Annual Research & Review in Biology 17(3): 1-7, 2017; Article no.arrb.36467 ISSN: 2347-565X, NLM ID: 101632869 Formulation and Evaluation of Natural Anti-Acne Cream Containing Syzygium samarangense Fruits Extract Mahendran Sekar 1* and Fouzia Hanim Abdul Halim 1 1 Faculty of Pharmacy and Health Sciences, Universiti Kuala Lumpur, Royal College of Medicine Perak, Ipoh - 30450, Malaysia. Authors contributions This work was carried out in collaboration between both authors. Author MS designed the study, performed the statistical analysis, wrote the protocol and wrote the first draft of the manuscript. Author FHAH managed the analyses of the study and literature searches. Both authors read and approved the final manuscript. Article Information DOI: 10.9734/ARRB/2017/36467 Editor(s): (1) George Perry, Dean and Professor of Biology, University of Texas at San Antonio, USA. Reviewers: (1) Muhammad Ali, Kano University of Science and Technology Wudil, Nigeria. (2) R. Jasmine, Bishop Heber College, India. (3) Renshan Sun, Third Military Medical University, China. Complete Peer review History: http://www.sciencedomain.org/review-history/21058 Original Research Article Received 29 th August 2017 Accepted 15 th September 2017 Published 19 th September 2017 ABSTRACT Pimple, acne, sunburn mark and pigmentation are issues that affected every individual at least once during life time. Consumers have begun to search for a product that can cure the skin issue and grant them with a good and healthy skin such as anti-acne cream. Nevertheless, most of the antiacne creams available in the market contain lots of chemicals that may have some kinds of side effects to the consumers. The present study was conducted to formulate and evaluate the anti-acne cream containing Syzygium samarangense fruits extract. The antibacterial activity of the extract and cream in different concentrations were investigated using two gram-positive bacteria (Staphylococcus aureus and Bacillus cereus) two gram-negative bacteria (Escherichia coli and Salmonella enterica) through disc diffusion method. The antibacterial potential of the extract was studied with four different concentrations (5, 10 and 25 mg/ml). The extract showed significant antibacterial activity against all the tested organisms. This activity was well maintained when the *Corresponding author: E-mail: mahendransekar@unikl.edu.my;
extract was incorporated into the cream formulation with the concentrations of 10, 25, 50 and 100 mg/ml. The formulated cream was also stable after two months. This formulated creams can be successfully used for skin infections which including acne vulgaris, after the confirmation of clinical and toxicity studies in future. Keywords: Acne vulgaris; antibacterial activity; Syzygium samarangense; red Jambu. 1. INTRODUCTION Acne vulgaris is a disturbing issue especially for teenagers and adolescence. According to global statistic, approximately 85% of population will suffer this skin issue at age around 12-25 years, nearly 8% adults at age 23-34 years old and only 3% of adults having acne vulgaris at age 35-44 years old. Although, some skin issue can be categorized as self-limiting, but it also can give negative psychosocial consequences to individual by having low self-esteem, social withdrawal due to embarrassment and at the worst scenario, it can lead to suicidal ideation [1]. Propionibacterium acnes and Staphylococcus aureus were responsible for acne since the presence of this bacterium in the sample isolated from acne patient. At the same time, the increasing number of resistance of acne inducing bacteria toward the antibiotic is a worrisome problem [2]. The alternative treatment of acne have been investigated and adopted. Among the alternate system of medicine, the usage of topical therapeutic agents is more convenient for application. Most of the people now more attracted toward the use of herbal formulations. According to WHO, there are four billion people use herbal medicine as a primary health care and convinced to be safe. Syzygium samarangense also known as Red jambu in Malaysia is a tropical tree growing to 16 to 50 ft. There are three varieties of red jambu fruits available in Malaysia which is pink, red and green. Moneruzzaman et al. [3], reported that phytochemicals in the Red Jambu fruits showed antibiotic action against Staphylococcus aureus, Candida albicans and Mycobacter smegmatis. This fruits were also reported as well to exhibit antibacterial, antidiabetic, antidiarrheal and immunostimulant activities [4-7]. However, there is no anti-acne cream formulation by using Syzygium samarangense extract though it was reported as potential antimicrobial activities [3]. Hence, in the present study we are interested to formulate anti-acne cream from Syzygium samarangense extract and study its antimicrobial potency. 