water (1:1) extract of the rhizomes of Acorus calamus (AC-002) increased the GSH content and GST activity in the corpus striatum while insignificant changes were observed in other parameters. Rats treated with ACR and AC-002 in combination had a lower incidence of paralysis (18%) compared with those treated with ACR alone on day 10 of the experiment. The rats also showed a partial recovery in other behavioural parameters. The levels of GSH content and GST activity increased in the corpus striatum, while the dopamine receptors decreased compared with the ACR treated rats. The results suggest that the neurobehavioural changes produced by ACR may be prevented following treatment with Acorus calamus rhizomes.

Study of antidiarrhoeal activity of four medicinal plants in castor-oil induced diarrhoea

A study was undertaken to evaluate the effect of aqueous and methanolic plant extracts of Acorus calamus rhizome, Pongamia glabra leaves, Aegle marmelos unripe fruit and Strychnos nux-vomica root bark for their antidiarrhoeal potential against castor-oil induced diarrhoea in mice. The methanolic plant extracts were more effective than aqueous plant extracts against castor-oil induced diarrhoea. The methanolic plant extracts significantly reduced induction time of diarrhoea and total weight of the faeces. The result obtained establish the efficacy of these plant extracts as antidiarrhoeal agents.

Effect of Acorus calamus (L.) (Araceae) essential oil vapours from various origins on Callosobruchus phaseoli (Gyllenhal) (Coleoptera: Bruchidae)

The toxic effects of vapours of essential oils of Acorus calamus (L.) rhizomes obtained from three countries on the adults and eggs of Callosobruchus phaseoli (Gyllenhal) reared on seeds of Lablab purpureus (Medik.) were studied. All oils were toxic to adults and eggs depending on exposure period and dose. Indian oil showed a strong latent effect, while an acute effect was more dominant in the Yugoslavian oil, followed by Russian oil. Significant reduction of oviposition was found in Yugoslavian and Russian oil vapours at 5 and 10 μl oil per 400 ml jar after 24 h exposure. Longer exposure periods increased the differences in egg production of treated and untreated insects. The fertility of the treated females was little affected. Newly-laid eggs were more susceptible than older ones, Indian oil vapours were the most effective.

Toxicity of Acorus calamus rhizome powder from Eastern Nepal to Sitophilus granarius (L.) and Sitophilus oryzae (L.) (Coleoptera, Curculionidae)

Powders prepared from sweetflag Acorus calamus rhizomes collected at both high (1700 m) and low (900 m) altitude in Eastern Nepal were admixed with wheat at concentrations in the range 0.05–2% w/w and the wheat infested with Sitophilus oryzae or S. granarius adults. Following a 7 day incubation, the mortality of both species was significantly lower at 20 °C than at 30 °C, and the material collected from high altitude was slightly less toxic than that from low altitude. The moisture content of the wheat (14 or 17%) and the part of the rhizome used to prepare the powder (young or mature growth) had no apparent effect on the toxicity of the preparation. Sitophilus granarius (L.) adults were more tolerant of the material than S. oryzae (L.), while the converse was true for larvae developing in treated grain. An admixture rate of approximately 2% w/w admixed rhizome powder was required to give complete kill of adults of both species following 7 days exposure at 20 °C, while a concentration of 1% w/w either prevented the emergence of adults or killed them rapidly following emergence when wheat containing eggs of either species was incubated for 7–8 weeks at 30 °C. The β-asarone content of the rhizome powders was determined by GC-MS as 6.4 and 4.7% w/w (mature section of rhizomes collected at high and low altitudes, respectively) and 3.6 and 4.0% w/w (young sections of rhizomes collected at high and low altitudes, respectively). It is anticipated that, under field conditions and in the temperature range 20–30 °C, the initial residue of approximately 1300 mg/kg β-asarone required to disinfest wheat containing these weevil species would decline rapidly due to evaporation.

Mobility inhibition and nematocidal activity of asarone and related phenylpropanoids on second-stage larvae of Toxocara canis.

The in vitro effect of asarone, the nematocidal principle of the rhizome of Acorus calamus, on second-stage larvae of Toxocara canis is composed of two independent actions: one is a fast acting inhibition of the larval mobility and the other is a slow acting larvicidal action. Mobility of the larvae was rapidly inhibited when they were incubated with asarone. Dye exclusion assay revealed that larvae were alive at this stage, and their mobility was restored after the first inhibition, suggesting that this effect was temporary and reversible. However, when the mobility decreased again during prolonged incubation, the cellular viability of larvae disappeared, showing that they were killed by the compound. The above two-stage effect of asarone was almost identical in two geometrical isomers ((E)- and (Z)-asarone). Di- and tri-methoxypropenyl or propylbenzenes carrying two methoxy groups at a vicinal position on a benzene ring showed, more or less, a two-stage effect of this type. These two actions were suggested to be separable by an appropriate modification of the structure.

