Inhibitory ability of extracts and compounds from leaves of cassia grandis l.f to propionibacterium acnes

AGU International Journal of Sciences – 2019, Vol. 7 (4), 1 – 11<br /> <br /> <br /> <br /> <br /> INHIBITORY ABILITY OF EXTRACTS AND COMPOUNDS FROM LEAVES<br /> OF CASSIA GRANDIS L.F TO PROPIONIBACTERIUM ACNES<br /> <br /> Ngo Quoc Luan1,2, Phung Thi Yen Thanh2, Tran Nhan Dung3, Huynh Van Ba4<br /> 1<br /> Vietnam Academy of Science and Technology<br /> 2<br /> School of Education, Can Tho University<br /> 3<br /> Biotechnology Research & Development Institute, Can Tho University<br /> 4<br /> Can Tho Dermatology Hospital<br /> <br /> Information: ABSTRACT<br /> Received: 070/8/2018<br /> Nine Propionibacterium acnes (P. acnes) strains resistant to both<br /> Accepted: 03/01/2019<br /> clindamycin and erythromycin were selected to investigate the inhibitory<br /> Published: 11/2019<br /> ability of extracts and isolated compounds from leaves of Cassia grandis L.f<br /> Keywords: to P. acnes. Ethyl acetate extract inhibited all P. acnes strains, with an<br /> P. acnes, Cassia grandis L.f, aseptic ring diameter of 15-25 mm at a concentration of 200 mg/mL and a<br /> quercitrin MIC value of 30 mg/mL. The fraction H:E 1:1 showed the strongest<br /> antibacterial activity in the three investigated fractions (H:E 1:1, H:E 1:3<br /> and E 100%) with an aseptic ring diameter of 19.3-23.7 mm at a<br /> concentration of 200 mg/mL (tested on three strongest resistance strains)<br /> and the MIC value was determined to be 0.3 mg/mL. In three pure<br /> compounds isolated from ethyl acetate extract, quercitrin strongest inhibited<br /> for P. acnes with a MIC value of 0.2 mg/mL.<br /> <br /> <br /> <br /> 1. INTRODUCTION al., 2014; Fu Y. et al., 2007) but no remarkable<br /> Propionibacterium acnes (P. acnes) is published announcement shows potential<br /> considered to be an important cause of acne. applications.<br /> However, the treatment with synthetic Traditionally, it is said that leaves of Cassia<br /> antibiotics in the long term also causes many grandis L.f can cure a number of skin diseases<br /> unwanted side effects. On the other hand, the such as Ringworm and Pityriasis Versicolor<br /> resistance of these bacterial strains to (Do Huy Bich et al., 2003) whose scientific<br /> antibiotics is increasingly trending. This basis can be the antimicrobial compounds<br /> increase is due to the mutation of the resistance found in leaves of Cassia grandis L.f, probably<br /> gene (Vu Thi Phuong Thao & Van The Trung, among the isolated compounds such as<br /> 2014). Therefore, the search for new sources of quercitrin, isoquercetin, afzelin, aloe-emodin,<br /> antibiotics to replace these synthetic antibiotics kaempferol, rutin, nicotiflorin (Ngo Quoc Luan<br /> is essential. Some authors have studied the et al., 2012; 2013). Therefore, this study aims<br /> antibacterial effects of P. acnes of some to explore the resistance to P. acnes strains that<br /> essential oils and herbal extracts (Charde YM et are resistant to the current common antibiotics<br /> <br /> <br /> <br /> <br /> 1<br /> AGU International Journal of Sciences – 2019, Vol. 7 (4), 1 – 11<br /> <br /> of extracting fragments and natural compounds 2.3 Equipment, instruments and chemicals<br /> from leaves of Cassia grandis L.f. Main equipment in the laboratory for natural<br /> 2. MATERIALS AND METHODS compound study such as rotary evaporator<br /> 2.1 Materials under low pressure, extraction equipment,<br /> chromatography column etc were supplied by<br /> The plant sample used was the Cassia grandis<br /> Department of Chemistry Education, Can Tho<br /> L.f collected in Vinh Thanh district, Can Tho<br /> University.