J Acupunct Meridian Stud 2011;4(1):75−79

B RI E F R EP ORT

Biological Activity and Phytochemical Analysis of Three Indonesian Medicinal Plants, Murraya koenigii, Syzygium polyanthum and Zingiber purpurea
Irawan Wijaya Kusuma1*, Harlinda Kuspradini1, Enos Tangke Arung1, Farida Aryani2, Yu-Hong Min3, Jin-Sook Kim4, Yong-ung Kim5*
Department of Forest Products, Faculty of Forestry, Mulawarman University, Samarinda, Indonesia Department of Forest Products, Polytechnic of Agriculture, Samarinda, Indonesia 3 Department of Herbal Skin Care, College of Bio-industry, Daegu Haany University, Gyeongsangbuk-do, Korea 4 Department of Secondary Special Education, College of Health and Therapy, Daegu Haany University, Gyeongsangbuk-do, Korea 5 Department of Herbal Pharmaceutical Engineering, College of Bio-industry, Daegu Haany University, Gyeongsangbuk-do, Korea
2 1

Received: Nov 15, 2010 Accepted: Jan 6, 2011 KEY WORDS:
antimicrobial activity; antioxidant activity; cytotoxicity; medicinal plants; phytochemical analysis

Abstract Extracts of Indonesian medicinal plants, Murraya koenigii, Syzygium polyanthum, and Zingiber purpurea were investigated for their biological activity. The presence of phytochemicals, cytotoxicity, and antimicrobial and antioxidant activities were investigated. Parts of M. koenigii, S. polyanthum, and Z. purpurea were extracted with ethanol. The extracts were evaluated for antimicrobial activity using the disc diffusion method, while antioxidant activity was determined with a 1,1-diphenyl2-picrylhydrazyl radical scavenging assay. Cytotoxicity was investigated using the brine shrimp lethality test, and phytochemical screening was performed using a standard method. M. koenigii leaf extract exhibited the most activity in the test microorganism activity index (AI), 0.38−1.25, when compared with standard drugs. S. polyanthum ripened fruit displayed significant antioxidant activity (90%) in comparison to ascorbic acid (95%). Z. purpurea rhizome extract possessed the highest cytotoxic effect with a LC50 of 52 μg/mL. Phytochemical analysis revealed that carbohydrate, tannin, alkaloid, steroid, triterpenoid, and flavonoid were present in the extracts of M. koenigii leaves and twigs, S. polyanthum leaves and ripened and unripe fruits, and Z. purpurea rhizome, while saponin was only present in the S. polyanthum ripened fruit extract. Our work revealed that the M. koenigii leaves, S. polyanthum ripened fruit, and Z. purpurea rhizome extracts have potential as sources of new antimicrobial, antioxidant, and cytotoxic compounds, respectively.

*Corresponding authors. Irawan Wijaya Kusuma, Department of Forest Products, Faculty of Forestry, Mulawarman University, Samarinda, Indonesia, 75118. Yong-ung Kim, Department of Herbal Pharmaceutical Engineering, College of Bio-industry, Daegu Haany University, Gyeongsangbuk-do, Korea. E-mail: kusuma_iw@yahoo.com OR ykim@dhu.ac.kr ©2011 Korean Pharmacopuncture Institute

