Investigation of in Vitro Anthelmintic Activity of Cinnamomum Camphor Leaves

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Investigation of in Vitro Anthelmintic activity of Cinnamomum Camphor Leaves

HAQUE RABIUL1, MONDAL SUBHASISH2, GHOSH PARAG2, *1Department of Biotechnology, Institute of Technology & Marine Engineering, D.H.Road, West Bengal, 2 Department of Pharmaceutical Technology, Jadavpur university,West Bengal, India.

C o Abstract Key words: v The aqueous extract of Cinnamomum camphorLeaves Cinnamomum camphor Leaves, , vermicidal, e r was investigated for an thelmintic activity using anthelmintic activity e d earthworms(Pheretima posthuma), tapeworms i (Raillietina spiralis) and rou ndworms (Ascaridia galli). n How to Cite this Paper:

O Various concentrations (10-70 mg/ml) of plant extract Haque Rabiu, Mondal Subhasish, Ghosh f were tested in the bioassay. Piperazine citrate (10 f Parag, “Investigation of in Vitro Anthelmintic i F mg/ml) was used as reference standard drug whereas c

u i activity of Cinnamomum Camphor Leaves”, Int. J.

a l distilled water as control.Determination of paralysis

l l

Drug Dev. & Res., Jan-March 2011, 3(1): 295-300

time and death time of the worms were recorded. L

e r Extract exhibited significant anthelmintic activity at the n o

h aqueous extract possesses ve rmicidal activity and found This is an open access paper distributed under the c t R to be effective as an anthelmintic. Therefore, the copyright agreement with Serials Publication, which o e anthelmintic activity of the aqueous extract of s f permits unrestricted use, distribution, and

E Cinnamomum camphorLeaves has been reported. a reproduction in any medium, provided the original l

r Introduction s

c e work is properly cited. h Infections with helminth are among the most v i

P widespread infections in humans and other domestic e a r animals affecting a large number of world population. Article History:------p ,

e T The majority of these infe ctions due to worms are Date of Submission : 23-10-2010 r h

generally restricted mainly to the tropical regions and Date of Acceptance : 25-01-2011 e

N the occurance is accelerated due to unhygienic lifestyle Conflict of Interest: NIL e and poverty also resulting in the development of Source of Support NONE t : h symtomps like anaemia, eosinophilia and pneumonia1. e r Parasitic diseases cause ru thless morbidity affecting l a principally in population. Introduction: n Synonym: Camphor tree, Gum camphor, camphor d s laurel.[1,2,3,45,] Botanical name: Cinnamomum

camphorScientific Classification:Kingdom- Planate *Corresponding author, M ailing address: Prof ( Dr) Rabiul Haque, Subkingdom- Tracheobionta, Superdivision- Institute of Technology & Marine Engineering. South 24 Parganas, Spermatophyta Division- Magnoliophyta, Class- West Bengal, Magnoliopsida ,Subclass –Magnoliidae, Order- India. Phone-+91 98317-85874. Laurales Family- Lauraceae, Genus- Cinnamomum E-mail- ruuubil_haque@y ahoo.com

Int. J. Drug Dev. & Res., Jan-March 2011, 3 (1): 295-300 Covered in Scopus & Embase, Elsevier 295 Haque Rabiul et al: Investigation of in Vitro Anthelmintic activity of Cinnamomum Camphor Leaves

,Species- Cinnamomum verum, Cinnamon is from a rough and fissured vertically.Camphor is a white tropical ever green tree of the laurel family growing crystalline substance, obtained from the tree

up to 7m(56feet) in itswild state. It has thick Cinnamomum camphora.Camphor has been[6,7,8,9] scabrous bark, strong branches, young shoots, used for many centuries as a culinary spice, a speckled greeny orange. The bark is smooth and component of incense, and as a medicine. Camphor yellowish. The leaves are ovate, petiolate,deeply is also an insect repellent and a flea-killing veined leaves that are dark green on top, lightgreen substance.Cinnamomum camphora is native to

C underneath. They become leathery when Taiwan, southern Japan, southeast China and o

v matureupper side shiny green, under side lighter. Indochina, where it is also cultivated for camphor e

r When bruished they smell spicy and have a hot taste. and timber production. The production and e

d The flowers are yellowish white with disaggreable shipment of camphor, in a solid, waxy form, was a

i odour that bears dark berries. The fruit is an oval major industry in Taiwan prior to and during the n

berry like an acron in its receptical is bluish when Japanese colonial era (1895–1945). It was used O f ripe with white spots on it, bigger than a medicinally and was also an important ingredient in f i F c blackberry.Chemical constituents: Cinnamaldehyde, the production of smokeless gunpowder and u i a l mannitol, coumarins, gum, tannins, essential oils celluloid. Primitive stills were set up in the l l

