ISSN: 0975-8585

Research Journal of Pharmaceutical, Biological and Chemical

Sciences

Development and Evaluation of Antiarthritic Herbal Ointment

Neel J Mehta1*, Neel D Patadiya1, Jay Patel1, Divyesh H Shastri1, and Pragna K Shelat1

1
Department of Pharmaceutics, KB Institute of Pharmaceutical Education & Research, Gandhinagar, Gujarat India-
382023

ABSTRACT

Formulation and evaluation of antiarthritic (anti-inflammatory) herbal preparation (Ointment) containing

Menthol and Methyl Salicylate. The Ointment formulation was designed by using five different ointment bases.
Various formulation batches containing 5% Menthol and 15% Methyl Salicylate were prepared. Clove Oil was used
as a penetration enhancer. The optimized herbal ointment formulations was subjected to stability studies and
evaluated for pH, Viscosity and Rheological studies, Spreadability, Washability. Final formulations along with
market preparation were evaluated for its topical anti-inflammatory activity against carrageenan induced Rat-paw
edema on wistar rats of either sex. Ten Ointment formulations were prepared successfully and evaluated for the
physical properties, skin feeling, stability study and anti-inflammatory activity. Formulation containing 5%
Menthol, 15 % Methyl salicylate and 1 % Clove oil and water miscible ointment base showed better % inhibition of
edema compared to others. Ointments containing 1 % Clove oil showed better Anti-arthritic (Anti-inflammatory)
activity in all five ointment base. From the present study we concluded that the water miscible base worked as the
best carrier for the drug (5% Menthol and 15% Methyl Salicylate) in concern with stability as well as for the anti-
inflammatory activity of the ointment. The optimized ointment formulation showed better inhibition of the
inflammation as compared to conventional marketed formulation.
Keywords: Herbal Antiarthritic ointment, Menthol, Methyl Salicylate, Clove oil, Rat paws edema method

*Corresponding author

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INTRODUCTION

Ointments are used topically for several purposes, e.g., as protectant, antiseptics,
emollients, antipruritic, kerotolytic, and astringents. The vehicle or base of an ointment is of
prime importance if the finished product is expected to function as any one of the above
categories. In the case of a protective ointment, it serves to protect the skin against moisture,
air, sun rays and other external factors. It is necessary that the ointment neither penetrates the
human skin barriers nor facilitates the absorption of substances through this barrier. An
antiseptic ointment is used to destroy or inhibit the growth of bacteria. Frequently bacterial
infections are deeply seated; a base which has the capacity to either penetrate or dissolve and
release the medication effectively is therefore desired. Ointments used for their emollient
effect should be easy to apply, be non-greasy and effectively penetrate the skin.

There are five types of ointment bases on the basis of their physical composition. Each
has different physical characteristics and therapeutic effectiveness [1]. i.e.

Oleaginous bases
Absorption bases
Water in oil emulsifying bases
Oil in water emulsifying bases
Water soluble or water miscible bases

It is the ability of Menthol to chemically trigger the cold-sensitive TRPM8 receptors in

the skin which is responsible for the well-known cooling sensation that provokes when inhaled,
taken orally or applied to the skin [2]. Similar to capsaicin, responsible for the spiciness of hot
peppers (which stimulates heat sensors, without causing a change in body temperature)
Menthol has analgesic properties that are mediated through a selective activation of -opioid
receptors [3]. Menthol also enhances the efficacy of ibuprofen in topical applications via
vasodilatation, which reduces skin barrier function [4].

In high concentration, Methyl salicylate is used as a rubefacient in deep

heating liniments (such as Bengay) to treat joint and muscular pain. Randomized double blind
trial reviews report evidence of its effectiveness that is weak, but stronger for acute pain than
chronic pain, and that effectiveness may be due entirely to counter-irritation. However in the
body, it metabolizes into salicylates [5].

The main chemical components of clove oil are eugenol, eugenol acetate, iso-eugenol
and caryophyllene (terpenes). Eugenol may reduce the ability to feel and react to painful
stimulation. Therefore, use of clove products on the skin with other numbing or pain-reducing
products such as lidocaine / prilocaine cream, theoretically it may increase effects. FT-IR and
partitioning studies reveal that the enhancement in the permeability coefficient of drug by
Eugenol is due to lipid extraction and improvement in the partitioning of the drug to the
systemic circulation [6-8]

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MATERIALS AND METHODS

MATERIALS

Carbopol 971 was gifted from Corel Pharma Chem, Ahmedabad. Menthol and Methyl
Salicylate were obtained from Chinubhai Motilal Saraiya, Ahmedabad. All other ingredients
used in the study were of analytical grade.

METHODS

Preparation of Ointment

5 Ointment bases were selected as per their performance properties for topical delivery.
They are prepared as per the formula mentioned in table: 1.

