Formulation and Evaluation of Antibacterial Herbal gels of Murraya koenigii Leaves Extract

 

Sandeep D. S*, Prashant Nayak, Jobin Jose, Rishal Relita M, Sumana D. R.

Department of Pharmaceutics, NGSM Institute of Pharmaceutical Sciences, Nitte University, Paneer, Deralakatte, Mangalore, Karnataka, India.

 

ABSTRACT:

Herbal medicine has become an item of global importance both medicinal and economical. Herbal remedies are getting increasing patient compliance as they are devoid of typical side effects of allopathic medicines. The present research has been undertaken with the aim to formulate and evaluate the herbal gels containing Murraya koenigii (curry leaves) plant leaf extract. The gel formulations were prepared by using Carbapol 940, Murraya koenigii leaf extract, propylene glycol, methyl paraben, propyl paraben, glycerine and required amount of distilled water. The skin pH (6.8-7) was maintained by drop wise addition of Tri-ethanolamine. The physical parameters of formulated gels like colour, homogeneity, pH, viscosity and spreadibility were evaluated. The gels were evaluated for antibacterial efficiency by agar diffusion method against some bacterial agents. The herbal gels showed that formulations containing Murraya koenigii leaves extract have better antibacterial activity.

 

 

 

 

INTRODUCTION:

Medicinal plants have been a major source of cure for human diseases since time immemorial. It is no wonder that the world’s one-fourth population i.e. 1.42 billion people, are dependent on traditional medicines for the treatment of various ailments. Recently considerable attention has been paid to utilize eco-friendly and biofriendly plant based products for the prevention and cure of different human diseases. It is documented that most of the world’s population has taken in traditional medicine, particularly plant drug for the primary health care. ¹

 

Antimicrobial properties of certain Indian medicinal plants were reported based on folklore information and only few reports are available on inhibitory activity against certain pathogenic bacteria and fungi.

 

Use of plants as source of medicine has been inherited and is an important component of the health care system in India. In these systems of Indian medicine, most practitioners formulate and dispense their own recipes; hence this requires proper documentation and research².

 

There is a continuous and urgent need to discover new antimicrobial compounds with diverse chemical structures and novel mechanism of action due to an alarming increase in the influence of new and reemerging infectious diseases and development of resistance to the antibiotics in current clinical use³.

 

Medicinal plants contain numerous biologically active compounds which are helpful in improving the life and treatment of disease. Compounds such as carbohydrates, proteins, enzymes, fats, oils, terpenoids, flavonoids, sterols simple phenolic compounds etc. Natural products are the source of synthetic and traditional herbal medicine and are still the primary health care system. The presence of various life sustaining constituents in plants made scientists to investigate these plants for their uses in treating certain infective diseases and management of chronic wounds.⁴

 

Murraya koenigii, belongs to the family Rutaceae, commonly known as curry-leaf tree, is a native of India, Sri Lanka and other south Asian countries. It is found almost everywhere in the Indian subcontinent, it shares aromatic nature, more or less deciduous shrub or tree up to 6 m in height and 15-40 cm in diameter with short trunk, thin smooth grey or brown bark and dense shady crown⁴.

 

The M. koenigii is having grey color bark, longitudinal striatations on it and beneath it white bark is present. Leaves are bipinnately compound, 15-30 cm long each bearing 11-25 leaflets alternate on rachis, 2.5-3.5 cm long ovate lanceolate with an oblique base. Margin is irregularly creatate, petioles 2-3 mm long, flowers are bisexual, white, funnel shaped sweetly scented, stalked, complete, ebracteate, regular with average diameter of fully opened flower being in average 1.12 cm inflorescence, terminal cymes each bearing 60-90 flowers. Fruits are ovoid to subglobose, wrinkled or rough with glands. It is having the size of 2.5 cm long and 0.3 cm in diameter and gets purplish black when ripen. Fruits are generally biseeded. Seeds generally occur in spinach green color, 11 mm long, 8 mm in diameter and weighs up to 445 mg. Most part of plant is covered with fine down and has a strong peculiar smell. M. koenigii is genus of tree, native to tropical Asia from Himalaya foothill’s of India to Srilanka eastward through Myanmar, Indonesia, Southern China and Hainan.⁵

 

Murraya koenigii tree leaves are commonly used as spice due to aromatic nature of leaves. Carbazole alkaloids, the major constituents of plant are known to have cytotoxic, antioxidant, antimutagenic and anti-inflammatory activities. The leaves are rich in mono-terpenoids and sesquiterpenoids which exhibited antimicrobial activities^{6, 7}.

 

In the present study, an attempt has been made to formulate and evaluate herbal gels containing M. Koenigii leaves extract for antimicrobial screening against pathogenic microorganisms.

