PHYSIO-CHEMICAL PROPERTIES OF TOPICAL MEDICATION FORMULATION MADE FROM SHEA BUTTER AND METHYL SALICY

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PHYSIO-CHEMICAL PROPERTIES OF TOPICAL MEDICATION FORMULATION MADE FROM SHEA BUTTER AND METHYL SALICYLATE Raphael Eze Nnam*1, Chidubem Theresa Chukwu2, Mabel Chidimma Nwachukwu3, Nkechi Jeniffer Emerenini4, Ifeanyi S E. Nwaorgu5 *1Department

of Food Technology, Akanu Ibiam Federal Polytechnic, Unwana Ebonyi State Nigeria. of Food Technology, Akanu Ibiam Federal Polytechnic, Unwana Ebonyi State Nigeria.

2,3,4,5Department

ABSTRACT In the present study, the use of locally sourced Shea butter as the oil base in the formulation of analgesic (Methyl salicylate) creams to be used as topical medication was investigated. Methyl salicylate (the active ingredient), Bees wax, water and Shea butter obtained from local sources and refined were used separately on two different types on emulsifying agents namely cetrimide and polysorbate 80, to formulate three different cream formulation. These formulations were labeled Y1, Y2 and Y3. Y1 contend Shea butter, Methyle salicylate, cetrimide, bee wax and water; Y2 contend Shea butter, Methyle salicylate, polysorbate 80, bee wax and water while Y3 as the standard control contains hydrous wool fat, Methyle salicylate, cetrimide, bee wax and water. The physiochemical properties of each formulation were evaluated to determine the PH, viscosity, spreadability, washability, rate of penetration through the rat skin, odour, homogeneity and colour. The results obtained showed different levels of acidity for each of the formulations. From the result obtained, the pH value for each of the formulation was 4.95, 6.01 and 5.33 for Y1, Y2 and Y3 respectively. This pH value falls within the range for human skin. It was also observed that the formulations with Shea butter as the oil base dissolved the methyl salicylate that served as the active ingredient more. They also had higher penetration rate compared to when the hydrous wool fat cream was used on the rat skin. Y1 and Y2 are better than the control in terms of homogeneity, spreadibility and viscosity while the control Y3 was the easiest to be washed-out among the three formulations. Keywords: Methyl salicylate, Shea butter, Anagelsic cream, Skin pH, Topical medication.

I.

INTRODUCTION

Topical applications from the cosmetic and pharmaceutical industries like creams and lotions are so rampant that visiting any cosmetic or pharmaceutical shop, one is sure to come out with a handful of varieties. These varieties come along with their varied functions as they can be used as protectants, antiseptics, emollients, antipruritics, keratolytics, or as astringents (Buhse et. al., 2005). As protectants, they serve to protect the skin against moisture, air, sun rays and other external factors; As antiseptics, they help destroy and inhibit the growth of bacteria on the skin; As emollients they are used to treat dryness of the skin; As antipruritic, they help to prevent or relieve the skin of itching; As keratolytics, they are used to improve the skin’s texture as they help to remove scales and flakes; As astringents, they are used to shrink and harden the protein of the skin, protecting it and healing scars on it. It can be seen that the preparation of topical creams cuts across both the cosmetic and pharmaceutical industries which are sub units within the chemical process industry. The conventional medications used on the skin surface are classified into three categories namely liquid, semisolid and solid topical medications. Liquid topical medication includes lotions, suspensions, and solutions that have low viscos emulsions. Examples of semi-solid topical dosage include collodions, foams, ointments, pastes, creams and gels while powders, patches, gauzes, tapes and sticks make up for the solid topical medication. These topical dosages vary widely according to their physical characteristics (Ueda et al. 2009; Murthy, and Shivakumar, 2010). Some centuries ago, apothecary owners in an attempt to solving stress-induced health issues, and also for cosmetic purposes, began to study the properties of various herbs, plants, and other organic matter. The knowledge from such research were used to help people soothe and treat common ailments like headaches, muscle pain, menstrual cramps, joint pain, skin diseases and a myriad of other complaints. Today, the use of natural, plant-based remedies for similar complaints has seen resurgence. People are gravitating towards products that are natural and revisiting the properties of those specific ingredients exclusive to nature.

