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Asian Journal of Pharmaceutical Science & Technology
e-ISSN: 2248 – 9185
www.ajpst.com
Print ISSN: 2248 – 9177FORMULATION AND IN-VITRO ASSESSMENT OF CREAM
PREPARED FROM
ALLIUM CEPA
L., BULB
E ESoje, J Muazu* and S J Madu
Department of Pharmaceutics and Pharmaceutical Microbiology, University of Maiduguri, Nigeria.
ABSTRACT
Onion (Allium cepa L.) is among the oldest cultivated plants which are used both as source of nutrition and for medicinal applications. The purpose of this study is to formulate herbal cosmetic cream containing the methanol extract of Allium cepa L. bulb and subject it to some quality assessment tests in order to ascertain the suitability of the formulation process. Methanol extract of Allium cepa L. was obtained by maceration. Subsequently, it was formulated to cream using the fusion method. The extract and the cream were then subjected to some quality assessment tests under different storage conditions of 5, 25 and 45°C at days five and seven consecutively. These tests include; enumeration of microbial count, physical appearance, odour, after feel, pH, spread ability and nature of the cream. Our findings demonstrates that values obtained from creams stored at room temperature(25°C) are within acceptable range for quality whereas, those stored at extreme temperatures (5and 45°C)had values outside the normal range (change in physical appearance, poor spread ability with altered nature of cream formulated). In conclusion, the fusion method of cream formulation using hard paraffin as a base can be suitably used to formulate methanol extract of Allium cepa L. cream and the formulation stored at room temperature.
Key words: Methanol extract, Allium cepa L., Cream, Assessment, Formulation.
INTRODUCTION
Onion (Allium cepa L., family Liliaceae) [1] has a long history of medicinal use. The fleshy bulb that grows below the ground is used medicinally as well as for food; other parts of the plant also have a place in traditional medicine. Onion is believed to have a positive effect on the circulatory system [2] and sexual debility [3]. It has been used as a diuretic to reduce swelling.Onion contains thiosulphinate, a compound that is effective in killing many common bacteria, including Salmonella typhi, Pseudomonas aeruginosa, and Escherichia coli[4]. Externally, fresh onion juice is used to prevent bacterial and fungal infections [5];it can also be applied to wounds and stings on the skin [6].
Creams are topical preparations usually for application to the skin. They are also applied unto the mucous membranes such as the rectum or vagina [7]. They are semi solid emulsions which are oil-in-water (O/W) or water-in-oil (W/O) type. Creams are often composed of two phases. Oil-in-water (O/W) emulsions are most useful as water washable bases, whereas water-in oil (W/O) emulsions are emollient and cleansing agents. An emulsifying agent is used to disperse the aqueous phases in the oily phase or vice versa[8].
The use of cosmetics not only developing an
attractive external appearance, but towards achieving longevity of good health by reducing skin disorders [9]. The demand for herbal cosmetic is rapidly increasing [9]. The plant parts used in cosmetic preparation should have varieties of properties like antioxidant, anti-inflammatory, antiseptic, emollient, antiseborrhatic, antikerolytic activity and antibacterial among others [9]. Cosmetic products are used to protect the skin against exogenous and endogenous harmful agents and enhance the beauty and attractiveness of skin [6]. Hence, the purpose of this study is to formulate herbal cosmetic cream containing the methanol extract of
Allium cepa L. bulb and subject it to some quality assessment tests in order to ascertain the suitability of the formulation process.
MATERIALS AND METHODS
Source of plant material, collection and proof of identity
Allium cepa L. bulbs acquired from Gamboru
market, Maiduguri, Borno state Nigeria was identified by a Plant Taxonomist from the Department of Biological Sciences, University of Maiduguri and a voucher specimen number PCG/H/2014/2015/001 was assigned and deposited in Department of Pharmacognosy Herbarium, University of
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Maiduguri.
Preparation of the methanol Allium cepa L. bulb extract The method used by Kavitha et al [7] was adopted with some modifications. Fresh healthy bulbs of Allium cepa L. were weighed and divided into three groups of 11 kg each. Subsequently, they were washed with distilled water and size reduced using a stainless steel knife prior to homogenization in a blender (Philips, Japan). The homogenized sample was drenched in 2L of 80% methanol which was allowed to stand for 24 hr with intermittent shaking. The resulting mixture was filtered and the filtrate concentrated using a rotary evaporator (PEC Medical, U.S.A). The concentrate was subsequently air dried and the yield determined. The sample was stored in a refrigerator for further use.
Enumeration of microbial count
Inoculation by pour plate method was carried out after 1 in 1000 serial dilutions of 1 g weight of the methanol extract Allium cepa L bulb. One milliliter of the diluted sample was then aseptically aspirated into the media (Nutrient Agar). The media was poured aseptically into a sterile petri dish at 40-45 °C then swirled and allowed to solidify for incubation (Uniscope, England) at 37 °C for 24 hr.
Typical colonies of microbial growth on plates were counted at the end of incubation and the result presented in colony forming unit (cfu/g) per gram[10].
Cream formulation
The fusion method utilised by Sanmathi et al [11] wasused to formulate the cream. About 24g of hard paraffin was weighed into a beaker and placed on a hot plate (Sea hot plate- scientific educational Aids ltd, England)which was allowed to melt; A 10g weight of petrolatum was added into the beaker to form the base. 5.5 g of the Allium cepa L. methanol extract was weighed into another beaker and 20ml of liquid paraffin was added with constant stirring until the extract has dissolved. Subsequently, the mixture containing the Allium cepa L. methanol extract was poured into the base with continuous stirring. Liquid paraffin was then added to make up to a volume of 60 ml. The mixture was
stirred in one direction until warm, Eucalyptus oil (0.5 ml) was added, stirred and the resulting cream transferred into a suitable container prior to quality assessment.
