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Shweta Paul *1, Karunanidhi Sharma 2
1Consultant, Shree Vishwapranda Ayurvedic Chikitsalya & Panchakarma center, Yermala, Kallam, Osmanabad,
Maharashtra, India
2Research officer, Multani Pharmaceutical Ltd., New Delhi, India
Received on: 12/04/19 Accepted on: 16/05/19
*Corresponding author
DOI: 10.7897/2277-4343.100364
Yavani or Trachyspermum ammi is an important drug used in traditional systems of medicines to treat gastrointestinal disorders like indigestion, colic,
and diarrhea. Yavani and its distillate (Arka) has been indicated in classics to treat Agnimandya (digestive impairment), Anidra (Insomnia), Atisara
(Diarrhea), Klaibya (Impotency) and Grahani (Irritable Bowel Syndrome). To evaluate its effect on smooth muscle of gastro intestine, the present study
has been planned. Isolated rat ileum was hanged and balanced in organ bath assembly; Tyrode solution (for nutrition), temperature of 37°C and Oxygen
(through aerator device) were maintained in the assembly (inner tube, where tissue was mounted), to keep the tissue alive. Contraction was induced by
Acetylcholine. Then the response of Yavani Arka was observed at the dose of 0.5 ml, 1 ml, 1.5 ml and 2 ml/ 40 ml Tyrode solution. Relaxation effect
or antispasmodic activity of Yavani arka was observed on all the contraction. The Effective Concentration for 50% effect (EC50) was found to be 0.0165
KEYWORDS: Ajawain, Arq-e-ajavain. Ajavayana Arka. Arka Kalpana, Smooth muscle.
Yavani (Trachyspermum ammi) is a well-known herb used in
kitchen to as spices and also as home remedy to treat common
abdominal problems1. The drug has also kept important place in
the traditional medicinal systems i.e. Ayurveda and Unani, where
it is used individually and in combinations in the form of powder,
Arka (Hydro distillate), Asava- Arishta (Fermented liquids),
tablets, oils etc2. Its constituents and oils are extracted and used
in modern system of medicine also3,4. In Ayurveda text it is
indicated in the treatment of Agnimandya (digestive impairment),
Anidra (Insomnia), Atisara (Diarrhea), Klaibya (Impotency) and
Grahani (Irritable Bowel Syndrome) etc5. Since it is an Aromatic
drug and having volatile oil as one of important active
constituent6, So Arka Kalpna, (the Hydro distillation) is very
much compatible to use of it, it is very easy to intake (higher
palatability). It is also indicated in Agnimandya (digestive
impairment), Trikashoola (pain in sacral region) and abdominal
discomforts7. Text of Ayurveda8 and Unani system of medicine9
both are having same type of indications, so for the present study
Yavani Arka has been selected for the study.
Isolated tissue bath assays are classical pharmacological tool for
evaluating dose concentration- response relationship in a myriad
contractile tissue10. It is isolated organ bath system, which is used
for over 150 years in the discipline of pharmacology. The
versatility of this system has allowed scientists across the world
to characterize receptors and receptor signal transduction, with
this knowledge forming the basis of therapies that have treated
millions of individuals with diseases or disorders such as
hypertension, heart failure, diabetes, gastrointestinal disease,
bladder dysfunction, asthma, and swallowing disorders, to name
just a few11. To this day, the isolated tissue bath remains an
important facet of drug development and basic research, as it
allows the tissue to function as a system. Due to all this, it comes
closer to examine that how drug would act in the body as whole.
Since the present study is aimed to evaluate the effect of Yavani
Arka on the smooth muscles, so it is planned to be carried out on
the isolated rat ileum.
Preparation and Analysis of Yavani Arka
Yavani Arka was prepared in the departmental lab, PG
Department of Rasashastra & Bhaishajya Kalpana, National
Institute of Ayurveda, Jaipur as per AFI12 (Fig. 1) and was
analyzed as per API (The Ayurvedic Pharmacopeia of
India).(Table 1). Study was conducted after clearance form
committee. The registration no. of study was Aca/863/14-15.
