Exploring Schleichera oleasa crude bark extract for its anti-helminthic potential

 

Jasmine Chaudhary¹*, Akash Jain¹, Anns Sabu¹, Priyanka Jha¹, Prerna Sarup²,

Vishal Sharma¹, Diksha Sharma³

¹MM College of Pharmacy, Maharishi Markandeshwer (Deemed to be University),

Mullana, Ambala-133207, Haryana, India.

²Swami Vivekanand Institute of Pharmacy, Banur, Punjab, India.

³Institute of Pharmaceutical Sciences, Kurukshetra University, Kurukshetra.

 

ABSTRACT:

Schleichera oleosa (Lour.) Oken or Kusum, belonging to family Sapindaceae is an important medicinal plant reported to possess various pharmacological activities viz. analgesic, antibiotic, anti-dysentery, anti-helminthic, antioxidant and anticancer activities. Scientific studies reporting antihelminthic properties of bark extract of this plant are still lacking, therefore, the present work has been designed to investigate the ethanolic and ethyl acetate extracts of bark for its antihelminthic potential.  Antihelminthic evaluation was done using Indian earthworms, Pheretima posthuma by determining time of paralysis and death. Phytochemical screening of the extracts revealed the presence of flavonoids, tannins and phenolic acids. The anti-helminthic activity of both extracts (0.1%) was found better than the standard drug Albendazole. Since, both extracts showed promising results, they can be further helpful in designing herbal formulations for commercial use.dry-powder inhaler (DPI) carries medication to lungs as a dry powder, useful against respiratory diseases.

 

 

 

INTRODUCTION:

Medicinal plants, the “backbone” of traditional medicines^1-4, have been found to be of immense global significance⁵ because of their efficacy in treatment of a variety of ailments⁶. Even today, majority of population is still dependent on traditional therapy as their primary healthcare^{7, 8}. Their high safety margin, easy availability, acceptability, affordability in comparison to allopathic drugs^9-12 and certain inevitable factors like population rise, scarcity in supply of drugs, development of resistance to currently used drugs, are further increasing their popularity and demand^{13, 14}.

 

Schleichera oleosa (synonym: kusum, lac tree, gum tree, Ceylon oak), a well known medicinal plant belonging to family Sapindaceae (soapberry) is used traditionally since ancient times for its spectacular medicinal properties viz. astringent, antimicrobial^{15, 16}, anti-inflammatory¹⁷, antimalarial¹⁸, antidiabetic¹⁹, anticancer²⁰, antihypertensive²¹, anti-oxidant^22-26 and antihelminthic²⁷. Also this plant has been used to treat arthritis²⁸, burns, various skin problems and alopecia^{29, 30} etc. It is a large deciduous tree of monotypic genus found naturally in foothills of Himalayas, Sri Lanka and China³¹. Kusum based herbal formulations are also available in global drug market as promising product³². Various phytoconstituents like triterpenoids, tannins, phenolic compounds, fatty acids, proteins, carbohydrates etc. have been reported in different plant parts³³, therefore, several pharmacological studies have been conducted to authenticate its use as a multi-purpose medicinal agent^{31, 34}.

 

 

Helminthiasis is also one of most prevalent worm infection posing a great threat to human health especially in rural areas which contribute to the prevalence of undernourishment, anaemia, ascariasis, lymphatic filariasis, onchocerciacis and schistosomiais^{35, 36}. Although, a large number of marketed drugs such as albendazole, piperazine, praziquantel and pyrantel are available for the treatment but their prolong use make the helminths resistant to the currently available anthelmintic drugs³⁷. Other limitations like cost, their common side effects like nausea, vomiting, headache, abdominal pain and diarrhoea etc. further leads to more interest of researchers towards assessment of medicinal plants as anthelmintic^38-39. Antihelminthic potential of S. oleosa leaves have already been explored^{34, 40-41}. Therefore, the present study has been designed to explore the bark extract of S. oleosa for its anti-helminthic potential.

 

MATERIALS AND METHODS:

Plant Material Collection and Identification:

The bark of Schleichera oleosa plant was collected from herbal medicinal garden of Maharishi Markandeshwar (Deemed to be University), Mullana, Ambala. The collected plant material was air dried and authenticated by CSIR-NISCAIR, New Delhi (Ref. No. NISCAIR/RHMD/Consult/2018/3158-07-02). All the chemicals and reagents used were of analytical grade. TLC plates were prepared with silica gel G and the spots were visualized using freshly prepared anisaldehyde reagent.