 

MATERIALS AND METHODS:

Collection of plant material:

The plant Murraya koenigii was collected from the surrounding agricultural area of paneer, NGSMIPS college campus, Mangalore. The fresh leaves were separated from the plant and used for extraction.

 

Preparation of leaves extract of Murraya koenigii:

The collected fresh leaves of Murraya koenigii were washed with water and dried in shade. After drying plant leaves were coarsely powderd and kept in well closed container. About 100gm of coarse powder of leaf was weighed and soaked in 500 ml of ethanol and left for maceration for about 4-5 days. After maceration the extract was concentrated and used for further formulations.⁸

 

Formulation of Placebo Gel (Control formulation):

For the preparation of gel formulation, firstly take carbopol 940 which was then dispersed in distilled water along with methyl paraben, propyl paraben and glycerine kept for overnight. Take the leaves extract of murraya koenigii in propylene glycol which was then added in polymer dispersion. Remaining quantity of water was then added and neutralized to pH 7 with triethanolamine by constant stirring for 10 minutes.⁹ The control batch formulation is shown in Table 1.

 

Development of Herbal gel formulations

For the preparation of gel formulation, firstly take carbopol 940 which was then dispersed in distilled water then methyl paraben, propyl paraben and glycerine were added and kept for overnight. Take the leaf extract of Murraya koenigii in propylene glycol which was then added in polymer dispersion. Remaining quantity of water was then added and neutralized to pH 7 with triethanolamine by constant stirring.⁹

 

The pictures of formulated gels are shown in below Fig. 1 and the various formulations of herbal gels are shown in Table 2.

 

Table 1: Control batch formulation of herbal gels

Ingredients	Quantity
Carbopol 940	1.0 gm
Propylene glycol	10 ml
Methyl paraben (0.5 %)	0.2 ml
Propyl paraben  (0.2 %)	0.1 ml
Glycerine	1 ml
Triethanolamine (to maintain pH)	q.s.
Distilled water	100 ml

 

Fig. 1:  Herbal gel formulations of Plant extract

 

Table 2: Development of Herbal gel formulations

Ingredients	F1	F2	F3	F4	F5
Murraya Koenigii Extract	0.5 gm	1.0 gm	1.5 gm	2.5 gm	3.0 gm
Carbopol 940	1.0 gm	1.0 gm	1.0 gm	1.0 gm	1.0 gm
Propylene glycol	10 ml	10 ml	10 ml	10 ml	10 ml
Methyl paraben (0.5 %)	0.2 ml	0.2 ml	0.2 ml	0.2 ml	0.2 ml
Propyl paraben (0.2 %)	0.1 ml	0.1 ml	0.1 ml	0.1 ml	0.1 ml
Glycerine	1 ml	1 ml	1 ml	1 ml	1 ml
Triethanolamine (to adjust pH)	q.s.	q.s.	q.s.	q.s.	q.s.
Distilled water	100 ml	100 ml	100 ml	100 ml	100 ml

 

Evaluation of Herbal gels:

Physical evaluation:

All the formulated herbal gels were checked for colour and homogeneity by visual observation.

 

pH:

The pH of all the formulated herbal gels was measured by using digital pH meter.¹⁰

 

Viscosity:

Viscosity of herbal gels was determined by using Brookfield rotational viscometer at 100 rpm using spindle no.64.¹⁰

 

Spreadability:

The spreadability of gel formulations was determined by measuring the spreading diameter of 1g of gel between two horizontal plates.¹¹

 

Antibacterial activity:

The antibacterial screening of herbal gels was done by disc diffusion method. The gels were tested against bacterial agents namely B. subtilus, P. aeruginosa,            S. aureus and E. coli. A loopful of the pure bacterial culture was suspended in nutrient broth and incubated for 24 hours. Nutrient agar media was sterilized and poured into petri plates. After solidification, 0.1ml of the inoculum was spread over the agar evenly using a rod. 6mm diameter cavity was prepared and formulated gel is placed in the cavity. A standard antibiotic was used as the control. The inoculated plates are incubated for 24 hours. Later, the zone of inhibition around the disc was measured and recorded.¹²

RESULTS AND DISCUSSION:

The results of physical parameters of formulated herbal gels like colour, homogeneity, pH, viscosity and spredibility were shown in below Table 3.

