Int. J. Pharm. Sci. Rev. Res., 42(2), January - February 2017; Article No. 39, Pages: 217-220

ISSN 0976 - 044X

Research Article

Phytochemical Analysis and Antimicrobial Activity of Caesalpinia bonducella Leaves

Nirmala R. Kakade^1*, Shirish S. Pingale², Manohar G. Chaskar³

¹Department of Chemistry, Dada Patil Rajale Arts and Science College, Adinath nagar, Ahmednagar, MS, India.

²P.G. Department of Chemistry, Gramonnati Mandal’sArts, Com. & Sci. College, Narayangaon, Pune. India.

³Department of Chemistry, Baburaoji Gholap College Sangavi, Pune, India.

*Corresponding author’s E-mail: [email protected]

Received: 30-12-2016; Revised: 30-01-2017; Accepted: 14-02-2017.

ABSTRACT

Medicinal plants are the main source of pharmaceuticals and healthcare products. In the present study, our goal was to investigate

the phytoconstituents in the leaf extracts of Caesalpinia bonducella and their antimicrobial activity. The phytochemicals analysis of

leaves powder of Caesalpinia bonducella was carried out by using extraction method. Caesalpinia bonducella leaves contain

Terpenoids &Phenolics - 4.490%, Alkaloids - 0.885%, Q. Alkaloid & N-oxides - 25.745 %, fats and waxes - 8.850 % and Fibers -

60.050%. Antimicrobial studies showed that the above extracts had considerable activities against Bacillus subtilis, Staphylococcus

aureus, Escherichia coli, Klebsiella aerogenes, Aspergillus niger and Penicillium chrysogenum. The percentage of fats and waxes,

terpenoids and phenolics, Q. Alkaloid & N - Oxides was observed good. Its beneficial effect on animals as well as human health may

be derived from its antibacterial and antifungal properties to afford protection against various diseases. It might be an alternate to

synthetic antibiotics available in the market.

Keywords: Caesalpinia bonducella, quaternary alkaloids and n-oxides, terpenoids and phenolics, phytochemical, antimicrobial

activity.

INTRODUCTION

increased on large scale and it is going to be a big

therapeutic problem. The problem can be solved with the

rom prehistoric times, it has been an aim to use

help of antibiotic resistance inhibition from plants^{13, 14}.To

herbal plants as drugs for human beings but only 5%

be safe from various pathogens, trees produce different

of the total 3, 00,000 plant species has been studied

compounds. Plant extracts are expected to be active

scientifically for their medicinal purposes^{1, 2}. More than

against drug resistance pathogens¹⁵. For hundreds of

30% of all prescription drugs are derived from plant origin

years, various plants have been used to cure various

in the industrialized countries³. Ayurveda, Unani,

human illnesses in many parts of the world traditionally.

Homeopathy and Siddha are the traditional systems of

So, phytomedicines and biologically active compounds

medicines which mainly depend on the natural

derived from plants which are used in herbal medicines

products⁴.People below poverty line in the undeveloped

have been focused by researchers^{16, 17}.

and developing countries of Asia and Africa are suffering

from health problems connected with the consumption of

Caesalpinia bonducellais classified under the family of

mycotoxin contaminated grains and cereals⁵.

Caesalpiniaceae. Commonly it is called as Fever Nut,

Bonduc Nut and Nicker Nut also¹⁸. Leaves, seeds, stem,

To prevent illnesses and to maintain human health,

bark, nuts and roots are useful as herbal medicines. It is

phytochemicals are important sources of food and

found in hotter parts of India especially in west Bengal

medicine⁶. Medicinal plant products are used as home

and the southern states of India. In conventional system

remedies to treat specific conditions as well as complex

of Indian medicine, Ayurveda, Caesalpinia bonducella

preparations to treat life threatening diseases^7-8. A

(roxb.) is largely used for its antiperiodic, antipyretic, anti-

mention has been made in various phytotherapy manuals

inflammatory, anthelmintic, antimalarial properties. It is

about the medicinal plants and their uses in the

also described to have antioxidant, antibacterial,

treatment of infectious diseases because of their

antitumor and antidiabetic activities¹⁹.

availability, less side effects and decreased toxicity⁹.