P (aldehyde,eugenol, pinene). mountainous areas in which the tree is usually e r n o [www.globalresearchonline.net] It is one of the major found. The wood was chipped; these chips were g d

t components in Ayurvedic medicine, which has been steamed in a retort, allowing the camphor to u h c

practiced in India since many centuries. The is an crystallize on the inside of a crystallization box, after t R o e ancient Indian cure-all due to its antibacterial, the vapour had passed through a cooling chamber. It s f

E antifungal, antiviral, antihistamine and antiseptic was then scraped off and packed out to government- a l r s properties.The Cinnamomum camphor leaves, run factories for processing and sale. Camphor was c e h v flowers, seeds, roots, bark and fruits are utilized to one of the most lucrative of several important i P e treat inflammation, infections, skin diseases. government monopolies under the Japanese.In the a r p , Antihelminthics or anthelmintics are drugs that ancient and medieval Middle East and Europe,

e T r

h expelparasitic worms (helminthes) from the body by camphor was used as ingredient for sweets but it is e either stunning or killing them. They may also be now mainly used for medicinal purposes. For

N called vermifuges (stunning) or vermicides (killing). example, camphor was used as a flavoring in e t

h The worms causing heliminthiasis are Round worms, confections resembling ice cream in China during the e Hook worms, Tape worms, Pin worms, Whip worms Tang dynasty (AD 618–907). An anonymous r l a e.t.c. Andalusian cookbook of the 13th century contains a n

d Cinnamomum camphora (commonly known as recipe for meat with apples, which is flavored with s [1]

Camphor tree, Camphorwood or camphor camphor and musk. A 13th century recipe for

laurel) is a large evergreen tree that grows up to "Honeyed Dates" is also flavored with Camphor.[2] By 20–30 metres tall. The leaves have a glossy, waxy the time of the Renaissance, camphor as a culinary appearance and smell of camphor when crushed. In ingredient had fallen into disuse in Europe.Today, spring it produces bright green foliage with masses of camphor is widely used in cooking (mainly for small white flowers. It produces clusters of black dessert dishes such as kheer or paal paayasam) in berry-like fruit around one centimetre in diameter. It India where it is known as pachha karpooram has a pale bark that is very (literally meaning "green camphor"). It is widely

Int. J. Drug Dev. & Res., Jan-March 2011, 3 (1): 295-300 Covered in Scopus & Embase, Elsevier 296 Haque Rabiul et al: Investigation of in Vitro Anthelmintic activity of Cinnamomum Camphor Leaves

available at Indian grocery stores and is labeled as called chemotypes, which are camphor, linalool, 1,8- "edible camphor". In Hindu poojas and ceremonies, cineole, nerolidol, safrole, or borneol. In China field camphor is burned in a ceremonial spoon or plates workers avoid mixing chemotypes when harvesting

for performing aarti. This type of camphor is also by their odour.[8][9] The cineole fraction of camphor sold at Indian grocery stores but it is not suitable for laurel is used in China to manufacture fake

cooking.The twigs and leaves of the camphor plant "Eucalyptus oil".[10] are used in the smoking and preparation of Zhangcha The chemical variants (or chemotypes) seem

C duck, a typical banquet and celebratory dish in dependent upon the country of origin of the tree. The o

v Szechuan cuisine tree is native to China, Japan, and Taiwan. It has e

r been introduced to the other countries where it has e

d In India's ancient Ayurvedic Medical texts it is been found, and the chemical variants are

i explained that every part of the : Cinnamomum identifiable by country. i.e., Cinnamomum camphora n

camphor tree has health promoting benefits. What is grown in Taiwan and Japan, (often commonly called O f clear from the above information is that the general "Ho Wood") is normally very high in Linalool, often f i F c population of India for, over 5000 years, has used : between 80 and 85%. In India and Sri Lanka the high u i a l Cinnamomum camphor safely and effectively. It is camphor variety/chemotype remains dominant. The l l

P also called Holy Tree (: Cinnamomum camphor In Cinnamomum camphora grown in Madagascar, on e r n o ancient times neem was the most celebrated the other hand, is high in 1,8 Cineole (averaging g d

t medicinal plant of India and found mention in a between 40 and 50%. The essential oil from the u h c

number of Puranic texts like the Atharava Veda, Madagascar trees is commercially known as t R o e Upanivahod, Amarkosha and Ghrysutra. They all Ravintsara.[11] s f