 

Preparation of Extract and Phytochemical Screening:

Dried bark powder of S. oleosa was sequentially extracted using different solvents such as chloroform, ethanol, ethyl acetate in increasing order of polarity by Soxhlet extraction. The extract was concentrated using rotatory evaporator and the percentage yield of all extracts were calculated. The dried extract was used for further studies.

 

Phytochemical screening plays important role in identifying new sources of active constituents present in the plant extracts⁴². The extract obtained was therefore, investigated for the presence of various bioactive compounds using different chemical tests^43-45.

 

Antihelminthic studies:

The antihelminthic activity was evaluated on Indian earthworm Pheretima postuma due to its easy accessibility and anatomical resemblance with the intestinal roundworm parasite of human being^46-48. Earthworms used in experimental protocol (4-5 cm in length and 0.1 - 0.2 cm in width) were obtained from moist soil of MM (DU), Mullana, Ambala.

 

Five earthworms of nearly equal size, washed with normal saline to remove all faecal material were placed in standard drug solution (Albendazole) and sample extracts solutions at room temperature. Earthworms tested in normal saline were taken as a control group. The standard as well as test extracts were dissolved in minimum quantity of dimethyl sulfoxide (DMSO) and volume was adjusted with normal saline solution to get concentration of 0.1% w/v. The extracts were evaluated by the time taken for complete paralysis (i.e. when no movement of any sort could be observed, except when the worm was shaken vigorously) and death (worms neither moved when shaken, nor when given external stimuli i.e. hot water 50 °C) of earthworms. Observations were made during the test period and extended up to 5 hours. The mean lethal time for extracts was recorded and compared with standard drug.

 

RESULTS AND DISCUSSION:

The powdered bark was subjected to sequential extraction in increasing order of their polarity and % extractive yield was calculated (Table 1).

 

Table 1. Percentage extractive value

S. No.	Solvent Used	% Extractive value
1.	Petroleum ether	1.14
2.	Chloroform	2.121
3.	Ethyl acetate	4.39
4.	Ethanol	7.31

 

Out of the four solvents used for determination of extractive value, the ethanolic extract gave the maximum yield followed by ethyl acetate, petroleum ether and chloroform ether. Preliminary phytochemical screening of the bark extracts were tested for the presence of primary and secondary metabolites like alkaloids, tannins, saponins, carbohydrates etc. The various components present in extracts are shown in Table 2. It was observed that no alkaloids, glycosides, saponins are present in any of the extract however carbohydrates and proteins are present in the ethanolic extract. The maximum amount is found for phenolic compounds, flavonoids, terpenoids and phytosterols indicating potency of these extracts for various biological properties.

 

Table 2. Qualitative analysis of S. oleosa extracts

Plant Constituents	Test performed	PE	CE	EAE	EE
Alkaloids	Dragondroff’s test	-	-	-	+
	Hager’s test	-	-	-	+
	Mayer’s test	-	-	-	+
Carbohydrates	Benedict’s test	-	-	-	+
	Molisch’s test	-	-	-	+
Glycosides	Legal’s test	-	-	-	+
	Keller killani	-	-	-	+
Proteins	Biuret’s test	-	-	-	+
Saponins	Foam’s test	-	-	-	-
Phenolic compounds and tannins	Ferric chloride test	+		+	++
Flavonoids	Alkaline reagent test	-	+	+	+
Terpenoids	LB test	+	+	+	+
Phytosterols	Salwoski’s test	++	+	+	+
Steroids	Libermann and Buchard test	++	+	+	+

(+ indicates the presence of constituents and ─ indicates the absence of constituents)

PE: Petroleum ether extract; CE: Chloroform extract; EAE: Ethyl acetate extract; EE: Ethanolic extract

 

Anti-helminthic activity:

The anti-helminthic activity of both selected extracts has been given in Table 3 and it reveals that both extracts show better activity than the standard drug Albendazole against Indian roundworm Pheretima postuma.

 

Table 3: Anthelminthic studies

Test Compound	Mean Paralysis Time (min.) ± SD	Mean Death Time (min.) ± SD
Standard (Albendazole) 0.1% (w/v)	111.67±29.94	177.83±26.98
Control (Normal saline) 0.9% (w/v)	NA	NA
Ethyl Acetate Extract 0.1% (w/v)	6.83±1.57	25.00±3.26
Ethanolic Extract 0.1% (w/v)	11.33±3.44	30.00±4.0

*NA indicates no activity

 

CONCLUSION:

The present work evidenced that the ethyl acetate as well as ethanolic bark extracts of Schleichera oleasa have promising antihelminthic activity which might be due to presence of flavanoids and triterpenoidal constituents present in them. The observed antihelminthic activity suggests that the phytoconstituents of S. oleosa should be isolated and further investigated in new detail as potential antihelminthic leads. The potential of this plant can be assessed for therapeutic application in the field of parasitology as this study has provided an insight into new resource for the development of antihelminthic drugs.