8% DW).8% DW) and twigs (8 g. cough. . 13. are three popular medicinal plants used in Indonesia.2]. 2. Despite the extensive use by Indonesian people. USA). purpurea rhizome has been found to possess inflammatory activity [5]. Faculty of Forestry. Kusuma et al sulfoxide). The rhizome of Z. are aromatic and commonly used in some Asian cooking. 2. This process was then repeated. and Z. acetic anhydride. UK). 14. We believe in Indonesia most research activity into natural products is still limited to the inventory of folkloric information and utilization of various plants and trees. S. polyanthum. Antimicrobial assays were conducted using the disc diffusion method as previously described by Kusuma et al [6]. the crude alcohol extract was rotoevaporated at 40ºC and put in a vacuum oven to near dryness to yield the extract of M.. purpurea (rhizome) were collected from Samarinda and Balikpapan. and as a cleansing solution for skin diseases. and 80 μg/10 μL. Japan). Following filtration of the suspension through Whatman filter paper no. is used as a culinary additive and also used for diabetes. and leaves (182 g. purpurea. Japan). Mulawarman University. Introduction Globally.. also known as Kare. in May 2010. UV absorption was measured on a Shimadzu UV-VIS 1240 spectrophotometer (Shimadzu Corp.2. koenigii leaves (46 g. polyanthum ripened fruits (87 g). In our investigation into biological activities of Indonesian plants. and phytochemical analysis of the extracts from parts of M. and Z. Germany) for 24 hours. Indonesia under the accession code KK1005-K009 and K010 for M. Syzygium polyanthum (Wight) Walp. and leaves (1395 g). polyanthum. unripe fruits (237 g). The leaves of S. S. Edmun Buchler. 9. Louis. Salmonella thypi. purpurea.. there have been only limited attempts to explore the biological properties of these plants in relation to their medicinal uses. Tricophyton mentagrophytes. Extraction Ground M. DPPH (1. 6. Kyoto.1. purpurea rhizome (17 g. MO. and Z. 1-naphtol and bismuth (III) nitrate were obtained from Sigma (St.7% DW). DMSO (dimethyl 2. Nutrient agar and potato dextrose agar were used in antibacterial and antifungal assays. cytotoxicity.4. S. 2. USA). and Candida albicans were used in all experiments.W. The plant materials were shade dried for 3 days and ground with a blender. Antioxidant assay The sample was first dissolved in DMSO and used at a 30 times dilution for the actual experiment. and Zingiber purpurea Rosc. koenigii. hydrochloric acid. Murraya koenigii (L) Spreng. koenigii.4]. Germany). potassium iodide and peptone were purchased from Merck (Darmstadt. and skin infection treatment [3. 5. and KK-1005-B004 for Z. KK-1005-S004 to S006 for S. we report here the antimicrobial and antioxidant potentials. Nutrient agar was obtained from Difco (Detroit. Antidiabetic and immunomodulatory effects of this plant have been reported [1. polyanthum (Myrtaceae). which was selected on the basis of our preliminary results showing the antimicrobial activity of plant extract tested simultaneously to correlate with the concentration of standard drugs used (10−20 μg/disk). meaning that obtaining scientific proof for their biological activity is still challenging. 60. Materials and Methods 2. the extract of Z. purpurea (Zingiberaceae). In fact.1-diphenyl-2-picrylhydrazyl) was purchased from Tokyo Kasei Kogyo (Tokyo. Activity index (AI) was calculated as the mean inhibition zone for test sample divided by the mean inhibition zone for the standard drug [7]. Indonesia. polyanthum (leaves. The DPPH radical scavenging method was performed as previously described by Arung et al [8]. is used in folk medicine for the treatment of conditions such as inflammation.7% DW). research into biologically active natural products from plants has attracted many natural product chemists. Plant materials and chemicals M. MI. sulfuric acid. and Z. diarrhea.76 I. koenigii leaves (679 g) and twigs (148 g). 2 (Maidstone. respectively.3. purpurea rhizome (125 g) were extracted with 95% ethanol at room temperature with continuous shaking on a shaker (7400 Tübingen. Other chemicals were of HPLC grade or the highest purity commercially available. Plant extracts were dissolved in acetone to obtain a concentration of 40. Antimicrobial assay Bacillus cereus. polyanthum ripened fruits (13 g. koenigii (Rutaceae). Zones of inhibition around the discs were measured in mm. ripened and unripe fruits). Voucher specimens were deposited in the Laboratory of Wood Chemistry. Various plants have been examined for their biological activities and in some cases active substances have been isolated and identified. also known as Salam.8% DW). koenigii (leaves and twigs). The leaves of M. unripe fruits (23 g. S. 1. also known as Bangle. 13% DW). Ascorbic acid.

41 at 80 μg/disk).50 0.46 0.92 0.00 IZ 8 10 11 6 9 11 4 8 11 11 12 11 0 0 0 0 7 11 NT NT 11 Ca AI 0.91 1. albicans.25 0.25 0. koenigii leaves.73 0.38 0. polyanthum leaves. and other Candida infection-caused diseases.00 IZ 13 13 12 11 12 12 5 8 8 12 13 16 5 9 11 6 9 9 26 NT NT Bc AI 0. Bc = B.46 0.5.Bioactivities of Indonesian medicinal plants 77 Salmonella thypi relative to erythromycin as a standard drug.0 1.19 0.50 0. thypi. mentagrophytes. MT = M.34 0.0 1. ER = erythromycin. and Z. and Z.00 MT SRF SUF SL ZR ER MZ CH 1.00 0 0 0 0 0.31−0.00 *IZ’s are presented as mean of triplicates and include the disc diameter (7 mm). Cytotoxicity assay The cytotoxicity assay was performed using the brine shrimp lethality test according to the method described in the literature [9].58 0. Tm = T. St = S.31 0.34 0. koenigii leaves. 2.00 1.83 0. SRF = S.75 0.67 0. The extracts from M.38 0.00 0. Canada).28 0.25 at 80 μg/disk) except with S.64 1.62 in comparison to that of erythromycin. T.41 0.35 0.34 0. polyanthum ripened fruits. skin and diaper rash. cereus. SUF = S.50 0.19 0. which was comparable to that of the positive control ascorbic acid (95% activity).25 0.28 0.11].25 1.42 0. Good activity of the plant extracts was also observed against Candida albicans (AI 1−1. polyanthum fruits. mentagrophytes (AI 0.36 0. At 80 μg/disk. purpurea rhizome.25 to Table 1 Antibacterial and antifungal activity of plant extracts Microorganisms tested Extract tested (μg/disk) IZ* ML 40 60 80 40 60 80 40 60 80 40 60 80 40 60 80 40 60 80 12 12 15 9 10 11 10 11 12 7 8 7 6 9 11 8 8 10 12 NT NT St AI† 1.83 0. koenigii.75 0.25 0. 2.00 1. Based on these results. . mentagrophytes growth.34 0. purpurea are presented in Figure 1. CH = chloramphenicol.67 0.58 0. Results and Discussion The results for antimicrobial tests of plant extracts are listed in Table 1. Analysis of the data was performed by probit analysis on a Finney computer program to determine LC50 (Biostat 2008.35 1.25 0. AI = activity index. polyanthum leaves.67 0. †AI = IZ of test sample divided by the IZ of a standard drug. The activities of these plant extracts were 86−90% at 100 ppm.31 0. koenigii leaves and S. SL = S.42 0.31 0. higher concentrations of the extract seemed to increase the inhibition of T.83 1. S.23 0. Phytochemical analysis One gram of the plant ethanol extracts was dissolved in 100 mL ethanol and subjected to preliminary phytochemical screening following standard methods [10.19 0.62 0.92 0. suggesting the possibility that the extracts may be useful for the treatment of vaginal yeast infection.25 0.92 1. AnalystSoft.00 0. 3.46 0. The extracts showed moderate activity against Bacillus cereus with an AI of 0.35 0.55 0.46 0. ZR = Z.25−0. purpurea rhizome appeared to be more active than other extracts. polyanthum. the extract of M. polyanthum unripe fruits.73 1.25 0. MZ = miconazole. Ca = C.00 IZ 11 12 12 8 9 8 6 11 12 10 11 13 8 8 9 8 8 8 NT 32 NT Tm AI 0.09 1.00 0. The plant extracts had low to moderate activity against a dermatophyte. IZ = inhibition zone.6. The results of antioxidant activity of M. polyantum ripened fruit showed good activity with an AI of 1−1. S. NT = not tested. koenigii twigs.50 0. ML = M.82 1. Vancouver.