E dealt with the outstanding qualities of the a l r s Cinnamomum camphor tree as a source of medicine Materials and methods c e h v pesticide.. Today, neem is once again steadily Plant Materials i P e becoming an agro-scientific celebrity. It has figured The Cinnamomum camphora tree was collected from a r p , as the priority in seminars and serious agricultural the nursery ( Amtala) south 24 paraganas district,

e T r

h workshops all over the world. Modern western West Bengal, India. The plant material was e medicine is finally discovering what the ancient taxonomically identified by the taxonomists of

N Indians have known for thousands of years: that the : Botanical Survey of India, Kolkata. A voucher e t

h Cinnamomum camphor tree has superb specimen has been preserved in our laboratory for e pharmaceutical and pesticide controlling qualities. future reference. The plant material was dried in r l a Its effectiveness, availability and safety have made shade, pulverized, passed through sieve no. 40 and n

d agro-scientists promote cultivation of: Cinnamomum stored in air tight container and used for further s

camphor forests. Camphor trees grow in full sun to extraction. partial shade. They tolerate clay, loam, sand, slightly

alkaline to acidic soils, and drought.[10,11,12,,13,] They Preparation of extract need to be well drained or they may suffer from root Aqueous extract (Maceration method) rot , Camphor laurel contains volatile chemical Mature green leaves,pulverized,root, wood are compounds in all plant parts, and the wood and collected and allowed to dry partially. These dried leaves are steam distilled for the essential oils. leaves , root,wood are then crushed and powdered. Camphor laurel has six different chemical variants The crushed leaves are then subjected to either

Int. J. Drug Dev. & Res., Jan-March 2011, 3 (1): 295-300 Covered in Scopus & Embase, Elsevier 297 Haque Rabiul et al: Investigation of in Vitro Anthelmintic activity of Cinnamomum Camphor Leaves

aqueous or organic solvent to get a concentrated The average size of round worm was 5-7 cm and extract. For making neem leaf extract, certain average size of tapeworm was 6-8 cm. Earthworm extraction process utilizes carbon dioxide at critical and helminths were identified in Dept. of Zoology, temperatures and pressures to extract the active Vivekananda College,Thakurpukur, Kolkata. ingredients of the leaf, the usual high temperatures or harsh chemicals are done away with, resulting in a Drugs & Chemicals better concentrated and potent extract. Piperazine citrate (Glaxo Smithkline) was used as

C The plant leaves were dried in shade, pulverized and standard anthelmintic during the experimental o

v then powdered material of Cinnamomum camphora protocol. e

r powder(500gm) were kept for maceration with 1000 e

d ml of double distilled water for 24 hours. The extract Anthelmintic activity

i was double filtered by using muslin cloth and The anthelmintic assay was carried out as per the n

Whatman no.1 filter paper and concentrated by method of Ajaiyeoba et al 4 . The assay was O f evaporation on water bath. The extract was dried and performed in vitro using adult earthworm f i F c used as a powder. The percentage yield of extract was (Pheretima posthuma) as it is having anatomical and u i a l found to be about 4%. physiological resemblance with the intestinal round l l

P worm parasites of human beings for preliminary e r

n 17,18,19] o Experimental Animals evaluation of anthelmintic activity [ . Use of g d

t Adult earthworms (Pheretima posthuma), Ascaridia galli and Raillietina species as a suitable u h c

Roundworm (Ascaridia galli) and Tapeworms model for screening of anthelmintic drug was t R o e (Raillietina spiralis) were used to evaluate advocated earlier [ 20,21]. s f

E anthelmintic activity in vitro. Earthworms were Test samples of the extract was prepared at the a l r s collected from moist soil and washed with normal concentrations, 10, 20,30,40 , 50,60,70, mg/ml in c e h v saline to remove all faecal matter were used for the distilled water and six worms i.e. Pheretima i P e anthelmintic study. The earthworms of 3-5 cm in posthuma, Ascaridia galli and Raillietina spiralis of a r p , length and 0.1-0.2 cm in width were used for all the approximately equal size (same type) were placed in

e T r

h experimental protocol. Roundworms and tapeworms each nine cm Petri dish containing 25 ml of above e were obtained from intestine of freshly slaughtered test solution of extracts. Piperazine citrate (10 mg