Table 1: Characters of Yavani Arka
Typical Yavani like
Clear watery
Floating oil drops
Specific Gravity
Refractive index
Preparation of physiological salt solution (Tyrode’s solution)
Tyrode solution was taken as physiological salt solution and used
to keep the ileum alive. It was prepared by mixing of salt and
glucose in distilled water as mentioned in Table 2. It was
composed as per previous studies13.
Table 2: Composition of Tyrode’s solution
16 g
0.4 g
1.5 g
2 g
0.1 g
4 g
0.4 g
Distilled water
2 L
Preparation of tissue
Ileum of wistar rat was isolated and kept in the oxygenated
Tyrode solution at room temperature. Longitudinal strips of
approx. 2-3 cm long were prepared from the ileum. (Fig. 2)
Preparation of organ bath assembly & conduction of study
In order to conduct the study first of all organ bath system was
assembled. Outer chamber was cleared and filled with warm
distilled water to maintain the temperature. Inner tissue bath is
filled with Tyrode solution and continuously oxygenated by
aerator; temperature of the chamber maintained at 37°C. The
frontal pointing liver was fixed on mantle rod of organ bath.
Kymograph paper was set on the rotating drum to record the
readings. A piece of ileum approx. 2 cm, with silk thread was tied
on the liver rod and suspended into inner chamber. The tissue was
then given 3 successive washings with fresh Tyrode solution an
allowed to be relax for obtaining a stable base line (Fig.3).
After setting of above assembly, Acetylcholine (AcH) in dose of
10µml was poured on the suspended tissue and contractions were
induced, the graph of readings was found on Kymograph paper,
place on rotating drum (Fig. 4). The procedure was repeated till
the ceiling effect was recorded (for 9 minutes maximum). Then
0.5 ml of Yavani Arka was poured on the tissue and readings were
noticed for 2.5 minutes (Fig. 5). In same manner the readings
were taken with the doses of 1.0 ml, 1.5 ml and 2.0 ml of Yavani
Arka against AcH induced contractions and the readings found on
kymographs was collected (Fig. 6, 7, 8). Three times washings
with Tyrode solutions were giving after every reading.
Data analysis
To analyze the data the reading was measured in centimeters.
Maximum response was considered as 100% response and
accordingly other readings were also calculated in percentage.
Then a graph (Graph no. 1) was plotted in between dose &
response (Dose-response curve; DRC). Finally, effective
concentration for 50% effect (EC50) was calculated from the
Fig. 1- Yavani Arka
Fig. 2- Isolated Rat ileum
Fig. 3- Organ Bath assembly
Fig. 4 Contraction induced by AcH
Fig. 5- Effect of Yavani Arka 0.5 ml/40 ml. against AcH induced
Fig. 6- Effect of Yavani Arka 1 ml/40 ml. against AcH induced
Fig. 7- Effect of Yavani Arka 1.5 ml/40 ml. against AcH induced
Fig.8- Effect of Yavani Arka 2 ml/40 ml. against AcH induced
Relaxations against Acetylcholine induced contractions were
noticed on all the dose of Yavani Arka. As per table 3 relaxation
of different intensity was noticed on different doses. The EC50
value of Yavani Arka was found as 0.0162 ml/ml.
Table 3: Response of different doses of Arka on isolated tissue
Dose of
Arka was
in cm
% Response
response %)
0.5 ml
1.0 ml
71 %
1.5 ml
82 %
2.0 ml
Graph 1: Response on smooth muscles of different doses
Present study was focused on evaluation of relaxant effect of
distillate of Trachyspermum ammi (Yavani Arka). Yavani is
indicated in Agnimandya (digestive impairment), Anidra
(Insomnia), Atisara (Diarrhea), Klaibya (Impotency) and Grahani
(Irritable Bowel Syndrome)14. Its distillate (Arka) is also
indicated in same15. Its major component is essence which is
mainly composed of thymol (49.0%), γ-terpinene (30.8%), p-
cymene (15.7), β-pinene (2.1%), myrcene (0.8%), and limonene
(0.7%)16. Along with the availability of modern medications, the
propensity toward the traditional medications is progressively
growing throughout the world17.