<br /> city in January 2015. The sample was identified<br /> by Dr. Dang Minh Quan and was stored at the Main equipment in the laboratory for<br /> Department of Biology - School of Education - microbiological research was supplied by the<br /> Can Tho University. The samples were treated Biotechnology Research and Development<br /> by washing, removing damaged parts, cutting, Institute, Can Tho University.<br /> drying, grinding into fine powder as raw Thin layer chromatography (TLC) was carried<br /> materials for extracting and isolating out on pre-coasted silica gel 60F254 (0.25 mm)<br /> compounds. alumium sheet (Merck). For common phase<br /> Source of P. acnes (24 strains) was isolated and column chromatography (CP-CC), silica gel 60<br /> identified from 40 medical waste samples (0.040-0.063 mm, Merck) with increasing<br /> which were collected from skin of patients with polarity solvent systems including n-hexane<br /> common acne at Can Tho Dermatology (H), EtOAc (E), CHCl3 (C), MeOH (M) and<br /> Hospital in February 2015. H2O (W) were used.<br /> <br /> 2.2 Methods Culture medium for TYEG agar (Trypticase-<br /> Yeast Extract-Heart Extract-Glycerol Agar)<br /> Method of extracting high modulation:<br /> prepared according to Kishishita M. et al.,<br /> marceration, liquid-liquid distributing extract<br /> 1980. Kovacs reagent used is produced by<br /> (Nguyen Kim Phi Phung, 2007).<br /> Merck. Antibiotic plates, gram-staining kit and<br /> Isolation of bacteria was followed by the other chemicals were provided by Nam Khoa<br /> method of Kishishita M. et al., 1980; Hans B.L. Company.<br /> and Kilian M., 2010: Identifying P. acnes<br /> 3. EXPERIMENTAL PROCEDURE<br /> strains of bacteria by biochemical tests and<br /> 16S-rARN sequencing (Rajakaruna L. et al., 3.1 Extraction and isolation of plant<br /> 2009). products<br /> <br /> Antibiogramme based on the diffusion of The leaves of Cassia grandis L.f were<br /> antibiotics on agar plates of Bauer A.W. et exhausted with 96% methanol. The sample<br /> al, 1966; Nguyen Thanh Ha, 1991. extract was evaporated to remove solvent and<br /> gain total extract. The methanol extract was<br /> Testing P. acnes’ antibacterial activity by<br /> dissolved in warm water and liquid-liquid<br /> diffusion method on agar plates of Murugan T.<br /> extracted with single solvents in order to<br /> and Saranraj P., 2011; Identifying the minimum<br /> increase polarity, resulting in n-hexane (H) and<br /> inhibitory concentration (MIC) by dilution<br /> ethyl acetate (E) extracts. The E extract<br /> according to Saising S. and Voravuthikunchai<br /> continued to be processed by column<br /> S.P., 2012.<br /> chromatography and distributed into fractions<br /> Statistical data processing using software: corresponding to the elution solvent systems<br /> Statgraphic centurion XVI.II, SPSS and H:E 1:1, H:E 1:3 and E 100%.<br /> Microsoft Excel 2007.<br /> <br /> <br /> 2<br /> AGU International Journal of Sciences – 2019, Vol. 7 (4), 1 – 11<br /> <br /> The process of isolation and structural All 24 isolated strains of P. acnes were fully<br /> identification of quercitrin, rutin and aloe- identified forms, biochemical and gene<br /> emodin was reported in previous publications expression 16S-rRNA, signed from Pa01 to<br /> (Ngo Quoc Luan et al., 2012; 2013). Pa24.<br /> 3.2 Isolation of P. acnes 3.3 Antibiogramming P. acnes strains<br /> Dilute the solution containing medical waste Antibiogramme of the P. acnes strain was<br /> samples in distilled water; extract 10 μL from investigated to identify its resistance to<br /> test tubes of 10-2, 10-3, 10-4 dilution; drip on antibiotics. Based on this result, the resistant<br /> medium TYEG agar with bromocresol purple strains were selected to conduct further steps.