 

CONFLICT OF INTEREST:

The authors have no conflicts of interest regarding this investigation.

 

ACKNOWLEDGMENTS:

The authors are thankful to management of MM College of Pharmacy, MM (Deemed to be University, Mullana (Ambala) for providing support and necessary facilities.

 

REFERENCES:

1.     Lone PA. Bhardwaj AK. Shah KW. Tabusam S. Ethnobotanical survey of some threatened medicinal plants of Kashmir Himalaya. Indian Journal of Medical Research. 2014; 8(47):1362-1373. doi: org/10.5897/JMPR2014.5649

2.     Ahvazi M. Sigaroodi FK. Charkhchiyan MM. Mojab F. Mozaffarian VA. Zakerif, H. Introduction of medicinal plants species with the most traditional usage in Alamut region. Iranian Journal of Pharmaceutical Sciences. 2012; 11(1):185–194.

3.     Singh R. Medicinal plants: A review. Journal of plant Sciences. 2015; 3(1-1):50-55. doi: org/10.11648/j.jps.s.2015030101.18

4.     Devprakash, Sumalatha BV, Srinivasan KK, Savitha. Tephrosia purpurea (Linn.)Pers.-A Comprehensive Review. Research Journal of Pharmacognosy and Phytochemistry. 2012; 4(2):134-143.

5.     Gangola S. Khati P. Bhatt P. Parul. Sharma A. India as the heritage of medicinal plant and their use. Current Trends in Biomedical Engineering & Biosciences. 2017; 4(4):50. doi: org/10.19080/CTBEB.2017.04.555641

6.     Shakya AK. Medicinal plants: Future source of new drugs. International Journal of Herbal Medicine. 2016; 4(4): 59-64. doi: org/10.13140/RG.2.1.1395.6085

7.     Karagoz A. Artun FT. Ozcan G. Melikoglu G. Sezin A. Kultur S. Sutlupınar N. In vitro evaluation of antioxidant activity of some plant methanol extracts. Biotechnology & Biotechnological Equipment. 2015; 29(6):1184-1189. doi: 10.1080/13102818.2015.1080600

8.     Jeyabaskar S, Viswanathan T, Mahendran R, Rathisre PR, Nishandhini M. Comparative quantitative screening of secondary phytoconstituents from the leaves extract of Sterculia foetida Linn. Research Journal of Pharmacy and Technology. 2017; 10(9):2907-2912. doi: 10.5958/0974-360X.2017.00513.3

9.     Philomena G. Concerns regarding the safety and toxicity of medicinal plants - An overview. Journal of Applied Pharmaceutical Sciences. 2011; 01(06):40-44. doi: org/10.1016/j.heliyon.2020.e04150

10.  Chattopadhyay RR. Bhattacharyya SK. Terminalia chebula: An update. Pharmacognosy Reviews. 2007; 1(1): 151-156.

11.  Bhargavi S, Swamy V, Kumari SM, Ushasri S, Kumari PM. Antimicrobial and wound healing activities of leaves of Solanum erianthum D. Don. Research Journal of Pharmacy and Technology. 2013;6(5): 531-536.

12.  Gowri SS. In vitro anticancer activity of Majidea zanquebarica J. Krik. ex Oliv. (Sapindaceae). Research Journal of Science and Technology. 2020;12(3):173-176. doi: 10.5958/2349-2988.2020.00023.6

13.  Oladeji OS. Oyebamiji AK. Stellaria media (L.) Vill.- A plant with immense therapeutic potentials: Phytochemistry and Pharmacology. Heliyon. 2020; 6(6):e04150.

14.  Atanasov AG. Waltenberger B. Wenzig EMP. Linder T. Wawrosch C. Uhrin P. Temml V. Wang L. Schwaiger S. Heiss EH. Rollinger JM. Schuster D. Breuss JM. Bochkov V. Mihovilovic MD. Kopp B. Bauer R. Dirsch VM. Stuppner H. Discovery and resupply of pharmacologically active plant- derived natural products: A review. Pharmaceutical Biology. 2015; 33(8):1582–1614. doi: org/10.1016/j.biotechadv.2015.08.001

15.  Srinivas K. Baboo RVC. GC-MS study of Schleichera oleosa. International Journal of Current Pharmaceutical Research. 2011; 2(2):1-11.