Quality assessment of the cream formulated Physical examination
Physical characters of the cream such as colour, physical state and odour were observed by randomly selected volunteers and their findings noted.
After feel
The cream was applied by ten (10) randomly selected volunteers to ascertain whether it’s greasy or non-greasy. Their observations were recorded.
Determination of pH
The pH meter (Hanna instruments, Italy) was calibrated using standard buffer solution. Similar to Kuntal
et al [12], 5 g weight of the cream was dispersed in 45ml distilled water to determine the pH of the suspension at 27oC using a pH meter.
Spreadability
The formulated cream (3g) was applied in between two glass plates and was compressed to uniform thickness by placing 1000 g weight on it for 5 min. Thereafter, weight (50g) was added to the pan and the top plate was subjected to pull with the help of string attached to the hook. The time in which the upper glass slide moves the lower plate to cover a distance of 10 cm was noted [13].
Stability test
The cream was exposed to altered temperatures of 5oC (refrigerator), 25oC (room temperature) and 45oC (oven) for a period of five days for batch A and seven days for batch B. In each case 100 g of cream was taken into 100 ml beaker. They were observed for any change in consistency, bleeding and phase separation [14].
RESULTS Percentage yield
The yield of methanol extract obtained from Allium cepa L bulbs was 42.2 %.
Table 1. Formula used in preparation of Allium cepa L. cream
INGREDIENT QUANTITY
Allium cepa L. methanol extract (% w/v) 5.50
Hard paraffin (g) 24.00
Petrolatum (g) 10.00
Eucalyptus (ml) 0.50
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Table 2. Enumeration of microbial count
Test Result Standard
Microbial load (cfu/g) 8.65x101 <500 for preparations to be applied near the eye < 1000 for preparations intended for other areas Key: < = less than; cfu/g = colony forming unit per gram
Table 3. Physicochemical Characters of Allium cepa L. cream at day five (5)
Parameter Monitoring phase Storage temperature (°C)
5 25 45
Physical appearance Initial Light brown colouration and smooth on application
Final Same colour but hardened Same colour but watery Odour Initial
Odour is masked and pleasant all through Final
After feel Initial Greasy Greasy Greasy
Final Greasy Greasy Greasy
pH Initial 6.00 6.00 6.00
Final 6.00 6.00 6.00
Spread ability Initial
< One minute over a distance of 10.00 cm at all temperatures (Good spread ability) Final
Nature Initial Smooth Smooth Smooth
Final Hardened Smooth Watery
Table 4. Physicochemical Characters of Allium cepa L. cream at day seven (7)
Parameter Monitoring phase Storage temperature (°C)
5 25 45
Physical appearance Initial Light brown in colour and smooth on application
Final Colour is the same but hardened Colour is the same but watery Odour Initial
Odour is masked and pleasant all through Final
After feel Initial Greasy Greasy Greasy
Final Greasy Greasy Greasy
pH Initial 6.00 6.00 6.00
Final 6.00 6.00 6.00
Spread ability Initial Good Good Good
Final Poor Poor Poor
Nature Initial Smooth Smooth Smooth
Final Hardened Smooth Watery
DISCUSSION
The yield of the methanol bulb extract of Allium cepa L. (spp. Red onion) was 42.20%. This yield is significantly higher than that of ethanol extract but however slightly lower in comparism to n-buthanol extract as reported by other authors [15].
As presented in Table 2, microbial load of the
Allium cepa L. methanol extract fall within the limit of bacterial prevalence unit of the British Pharmacopoeia (BP) and the United States Pharmacopoeia (USP) of 102 cfu/g used in creams. Our findings are also lower compared to studies[16, 17]thus implying that the formulation may not require preservative due to its proven antimicrobial activity [18]which helps in wound healing and other dermatological infections.
The physical appearance of the cream at various temperatures and duration of storage demonstrated no difference as there was no change in colour nor was there
cracking or separation of phases of the cream. These findings (Tables 3 and 4) were in concordance with the results obtained in literature [17]. In a like manner, odour emitted by thiosulphate gasses in Allium cepa L. was masked with the addition of eucalyptus oil.
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creams. Therefore, the cream can stay long enough to release the medicaments before been washed out [19].
In concordance with Khan et al [20] who evaluated cream containing diclofenac sodium and Curcuma longi, the pH of the cream with the extract was found to be 6.00 in all the instances. This is suitable for topical application because the pH of the skin is between 4.50-6.00 [9, 20].
Spread ability of the cream formulated decreases with increase in duration of storage and also alteration of storage temperature similar to other studies [20], it was observed that the samples at day 5 spread to a distance of 10 cm in less than a minute indicating good spread ability while on day 7 a higher time was used to cover similar distance (10 cm). The effect of varying temperatures clearly demonstrates the effect different storage conditions would have on the cream using hard paraffin as a base, it has poor spread ability at 5 and 45 °C hence it is not stable for storage at these temperatures. These findings are similar to the studies carried out using stearic acid as a base, however, formulation using natural palm oil as base showed good
spread ability at 5, 25 and 45°C irrespective of the duration of storage[19].
CONCLUSION
Conclusively, the methanol extract of Allium cepa
L. bulb obtained via maceration and formulated into cream passed all the quality assessment tests when subjected to room temperature. However, samples stored at low (5°C) and high (45°C) temperatures deviated from the range of normal values of the quality assessment test. Therefore, the fusion method of cream formulation using hard paraffin as a base can be suitably used to formulate methanol extract of
Allium cepa L. cream and the formulation stored at room temperature.
ACKNOWLEDGEMENT: None
CONFLICT OF INTEREST:
The authors declare that they have no conflict of interest.
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