CHARACTERIZATION AND ANTIMICROBIAL ACTIVITIES OF CINCHONA (CINCHONA LEDGERIANA MOENS EX TRIMEN) HONEY AS ACTIVE SUBSTANCE CANDIDATE FOR ANTIACNE

• Main Author: Andam Dewi, Mira
• Format: Dissertations
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/53017
• Institution: Institut Teknologi Bandung

Currently, many studies have reported the therapeutic properties of honey, including manuka honey whose activity has been confirmed against about 60 types of bacteria, including aerobic and anaerobic, as well as Gram positive and negative. Manuka honey has been reported to be effective in treating skin infections that are resistant to antibiotics and gastrointestinal disorders and is recognized as having a role and potential as a topical active ingredient for post-surgical treatment or wound infection. Besides being consumed, manuka honey is often used as a mask to moisturize the skin and relieve acne inflammation. This fact has encouraged research to test and prove the antimicrobial effect of other types of honey that are specific, such as manuka honey. A recently published research report on honey disseminates the importance of continued research activities to encourage new findings on the benefits and uses of honey as a functional food and medical application. In line with this, this study carried out the characterization and antimicrobial activity test of cinchona (Cinchona ledgeriana Moens ex Trimen) honey as a candidate substance for antiacne. The research begins with the characterization of cinchona honey using the Fouriertransform Infrared-Attenuated Total Reflectance (FTIR-ATR) method. A total of seventeen honey samples were measured to obtain the infrared spectrum of each honey. The wave number and percent transmittance data obtained were then statistically processed using the principal component analysis (PCA) method to determine the similarities in the characteristics of the honey samples. The results of characterization for cinchona honey S7, S8, and S9 show the proximity of the distance and high similarity values based on dendrogram data, so that it is predicted to have constituent components with similar structures and physicochemical properties. Furthermore, the cluster analysis showed that the honey samples of cinchona S7, S8, and S9 were in the same quadrant, so that besides having similar physicochemical properties, they were also thought to come from the same flower nectar source. In the next stage of research, the botanical determination of cinchona planted in beekeeping location was carried out. It was confirmed that the cinchona was a grafting combining Cinchona pubescens Vahl., known also as Cinchona succirubra Pav. ex Klotzsch which is used as the rootstock, while Cinchona officinalis L. or known as Cinchona ledgeriana Moens ex Trimen which is used at the top. Characterization of flower herb includes determination of water content, water soluble extract content, 96% ethanol soluble extract content, loss on drying, total ash content, water soluble ash content, and acid insoluble ash content, giving the following determination results 4,6 ± 1.15% v/w, 48.7 ± 0.59% w/w, 70.9 ± 0.95% w/w, 6.6 ± 0.19% w/w, 2.9 ± 0.25% w/w, 1.6 ± 0.02% w/w, and 3.0 ± 0.04% w/w, respectively. The characterization of cinchona flowers herb has neither been listed in Materia Medika Indonesia (MMI) and nor was reported in other literature sources. The preparation of cinchona flowers extract was conducted applying cold extraction method maceration to minimize the deterioration of thermolabile metabolite compounds. To facilitate evaporation, methanol which is a polar organic solvent with relatively low boiling point was used as extracting solvent. The method gave a yield of 39% w/w. Based on the results of FTIR-ATR characterization combined with the chemometrics method, it shows that S7, S8 and S9 have similar physicochemical properties and are thought to originate from the same flower nectar source, so that further testing the quality of honey is carried out on S7, S8 and S9 to confirm whether the cinchona honey from the type of Cinchona ledgeriana Moens ex Trimen that is used meets the requirements of SNI 3545: 2013 concerning honey. Organoleptic tests include odor and taste tests, while physicochemical tests include