2. MATERIALS AND METHODS 2.1 Collection and Authentication 4 kg of red jambu fruits (Syzygium samarangense) were purchased from local market at Bagan Serai, Perak, Malaysia (Fig. 1) and authenticated by a botanist (UniKL/FPHS/2017/23). 2.2 Extraction The collected fruits were washed thoroughly in distilled water to remove the contaminants. The fruits were chopped into small pieces and dried under shade for 1 week; the dried materials were coarsely powdered using mechanical blender. The powdered fruits of Syzygium samarangense with total weight of 200 g were extracted by maceration method using 2.5 L of 70% ethanol as a solvent for about 1 week with occasional shaking. The ethanolic extract of Syzygium samarangense fruits was filtered and concentrated to dryness under reduced pressure and controlled temperature using rotary evaporator. The extract was stored in air-tight containers in a refrigerator at 4 C until further use. 2.3 Formulation of Anti-Acne Cream The composition of anti-acne cream was shown in Table 1. The oil phase consists of stearic acid and other oil soluble component such as cetyl alcohol and liquid paraffin were dissolved in the oil phase. The oil phase was placed inside the beaker in the water bath. The temperature of water bath was set to 75 C during the heating time. The water soluble components and preservatives (glycerine, methyl paraben and thiethanolamine) were dissolved in the aqueous phase and heated in the same water bath at temperature 75 C. After heating, the aqueous phase was added in portions to the oil phase with continuous stirring until the cooling of emulsifier took place. Different proportion of Syzygium 2
samarangense fruits extract (5% and 10%) was mixed with the base along with fragrance and named as F1 and F2, respectively (Fig. 2). 2.4 Evaluation of Anti-acne Cream [8] The following parameters weree checked to evaluate the anti-acne cream: 2.4.1 Determination of the type of emulsion A scarlet red dye was mixed with the cream. A drop of the cream was placed on microscopic slide and examined under a microscope. If the disperse globules appear red the continuous phase colourless, the cream is oil in water (o/w) type. The reverse condition is occurs water in oil (w/o) type cream. 2.4.2 ph of the cream Fig. 1. Syzygium samarangense Table 1. Composition of the anti-acncontaining Syzygium samarangense cream extract Components Amount (%w/w) Active ingredients Syzygium samarangense 5% (F1) 10% (F2) Oily phase Stearic acid 10% 20% Cetyl alcohol 4% 8% Liquid paraffin 4% 8% Aqueous phase Glycerine 5% 10% Methyl paraben 0.05% 0.05% Thiethanolamine 0.05% 0.05% Distilled water q.s Upto 100% Upto 100% The ph meter was calibrated using standard buffer solution. About 0.5 g of the cream was weighed and dissolved in 50 ml of distilled water and its ph was measured. 2.4.3 Homogeneity The formulation was tested for homogeneity by visual appearance and touch. 2.4.4 Appearance The appearance of the cream was judged by its color, pearlescence and roughnesss and graded. 2.4.5 After feel Emolliency, slipperiness and amount of residue left after the application of fixed amount of cream were checked. Fig. 2. Formulated anti-acne cream 3
2.4.6 Type of smear After application of cream, the type of film or smear formed on skin were checked. 2.4.7 Removal The ease of removal of the cream applied were examined by washing the applied part with tap water. 2.4.8 Stability study The stability study was carried out by storing the anti-acne cream at three different temperatures which are 8 C, 27 C and 40 C for 2 months. 2.5 Antibacterial Screening 2.5.1 Test microorganisms [9] Four common pathogenic microorganisms were used in this study, two gram positive bacteria which are Staphylococcus aureus and Bacillus cereus while Escherichia coli and Salmonella enterica were gram negative bacteria. All the four bacterial strains obtained from Microbiology and Cell Culture Department, Royal College of Medicine Perak, Perak, Malaysia, were used for evaluating antibacterial activity. 