Antibiotics which have been effectively used against

The phytochemical analysis of Caesalpinia bonducella

bacterial infections are very significant things that have

leaves powder was carried out by using extraction

been discovered in the twentieth century. But these

method and leaves extracts were screened for

synthetic drugs are used to treat only one third of the

antimicrobial

activity

against

Bacillus

subtilis,

known infectious diseases¹⁰. It is so because of

Staphylococcus aureus,

Escherichia coli, Klebsiella

indiscriminate and continuous use and misuse of

aerogenes,

Aspergillus

niger

and Penicillium

antibiotics and consequently the resistant pathogens

chrysogenum. It may provide scientific justification for

have emerged^{11, 12}. Recently, antibiotic resistance has

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Int. J. Pharm. Sci. Rev. Res., 42(2), January - February 2017; Article No. 39, Pages: 217-220

ISSN 0976 - 044X

their use in indigenous communities and broaden the

disc was placed near the edge of the agar surface of the

scope of existing drug discovery programs.

inoculated plate. The incubation of the plates was carried

out at 37°C for 24 hours. The transparent meter rule was

MATERIALS AND METHODS

used to measure the zones of inhibition. Chloramphenicol

Preliminary Phytochemical Analysis^28-29

with concentration 10 µg/ml was used as standard.

To identify the phytochemical constituents present in C.

RESULTS AND DISCUSSION

bonducella leaves, a preliminary analysis of water and

Phytochemical analysis and antimicrobial activity of

ethanol extracts was done by using different testing

Caesalpinia bonducella were carried out in this research.

methods like Frothing test, Mayer’s test, Hager’s test,

The preliminary phytochemical screening revealed the

Foam formation test, Lead acetate test, Molisch’s and

presence of saponins, tannins, alkaloids, flavonoides,

Felhing’s test and Ferric Chloride test.

carbohydrates, proteins, phenolic compounds and

Phytochemical analysis²⁰

quinine (Table 3). Saponins were not detected in the

ethanol extract. These constituents present in the leaves

Accurately weighed 20 gm of the Caesalpinia bonducella

extracts have good therapeutic values.

leaves powder was continuously extracted in Soxhlet

apparatus for 16 hrs. The 500 ml mixture of methanol and

Table 1: Bacterial culture

water in volume ratio 4:1 was used as extractant. The

S.No.

Name

Type

ATCC No.

extract was cooled and filtered by using Whatman filter

paper no.41 into a dry and preweighed beaker. From the

Bacillus subtilis

Gram positive

ATCC 2239

residue, fats and waxes were separated firstly by using

Staphylococcus

Gram positive

ATCC 2178

ethyl acetate solvent. The further separation was made

aureus

by using separating funnel. The filtrate was acidified with

Gram

the help of 2M H₂SO₄.The acidified filtrate was again

Escherichia coli

ATCC 25744

negative

extracted by using 150 CC (3 x 50 CC) chloroform in a

Klebsiella

Gram

separating funnel. The chloroform layer obtained was the

ATCC 2239

aerogenes

negative

moderately polar extract. It consists of terpenoids. The

aqueous layer obtained was basified to pH-10 with 2M

NaOH. It was again extracted with 120 CC (2 x 60 CC)

Table 2: Fungal cultures

chloroform and methanol in volume ratio 3:1 followed by

Sr. No.

Name

ATCC No.

extraction with

80 CC (2 x

40 CC) chloroform in a

Aspergillu sniger

ATCC 504

separating funnel. The aqueous basic layer was collected

in a dry preweighed beaker. The methanol layer contains

Penicillium chrysogenum

ATCC 709

quaternary alkaloids and N- oxides and the chloroform

Table 3: Preliminary Phytochemical Analysis of extracts of

extract was the basic extract. It consists of alkaloids.