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The use of skin cream as topical medications for the treatment of dislocation, joint-ache, back-pain, and for body massage has become popular in today’s society as its use cuts across tribes, nations, languages and races. These medications many times, come in the form of creams and ointments (sometimes refer to as balms) and are termed pain relief creams or ointments. These topical pain relief creams or ointment work by penetrating the skin and thereby transporting herbal extracts or ingredients through the skin into the underlying tissue and joints where they give relief to the aching area. The form in which such pain relief medications are produced whether as creams or as ointments(balms), becomes very important as it dictates the way these herbal ingredients are transported to their sites of function. According to Allen, and Popovich, (2005), a pharmaceutical cream can be defined as a semisolid preparations containing one or more medicinal agents dissolved or dispersed in either water-in-oil (W/O) emulsion or oil-inwater (O/W) emulsion or in another type of water-washable base. Another type of topical dosage that shares similar characteristics with creams is lotion. According to Ueda, C.T., et al. 2009, lotions are low viscosity emulsions that are used for external application to the skin. An emulsion is a two-phase system consisting of at least two immiscible liquids, in which one phase is dispersed throughout a vehicle. According to Aulton, (2002); Benson, and Watkinson, (2012); Israelachvili, (1994); Ueda, C.T., et al. (2009), there are two types of emulsions namely O/W and W/O. FACTORS AFFECTING SUITABILITY OF CREAM AS A TOPICAL MEDICATION Today's cosmetic emulsions are not simply mixtures of oil, water, and emulsifier, but they also contain a lot of active ingredients whose function is to ameliorate the condition of the skin. Most of these ingredients according to TH. Forster et al (1999) will work only if they penetrate into the stratum corneum or the deeper layers of the epidermis. TH. Forster et al (1999) further noted that the rate of penetration of an active ingredient from a topical formulation depends on its passive diffusion into the skin. This in principle obeys Fick's law of diffusion. Many cosmetic products represent non-Newtonian liquids, which mean that there is dependence on many measurement conditions: load, viscometer type, force etc. Rotational viscometers are used for study of the nonNewtonian liquids (Viscometer system of coaxial cylinders or Viscometer with cone-plate system). There are pseudo elastic liquids, like soap, which lose their viscosity under the increasing influence. Therefore, to make cosmetics stay on the body skin and face the products should have more viscosity, no matter, the type of product it is. To compare different non-Newtonian liquids it is necessary to create absolutely similar condition, because the change of just one component may give an absolutely wrong result. According to Yao and. Patel (2001), creams and lotions require a complex both rheological and thermal testing in order to properly evaluate, characterize and control the quality for the end user satisfaction. There is need for creams, when applied to the human body, to spread easily on the skin without feeling greasy or sticky. Yao and. Patel (2001), further argued that during the shelf storage of the cream, the ingredients of the products should be able to remain intact and not separate or settle in the container. This will make the cream not to feel lumpy or grainy when it is applied on the skin. It therefore means that the product has to be formulated with the aim of meeting the standard and satisfaction of the consumers. Babcock (1931) noted that the physical state of the butterfat contained in the cream affects the viscosity of the cream. Homogenization increases the viscosity of cream. Laba (1993) listed the rheological measurements required in various pharmaceutical and cosmetics industries to include quality control, storage stability under different weather and transportation conditions, correlation with sensory assessment and consumer evaluation, effects of formulation on consistency and prediction of flow behavior under manufacturing or production environment conditions. According to Ali and Yosipovitch (2013), the skin pH is an important factor in the penetration of cream into the skin. Behne et al. (2005); Fluhr et al (2001); Gunathilake et al. (2009); Hachem et al. (2003); Hachem et al (2005); Rippke et al 2009, has shown that skin pH majorly affect the barrier homeostasis, antimicrobial defense mechanisms, SC integrity and cohesion. According to Blaak et al. (2011) studies in recent times have provided insight on how the skin surface pH perform key role in SC functions as permeability barrier homeostasis and SC integrity/cohesion. The worked showed that the Permeability barrier homeo-stasis depend majorly on the pH value different throughout the SC when seriously unsettled skin sites are exposed to a neutral pH buffer (Mauro et al 1998) or to “superbases” Hachem et al. 2003), structure healing is delayed. These pH-induced barrier abnormalities are associated with an inhibition of two lipid processing en-zymes, β-glucocerebrosidase (βGlcCer’ase) and acid sphingomyelinase (aSMase), which exhibit low pH op-tima and transfer polar lipids as,