hydroxymethylfurfural (HMF) levels, diastase enzyme activity, water content, glucose levels, sucrose levels, acidity tests, water insoluble solids, ash content, and heavy metal contamination, chloramphenicol test, and microbial contamination test. The test results on the S8 and S9 cinchona honey showed that the HMF content met the requirements, except for the S7 cinchona honey, the HMF content did not meet the requirements, namely 52.3 mg / kg, while the requirements of SNI 3545: 2013 should not be more than 50 mg / kg. However, according to Codex Alimentarius (2000) standards, for tropical areas, HMF levels are determined not to exceed 80 mg / kg. Extraction of cinchona honey was also carried out by maceration method using methanol as solvent, and the yielded methanol extract of cinchona honey of 83, 90 and 87% for E7, E8 and E9, respectively. Phytochemical screening was carried out on cinchona flower simplicia, cinchona flower methanol extract, cinchona honey S7, S8, S9 and methanol extract of cinchona honey E7, E8, E9, to see the correlation of secondary metabolite content of cinchona flower and cinchona honey. The results of phytochemical screening of cinchona flower simplicia and cinchona flower honey showed similarities in the secondary metabolites, namely flavonoid, polyphenols, quinones, and monoterpenoids-sesquiterpenoids. The results of research on honey content by several researchers reported that the secondary metabolites that were commonly detected in honey were the flavonoids and polyphenols. Acne is a widespread and severe inflammatory skin condition that affects approximately 80% of adolescents and young adults in the 11-30 age range with a percentage of about 43% in men and 51% in women in their twenties. Propionibacterium acnes (P. acnes), Staphylococcus epidermidis (S. epidermidis), and Staphylococcus aureus (S. aureus) are the main bacteria that can cause acne on the skin. Exploration of the antibacterial function of cinchona honey (Cinchona ledgeriana Moens ex Trimen) and its methanol extract was carried out to study its inhibitory activity against acne-causing bacteria. Based on the results of honey quality testing, only honey S8 and S9 meet the requirements of SNI 3545: 2013, therefore in vitro testing for the antibacterial activity of cinchona honey was only carried out on S8 and S9, and the methanol extract E8 and E9 used the liquid microdilution method and reported as the value of the minimum inhibitory concentration (MIC). Furthermore, the MIC value obtained was confirmed for its growth on agar media that had been added with the test microbes and the results were reported as the minimum bactericidal concentration (MBC). The sample test results were compared with the comparable medical honey manuka MGO 550+, manuka MGO 400+ and the antibiotic tetracycline HCl, erythromycin stearate. Samples of cinchona honey S8, S9, manuka honey MGO 550+, and manuka honey MGO 400+ showed an inhibitory effect against P. acnes with MIC and MBC values, respectively 32, 32, 32, 32 ?g/mL and 512, 512, 512, 512 ?g/mL. The same sample showed an inhibitory effect on S. epidermidis and S. aureus with the same MIC values, 128, 128, 128, 128 ?g/mL, respectively, and MBC values > 512,> 512,> 512,> 512 ?g/mL. Methanol extract of cinchona honey (E8, E9), MGO 550+ manuka honey, 400+ manuka honey showed an inhibitory effect against P. acnes and S. epidermidis, with the same MIC and MBC values, 0.0625, 0.0625, 0.0625, 0.0625 ?g/mL, respectively. The same sample showed an inhibitory effect against S. aureus with MIC and MBC values of 2, 32, 0.0625, 2 ?g/mL and 4, 32, 0.0625, 2 ?g/mL, respectively. The comparative antibiotic tetracycline HCl and erythromycin stearate showed an inhibitory effect on P. acnes with MIC and MBC values of 1.1 ?g/mL and 32, 256 ?g/mL, respectively, then S. epidermidis showed MIC and MBC values, respectively. 32, 64 ?g/mL and 256, 512 ?g/mL, whereas S. aureus showed MIC and MBC values of 32, 128 ?g/mL and 128, 512 ?g/mL, respectively. Cinchona honey, and methanol extract of cinchona honey show strong antimicrobial activity because the MIC value is in the range of 50-500 ?g/mL, so it has strong antiacne potential, especially against P. acnes, therefore it can be used as a candidate substance for antiacne.