Caesalpinia bonducella

Those extracts like methanol crude extract, fats & waxes,

terpenoids, quaternary alkaloids & N- oxides, alkaloids

Phytochemical

Water

Ethanolic

were screened for antimicrobial activities.

S. No.

constituents

extract

Procurement of cultures

Saponins

All the microbial cultures (Table 1 and 2) were procured

from National Collection of Industrial Microorganisms

Tannins

(NCIM), National Chemical Laboratory (NCL), Pune.

Antimicrobial Activity^{21, 22}

Alkaloids

To carry out antimicrobial activity, agar diffusion method

Flavonoids

was used and the diameter of growth inhibition zone

surrounding the antibiotic disc was measured to

Carbohydrates

determine it. At first the sterilized Muller- Hinton agar

medium 20 ml was poured into a sterile petriplate. Then

proteins

the plate was covered and allowed to gel. The sterile

cotton swab was dipped into the culture suspension of

Phenolic

bacteria. The agar surface of each plate was inoculated by

compounds

using the swab and ensuring the even distribution of the

organism over the agar surface. The agar surface was

quinine

allowed to dry for ten minutes. A sterile filter paper disc

was picked up by the outer edge with sterile forcep and

= High concentration;

Low concentration;

dipped the opposite edge of the disc in the prepared

- = absent

solution of the extract with concentration 100 µg/ml. The

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Int. J. Pharm. Sci. Rev. Res., 42(2), January - February 2017; Article No. 39, Pages: 217-220

ISSN 0976 - 044X

Secondary metabolites of the plants like alkaloids,

8.850 % fats and waxes and 60.050% fibers observed in

terpenoids and glycosides play the role of protective

this plant extract (Table 4). Lipids are useful in nutrition

agents against different pathogens like insects, fungi or

and dietary, food science, cosmetics, pharmaceuticals,

bacteria. They also function as growth regulatory

paints and varnishes, detergents in human society²⁷.

molecules such as hormone like substances.

In the anti-microbial studies (Table 5), all the extracts

Consequently, they are used as potential anticancer drugs

exhibited larger zones of inhibition against bacterial

by direct cytotoxic activity against cancer cells or by

species like Bacillus subtilis, Staphylococcus aureus,

reducing the tumor development process²³.The

Escherichia coli and Klebsiella aerogenes as well as fungal

phytochemical analysis study showed the presence of

species like Aspergillus niger and Penicillium

4.490%terpenoids &phenolics (Table 4).In Pharmaceutical

chrysogenum.

and food industries, terpenes are used as medicines and

flavor enhancer because of their potentials and

Table 4: Phytochemical Analysis of Caesalpinia bonducella

effectiveness. As antibiotic resistant bacteria are being

Net wt. of

increased globally, terpenes are important²⁴.The group of

S. No.

Parameters

Content (gm)

Composition

terpenoids exhibits various pharmacological activities like

Fats and waxes

1.770

8.850 %

anti-viral, anti-malarial, anti-inflammatory and anti-cancer

activities. It also inhibits cholesterol synthesis²⁵.

Terpenoids and

0.898

4.490%

phenolics

The extract showed the presence of 0.885% alkaloids and

Q. Alkaloid & N -

25.745

% Q. alkaloid & N-oxides

(Table

4).Alkaloids

5.149

25.745 %

Oxides

govern plant growth. Different alkaloids show different

medicinal properties such as Caffeine is stimulant;

Alkaloids

0.177

0.885%

Codeine is cough medicine and analgesic; Quinidine is

Fibers

12.010

60.05 %

anti-arrhythmic; Quinine is antipyretics and antimalarial;

Reserpine is antihypertensive etc.²⁶

Table 5: Inhibition zones in mm of Caesalpinia bonducella against tested Microorganisms

Gram Negative bacteria

Gram Positive bacteria

Fungal Species

Sr. No

Compounds Name

E-coli

K.aerogens

B.subtilis

S.aureus

A.niger

P.chrysogenium

Crude Methanol Extract

6.75

7.25

8.5

7.5

Fats & Waxes

7.5

9.5

Terpenoids

7.5

Quaternary Alkaloids & N-Oxides

7.5

7.75

Alkaloids

6.75

7.25

8.5

7.5

Standard (Chloramphenicol)

CONCLUSION

investigate the plant in detail, the present work may be

used as a useful supplement.