Aqueous phase (distilled water at 60-70C) was taken in a 250 ml glass beaker with
continuous stirring by using propeller stirrer. An accurate quantity of water-soluble material
was dispersed when vortex is formed. Oils and other oil soluble material were melted at 60-
70C.
Table 1: Base formulation A to E
INGREDIENTS AMOUNT (%)
A B C D E
Bees Wax 8
Borax 0.5
Carbomer 971 1
Castor Oil 5
Cetomacrogol 6
Cetyl Alcohol 0.5 1 1.5
Dimethicone 1
Glycerin 18 3 1.5
Lanolin 1
Liquid Paraffin 50 15
Methyl Paraben 0.2 0.2 0.2 0.2
Paraffin Wax 12
Polyethelene Glycol 30
4000
Polyethelene Glycol 400 50
Potassium Hydroxide 0.52
Sodium Lauryl Sulphate 0.5
Stearic Acid 6
Stearic Acid 18
Triethanolamine 4.5 Q.s.
Purified Water Q.s to 100 Q.s to 100 Q.s to 100 Q.s to 100

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Both the phases were mixed using propeller stirrer by maintaining the temperature. 5%
Menthol was dissolved in 15% Methyl Salicylate in separate container and clear solution was
obtained. Clove oil was added up to 1 % in the respective formulation. This clear solution was
added to biphasic mixture of oil and water when it was hot in the propeller stirrer. It was
allowed to cool at room temperature until uniform consistency obtained. The final formulation
was pecked in the wide-mouth jar, labelled and evaluated.

Table 2: Formulation code

Formulation Base code Amount (%)

Code Menthol Methyl salicylate Clove oil
I A 5% 15 % -
II A 5% 15 % 1%
III B 5% 15 % -
IV B 5% 15 % 1%
V C 5% 15 % -
VI C 5% 15 % 1%
VII D 5% 15 % -
VIII D 5% 15 % 1%
IX E 5% 15 % -
X E 5% 15 % 1%

EVALUATION OF OINTMENTS [9]

Organoleptic Parameters

Herbal ointment formulations were evaluated based on their appearance, texture and
consistency. Texture was determined on the basis of grittiness / smoothness. Texture should be
smooth so it can be spreadable and washable easily. It should penetrate through the skin.

pH

10% W/V homogenous solution of ointment was prepared and then pH was calculated by pH
meter.

Effect on skin

Spreadability

Spreadability was determined by applying the lotion slowly on the skin.

Skin feeling (Oily/Greasy)

Skin feeling was noted as either Greasy or not.

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Film formation

Film formation was determined on the basis of its uniformity (continuous/not).

Washability

Washability was checked by keeping applied skin area under the tap water for about 10 min.

Cooling effect

Cooling effect was determined manually.

Stability study

The optimized formulations along with marketed formulation were evaluated for their
stability at an ambient condition of pressure and temperature for two weeks. Formulations
were observed for phase separation and particle agglomeration.

Pharmacological screenings by rat paw edema method

Animal study was performed on the developed herbal ointment formulations along with
marketed formulation. The animal experiments were performed according to CPCSEA
guidelines and after the approval from Institutional Animal Ethics Committee (I.A.E.C.), PROJECT
NO. KB/11/227, K. B. Institute of pharmaceutical education and research (KBIPER),
Gandhinagar, Gujarat, India. Experiments were conducted in accordance with the standard
guidelines. The detailed procedure is discussed as below.

Animal used

Albino rats (Wistar strain) of either sex (150- 180 g) were obtained from the animal
house of K. B. Institute of pharmaceutical education and research, gandhinagar, Gujarat.
Animals were kept in animal caging system (four rats per cage on beds of sawdust) under the
laboratory conditions (25 2C, 12 h light). They were provided with animal feed pellets
manufactured by Hindustan Lever (India) Ltd. Mumbai. Animals were randomly selected for
different experimental groups and used for the in vivo determination of anti-inflammatory
activity. During the course of the experiment the animal behavior was normal.

Drug/Formulation
Developed Ointment formulations, market preparation (ointment)

Experimental Method

Anti-inflammatory activity was evaluated using carrageenan induced rat paw edema
method. Carrageenan (0.1 ml of 1% w/v suspension) was injected into the sub plantar region of

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the both the hind paw of each rat. The right hind paw was kept as control and left hind paw was
considered as test one. After the carrageenan injection, the paw volumes were measured at 15
min, 30 min, 1 hour, 2 hour, 4 hour by using a plethysmometer (Model 7150, UGO Basile, Italy).
Edema was expressed as the mean increase in paw volume relative to control. The percentage
inhibition of edema was calculated by the following equation:

% inhibition of edema = 100 [1-(Vt/Vc)]

Where, Vc = edema volume in the control

Vt = edema volume in test.

RESULTS AND DISCUSSION

As mentioned in table no. 2 different ointment formulations were prepared by using five
ointment bases, 5 % Menthol, 5 % Methyl salicylate and 1 % Clove oil. Among them five
ointment formulations (formulation code II, IV, VI, VIII, X) contained 1 % Clove oil.