2.5.2 Disc-diffusion method A suspension of the tested microorganisms in standard quantity was uniformly swabbed on nutrient agar plates using sterile cotton swabs. Sterile blank discs were individually impregnated to the different concentration of formulated cream (10, 25, 50 and 100 mg/ml) and extracts (5, 10 and 25 mg/ml) were placed onto the inoculated agar plates. The plates were inverted and incubated at 37 C for 24 h for bacteria growth. The antibacterial activity was evaluated by measuring diameter of the resulting zone of inhibition against tested microorganisms. The positive control used was Vancomycin due to its activity against a wider number of bacterial types and distilled water used as negative control. All disc diffusion experiments were performed in two separate experiments and antibacterial activity was expressed as Mean ± Standard Error Mean [9]. 3. RESULTS AND DISCUSSION The dye test was confirmed that both F1 and F2 formulations were o/w type of emulsion cream. The ph of the formulated cream was found to be in range 4.6 to 4.8 which is good and recommended ph for the skin. The formulated anti-acne cream was evaluated for several physicochemical tests and the results were shown in Table 2. The type of smear formed on the skin was not greasy after the application of both creams. The creams were easy to remove after application by washing with water. The formulations were able to produce uniform distribution of extracts in the cream. This was confirmed by visual examination and by touch. There were no changes in term of colour of the cream even it was kept for a long period of time. After feel test showed that the creams were emollient and slipperiness. All the physicochemical parameters were maintained during the accelerated stability studies at temperatures 8 C 0.1 C in refrigerator and at 25 C ± 1 C, 40 C 1 C in incubator for 8 weeks. The results of accelerated stability test showed that there were no any changes in the colour of the cream. The ethanolic extract of Syzygium samarangense fruits showed the antibacterial Table 2. Physicochemical evaluation of the formulated anti-acne cream Parameter Result F1 F2 Homogeneity Good Good Appearance No change in colour No change in colour Odour Good Good Spreadability Good Good After feel Emollient and slipperiness Emollient and slipperiness Type of smear Non-greasy Non-greasy Removal Easy Easy 4
Table 3. Antibacterial activity of the ethanolic extract of Syzygium samarangense fruits Concentration Gram positive Gram negative (mg/ml) Staphylococcus aureus Bacillus cereus Escherichia coli Salmonella enterica Zone of inhibition (mm) 5 8.73±0.90 8.93±1.03 13.80±0.93 5.00±0.58 10 11.80±0.65 7.27±0.54 12.30±0.44 11.20±0.61 25 13.50±0.29 6.93±0.23 15.00±0.58 13.30±0.50 Negative control Nil Nil Nil Nil Positive control (Vancomycin) 17.20±1.30 13.10±0.21 13.50±1.27 12.90±1.00 Values are expressed as mean ± SEM (n=3). P<0.05 between negative control disc and extract treated disc. P<0.05 between negative control disc and standard treated disc activity against Escherichia coli as seen by the zone of inhibition ranges from 12.30±0.44 mm to 15.00±0.58 mm (Table 3). The zone of inhibition against Staphylococcus aureus, Salmonella enterica and Bacillus cereus were 8.73±0.90 mm to 13.50±0.29 mm, 5.00±0.58 mm to 13.30±0.50 mm and 8.93±1.03 mm to 6.93±0.23 mm, respectively. All the extracts were showed a significant (p<0.05) zone of inhibition when increasing the concentration of the extract except for extract tested on gram positive Bacillus cereus. However, the standard vancomycin showed significantly increased zone of inhibition against the entire tested organism when compared to tested extract. The antibacterial potential of the formulated creams (F1 and F2) were studied with four different concentrations (10, 25, 50 & 100 mg/ml). F1 and F2 showed significant antibacterial activity against all the tested organisms and the zone of inhibition was increased when increasing the concentration of the formulated cream (Tables 4 and 5). However, the standard vancomycin showed significantly increased zone of inhibition against the entire tested organism when compared to F1 and F2. Syzygium samarangense is widely used as medicinal plant especially for the treatment of bronchitis, diabetes mellitus and inflammation syndrome. According to Moneruzzaman et al., 2015 [3], the phytochemicals present in Syzygium samarangense extract such as anthocyanin, flavonoids, phenolic acid and tannins are responsible against microbial infection. Besides that, Syzygium samarangense is also used to cure the skin infection due to its ability to treat the inflamed skin. Table 4. Antibacterial activity of formulated anti-acne cream (F1) F1 Gram positive Gram negative Concentration (mg/ml) Staphylococcus aureus Bacillus cereus Escherichia coli Salmonella enterica Zone of inhibition (mm) 10 1.83±1.01 10.00±1.15 1.50±0.76 9.17±1.30 25 7.73±0.37 11.90±0.52 6.17±1.01 10.30±0.37 50 10.20±0.44 12.50±0.764 9.17±0.44 10.70±0.82 100 13.80±0.72 9.07±0.58 7.93±0.23 14.00±0.61 Negative control Nil Nil Nil Nil Positive control (Vancomycin) 14.40±0.35 13.10±0.58 12.80±0.42 20.00±0.84 Values are expressed as mean ± SEM (n=3). P<0.05 between negative control disc and extract treated disc. P<0.05 between negative control disc and standard treated disc 5