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glucosylceramide and sphingomyelin, to the non-polar barrier organization (Holleran et al 1992; Jensen et al 1999; Schmuth et al 2000; Takagi et al. 1999). The pH of the Skin is ordinarily acidic with a range between 4–6 and that of the internal system of the body maintains a near-neutral pH that is between 7–9. This variation creates a pH difference or gradient of 2–3 units between the SC and the underlying epidermis and dermis. The factors that affect skin pH include age, anatomic site, genetic predisposition, ethnic differences, sebum, skin moisture and sweat (Ali and Yosipovitch 2013). A. SHEA BUTTER AND HEALTH BENEFITS According to Alonge and Olaniyan (2007), Shea butter is the oleaginous material obtained from the kernel of the shea nut tree Vitellaria paradoxa. Shea butter has been described as a vegetable fat extracted from the kernels of the fruit of V. paradoxa, Sapotaceae by many authors (Hall et al., 1996; Pontillon, 1996; Kengue and Ndo, 2003; Elias and Carney, 2004; Schreckenberg, 2004). Furthermore, Abdul–Mumeen et al., (2013) described Shea butter as a yellowish– grey solid material or yellowish white in colour with a strong smell extracted as fat from the kernels of the Shea nut fruit. Abdul–Mumeen et al., (2013) further noted that Shea butter when extracted from Shea kernels is raw which can be then refined into a purer product. Shea butter is renowned for its use as an ingredient in the formulation of cosmetics (Abbiw D K 1990; Akihisa et al. 2010; Malachi, 2014), substitute for Cocoa butter in chocolate industries (Ogbonnaya and Adgidizi 2008). When used as a Coco butter substitute, the taste is noticeably different (Fold N 2000). Shea butter has also been used as raw material in the making margarine, soap, detergent and candle (Russo and Etherington 2001). It can be used as waterproof to protect the walls of the house during the rainy season when low quality butter and byproducts of processed nuts are used to coat earthen walls of houses (Fluery 1981). In African pregnant women, Shea butter has been used to prevent stretch marks. It has also been used for different purposes like soothing and accelerated healing after circumcision, repelling insect and protection against Simulium infection (Goreja WG 2004). They have not been reported of people being allergic to the consumption of Shea butter or products made from it. According to Essengue et. al, (2009); Kanwaljit et. al., (2010), for centuries, Shea butter has been used as a cosmetic ingredient because of its high vitamins content and fatty acids coupled with its easy-to-spread consistency. These attributes of Shea butter has made it a great product for smoothing, soothing, and conditioning of skin. It has been used by local healers for the treatment of rheumatism, inflammation of the nostrils, nasal congestion, leprosy, cough, and minor bone dislocation (Tella A 1979; Badifu 1989; Goreja 2004; Olaniyan and Oje 2007). As a plant fat, Shea butter contains majorly 90% or more of triglycerides namely palmitic, stearic, oleic, linoleic and arachidic and a minor unsaponifiable fraction. Triglycerides present in Shea butter are responsible for its emollient properties. The unsaponifiable fraction of the Shea butter is where the bioactive substances are found and thus responsible for the Shea butter’s medicinal properties (Esuoso et al., 2000; Maranz et al 2004). These bioactive substances include hydrocarbons, tocopherols, sterols, and alcohols. In the work of Honfo et al, (2014) it was further reported that the most representative tritrepene alcohols found in the unsaponifiable part of Shea butter were α-amyrin (26.5%), β-amyrin (10.2%), lupeol (21.7%), and butyrospermol (25%), most of which occur as acetic acid and cinnamic acid ester. The α-amyrin content was 40–50%, the β-amyrin content 5–10%, the lupeol content 10–20%, and the butyrospermol content 15–25%.

II.

METHODOLOGY

a. PURIFICATION OF SHEA BUTTER Solid Shea butter used were locally sourced from Kogi state and heated to melting point. This liquid was mixed with activated charcoal to remove the characteristic odour of Shea butter. The resulting mixture was heated over a water bath for thirty minutes; filtered and allowed to cool. The other ingredients such as bees wax, cetrimide, methyl salicylate, hydrous wool fat and polysorbate 80 used for the formulation were of analytical grade. b. PRE-TREATMENT OF THE RAT (ALBINO MOUSE) SKIN Albino mouse was butchered with its skin removed and treated to make it suitable for the rate of penetration (mass transfer) test. After butchering the rat its skin was firstly, separated and washed with water, then all the