Isolated organ bath study technique was elected for the study
design. The primary advantage of this technique is that the tissue
is living and functions as a whole tissue, with a physiological
outcome (contraction or relaxation) that is relevant to the body. It
is a synthesis of steps i.e. drug-receptor interaction, signal
transduction, second messenger generation, change in smooth
muscle excitability, and change in tissue function. While other
techniques allow study of each of these steps (e.g. radio ligand
binding for drug affinity, measurement of second messengers),
the isolated tissue bath technique allows for integration of all
these steps18. Another advantage is that retaining tissue function
permits calculation of important pharmacological variables that
are more meaningful in a tissue vs a cellular setting; it comes
closer to how the drugs examined would work in the body as a
As the result of the study says that the Yavani Arka relaxed the
contractions induced by the Ach and showed antispasmodic
effect. The main constituent of T. ammi is thymol20 and in
previous studies same effects were noticed by using different
concentration of thymol21 and by aqueous & ethanolic
extracts22,23. There are various mechanism involved for relaxation
of gastrointestinal smooth muscles within the human body; these
may be blocking action on excitatory pathways, such as
cholinergic24 and histaminergic25 or may be through agonistic
actions on inhibitory modulators such as adrenergic26,
purinergic27, GABA regic28, and nitric oxide29. Thymol, which is
reported to be present in the essential oil and Yavani Arka also
contains essential oil. Anticholinergic activity of Thymol has
been reported in earlier studies30, where it showed significant
relaxant effect on the smooth muscles. That may be most probable
reason for the same effected, observed in the present study. Other
probable reasons may be due to antihistaminic activity or calcium
channel clocking activity of some active part in Yavani Arka.
Some reports are available in the favor of this, where thymol is
reported for relaxant effect on isolated rat aorta31 and ventricular
myocardium32, may be due to blocking of calcium channels.
It can be concluded by the current study that Yavani Arka has
relaxant or antispasmodic effect on the smooth muscles. It relax
the GIT muscles against contraction induced by Ach. Results
supports that it can be used to treat abdominal discomfort, colic
pain etc. Further clinical trials on human subjects are suggested
on the basis of the current study, it can confirm the effects in more
validated way.
Head, Professors and all the team members, PG Department of
Rasashastra & Bhaishajya Kalpana, National Institute of
Ayurveda, Jaipur. Dean & Director National Institute of
Ayurveda, Jaipur (RAJ.).
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Cite this article as:
Shweta Paul and Karunanidhi Sharma. Effect of yavani arka
(hydro distillate of Trachyspermum ammi) on rat ileum, against
acetyl choline induced contractions. Int. J. Res. Ayurveda Pharm.
Source of support: National Institute of Ayurveda, Jaipur, India, Conflict of interest: None Declared
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Full-text available
Isolated tissue bath assays are a classical pharmacological tool for evaluating concentration-response relationships in a myriad of contractile tissues. While this technique has been implemented for over 100 years, the versatility, simplicity and reproducibility of this assay helps it to remain an indispensable tool for pharmacologists and physiologists alike. Tissue bath systems are available in a wide array of shapes and sizes, allowing a scientist to evaluate samples as small as murine mesenteric arteries and as large as porcine ileum - if not larger. Central to the isolated tissue bath assay is the ability to measure concentration-dependent changes to isometric contraction, and how the efficacy and potency of contractile agonists can be manipulated by increasing concentrations of antagonists or inhibitors. Even though the general principles remain relatively similar, recent technological advances allow even more versatility to the tissue bath assay by incorporating computer-based data recording and analysis software. This video will demonstrate the function of the isolated tissue bath to measure the isometric contraction of an isolated smooth muscle (in this case rat thoracic aorta rings), and share the types of knowledge that can be created with this technique. Included are detailed descriptions of aortic tissue dissection and preparation, placement of aortic rings in the tissue bath and proper tissue equilibration prior to experimentation, tests of tissue viability, experimental design and implementation, and data quantitation. Aorta will be connected to isometric force transducers, the data from which will be captured using a commercially available analog-to-digital converter and bridge amplifier specifically designed for use in these experiments. The accompanying software to this system will be used to visualize the experiment and analyze captured data.