16.  Ghosh P. Chakraborty P. Mandal A. Saha A. Triterpenoids from Schleichera oleosa of Darjeeling foothills and their antimicrobial activity. Indian Journal of Pharmaceutical Sciences. 2011; 73(2):231-3. doi: 10.4103/0250-474x.91568

17.  Khan MJ. Saraf S. Saraf S. Anti-inflammatory and associated analgesic activities of HPLC standardized alcoholic extract of known ayurvedic plant Schleichera oleosa. Journal of Ethnopharmacoogy.. 2017; 197:257–65. doi: org/10.1016/j.jep.2016.08.021

18.  Mahaptma SP. Sahoo HP. An ethno-medico-botanical study of Bolangir, Orissa, India: Native plant remedies against gynaecological diseases. Ethnobotanical Leaflets. 2008; 12:846-50.

19.  Goswami S. Singh RP. Ayurvedic, phytochemical and pharmacological review of Schleichera oleosa (Lour.) Oken: A traditional plant with enormous biological activity. World Journal of Pharmaceutical Research. 2017; 6: 295-309. doi: org/10.20959/wjpr201710-9370

20.  Chan MK. Khine ZWL. Nwet NW. Hla N. Investigation of phytochemical constituents and some biochemical properties of the bark of Schleichera oleosa (Lour.) Oken. (GYO). Journal of the Myanmar Academy of Arts and Science. 2019; XVII(1A):477-495.

21.  Jose S. Dandapat S. Sinha MP. Anti-hypertensive activity of aqueous and methanolic leaf extracts of Schleichera oleosa (Lour.) Merr. Journal of Analytical & Pharmaceutical Research. 2019; 8(3):94-97. doi: org/10.15406/japlr.2019.08.00320

22.  Hobanthad T. Maneetong, S. Simple extraction for the scanning of antioxidant activity of vegetables and fruits in Buriram, Thailand by DPPH, ABTS and FRAP assays. SNRU Journal of Science and Technology. 2019; 11(3):114-121.

23.  Toobpeng N. Powthong P. Suntornthiticharoen P. Evaluation of antioxidant and antibacterial activities of fresh and freeze-dried selected fruit juices. Asian Journal of Pharmaceutical and Clinical Research. 2017; 10(9):156-160.

24.  Khandekar U. Bobade A. Ghongade R. Evaluation of antioxidant activity, in-vitro antimicrobial activity and phytoconstituents of Schleichera oleosa (Lour.) Oken. International Journal of Biological & Pharmaceutical Research. 2015; 6(2): 137-143.

25.  Srinivas K. Baboo RVC. Antioxidant activity of ethanolic extract of stem bark of Schleichera oleosa (Lour. Oken). International Journal of Pharmacotherapeutics. 2013; 3(1):12-14.

26.  Thind T. Agrawal S. Saxena A. Arora S. Evaluation of cytotoxic and radical scavenging activities of root extracts of Schleichera oleosa (Lour.) Oken. Natural Product Research. 2015; 26(18): 1728–31. doi: org/10.1080/14786419.2011.606547

27.  Prasad B. Subedi L. Medicinal plant diversity and their pharmacological aspects of Nepal Himalayas. Pharmacognosy Journal. 2011; 3:6-17. doi: org/10.5530/pj.2011.25.2

28.  Santha ML. Kanchana P. Shakeela SK. Anti-inflammatory and anti-arthritic activity of Scheilchera oleosa (Lour.) Oken bark. International Journal of Pharmaceutical Sciences Review and Research. 2017; 46(1):79-84.

29.  Bhatia H. Kaur J. Nandi S. Gurnani V. Chowdhury A. Reddy PH. Vashishtha V. Rathi B. A review on Schleichera oleosa: Pharmacological and environmental aspects. Journal of Pharmaceutical Research. 2013; 6(1):224-229. doi: org/10.1016/j.jopr.2012.11.003

30.  Kirtikar KR. Basu BD. Indian medicinal plants. 1981; 1:794-795. Lalit Mohan Basu Publishing, Allahabad.

31.  Mall TP. Tripathi SC. Kusum- a multipurpose plant from Katarniaghat wildlife sanctury of Bahraich (UP) India-a review. World Journal of Pharmaceutical Research. 2017; 6(4):463-477. doi: org/10.20959/wjpr20174-8082

32.  Meshram N. Ojha M. Singh A. Alexander A. Ajazuddin. Sharma M. Significance and traditional medicinal properties of Schleichera oleosa. Asian Journal of Pharmaceutical Research. 2015; 5(1): 61-64. doi: org/10.5958/2231-5691.2015.00009.X

33.  Soundararajan M. Sivakkumar T. Pharmacognostical investigation and phytochemical studies of Schleichera oleosa. International Journal of Research in Pharmaceutical Sciences. 2017; 8(2):109-119.