Effect of Acorus calamus (L.) (Araceae) oil and its main compound β-asarone on Prostephanus truncatus (Horn) (Coleoptera: Bostrichidae)

The essential oil of Indian Acorus calamus (L.) shows potential for stored-product pest control. Its active ingredient, β-asarone, has toxic and sterilizing effects. Prostephanus truncatus (Horn) was imported to Africa in the 1970s from Central America. Now, the beetle is widely spread in Africa where a large amount of harvested maize is attacked. An environmentally acceptable and economic control method is sought. For this purpose Acorus calamus oil and β-asarone were tested as possible control agents. For direct treatment, corn grains were covered with acetone solutions of the oils. After evaporation of the solvent, the grains were apportioned and infested with beetles. Additionally, grains were treated with rhizome powder of A calamus. The damage was calculated on the weight of corndust received from the infested grains. A treatment with only 0.01% oil reduced grain loss significantly within 21 days, the amount of feeding being lowered to 50% compared to the control. After treatment with β-asarone vapours a strong depression of feeding was observed between 21 and 42 days after treatment. The temperature was very important and this effect was observed at 30°C but not 25°C. An admixture of Acorus calamus rhizome power to te corn reduced feeding activity by up to 83%.

Toxicity of vapours of Acorus calamus L. oil to the immature stages of some stored-product Coleoptera

The toxicity of the vapours of Acorus calamus rhizome oil against the immature stages of Sitophilus granarius (L.), Sitophilus oryzae (L.), Tribolium confusum J. du V. and Callosobruchus chinensis (L.) was studied. The eggs of C. chinensis were the most susceptible, S. granarius eggs were slightly more susceptible than those of S. oryzae, whilst those of R. confusum were not affected. In all cases, the younger embryonic stages were more susceptible than later stages. Larvae and pupae did not show any appreciable susceptibility to the vapours.

Central nervous system studies on an ethanol extract of Acorus calamus rhizomes

An ethanol extract of Acorus calamus rhizomes was screened for CNS effects using a battery of 20 tests in rats and mice. The extract exhibited a large number of actions similar to α-asarone (an active principle of A. calamus) but differed from the latter in several other respects including the responses to electroshock, apomorphine- and isolation-induced aggressive behaviour, amphetamine toxicity in aggregated mice, behavioural despair syndrome in forced swimming, etc. These differences could be due to chemical substances yet to be isolated from the plant extract.

Gas chromatographic and mass spectrometric studies of the constituents of the rhizome of calamus : I. The volatile constituents of the essential oil

The composition of the volatile oil of European calamuse ( Acorus calamus L.) was investigated by liquid—solid chromatography and gas chromatography—mass spectrometry (GC—MS). A total of 184 volatile components were detected, including 67 hydrocarbons, 35 carbonyl compounds, 56 alcohols, 8 phenol, 2 furans and 4 oxido-compounds. The essential oil of Indian calamus was also analyzed by GC—MS and 93 volatile components were detected, β-asarone being the major component. A probable oxidation scheme for β-asarone is shown and the importance of minor components for the development of calamus aroma is also discussed. Of the 125 and 79 components identified in the European and Indian oils, respectively 72 and 39 had not been reported previously in calamus oil.

Gas chromatographic and mass spectrometric studies of the constituents of the rhizome of calamus : II. The volatile constituents of alcoholic extracts

An alcoholic extract of European calamus ( Acorus calamusL.) was preseparated by liquid—solid chromatography and analyzed by gas chromatography—mass spectrometry. Of the 243 volatile components detected, 45 are newly identified in this extract. The occurrence of β-asarone oxidation products and the formation of artifacts are also discussed.

Extraktion labiler Naturstoffe mit überkritischen Gasen

The thermolabile acoragermacrone and other sesquiterpenes could be extracted with supercritical carbon dioxide from beta-asarone free calamus rhizomes ( Acorus calamus L. var. americanus Wulff. A usual steam distillation lets this substance partly decompose into shyobunone. By using high pressure extraction with fractionated separation a higher yield of the bitter principles of calamus, acorone and isoacorone, is obtained.

Zusammensetzung des ätherischen Öles von β-asaronfreiem Kalmus

The study of the essential oil extracted from american diploid calamus rhizomes proved that this variety does not contain the cancerogenic cis-isoasarone (beta-asarone) nor other phenylpropane derivatives which are believed to be toxic. Typical compounds of the examined oil are sesquiterpeneketones, e. g. shyobunones and acorones which pharmaceutically used calamus oils also contain. We believe the isoasarone-free calamus rhizome to be an alternative to the triploid calamus variety often used in todays phytotherapy.