<br /> 0.002%, 5 drops on each concentration; The bacterial suspension of strains one with a<br /> anaerobically incubate in a desiccator at 37°C density of about 108 cells/mL (equivalent to Mc<br /> for 5-7 days (Kishishita M. et al., 1980). Farland turbidity 0.5) was placed on the agar<br /> Yellow colonies forming and emerging over plates (TYEG agar, 4 mm thick); place them the<br /> agar plates were selected, and once the medium antibiotics in turn so that the sides of the paper<br /> color was changed from purple into yellow, are close to the medium, the antibacterial wells<br /> streaking in the plate of TYEG agar for strain are 20 mm apart; anaerobically incubate for 48<br /> separation. Observe under microscopic and hours at 37°C. The diameter of aseptic ring was<br /> select the rod-shape strain and store in TYEG measured and followed according to standards<br /> broth with supplement of 20% glycerol at -40 of Clinical and Laboratory Standards Institute<br /> °C as the source of material for further (2009) (Table 1) to assess the antibiotic<br /> experiments (Hans B.L. and Kilian M., 2010). sensitivity of the isolated bacterial strains.<br /> Table 1. Standard for sensitivity evaluation of P. acnes to some synthetic antibiotics<br /> <br /> Diameter of aseptic ring (mm)<br /> No Antibiotic type Sign Concentration/well (μg)<br /> Sensitive Medium Resistant<br /> <br /> 1 Tetracycline Te 30 ≥31 24-30 ≤23<br /> 2 Cefuroxime Cu 30 ≥36 27-35 ≤26<br /> 3 Levofloxacin Lv 5 ≥31 25-30 ≤24<br /> 4 Erythromycin Er 15 ≥31 22-30 ≤21<br /> 5 Clindamycin cL 2 ≥31 24-30 ≤23<br /> <br /> <br /> Then the results were counted according to the extracts (or compound) with the highest<br /> number of strains at 3 levels of susceptibility: antibacterial activity and resistant was selected<br /> Sensitive, Medium, Resistant. to conduct the next experiment.<br /> 3.4 Investigating the inhibitory ability of After spreading the bacterial suspension on the<br /> extracts, fractions and compounds to P. agar plate (TYEG agar); drain; 4 wells with a<br /> Acnes diameter of about 6 mm was created so that<br /> The extracts (or compound) from the leaves of each well is about 2-3 cm apart. 30 µL of the<br /> Cassia grandis L.f was identified to inhibit P. agar was added into each well of compounds,<br /> acnes. From the experimental results, the negative control and positive control<br /> <br /> <br /> 3<br /> AGU International Journal of Sciences – 2019, Vol. 7 (4), 1 – 11<br /> <br /> respectively. Clindamycin (2 μg/mL) was used compounds: Cassia grandis L.f extracted in 3<br /> for the positive control and DMSO 1% was types of solvents: n-hexane, ethyl acetate and<br /> used for the negative control. Proceed in turn at methanol at concentrations of 200 (Figure 1),<br /> different concentrations; Anaerobically 100 and 50 mg/mL; ethyl acetate extract in E<br /> incubate them at 37 °C for 48 hours. Measure 100%, H:E 1:3 and H:E 1:1 fractions at<br /> aseptic ring diameter. concentrations of 200, 100 and 50 mg/mL;<br /> Conduct an experiment to test the antibacterial quercitrin, rutin and aloe-emodin compounds at<br /> activity of the following extracts and concentrations of 10, 5 and 2.5 mg/mL.