In conclusion, the results of the present study showed

that the crude ethanolic and aqueous extracts of

REFERENCES

Caesalpinia bonducella contain saponins, tannins,

1. Olutayo O, Michael A, John AA, Olusola A, Antimicrobial

alkaloids, flavonoids, carbohydrates, proteins, phenolic

activity and Elemental analysis of Cassia siberiana leaves

compounds and quinines. The percentage of fats and

Using Atomic Absorption Spectrometer, Journal of Natural

waxes, terpenoids and phenolics, Q. Alkaloid & N -

Product and Plant Resources, 2(1), 2012, 9-18.

Oxides was observed good. It showed good antibacterial

2. Babu K, Poornima N, Umarajan KM, Anoop Austin, Studies

and antifungal activities. It might be an alternate to

on Histology and Phytochemical Evaluation of Wattakaka

synthetic antibiotics available in the market. So, its

volubilis

(L.

f.)

Stapf, International Journal of

beneficial effect on animals as well as human health

Pharmaceutical Sciences Review and Research,

41(1),

may be derived from its antibacterial and antifungal

2016, 99-103.

properties to afford protection against various diseases.

3. Iwu MM, Angela RD, Chris O, New microbials of plant

The authenticity of this plant can be detected with the

origin in J. janick(ed) perspective on crops and their uses,

help of the parameters laid down by the findings of the

ASHS press, Mexandrria, 1999, 457 - 462.

present study. Preliminary phytochemical and

4. Gopalakrishnan S, In: Rakesh K Sharma, Rajesh Arora,

phytochemical analysis reports can be useful to

Herbal Drugs: A Twenty First Century Prospective, 1st ed.,

reinforce and authenticate the drug. To evaluate and

International Journal of Pharmaceutical Sciences Review and Research

219

Available online at www.globalresearchonline.net

Int. J. Pharm. Sci. Rev. Res., 42(2), January - February 2017; Article No. 39, Pages: 217-220

ISSN 0976 - 044X

Jaypee Brothers Medical Publishers, New Delhi, 2006, 387-

in Indian herbal medicine, International Journal of green

88.

Pharmacy, 10, 2010, 22-28.

5. Majumder UK, Gupta M,Mukhopadhyay DK,Effect of

17. Sen A, Batra A, Evaluation of antimicrobial activity of

mycotoxins isolated from Penicillium nigricans on glucose-

different solvent extracts of medicinal plant Melia

6-phosphate dehydrogenase, Indian Journal of

azedarach,

International

Journal

Current

Experimental Biology, 35, 1997, 1233-1236.

Pharmaceutical Research, 4(2), 2012, 67-73.

6. Aliyu AB, Musa AM, Oshaniyi JA, Phytochemical Analysis

18. Khare CP, Indian Medicinal Plants: An Illustrated

and Mineral Composition Analysis of Some Medicinal

Dictionary, Springer Science+ Business Media, New York,

Plants of Northern Nigeria, Nigerian Journal of

2007, 107- 108.

Pharmaceutical Sciences, 7(1), 2008, 119.

19. Ali A, VenkatRao N, Md. Shalam, Shivaraj Gouda T,

7. Cano JH, Volpato G, Herbal mixtures in the traditional

Shantakumar SM, Anticonvulsive Effect of Seed Extract of

medicine of Eastern Cuba, Journal of Ethnopharmacology,

Caesalpinia bonducella

(Roxb.), Iranian Journal of

90, 2004, 293-316.

Pharmacology & Therapeutics, 8(2), 2009, 51-55.

8. Ndhala AR, Stafford GI, Finnie JF, Van Staden J,In vitro

20. Harborne JB, Phytochemical Methods: A guide to modern

pharmacological effects of manufactured herbal

techniques of plant analysis Sixth Indian reprint, Springer

concoctions used in Kwazulu-Natal South Africa, Journal of

International Edition, 2010, 5-9.