Table 5. Antibacterial activity of formulated anti-acne cream (F2) F2 Gram positive Gram negative Concentration (mg/ml) Staphylococcus aureus Bacillus cereus Escherichia coli Salmonella enterica Zone of inhibition (mm) 10 3.83±0.72 11.60±1.07 4.07±1.10 8.53±1.74 25 8.27±1.51 13.00±1.08 10.10±0.62 11.90±0.19 50 12.60±0.95 13.50±0.55 9.07±0.52 12.00±0.58 100 14.20±1.42 13.60±0.70 10.30±0.67 15.30±1.20 Negative control Nil Nil Nil Nil Positive control (Vancomycin) 17.10±0.64 15.10±0.70 16.90±0.75 20.20±0.49 Values are expressed as mean ± SEM (n=3). P<0.05 between negative control disc and extract treated disc. P<0.05 between negative control disc and standard treated disc The antibacterial potential of ethanolic extract of Syzygium samarangense along with two formulated anti-acne cream were determined by using disc diffusion method. Four bacteria that involved in the test were Staphylococcus aureus, Bacillus cereus, Escherichia coli and Salmonella enterica. From the result, the antibacterial activity of the formulated cream showed greater zone of inhibition toward all bacteria especially Salmonella enterica, Staphylococcus aureus and Escherichia coli. Therefore, the formulated antiacne cream has a potential in the treatment of acne because Staphylococcus aureus has been reported as common bacteria in causing acne while Escherichia coli is one of the bacteria that responsible in causing skin infections [10,11]. 4. CONCLUSION The observed potential antibacterial activity of the formulations may be due to the presence of active constituents in the ethanolic extract of Syzygium samarangense. The antibacterial activity was well maintained when it was converted into cream formulation. This was good sign for further studies to make this product into commercial standards. We recommended that the formulated cream can be successfully used for skin infections which including acne vulgaris, after the confirmation of clinical and toxicological studies in future. COMPETING INTERESTS Authors have declared that no competing interests exist. REFERENCES 1. Aditi V, Pranav S. Formulation and evaluation of topical anti acne formulation of coriander extract. International Journal of Pharmaceutical Sciences Review and Research. 2012;16(2):97-103. 2. Srivastava A, Alam S, Shams S, Tiwari M, Mittal A, Sanjeev C. Formulation and evaluation of antiacne cream containing Withania somnifera. Journal of Pharmaceutical and Scientific Innovation. 2014;3(4):348-352. 3. Moneruzzaman KM, Jahan SM, Nashriyah M, Boyce AN. Bioactive constituents, antioxidant and antimicrobial activities of three cultivars of wax apple (Syzygium samarangense L.) fruits. Research Journal of Biotechnology. 2015;10(1):7-16. 4. Tina P, Padmavathi D, Jasmin Sajini R, Sarala A. Syzygium Samarangense: A review on morphology, phytochemistry & pharmacological aspects. Asian Journal of Biochemical and Pharmaceutical Research. 2011;4(1):155-163. 5. Venkata Ratnam K, Venkata Raju RR. In vitro antimicrobial screening of the fruit extracts of two Syzygium species (Myrtaceae). Advances in Biological Research. 2008;2:17-20. 6. Consolacion YR, Francisco CF, Dennis DR, Chien-Chang S. Chemical constituents of Syzygium samarangense. Der Pharma Chemica. 2014;6(3):256-260. 7. Rajib M, Nur Hasnat E, Ashraf UZ, Badrul A. In vivo evaluation of the pharmacological activities of Syzygium samarangense (Blume) Merr. & L.M. Perry. Advances in Biological Research. 2014; 8(3):107-115. 8. Mahendran S, Pavitra S, Afzan M. Formulation and evaluation of novel antiaging cream containing rambutan fruit extract. International Journal of 6
Pharmaceutical Sciences and Research. 2017;8(3):1056-1065. 9. Mahendran S, Fatin Nabila AJ, Nur Hasyimah Z, Nur Izzati M, Nurul Ain Z, Ramizatun Annisa K, Syafiq A. Comparative evaluation of antimicrobial properties of red and yellow rambutan fruit peel extracts. Annual Research & Review in Biology. 2014;4(24):3869-3874. 10. Mullaicharam AR, Fatma AE. Evaluation of anti-acne property of poly herbal formulation. Journal of Biomedical and Pharmaceutical Research. 2012;1(3):28-35. 11. Panupon Khumsupan WG. Anti-acne activity of Garcinia mangostana L.: A review. Plant Science Today 2014; 1(3):147-150. 2017 Sekar and Halim; This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Peer-review history: The peer review history for this paper can be accessed here: http://sciencedomain.org/review-history/21058 7