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hair follicles on the skin and the adipose tissue layer were thoroughly scraped off using a sharp razor blade after which it was washed with acetone and sodium hydroxide to remove any remnant tissue impurity. Secondly, a buffer solution was prepared from dissolving a mixture of 0.5g of potassium hydrogen tetraoxophosphate and 0.3g of sodium hydrogen tetraoxophosphate in 1 liter of water. The skin was further treated by soaking in the buffer solution for about 24hrs to give the skin a balanced PH before the test, to enable easy penetration and to preserve the skin. c. CREAM FORMULATION Three different formulations were made and accordingly labeled as Y1, Y2 and Y3 respectively. Y1 consists of Shea butter, bees wax, cetrimide and water; Y2 consists of Shea butter, bees wax, polysorbate 80 (2%) and water. Y3 consists of bees wax, cetrimide (2%), hydrous wool fat and water; it was used as the standard for comparison. Shea butter and bees wax were mixed and heated over a water bath till they melted at a temperature of 750C. To form the water phase of the formulation, water was heated over a water bath and cetrimide was measured and added when the temperature was 750C. It was stirred continuously to make sure the emulgent totally dissolved. After preparing the oil and water phases separately, the water phase (containing the emulgent) was added into the oil phase in a thin, steady stream with continuous stirring using stirrer from side to side for about 5 to 10 minus. The mixture was allowed to cool while stirring until the temperature falls to below 400C and at this stage. The active ingredient (methyl salicylate) was added when the temperature was reduced to 40 . The mixture was continuously stirred until the cream was formed. The formed cream was allowed to cool and packaged in a dispensing bottle and labeled Y1. This method was used for each of formulation Y2 and Y3. d. pH DETERMINATION The determination of the cream formulation PH was done using a PH meter (3 PSC model). A 3.1% aqueous solution of each of the formulated creams (Y1, Y2, Y3) was prepared and their PH values taken at a constant temperature of 25⁰C. e. DETERMINATION PENETRATION RATE This was achieved with the pretreated skin of an albino mouse using an improvised Franz diffusion cell and a dissolution chamber. The receptor compartment contained the prepared phosphate buffer used to treat the skin in an amount close to 100ml. 5g of the test formulation was applied over a small area of the skin across the donor compartment. The temperature of this diffusion medium was maintained at 37⁰C while the buffer solution was stirred continuously with a Teflon coated magnetic bar at 500 rpm.5 ml of each sample were withdrawn from the release medium at time intervals of 5minus until 60minus. The samples were analyzed using a UV spectrophotometer at 278nm against their respective blank.

III. Formulation Code

FC

RESULTS AND DISCUSSION

a. RESULTS Table 1: Composition and Concentration of the formulations MethylSheaBees Hydrous Cetrimide Polysorbate butter wax wool fat (g) 80 (ml) salicylate (g) (g) (g) (ml)

Y1

6.4

12.5

8.3

Y2

6.4

12.5

8.3

Y3

6.4

-

8.3

FH

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-

12.5

Water (H₂O) (ml)

2

-

20

-

2

20

2

-

20

Table 2: Physiochemical Attributes of Shea butter analgesic cream FpH FW FCO FO

FS (cm/s)

FV(secs)

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Y₁

extremely good

4.95

Moderate

Milky

Shea butter and methyl salicylate

3.5

55.81

Y₂

Very Good

6.01

Not washable

Deep tan

Shea butter and methyl salicylate

7.66

60.3

Y₃ Good 5.33 Easy Deep tan methyl salicylate only 3.3 52.62 Where FC= Formulation Code; FH= Formulation Homogeneity; FpH= Formulation pH Value; FW= Formulation Washability; FCO= Formulation Colour; FO= Formulation Odour; FS= Formulation Spreadability; FV= Formulation Viscosity. Table 3: Values for Penetration Rate at different time variations for each formulation Time Y1 Penetration Y2 Penetration Rate Y3 Penetration (minus) Rate Rate 5 0.513 0.436 0.210 10 0.534 0.453 0.222 15 0.551 0.480 0.231 20 0.588 0.508 0.239 25 0.605 0.532 0.253 30 0.628 0.556 0.273 35 0.643 0.569 0.291 40 0.663 0.576 0.309 45 0.680 0.590 0.316 50 0.701 0.612 0.323 55 0.724 0.621 0.331 60 0.741 0.638 0.339

70 60 50 Y3 Penetration Rate

40

Y2 Penetration Rate

30

Y1 Penetration Rate

20

Time (minus)

10 0 1

2

3

4

5

6

7

8

9 10 11 12

Fig 1: Penetration Rate for the three formulations.