Ethnopharmacology, 122, 2009, 117-122.

21. Aneja KR, Experiments in Microbiology, Plant Pathology

9. Lee SB, Cha KH, Kim SN, Altantsetseg S,Shatar S,

and Biotechnology, New Age International, New Delhi,

Sarangerel O, Nho CW, The Antimicrobial Activity of

2003, 390 - 391.

Essential Oil from Dracocephalum foetidum Against

22. Vlietinck AJ, Van Hoof L, Totte J, Lasure A, Vanden Berghe

Pathogenic Microorganisms, Journal of Microbiology, 45,

D, Rwangabo PC, Mvukiyumwami J, Screening of a

2007, 53-57.

hundred Rwandese medicinal plants for antimicrobial and

10. Sharma A, Antibacterial activity of ethanolic extracts of

antiviral properties, Journal of Ethnopharmacology, 46(1),

some arid zone plants, International Journal of Pharm

1995, 31-47.

Tech Research, 3(1), 2011, 283-286.

23. Nassar Z, Aisha A, Abdul Majid A, The Pharmacological

11. Enne VI, Livermore DM, Stephens P, Hall LMC, Persistence

Properties of Terpenoids from Sandoricum Koetjape,

of sulphonamide resistance in Escherichia coli in the UK

Webmed Central COMPLEMENTARY MEDICINE,

1(12),

despite national prescribing restriction, The Lancet, 28,

2010.

2001, 1325- 1328.

24. Zwenger S,Basu C,Plant terpenoids: applications and

12. Westh H, Zinn CS, Rosdahl VT, An international

future potentials, Biotechnology and Molecular Biology

multicenter study of antimicrobial consumption and

Reviews, 3(1),2008, 001-007.

resistance in Staphylococcus aureus isolates from

25. Mahato SB, Sen S, Advances in triterpenoid research,

hospitals in 14 countries, Microbial Drug Resistance, 10,

1990-1994, Phytochemistry, 44(7), 1997, 1185-236.

2004, 169-176.

26. Babbar N, An introduction to alkaloids and their

13. Kim H, Park SW, Park JM, Moon KH, Lee CK, Screening and

applications in pharmaceutical chemistry, The Pharma

isolation of antibiotic resistance inhibitors from herb

Innovation Journal, 4(10), 2015, 74-75.

material Resistant Inhibition of 21 Korean plants, Natural

Product Sciences, 1, 1995, 50 - 54.

27. Antonio M, Rabasco Alvarez, Maria Luisa Gonzalez

Rodriguez, Lipids in pharmaceutical and cosmetic

14. Alagesaboopathi C, Antimicrobial Potential and

preparations, Grasas y Aceites, 51, 2000, 74-96.

Phytochemical screening of Andrographis Affinis Nees an

endemic medicinal plant from India, International Journal

28. Olusola A, Olutayo O, Michael A, Olakunle F, Edah AO,

of Pharmacy and Pharmaceutical Sciences, 3(2), 2011, 157-

Elemental analysis and Anti-microbial potentials of the leaf

159.

extract of Cassia arereh Del, International Research of

Pharmacy and Pharmacology, 1(8), 2011, 188-193.

15. Ahmad I, Beg AZ, Antimicrobial and phytochemical studies

45 Indian medicinal plants against multiple drug

29. Olutayo O, Michael A, John AA, Olusola A, Antimicrobial

resistant

human

pathogens,

Journal

activity and Elemental analysis of Cassia siberiana leaves

Ethnopharmacology, 74, 2001, 113-123.

Using Atomic Absorption Spectrometer, Journal of Natural

16. Pavithra PS, Janani VS, Charumathi KH, Indumathy R,

Product and Plant Resources, 2(1), 2012, 9-18.

Potala S, Verma RS, Antibacterial activity of the plant used

Source of Support: Nil, Conflict of Interest: None.

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