Table 3 showed the comparative evaluation of the prepared ointments. pH,

spreadability, consistency, washability, film formation, irritation, cooling effect and Avg. globule
size were evaluated for all the ten ointment formulations. Alongside, stability study was
performed and finally rat paw edema method was carried out for the Anti-arthritic (anti-
inflammatory) activity. Nine ointments were found to be stable at ambient condition, expect
the ointment base D was found to be incompatible with clove oil. As indicated in table 3, the
ointment base E (formulation ointment IX and X) provide good physical properties as well as
good skin feeling effect.

Comparison of the market ointment preparation (MOOV and FAST RELIEF) was
performed with the developed ointments. Table 4 showed the higher % inhibition of rat paw
edema in ointment base E (formulation code IX and X) and it is quite identical with the market
preparation. Onset of action and duration of action could be determined by the table 4.
Comparison of formulations (e.g. formulation code I-II, III-IV, V-VI, VII-VIII, IX and X) explained
that the presence of Clove oil (1 %) showed better inhibition of paw edema and that confirmed
the activity of Clove oil as a penetration enhancer.

CONCLUSION

The ointments prepared by using 5 % menthol and 15 % methyl salicylate as an Active,

showed similar effectiveness as the market product did. The addition of clove oil (1%) showed
better inhibition of the inflammation with compared to the normal ointment. The Ointments
provided approx. 35 % of inhibition and that seemed to be the evidence for the anti-
inflammatory activity of the ointment. Water miscible base worked as the best carrier for the
drug in concern with stability as well as for the anti-inflammatory activity of the ointment.

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Table 3: Evaluation parameters of developed Ointment formulations

Formulation pH Spreadability Consistency Washability Film Irritation Cooling Avg.

Code Formation Globule
Size ()
I 7.4 ++* ++ + Greasy No Yes 55.4
II 7.6 ++ ++ + Greasy No Yes 56.8
III 6.8 ++ ++ + Poor No Yes 59.6
IV 6.8 +++ ++ + Poor No Yes 61.9
V 6.9 ++ ++ ++ Good No Yes 65.1
VI 6.4 ++ ++ ++ Good No Yes 66.7
VII 7.2 +++ ++ ++ Good No Yes 38.5
VIII 7.3 -- -- -- -- No No 41.9
IX 7.3 +++ +++ +++ Conti. No Yes 39.4
X 7.1 +++ +++ +++ Conti. No Yes 42.5
*Indicated Good: +++, Moderate: ++, Poor: +

Table 4: Percent Inhibition on carrageenan induced Rat paw edema

Formulation % Inhibition of edema with Time (hr)

Batch code 15 min 30 min 1 hr 2 hr 4 hr
I 9.24 11.5 14.25 17.12 18.92
II 12.83 15.21 17.8 19.72 20.26
III 13.31 14.55 16.72 19.92 22.71
IV 15 18.3 19.5 21.13 25.5
V 13.61 15.62 20.97 24.2 26.11
VI 16.22 19.03 22.18 25.67 27.47
VII 20.6 22.52 27.8 29.92 30.11
VIII 21.2 24.56 29.33 31.5 39.54
IX 24.67 28.45 33.49 38.92 41.23
X 25.82 32.91 38.28 40.65
M1* 18.25 22.98 31.14 39.43 42.68
M2* 28.51 32.1 41.23 37.92 28.12
*= Marketed formulation (M1 MOOV, M2 - FAST RELIEF)

ACKNOWLEDGEMENT

The assistance and support from Mr. Mehul Chorawala, Lecturer, and Ms. Gopi Talati,
Research scholar, Department of Pharmacology, K. B. Institute of Pharmaceutical Education and
Research, Gandhinagar is gratefully acknowledged. The Authors are highly thankful to the staff
of Department of Pharmaceutics and Pharmacology, KBIPER for the assistance of experiment
during the project work.

REFERENCES

[1] Gennaro AR. Remington's science and practice of pharmacy, 18th edition, 1309-1314.

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[2] Eccles R. J Pharm Pharmacol 1994; 46 (8): 618-630.

[3] Galeottia N, Mannellia LDC, Mazzantib G, Bartolinia A, Ghelardini C. Neuroscience
Letters 2002; 322 (3): 145-148.
[4] Braina KR, Greena DM, Dykesb PJ, Marksb R, Bola TS. Skin Pharmacol Physiol 2006; 19:
17-21.
[5] Mason L, Moore RA, Edwards JE, McQuay HJ, Derry S, Wiffen PJ. BMJ 2004; 328 (7446):
995.
[6] Zhao K, Singh J. Mechanisms of percutaneous absorption of tamoxifen by terpenes :
Eugenol, D-limonene and menthone Department of Pharmaceutical Sciences, College of
Pharmacy, North Dakota State University, Fargo.
[7] Lahora D, Chaudhary V, Swami G, Chaudhary G, Saraf SA. IJPRD 2011; 2; 39-45.
[8] Singla V, Saini S,Singh G, Rana A. IRJP 2011; 2(12), 32-36
[9] Patel PM, Mehta NJ, Limbachiya S, Dhandhalia M, Shastri DH, Shelat PK. Development
and evaluation of anti-inflammatory activity of herbal oil. JSRP 2012, 1(2).

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