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100% 90% 80% 70% 60%

Y3 Penetration Rate

50% 40% 30% 20% 10% 0%

Y2 Penetration Rate Y1 Penetration Rate Time (minus)

1 2 3 4 5 6 7 8 9 10 11 12 Fig. 2: Penetration Rate for the three formulations b.

DISCUSSION

From table 2, Y1 and Y3 formulated using cetrimide (cationic surfactant) as the emulgent showed different level of acidity from Y2 formulated using polysorbate 80 (nonionic surfactant) as the emulgent. While the pH of Y 1 and Y3, showed high level of acidity (4.95 and 5.33) respectively, that of Y2 gave a pH of 6.01 depicting weak acidity. It can therefore be inferred that the type of emulgents used in cream formulation becomes very significant in preparing a cream with a good pH. However, pH values of all the cream formulated were within the pH value allowed for the human skin (i.e. a pH between 4 - 6) (Wohlrab and Gebert 2018; Zlotogorski 1987; Dikstein and Zlotogorski 1994; Rippke et al. 2002). Blaak et al. (2011) noted that the physiological skin surface pH is just below 5. However, with age the skin surface pH value increases up to 6. An increased pH relates with reduced barrier integrity or cohesion. Haut (NachGaskettenmessung) (1928); Öhman and Vahlquist (1994); Elias (2005) further noted that the acid pH value of the skin surface, known as the acid mantle and the pH gradient of the stratum corneum (SC), regulates at least three epi-dermal functions: antimicrobial barrier, permeability barrier and barrier integrity/cohesion. It can therefore be said that all the formulation’s pH is ‘safe for consumption’ (Holland et al. 1978; Korting, 1990; Kurabayashi et al. 2002; Korting et al. 1992; Buraczewska and Lodén 2005). It can also be seen from table 2 that both Y1 and Y2 are more homogenous and viscose than the control Y3 with Y1 most homogenous amongst the three formulations. Hence, it can be deduced that creams made with Shea butter are more spreadable than creams formulated from hydrous wool fat. Rate of skin Penetration (in vitro release) From Table 3 and Fig. 2, it is seen that the absorbance or rate of penetration of each of the formulations increased with time. With Y1 having the highest penetration per time. For the first 30mins, there was a uniform increase with time, after which a uniform jump in the time rate of increase in absorbance for the next 10 mins, and then finally the rate of penetration increase with time slows down uniformly. Comparatively, the formulation with Shea butter performed better in breaking down the barrier homeostasis, antimicrobial defense mechanisms and SC integrity and cohesion of the skin than the formulation with hydrous wool fat. This is in agreement with Hee (2011); Kraft and Lynde (2005). From fig. 2, i.e. the plot of the absorbance of M 2 against time, it was also observed that the absorbance of M 2 increased with time. For the first 20 mins, the absorbance rate increased almost uniformly and just like M1, there was a uniform jump in the increasing rate of absorbance for the next 10 mins and then finally the time rate of increase slows down uniformly. The plot depicts a clear picture showing that Y 1 and Y2 had a higher and better penetration rate than Y3. It can therefore be said, that the concentration of methyl salicylate that diffused

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through the skin, within the same time interval was higher for Y1 and Y2 than for Y3. From the above results, it can therefore be deduced that Shea butter as the oil (carrier) base gave a cream that contained more of the methyl salicylate thereby giving easier and faster absorbance through the skin of the rat as compared to the use of hydrous wool fat.

IV.

CONCLUSION

This study has shown that physio-chemical properties of Shea butter as a base for the formulation of analgesic cream (Methyl 2-hydroxybenzoate) in the presence of either cetrimide or Polysorbate 80 as the emulgent. The result from the formulation has shown that Shea butter can be used as the oil base for the formulation of the cream under consideration. The result also show that the creams formulated with Shea butter penetrated more into the skin of the rate than that of hydrous wool fat cream. It therefore can be said that creams made from Shea butter has more of the active ingredient (methyl salicylate) as shown by its absorbance rate when compared to the hydrous wool fat cream. The formulations’ pH values were well within the pH values allowable for the human skin. This shows that the creams can be used on the human body without been worried about skin reaction but rather it will effectively moisturize the skin. Therefore from the forgoing, Shea butter can be effectively incorporated into the production of creams to replace the costly hydrous wool fat.

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