<br /> <br /> <br /> Pa01 Pa22<br /> <br /> <br /> <br /> 4<br /> 4<br /> <br /> 3<br /> 2 3<br /> 2<br /> <br /> <br /> 1<br /> 1<br /> <br /> <br /> Figure 1. The ability to inhibit P. acnes of leaves of Cassia grandis L.f extracts<br /> at a concentration of 200 mg/mL<br /> Notes: (1) Ethyl acetate extract, (2) n-hexane extract, (3) Methanol extract, (4) Control (-) DMSO<br /> 3.5 Determining the minimum inhibitory hours at 37 °C. The negative control was<br /> concentration (MIC) of extracts, DMSO 1%. Each concentration of the survey<br /> fractions and compounds was diluted and spread to TYEG agar,<br /> Based on the results obtained from the anaerobically incubate them at 37 °C for 48<br /> experiment on inhibitory ability of the bacteria hours, then see the results. The MIC value was<br /> strains of the extracts (or compounds), select defined as the minimum concentration of<br /> one extract (or compound) with the highest extracts (or compounds) where no P. acnes<br /> resistance to P. acnes and concentration range colonies appeared.<br /> containing the MIC value for the experiment. 4. RESULTS AND DISCUSSION<br /> Add 100 μL of extract (or compound) to 4.1 Antibiogramme results<br /> eppendorf tube with different concentrations, Investigate the susceptibility of 24 strains of P.<br /> with a difference of 5-10 mg/mL in each acnes to synthetic antibiotics. Antibiogramme<br /> concentration; add 500 μL of P. acnes results of P. acnes for 5 common antibiotics<br /> suspension (density 108 cells/mL) and 400 μL were presented in Table 2.<br /> of medium; Anaerobically incubate them for 24<br /> <br /> <br /> <br /> <br /> 4<br /> AGU International Journal of Sciences – 2019, Vol. 7 (4), 1 – 11<br /> Table 2. Antibiogramme results of P. acnes for some common antibiotics<br /> <br /> Susceptible Medium Resistant<br /> <br /> Antibiotics Number Number<br /> Number of of Rate<br /> of Strains Rate (%) Strains Rate (%) Strains (%)<br /> Tetracycline 10 41,7 14 58,3 0 0,0<br /> Cefuroxime 13 54,2 11 45,8 0 0,0<br /> Levofloxacin 13 54,2 11 45,8 0 0,0<br /> Erythromycin 0 0,0 13 54,2 11 45,8<br /> Clindamycin 0 0,0 5 20,8 19 79,2<br /> Erythromycin & Clindamycin 9 37,5%<br /> <br /> <br /> 24 investigated strains of P. acnes had the rate produced in many forms: topical application in<br /> of resistance to antibiotics as follows: single or combined form in acne treatment.<br /> Clindamycin 79.2%; erythromycin 45.8%; Very few tetracycline group topical products<br /> tetracycline 0%; levofloxacin 0%; cefuroxime are currently used in the treatment of acne due<br /> 0% and 9/24 P. acnes lines were resistant to to many side effects.<br /> both clindamycin and erythromycin antibiotics From this result, 9 strains of P. acnes resistant<br /> (accounting for 37.5%). to both clindamycin and erythromycin<br /> This result was consistent with studies of Tan et antibiotics were selected to test the inhibitory<br /> al., 2007 in Singapore; Zandi et al., 2011 in ability of extracts from leaves of Cassia grandis<br /> Iran; Luk et al., 2013 in Hong Kong; Nguyen L.f (Table 3).<br /> Thanh Hung and Nguyen Tat Thang, 2013 in 4.2 The results of the survey on the inhibitory<br /> Vietnam. ability to P. acnes of extracts from the<br /> Thus, the resistance to antibiotics such as leaves of Cassia grandis L.f<br /> clindamycin and erythromycin of P. acnes, Experimental results (Table 3) showed that:<br /> which caused acne, was very high. This Leaves of Cassia grandis L.f extracted in ethyl<br /> increasing resistance may be for the acetate solvent at concentrations of 200, 100<br /> prescription used a combination of oral and and 50 mg/mL inhibited all 9 examined strains<br /> topical antibiotics that were the same types of of P. acnes, while n-hexane and methanol<br /> drugs rather than different from each other. In extracts did not show antibacterial activity at<br /> Vietnam as well as Asian countries, many tested concentrations.