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Ethnobotanical Leaflets 13: 532-39, 2009. Anthelmintic Activity of Trikatu Churna
and its Ingredients B. Uma
Reddy and Y. N. Seetharam Department of
Botany, E-mail: drumareddy11@yahoo.co.in Issued
Abstract The alcoholic extract of Trikatu churna and its ingredients were evaluated for anthelmintic activity. The dried fruits of Piper nigrum L. (Piperaceae), Piper longum L. (Piperaceae) and rhizome of Zingiber officinale Roscoe. (Zingiberaceae) were powdered and mixed together in equiproportions to get a polyherbal formulation, Trikatu churna. All these three ingredients are spicy, commonly used in our daily diet, also well known for their tremendous therapeutic potential, since from the Vedic period. The alcoholic extract of Trikatu churna and its ingredients were screened for preliminary phytochemical studies and also tested for anthelmintic activity against Pheritima posthuma and recorded the time taken for induction of paralysis and death. Piperazine citrate (10 mg/ml) was included as standard reference and distilled water as control. The results demonstrated that, the extracts of Trikatu churna and its plant ingredients showed the presence of alkaloids, flavonoids, tannins, lignins and steroids, these test samples were also exhibited potent anthelmintic activity, but the highest activity was noticed in Trikatu churna, this might be due to the multifunctional effect of all the three plant ingredients of Trikatu churna. Based on the above results, it is confirmed that, combination of Piper nigrum, Piper longum and Zingiber officinale in Trikatu churna offered promising anthelmintic effect than using the ingredients alone. Keywords:
Trikatu Churna, Piperazine citrate, Pheritima posthuma. Introduction Trikatu churna is one of the traditional polyherbal
preparations, made up of combination of three important spicy materials, such
as Piper nigrum
L (Piperaceae), Piper longum L.
(Piperaceae) and Zingiber officinale Roscoe. (Zingiberaceae). All these plant materials are used
world wide as spices. They are also used as important ingredients in folklore
medicine in many Asian countries. However, the consumption of these spices
would exert several health beneficial effects by the virtue of their
innumerable therapeutic potentials, such as fever, asthama,
cold, cough and other general health disorders
(Anonymous, 1985; Chopra, et al, 1992;
George watt, 1972; Rakesh and Sushil,
2003; Namjoshi, 1976; Sivarajan
and Indira Balachandran,
1996) There are
voluminous research carriedout in this particular
stream, a brief review of the notable work is highlighted, such as Krimikuthar Ras, Sanjivani vati, Kumari asava, Bhallakasava, Vidanasava and
the combination of two or more these formulations were tested for anthelmintic activity against Pheritima posthuma. (Nirmal et al, 2008), aqueous extract of fruits of Terminalia chebula Retz.
(Dwivedi et
al, 2008). But so far no
clinical trials are made on this compound polyherbal
formulation, Trikatu churna.
Hence, the present study was undertaken to explore the anthelmintic
activity of Trikatu churna
and compared its effects to its individual ingredients. The present study is
also aimed to establish its clinical validity. Materials and Methods The anthelmintic
activity was tested on earth worms (Pheretima posthuma) using ethanolic
(95%) extracts of Piper longum, Piper nigrum, Zingiber officinale and Trikatu churna. The crude extracts
of Trikatu churna and its
ingredients were also screened for preliminary phytochemical
studies to find out the occurrence of possible major chemical groups in the
given extracts. Collection of Plant materials
The plant materials of Piper nigrum L, Piper longum L.
along with the fruits were collected from the Preparation of the Trikatu churna The Trikatu churna is a fine powder of drugs. It is prepared by mixing equal quantities of the powder of the dried fruits of Piper nigrum, Piper longum and rhizomes of Zingiber officinale and then sieved through muslin cloth. This churna is stored in airtight container for further processing (Rakesh and Sushil, 2003). Preparation of the extract The 100g of Trikatu churna and its ingredients were extracted in 95% ethanol at 50 - 60°C in a soxhlet apparatus separately. The extract was concentrated to dryness in a flash evaporator (Buchi type) under reduced pressure and controlled temperature (50 -60°C). The dried 95% of the ethanolic extracts weighed in a required dose and dissolved in known volume of distilled water, separately for further treatment. Phytochemical Evaluation of the Crude Extracts Phytochemical
screening of the extracts for the presence of secondary metabolites were
performed using the following reagents and chemicals: for alkaloids with
Mayer’s, Wagner’s and Dragendroff’s reagents, for flavonoids with the use of Mg and HCl,
tannins with 1% gelatin and 10% NaCl solutions, for
saponins with distilled water (Harborne, 1998; Sadasivam and Manickam, 1992; Ogbonnia, et al, 2008; Nooman,
et al, 2008; Mohd.