polyanthum was the most active with 90% radical scavenging activity which was clearly more active than the unripe fruit. respectively). This observation may be related to the decomposition of chlorophyll into non fluorescing chlorophyll catabolites which is an effective antioxidant present during fruit ripening [12]. purpurea rhizome. Flavonoids have been reported to possess antibacterial. the M. and other cancer cells [13]. SUF = S. and therefore. polyanthum ripened fruits. SL = S. and 51. S. and Z.W. MT = M. polyanthum leaves. purpurea extracts. Kusuma et al 88 82 83 84 88 88 90 83 86 95 100 Figure 1 DPPH (1. carbohydrate. purpurea may open possibilities of these plant extracts possessing anticancer activity. The presence of biologically important phytochemicals in the M. koenigii. Ascorbic acid was 100 ppm. polyanthum. anti-inflammatory.85 > 1000 747. as tested for in this study. . The ripened fruit of S.58 μg/mL. contribute to their medicinal value. alkaloid.45 > 1000 51. ML = M. polyanthum leaves and fruit extracts were shown to be inactive (LC50 > 1000). The phytochemicals tested are known to exhibit medicinal activity and physiological activity. while M. P388 (murine leukemia). antimutagenic. Cytotoxicity against brine shrimp shows strong correlation with cytotoxicity towards 9 KB (nasopharynx cancer).85 μg/mL. koenigii leaf and twig extracts and Z. Saponins showed hypocholesterolemic and antidiabetic properties.78 ZR 50 ZR 100 SL 50 SL 100 SUF 50 Plant extract (ppm) SUF 100 SRF 50 SRF 100 MT 50 MT 100 ML 50 ML 100 Ascorbic acid DMSO 0 0 20 40 60 DPPH scavenging activity (%) 80 40 65 70 I. koenigii twigs. Therefore. Plant extracts tested for cytotoxicity using the brine shrimp lethality test are presented in Table 2.84 187. tannin. cytotoxic properties of M.1-diphenyl-2-picrylhydrazyl) scavenging activity of plant extracts. koenigii leaf and twig extracts and Z. purpurea rhizome extract showed significant activity (LC50 197. polyanthum unripe fruits. antioxidant. while steroids and triterpenoids displayed analgesic properties [15−17]. polyanthum. and flavonoid. SRF = S. steroid. Further investigation into isolation of antioxidant compounds from the active extracts is in progress. S. koenigii and Z. while saponin was only present in S. purpurea rhizome have potential to be candidates for investigation as cytotoxic compounds. The search for antioxidants from natural sources has received much attention and efforts have been put into identifying compounds that can act as suitable antioxidants to replace synthetic ones. Therefore.58 Murraya koenigii Syzygium polyanthum Zingiber purpurea Phytochemical screening of the plant extracts (Table 3) revealed that the crude extracts contained alkaloid. triterpenoid. ZR = Z. and vasodilatory activity [14]. point to potential sources for useful drugs. Table 2 Plant Cytotoxicity of the plant extracts in the brine shrimp lethality test Part used Leaves Twigs Leaves Ripened fruit Unripe fruit Rhizome LC50 (μg/mL) 197. 187. antiallergic.84 μg/mL. koenigii leaves.

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