<br /> clindamycin and erythromycin products are<br /> <br /> <br /> <br /> <br /> 5<br /> AGU International Journal of Sciences – 2019, Vol. 7 (4), 1 – 11<br /> Table 3. The ability of extracts to inhibiting P. acnes by diffusion method on agar plates<br /> <br /> Diameter of aseptic ring (mm)<br /> Extract (mg/mL) Control (+)<br /> Strains Clindamycin<br /> n-Hexane Ethyl acetate Methanol<br /> 200 200 100 50 200 2µg/well<br /> Pa01 0a 15,3de 11,7d 6,0de 0a 1,0d<br /> Pa05 0a 23,3b 17,7a 12,3ab 0a 14,0b<br /> Pa08 0a 15,0e 11,0d 5,0e 0a 5,0c<br /> Pa11 0a 20,3c 16,3ab 13,0a 0a 14,0b<br /> Pa15 0a 20,7c 15,7bc 10,7bc 0a 14,0b<br /> Pa17 0a 19,3c 14,3c 10,3c 0a 14,0b<br /> Pa19 0a 22,7b 17,3ab 10,7bc 0a 18,0a<br /> Pa22 0a 17,0d 12,3d 7,7d 0a 2,0d<br /> Pa23 0a 25,0a 17,3ab 11,3abc 0a 8,0b<br /> Note: The data of aseptic ring diameter was the average value of the three replicates. In the same column, numbers<br /> with at least 1 same digit following were not different in the 5% sense. Control (-): DMSO resulted in zero as the<br /> diameter of resistance ring.<br /> At a concentration of 200 mg/mL, ethyl acetate resistance ring: 11.3-13 mm) but the resistance<br /> extract strongly inhibited P. acnes to create an activity decreased significantly, having the<br /> aseptic ring with a diameter of 15-25 mm. This smallest diameter of the resistance ring,<br /> result showed that the inhibitory ability to P. especially in strain Pa08 (5 mm). At this<br /> acnes strain of ethyl acetate extract at this concentration, the antibacterial activity of ethyl<br /> concentration was higher than clindamycin acetate extract was higher than that of the<br /> (positive control). positive control on Pa01, Pa08 and Pa22 strains<br /> At a concentration of 100 mg/mL, ethyl acetate but lower than the others.<br /> extract created a diameter of P. acnes-resistance Based on the results of the above experiment, it<br /> rings from 11.0 to 17.7 mm. The diameter of showed that the Pa08 strain has the smallest<br /> the aseptic ring reached the largest (16.3-17.7 antibacterial ring diameter, which means that<br /> mm) on Pa11, Pa19, Pa23, Pa05 strains and it this bacterium has a higher activity and the<br /> had the smallest size (11-12.3 mm) for strains inhibitory ability of ethyl acetate extract to<br /> Pa08, Pa01 and Pa22. At this concentration, the Pa08 was lower than that of other bacteria<br /> antibacterial activity of ethyl acetate extract strains. Therefore, the Pa08 strain was chosen<br /> was generally higher than that of the positive to determine the minimum inhibitory<br /> control. concentration of ethyl acetate extract (because<br /> At a concentration of 50 mg/mL, the ethyl if it can inhibit the strongest strain, all it can<br /> acetate extract was the strongest inhibitory with also inhibit the other strains), the results were<br /> strains Pa11, Pa05 and Pa23 (diameter of presented in Table 4.<br /> <br /> <br /> <br /> <br /> 6<br /> AGU International Journal of Sciences – 2019, Vol. 7 (4), 1 – 11<br /> <br /> Table 4. Results of determining the MIC value of ethyl acetate extract on P. acnes by dilution method<br /> <br /> Concentration<br /> No P. acnes Colony<br /> (mg/mL)<br /> 1 50 -<br /> 2 40 -<br /> 3 30 -<br /> 4 20 +<br /> 5 10 +<br /> 6 1 +<br /> 7 0,9 +<br /> 8 0,8 +<br /> 9 0,7 +<br /> 10 0,6 +<br /> 11 Control (-) DMSO 1% +<br /> Note: This was the average value of the three replicates; (-) no colonies appear; (+) colonies appear.