N fowls. Infested intestines of fowls were collected from /ml) was used as reference standard was advocated e t 20,21 h the local slaughter house and washed with normal earlier .Test samples of the extract was prepared e saline solution to remove all the faecal matter. These at the concentrations, 10, 20,30,40 , 50, 60 and 70 r l a intestines were then dissected and double distilled mg/ml in distilled water and six worms i.e. n ,[13,14,15,16] d water as control . This procedure was Pheretima posthuma, Ascaridia galli and Raillietina s

adopted for all three different types of worms. All the spiralis of approximately equal size (same type) were test solution and standard drug solution were placed in each nine cm Petri dish containing 25 ml of prepared freshly before starting the experiments. above test solution of extracts. Piperazine citrate (10 Observations were made for the time taken for mg /ml) was used as reference standard and double

paralysis was noted when no movement of any sort distilled water as control [14,15,16,17] . This procedure could be observed except when the wormsand worms was adopted for all three different types of worms. All were collected and kept in normal saline solution. the test solution and standard drug solution were prepared freshly before starting the experiments.

Int. J. Drug Dev. & Res., Jan-March 2011, 3 (1): 295-300 Covered in Scopus & Embase, Elsevier 298 Haque Rabiul et al: Investigation of in Vitro Anthelmintic activity of Cinnamomum Camphor Leaves

Observations were made for the time taken for[ shaken vigorously nor when dipped in warm water

22,23,24,25,26,27,,28] paralysis was noted when no (500 C). movement of any sort could be observed except when All the results were shown in Table.1 and expressed the wormsTime for death of worms were recorded as a mean ± SEM of six worms in each group. after ascertaining that worms neither moved when

Table – 1: Anthelmintic activity of Cinnamomum camphora aqueous extract

C ± SEM (N=6) Each value represents mean o

v (P) in min.

e (Mean & Time taken Time Time taken Time Time taken r SEM)

e for taken for for taken for for Time d paralysis death (D) paralysis death (D) paralysis GROUP taken for (P) in min. in min. (P) in min. in min. (P) in min. i death (D) n (Mean & (Mean & (Mean & (Mean & (Mean & in min.

SEM) SEM) SEM) SEM) SEM) O (Mean &

f SEM) f Control i F 01 ……….. ………. ……….. ……… …………… ……… ……….. c (Water Only u i 32±0.33 28±0.37 a l 53±0.23 55±0.57 61±1.45 l

l 10 34±1.65

P 25±0.25 23±0.78 46±0.27 52±0.43 47±0.54 e

r 20 24±0.22

n o 19±0.55 14±0.29

g 30±0.64 38±0.56 40±1.18 d 30 20±1.67

t u

h 17±0.38 14±0.55 31±0.72 39±1.25 41±0.50 c

40 19±0.59 t R 02 Cinnamomum camphora 17±0.11 13±0.32

o 32±0.25 39±1.101 41±0.11 e 50 19±0.15 s f

e E

a 22±1.17 13±1.58 62±0.77 38±1.19 53±0.14 l

r 60 26±0.56 s c e 19±0.56 24±0.77

h 45±0.28 38±0.28 49±0.17 v 70 26±0.53

P e a

r 22±1.10 60±0.75 12±1.50 38±1.10 24±0.50 52±0.10 03 Piperazine citrate 10 p ,

e T Each value represents mean ± SEM (N=6) r h

N RESULTS AND DISCUSSIONS conclude that Neem which is used traditionally to e

t From our observations, higher concentration of treat intestinal worm infections, exihibited h

e extract produced paralytic effect much earlier and the significant anthelmintic activity. Therefore, further r

l time taken for death was shorter for all types of study must be carried out so that the general people a n worms. Aqueous extract of Cinnamomum camphora can get actual benefit from this important medicinal d

s exhibited anthelmintic activity in dose-dependent plant. manner showing maximum efficacy at 50 mg/ ml concentration for all three types of worms. Our plant Acknowledgments extract exhibited more potent activity at lowest We wish to thank the Director Dr Professor T.K Basu concentration (10 mg/ml) against (roundworm) of my institute The Institute of Technology & Marine Ascaridia galli. Anthelmintic activity of the extract Engineering for his moral support and blessings and was compared with the standard drug Piperazine I also express my heartful thanks to him for citrate (Table.1). From the above results, we can motivating me with strength and spirit.

Int. J. Drug Dev. & Res., Jan-March 2011, 3 (1): 295-300 Covered in Scopus & Embase, Elsevier 299 Haque Rabiul et al: Investigation of in Vitro Anthelmintic activity of Cinnamomum Camphor Leaves

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