Nawagish et
al, 2007). Preparation of Standard Solution and
Control Piperazine
citrate powder standard drug [Adani
Pharmachem Pvt. Ltd, Experiment Design Adult earth worms (Pheretima posthuma) were collected (due to their
anatomical and physiological resemblance with the intestinal round worm
parasites of human beings) Earth worms were thoroughly washed with normal
saline to remove the adhering material. Petridishes
of equal size were collected and 20ml of normal saline alone was poured in
the first petridish, 20ml of Piperazine
citrate solution of concentration 1, 2 and 4mg/ml were poured in second,
third and fourth petridishes, respectively. Then
20ml (4mg/ml) of the test solutions that is, the ethanolic
extracts of Piper nigrum,
Piper longum, Zingiber officinale and Trikatu churna were taken in fifth, sixth, seventh and eighth petridishes, respectively. Placed six earth worms of
nearly equal size in each petridish and time taken
for the induction of paralysis (motion less) and complete death of earth
worms was noted. The experiment was repeated thrice and confirmed the readings (Dwivedi et al, 2008). Statistical Analysis All the data are expressed as mean ± S.E.M. (standard error of the mean). The significance level was determined using the Student’s ‘t’ test. A p value of <0.05 was considered statistically significant. Results and Discussion The results of the above studies demonstrated that, the alcoholic extract of Trikatu churna and its individual ingredients possess potent anthelmintic activity with varying magnitudes. But the extract of Trikatu churna showed highest activity, which is almost equal in effectiveness to standard Piperazine citrate. The difference in the time taken for induction of paralysis in both Piperazine citrate and Trikatu churna was insignificant or almost same. However, significant difference was observed when compared the induction of paralysis time of Piperazine with ingredients of churna alone. The mode of action for the piperazine is generally by paralysing parasites, which allows the host body to easily remove or expel the invading organism (Table 1). The preliminary phytochemical observations of the alcoholic extracts of four different test samples such as Trikatu churna and its plant ingredients have shown the occurrence of alkaloids, flavonoids, tannins, lignins and steroids (Table 2). It indicates that, the Trikatu churna is a mixture all these phytoconstituents and interaction all these chemicals might be resulted in synergistically enhanced therapeutic efficacy of anthelmintic activity. Table 1. Anthelmintic activity of ethanolic (95%) extracts of Piper longum, P.nigrum, Zingiber officinalis, Trikatu churna and Piperazine citrate.
Table 2. Distribution of primary and secondary metabolites in Trikatu churna and its ingredients.
The main finding from this
investigation is that the alcoholic extracts from all four test samples
produced paralysis of earth worms after 30-120 minutes when treated 4mg/ml concentration of the Trikatu churna and individual
extracts. Moreover earthworms did not get recovery from paralysis even after
2-3 hours of post treatment period. Chemotherapeutic drugs against helminthes
infection act mainly through three different mechanisms, such as, disruption
of the neuromuscular physiology, blocking the energy metabolism, disrupting
the highly efficient reproductive system of the parasites (Geary et al,
1992). Several important anthelmintics cause
paralysis by disrupting one or the other aspect of neuromuscular system (Loukas and Hotez, 2005). The
paralytic effect of alcoholic extract of Piper
longum on Gigantocotyle explanatum by
progressive reduction in the spontaneous muscular activity, which may be associated with
their inhibitory effect on the neuromuscular system of the amphistome (Singh et
al, 2008). The anthelmintic activity of Zingiber officinale and
Piper longum a chief ingredients in
Krimikuthar Ras, Sanjivani vati and Vidangasava, an Ayurvedic
formulations were reported as potent anthelmintic
agents (Nirmal et
al, 2008). In vivo anthelmintic activity of
ginger was confirmed against gastrointestinal nematodes of sheep, thus
justifying the age-old traditional use of this plant in helminth
infestation (Iqbal et al, 2006). The oil of Piper
longum elicited the pronounced effect on the
rhythmic movements of Ascaris
as evidenced from resulting paralysis that occurred between 12 to 15 minutes
of exposure with 1:1000 v/v concentration of oil. The essential oil of Piper longum and
its non-phenolic fraction did not significantly
differ in their ability to inhibit the rhythmic motions, even though; minimal
motions persisted in case of non-phenolic fraction
(Kokate et al,
1980). Thus, it has happened obviously that, Trikatu
churna is a mixed preparation of all these useful phytoconstituents, perhaps the synergistic interaction of
alkaloids, flavonoids, tannins, lignins,
steroids and other constituents in the extract may impart strong anthelmintic activity to the poly herbal preparation. But
the mechanism involved in the interaction between the different plant
extracts remain unclear and should be further evaluated. Conclusion
Trikatu churna was
found to possess higher the rate of phytoconstituents
and promising anthelmintic activity. It is also
confirmed that, these spicy products triggers natural immune system to fight
against various parasites as well as helminthes. This study would provide the
preliminary scientific evidence for the folkloric, ethno-botanical and
traditional use of this churna for destruction of helminthes
/ parasites and eliminates from host body and other health benefits. References Anonymous. (1985). Indian
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