<br /> Experimental results showed that 30 mg/mL is Statistical result (Table 5) showed the ability to<br /> the lowest concentration of ethyl acetate extract inhibit P. acnes strains of ethyl acetate extract<br /> capable of inhibiting P. acnes (Pa08 strain). and its fractions: the lowest antibacterial<br /> Highly ethyl acetate effectively inhibiting P. activity was ethyl acetate extract, up to 2<br /> acnes bacteria was divided into segments by fractions E 100% and H:E 1:3 (non-significant<br /> solvent polarity to examine in more detail difference) and the highest was the H:E 1:1<br /> which segment contains active compounds. fraction.<br /> <br /> 4.3 Results of the investigation of the ability<br /> to inhibit P. acnes of ethyl acetate extract<br /> and its fractions<br /> Table 5. The ability of ethyl acetate extract and fractions to inhibiting P. acnes strains<br /> by the diffusion method on agar plates<br /> <br /> Diameter of aseptic ring (mm)<br /> Extracts &<br /> 200 mg/mL 100 mg/mL 50 mg/mL<br /> fractions<br /> Pa22 Pa01 Pa08 Pa22 Pa01 Pa08 Pa22 Pa01 Pa08<br /> Ethyl acetate 17,0d 15,3c 15,0c 12,3c 11,7d 11,0c 7,7b 6,0b 5,0c<br /> H:E 1:3 19,3c 17,7b 17,3b 14,3c 14,0c 13,0b 8,0b 7,3b 6,3bc<br /> E 100% 21,7b 18,3b 17,7b 16,7b 15,3b 14,7b 8,0b 7,7b 6,7b<br /> H:E 1:1 23,7a 22,0a 19,3a 19,0a 18,3a 17,7a 13,3a 13,0a 12,7a<br /> Note: The data of aseptic ring diameter was the average of the three replicates. In the same column, numbers with at<br /> least 1 similar letter following are not different in the 5% sense. Control (-): DMSO resulted in zero as the resistance<br /> diameter.<br /> <br /> <br /> <br /> <br /> 7<br /> AGU International Journal of Sciences – 2019, Vol. 7 (4), 1 – 11<br /> <br /> In general, at the three concentrations tested, and the remaining fractions increased with<br /> the difference in the diameter of antibacterial decreasing concentration.<br /> rings between ethyl acetate extract and its The H:E 1:1 fraction continued to be<br /> fractions had a significant difference of 5%. investigated for MIC value, the results were<br /> The difference in antibacterial activity of H:E shown in Table 6.<br /> 1:1 fraction compared with ethyl acetate extract<br /> Table 6. Results of determining MIC value of H:E 1:1 fraction on P. acnes by dilution method<br /> <br /> Extract concentration<br /> No H:E (1:1) (mg/mL) P. acnes Colony<br /> 1 50 -<br /> 2 40 -<br /> 3 30 -<br /> 4 20 -<br /> 5 10 -<br /> 6 1 -<br /> 7 0,9 -<br /> 8 0,8 -<br /> 9 0,7 -<br /> 10 0,6 -<br /> 11 0,5 -<br /> 12 0,4 -<br /> 13 0,3 -<br /> 14 0,2 +<br /> 15 Control (-) DMSO 1% +<br /> Note: This is the result of three repetitions; (-) no colonies appearing; (+) colonies appearing.<br /> The results above showed that 0.3 mg/mL is the of Combretaceae family) showed MIC value of<br /> lowest concentration of the extract from leaves 0.315 mg/mL on P. acnes bacteria.<br /> of Cassia grandis L.f in ethyl acetate in H: E 1: 4.4 Results of investigating the resistance of<br /> 1 fraction capable of inhibiting P. acnes pure compounds to P. acnes<br /> bacteria.<br /> From the experimental results (Table 7), the<br /> The results of this experiment can be compared quercitrin showed a higher ability to inhibit P.<br /> with that of Vijayalakshmi et al., 2011 that the acnes (Pa08) than rutin and aloe-emodin, the<br /> flavonoid segments from ethyl acetate extract difference was significant at 5%.