Full-text available
There are many biological investigations that have been done to determine cure for the dysfunction of GIT, using herbal medicine. It has been reported that Carum copticum is a plant in Umbelliferae family grows as herb with feliciaform roots, possesses bactericidal, anticholienergic and antistaminic activities. In addition, it has also Beta-adrenergic stimulatory effects. However, these effects of Carum copticum are not yet identified with respect to mechanical activities of isolated tissues. Therefore, the present study has been designed to find out the specific effects of Carum copticum on mechanical activity of ileum, both qualitatively & quantitatively. In this study anaesthetized rats (male albino rat) will be used in experiments for mechanical recording through isolated organ bath and oscillograph. The affect of Carum copticum obtained on intestinal motility will also be tested for receptors identification and differentiation with cholinergic and adrenergic agents.The results showed the effective concentrations of acetyl choline causing 50% of maximum response (EC50) obtained in the presence of 0.01 extracts in all five sets of experiments were significantly higher than those of saline (P = 0.000) and also the maximum response to acetyl choline obtained in the presence of extracts were lower (P = 0.000) The results of this study indicated a competitive antagonism effect of Carum copticum at acetyl choline receptors. An anti-cholinergic property of the plant were also suggested
Full-text available
There are many biological investigations for determining an effective cure for the dysfunction of gastrointestinal tracts, using herbal medicine. It has been reported that Carum Copticum is a bactericidal agent and possesses anticholinergic, antihistaminic and b-adrenergic stimulatory effects in some tissues. However, these effects of Carum Copticum on mechanical activities of isolated intestine are not clearly identified yet. The present study has been designed to find out the specific effects of Carum Copticum on mechanical activity of isolated rat's ileum. In this study rat's ileum contraction was recorded through an isolated tissue chamber in an organ bath by using isotonic transducer and oscillographic device. The effect of Carum Copticum extract on acetylcholine induced contraction in isolated rat's ileum was evaluated. Our findings showed that 1% aqueous extract of Carum Copticum reduces the basal contractile activity of rat's ileum. The extract also reduced acetylcholine induced contraction to 40% of its maximum response. The inhibitory action of Carum Copticum extract on acetylcholine induced contraction was similar but slower than that of atropine sulfate. The results of this study showed an inhibitory effect of Carum Copticum extract on acetylcholine induced contraction in rat's ileum.
Various essential oils are rich in carvacrol, a monoterpenic phenol isomeric with thymol. This study was undertaken to assess the vasorelaxant effects of thymol and carvacrol in rat isolated aorta and the putative mechanisms underlying these effects. Thymol and carvacrol produced a concentration-dependent relaxation on the aortic ring preparations pre-contracted using KCl (IC50 value of 64.40 ± 4.41 and 78.80 ± 11.91 μm, respectively) or using phenylephrine (PHE, 0.1 μm) (IC50 value of 106.40 ± 11.37 and 145.40 ± 6.07 μm, respectively) and inhibited the concentration-response curves of aortic rings to PHE or KCl. In Ca2+-free medium with ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid (2 mm), thymol and carvacrol both at 1000 μm completely abolished the phasic component of PHE-induced endothelium-containing ring contractions. At 400 μm, thymol and carvacrol significantly reduced the CaCl2-induced contractions in Ca2+-free medium. Furthermore, both thymol and carvacrol (300 and 1000 μm) significantly reduced the contraction evoked by phorbol dibutyrate (1 μm), an activator of protein kinase C. Magnitude of this inhibitory effect was enhanced in the presence of the Ca2+ pump inhibitor, thapsigargin (1 μm). At 1000 μm, neither thymol nor carvacrol altered the resting potential of vascular smooth muscle cells. In conclusion, thymol and carvacrol induced an endothelium-independent relaxation in rat isolated aorta, an effect that seems mediated through some mechanisms probably involving a transduction pathway between Ca2+ release from sarcoplasmic reticulum and/or regulation of the Ca2+ sensitivity of the contractile system. Moreover, it’s conceivable that thymol and carvacrol, at low concentrations, block the Ca2+ influx through the membrane.