 

The spredibility of formulated herbal gels is shown in below Fig 2

 

Fig 2: Spreabability of  formulated herbal gels

 

The results of antibacterial activity of all formulated herbal gels against some pathogenic microorganisms is shown in below Table 4 and the results of zone of inhibition of all formulated herbal gels against the pathogens is represented graphically in below Fig 3

 

 

 

Table 3: Results of physical parameters of all formulated herbal gels

Formulation Code	Colour	Homogeneity	pH	Viscosity (cp)	Spreadibility (mm)
F1	Light green	Good	6.4	4530	36
F2	Greenish	Good	6.4	4656	38
F3	Dark greenish	Good	7.1	4786	40
F4	Dark greenish	Good	6.8	4833	44
F5	Dark greenish	Good	6.8	4875	48

 

Table 4: Antibacterial activity of herbal gels

Micro-organism culture	Zone of inhibition of Herbal gels (mm)
	Standard drug	F1	F2	F3	F4	F5
S. aureus	28	11	14	17	21	25
P. aeruginosa	33	8	12	15	22	26
E. coli	30	10	13	16	20	24
B. subtilus	36	12	18	24	28	32

 

Fig. 3: Antibacterial activity of formulated herbal gels

 

DISCUSSION:

The colour of all the formulated herbal gels was greenish to dark greenish and all the herbal gels were good in homogeneity. The pH of all the formulated gels was in the range of 6.4- 7.1 matching with skin pH range. Viscosity of all the herbal gels was ranging from 4530- 4875 cp at 100 rpm measured with Brookfield viscometer. The spreadability of all herbal gels was in the range of 36-48 mm. The antibacterial activity of all the formulated herbal gels showed good results of zone of inhibition against skin pathogens.

 

CONCLUSION:

From the present investigation, it has been revealed that herbal gels of plant Murraya koenigii leaves extract can be formulated using carbopol 940 as polymer with other ingredients and the evaluation of physical parameters shown satisfactory results. From the antibacterial activity it was found that prepared herbal gels of Murraya koenigii leaves extract were significantly active against tested pathogens which was comparable with standard antibiotic. Hence, from the overall results, finally it was concluded that the formulated herbal gels have significant antimicrobial properties and hence will be better, safe and effective than allopathic medications.

 

ACKNOWLEDGEMENT:

The authors would like to thank authorities of NGSM Institute of pharmaceutical sciences, Nitte University, Deralakatte, Mangalore for providing necessary facilities and support for carrying out this project work.

 

REFERENCES:

1.       Bauer J, Rojus R, Bustamate B. Antimicrobial activity of selected peruvian medicinal plants. J Ethnopharmacol. 88; 2003: 199-204.

2.       Sandeep DS, Narayana CR, Prashant N, Aliss M. Formulations of antimicrobial polyherbal hand wash. Res J Pharm Tech. 9 (7); 2016: 864-866.

3.       Shivanand Nayak. Influence of ethanol extract of vinca rosea on wound healing in diabetic rats. J Biologi Sci. 6 (2); 2006: 51-55.

4.       Kaur K, Aravind Kumar G, Sayyed Ahmad. Pharmacognostic studies on bark of Murraya koenigii spreng. Int J Res Pharma Biomedical Sci. 10 (1); 2011: 1-4.

5.       Harish KH, Anup P, Shruthi SD. A review on Murraya koenigii: Multipotential medicinal plant. Asian J Pharm Clini Res. 5 (4); 2012: 5-14.

6.       Baskaran C, Rathabai V, Kanimozhi D. Screening for antimicrobial activity and phytochemical analysis of various leaf extract of Murraya koenigii. Int J Res Ayur Pharm. 2 (6); 2011: 1807-1810.

7.       Malwal M, Sarin R. Antimicrobial efficacy of Murraya koenigii (Linn.) spreng root extracts. Ind J Nat Produ Resour. 2 (1); 2011: 48-51.

8.       Rajesh B, Saumya D, Dhramjit P, Pavani M. Formulation design and optimization of herbal gel containing Albizia lebback bark extract. Int J Pharm Pharm Sci. 6 (1); 2014: 111-114.

9.       Jadhav VD, Talele SG, Bakliwal AA, Chaudhari GN. Formulation and evaluation of herbal gel containing Tridax procumbens. J Pharma Bio Sci. 3 (2); 2015: 165-172.

10.     Deepak PA, Prashanth BS. Formulation and evaluation of herbal gel containing Lanata camara leaves extract. Asian J Pharm Clini Res. 6 (3); 2013: 122-124.

11.     Ajinkya MB, Manjusha ND. Formulation and evaluation of herbal antimicrobial gel containing Musa acuminata leaves extract. J Pharamcog Phytochem. 5 (1); 2016: 1-3.

12.     Vidya V, Aleykutty NA, Jayakar B, Subin MZ. Development and evaluation of antimicrobial herbal formulations containing the methanolic extract of Samadera indica for skin diseases. J Adv Pharm Technol Res. 3 (2); 2012: 106-111.

 

 

 

Accepted on 28.04.2017        © RJPT All right reserved