<br /> from Terminalia arjuna bark (a flowering plant<br /> <br /> <br /> <br /> <br /> 8<br /> AGU International Journal of Sciences – 2019, Vol. 7 (4), 1 – 11<br /> Table 7. The ability of quercitrin, rutin and aloe-emodin to inhibit P. acnes strains by diffusion<br /> method on agar plates<br /> <br /> Diameter of aseptic ring (mm)<br /> No Compounds<br /> 10 mg/mL 5 mg/mL 2,5 mg/mL<br /> <br /> 1 Quercitrin 16,3a 13,7a 5,7a<br /> 2 Rutin 7,0b 0b 0b<br /> 3 Aloe-emodin 5c 0b 0b<br /> Note: The data of aseptic ring diameter is the average value of the three replicates. In the same column, the numbers<br /> with at least 1 similar letter following are not different in the 5% sense. Control (-): DMSO resulted in zero as the<br /> resistance diameter.<br /> At a concentration of 10 mg/mL, the quercitrin Based on this result, the quercitrin compound<br /> compound created a resistance diameter of 16.3 with the highest antimicrobial activity to P.<br /> mm that was significantly different from the acnes (Pa08 strain) was selected for the<br /> other two compounds on the P. acnes strains experiment to determine the minimum<br /> studied. Diameter of resistance ring made by inhibitory concentration.<br /> rutin and aloe-emodin were 7 and 5 mm,<br /> respectively (Table 7).<br /> Table 8. Results of determining the MIC value of quercitrin on P. acnes by dilution method<br /> <br /> No Quercitrin concentration (mg/mL) P. acnes colony<br /> 1 5 -<br /> 2 1 -<br /> 3 0,9 -<br /> 4 0,8 -<br /> 5 0,7 -<br /> 6 0,6 -<br /> 7 0,5 -<br /> 8 0,45 -<br /> 9 0,4 -<br /> 10 0,35 -<br /> 11 0,3 -<br /> 12 0,25 -<br /> 13 0,2 -<br /> 14 0,15 +<br /> 15 0,1 +<br /> 16 0,05 +<br /> 17 Control (-) DMSO 1% +<br /> Note: This was the result of three replicates; (-) no colonies appeared; (+) colonies appeared<br /> <br /> <br /> <br /> <br /> 9<br /> AGU International Journal of Sciences – 2019, Vol. 7 (4), 1 – 11<br /> <br /> Experimental results showed that 0.2 mg/mL Medicinal plants and animals in Vietnam,<br /> was the lowest concentration of quercitrin Part II. Science and Technology Publisher:<br /> compound capable of inhibiting P. acnes (Table 495-496 (in Vietnamese).<br /> 8). Fu Y., Zu Y., Chen L., Efferth T., Liang H. and<br /> Quercitrin belongs to the flavonoid group Liu Z. (2007). Investigation of antibacterial<br /> isolated from H:E 1:1 fraction of ethyl acetate activity of rosemary essential oil against<br /> extract. In the H:E 1:1 fraction, the MIC value Propionibacterium acnes with atomic force<br /> for P. acnes was 0.3 mg/mL, then the pure microscopy. Planta Medica, 73(12), 1275-<br /> quercitrin compound had a lower MIC value of 1280.<br /> 0.2 mg/mL. From this result, it can be deduced Hans B.L. and Kilian M. (2010). Population<br /> that quercitrin was a very important compound genetic analysis of Propionibacterium acnes<br /> contributing to the inhibitory ability to P. acnes identifies a subpopulation and epidemic<br /> of ethyl acetate extract, namely H:E 1:1 clones associated with acne. PLoS One,<br /> fraction. 5(8), e12277.<br /> 5. CONCLUSION Kishishita M., Ushijima T., Ozaki Y. and Ito Y.<br /> The study investigated the inhibitory ability of (1980). New Medium for Isolating<br /> extracts and compounds from leaves of Cassia Propionibacteria and its application to assay<br /> grandis L.f on 9 strains of P. acnes. The H:E of normal flora of human facial skin.<br /> 1:1 fraction and quercitrin from ethyl acetate Applied and environment Microbiology,<br /> extract have antimicrobial effect against P. 40(6), 1100-1105.<br /> acnes with MIC values of 0.3 mg/mL and 0.2 Luk N.M., Hui M., Lee H.C., Fu L.H., Liu<br /> mg/mL, respectively. This result can be Z.H., Lam L.Y., Eastel M., Chan Y.K., Tang<br /> reference for subsequent studies aimed at L.S., Cheng T.S., Siu F.Y., Ng S.C., Lai<br /> applying natural products from leaves of Cassia Y.K. and Ho K.M. (2013). 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