Essential oil of Carum copticum cultivated in Iran was obtained by hydrodistillation and supercritical (CO2) extraction (SFE) methods. The oils were analysed by capillary gas chromatography, using flame ionization and mass spectrometric detection. The compounds were identified according to their retention indices and mass spectra (EI, 70 eV). The effects of different parameters, such as pressure, temperature, modifier volume and extraction time, on the supercritical fluid extraction of C. copticum oil were investigated. The results showed that, under pressure of 30.4 MPa, temperature 35 °C, methanol 0% and dynamic extraction time of 30 min, the method was most selective for the extraction of thymol. Eight compounds were identified in the hydrodistilled oil. The major components of C. copticum were thymol (49.0%), γ-terpinene (30.8%), p-cymene (15.7), β-pinene (2.1%), myrcene (0.8%) and limonene (0.7%). However, by using supercritical carbon dioxide under optimum conditions, only three components constituted more than 99% of the oil. The extraction yield, based on hydrodistillation was 2.8% (v/w). Extraction yield based on the SFE varied in the range of 1.0–5.8% (w/w) under different conditions. The results show that, in Iranian C. copticum oil, thymol is a major component.
To investigate the chemical composition and vasorelaxant effect of the essential oil of Lippia alba (EOLA) in rat mesenteric artery. Chemical composition of EOLA was investigated by gas chromatography-mass spectrometry (GC/MS). Vasorelaxant effect was evaluated in vitro in rat superior mesenteric artery rings. GC/MS analysis revealed the presence of 19 compounds, with geranial (48.58%) and neral (35.42%) being the major constituents. In intact rings precontracted with phenylephrine (Phe: 1 μM), EOLA (100-1000 μg/mL) induced relaxation, where the maximal effect (Emax) was 110.8 ± 10.8%. This effect was not modified after endothelium removal (Emax = 134.8 ± 16.5%), after tetraethylammonium (TEA) (Emax = 117.2 ± 4.96%), or in rings precontracted with KCl (80 mM) (Emax = 112.6 ± 6.70%). In addition, EOLA was able to inhibit the contraction caused by CaCl(2) and produced a small but significant (P<0.05) additional effect (from 70.5 ± 3.4 to 105.3 ± 13.5%, n = 5) on the maximal relaxation of nifedipine (NIF: 10 μM). The results demonstrated that EOLA induces endothelium-independent vasorelaxation, which appears to be caused, at least in part, by blocking Ca(2+) influx through voltage-operated Ca(2+) channels.
Functional and molecular approaches were used to characterize the β-AR subtypes mediating relaxation of rat ileal smooth muscle. In functional studies, (−)-isoprenaline relaxation was unchanged by CGP20712A (β1-AR antagonist) or ICI118551 (β2-AR antagonist) but shifted by propranolol (pKB=6.69). (±)-Cyanopindolol, CGP12177 and ICID7114 did not cause relaxation but antagonized (−)-isoprenaline relaxation. BRL37344 (β3-AR agonist) caused biphasic relaxation. The high affinity component was shifted with low affinity by propranolol, (±)-cyanopindolol, tertatolol and alprenolol. CL316243 (β3-AR agonist) relaxation was unaffected by CGP20712A or ICI118551 but blocked by SR58894A (β3-AR antagonist; pA2=7.80). Enhanced relaxation after exposure to forskolin and pertussis toxin showed that β3-AR relaxation can be altered by manipulation of components of the adenylate cyclase signalling pathway. The β1-AR agonist RO363 relaxed the ileum (pEC50=6.18) and was blocked by CGP20712A. Relaxation by the β2-AR agonist zinterol (pEC50=5.71) was blocked by SR58894A but not by ICI118551. In rat ileum, β1-, β2- and β3-AR mRNA was detected. Comparison of tissues showed that β3-AR mRNA expression was greatest in WAT>colon=ileum>cerebral cortex>soleus; β1-AR mRNA was most abundant in cerebral cortex>WAT>ileum=colon>soleus; β2-AR mRNA was expressed in soleus>WAT>ileum=colon>cerebral cortex. These results show that β3-ARs are the predominant β-AR subtype mediating rat ileal relaxation while β1-ARs may produce a small relaxation. The β2-AR agonist zinterol produces relaxation through β3-ARs and there was no evidence for the involvement of β2-ARs in relaxation despite the detection of β2-AR mRNA. British Journal of Pharmacology (1999) 127, 949–961; doi:10.1038/sj.bjp.0702605