34.  Goswami S. Singh RP. In vitro assessment of anthelmintic and alpha-amylase inhibition of Schleichera oleosa (lour.) Oken leaf extracts. Asian Journal of Pharmaceutical and Clinical Research. 2018; 11(9):487-491. doi: org/10.22159/ajpcr.2018.v11i9.28338

35.  Blair P. Demeart D. Update on prevention and treatment of intestinal helminth infections. Current Infectious Disease Reports. 2015; 17(3): 465. doi: org/10.1007/s11908-015-0465-x.

36.  Harindran J, Rajalakshmi GR. In vitro anthelmintic activity of Samadera indica. Research Journal of Pharmacy and Technology. 2011;4(10): 1593-1595.

37.  Kousalya M, Geetha P, Jesuraja A, Vinoth KM. In-Vitro study of anthelmintic activity of  Eclipta prostrata (L) y various Extracts. Research Journal of Pharmacy and Technology. 2017; 10(1): 58-60.doi: 10.5958/0974-360X.2017.00014.2

38.  Shabi RR, Ajina S. Qualitative phytochemical screening and in-vitro anthelmintic activity of Adhatoda vasica (Acanthaceae). Research Journal of Pharmacy and Technology. 2017; 10(2):414-420. doi: 10.5958/0974-360X.2017.00083.X

39.  Siju EN, Rajalakshmi GR, Hariraj N, Sreejith KR, Sudhakaran S, Muneer EK, Premalatha K. In-vitro anthelmintic activity of Mussaenda frondosa. Research Journal of Pharmacy and Technology. 2010; 3(1):151-153.

40.  Dubey RS. Shukla R. Pandey V. Shukla AK. Investigations on Drosera regia and Schleichera oleosa leaves for anthelmintic activity. Advance Pharmaceutical Journal. 2019; 4(4):97-99. doi:  org/10.31024/apj.2019.4.4.1

41.  Goswami S, Singh RP, Gilhotra RM. HPTLC Fingerprinting of Quercetin and comparative assessment of Anthelmintic potency against Eudrilus eugeniae of Schleichera oleosa (Lour) Oken and Tagetes eretca Linn. Research Journal of Pharmacy and Technology. 2020; 13(12):5699-5704. doi: 10.5958/0974-360X.2020.00992.0

42.  Kokate CK. Pharmacognosy. 2004: 109-113. Nirali Prakashan Publishing, Pune, India.

43.  Jork H. Funk W. Fischer W. Wimmer H. Thin layer chromatography: reagent and detection methods. 1990: 195. Weinhein: Weinheim, Basel (Switzerland) Publishing, Cambridge, New York.

44.  Khandelwal KR. Kokate CK. Pawar AP. Gokhale SB. Practical Pharmacognosy. 1988: 31-40. Nirali Prakashan Publishing, Pune, India.

45.  Harbone JB. A guide to modern techniques of plant analysis. 1981:1. Chapman & Hall Publishing, London.

46.  Kumar KIA. Manickam MS. Sreejith M. Sebastin V. In vitro antioxidant and anthelmintic activity of the extracts of whole plant Crotalaria biflora (L). Annals of the Romanian Society for Cell Biology. 2021; 25(4):150-161.

47.  Indirayati N. Nisa K. Kurang RY. Tarmo NC. Adang KTP. Radical scavenging activity and total phenolic content of seven tropical plants. IOP Conference Series: Earth and Environmental Science. 2020; 462:1-7. doi: org/10.1088/1755-1315/462/1/012043

48.  Ajaiyeoba EO. Onocha PA. Olarenwaju OT. In vitro anthelmintic properties of Buchholzia coriaceae and Gynandropsis gynandra extract. Pharmaceutical Biology. 2001; 39:217-20. doi: org/10.1076/phbi.39.3.217.5936

 

 

 

Accepted on 11.02.2022           © RJPT All right reserved

Research J. Pharm. and Tech 2022; 15(11):5168-5171.