Bandung Collage of Pharmacy (STFB) Lia Marliani, Winasih Rahmawati, Anju Sinurat

Transcription

1 ANTIOXIDANT ACTIVITY AND TOTAL PHENOLIC CONTENT OF BANGLE (Zingiber cassumunar Roxb.) RHIZOME Aktivitas Antioksidan dan Kandungan Fenol Total Rimpang Bangle (Zingiber cassumunar Roxb.) Lia Marliani, Winasih Rahmawati, Anju Sinurat Bandung Collage of Pharmacy (STFB) ABSTRAK Bangle (Zingiber cassumunar Roxb.) merupakan salah satu tanaman aromatik yang telah lama digunakan sebagai obat tradisional. Rimpang bengle memiliki banyak senyawa aktif yang prospektif menjadi salah satu pengobatan komplementer dan alternatif tradisional, antara lain minyak esensial, terpenoid, kurkuminoid, dan senyawa fenolik yang memiliki antioksidan, aktivitas anti-inflamasi dan anti-bakteri. Tujuan dari penelitian ini adalah untuk menentukan aktivitas antioksidan dan kandungan total fenolik dari rimpang bengle. Penentuan aktivitas antioksidan dilakukan dengan metode radikal bebas DPPH. Total kandungan fenolik ditentukan dengan menggunakan Spektrofotometri UV-Vis menggunakan Folin Cio-calteu reagen. Hasil penelitian menunjukkan bahwa rimpang bangle memiliki aktivitas antioksidan sedang (IC mg / ml). Total kandungan fenolik dari Bangle Roxb. rimpang adalah 2.71 g GAE / 100 g ekstrak atau 0,30 g GAE / 100 g simplisia. Kata kunci: Zingiber cassumunar, antioksidan, DPPH, kandungan fenol total. ABSTRACT Bangle (Zingiber cassumunar Roxb.) is one of aromatic plants that has long been used as traditional medicine. Cassumunar rhizome has many active compound which are prospective to be one of traditional complementary and alternative medicine. Those active compounds such as essential oils, terpenoids, curcuminoids, and phenolic compounds that has antioxidants, anti-inflammatory and anti-bacterial activity. The aims of this study was to determine antioxidant activity and total phenolic content of Z. cassumunar Roxb. rhizome. Determination of antioxidant activity was done by DPPH free radical scavenging method. Total phenolic content was determined using Spectrophotometry UV-Vis used the Folin Cio-calteu reagent. The results showed that Z. cassumunar Roxb. rhizome has moderate antioxidant activity (IC μg/ml). Total phenolic content of Z. cassumunar Roxb. rhizome was 2.71 g GAE/100 g extract or 0.30 g GAE/ 100 g crude drugs. Keywords: Zingiber cassumunar Roxb., antioxidant, DPPH, total phenolic content Volume 7, No.2, Desember

2 INTRODUCTION Zingiber cassumunar Roxb. (synonyms: Amomum montanum Koenig, Zingiber montanum (Koenig) Dietrich, Zingiber purpureum Roscoe) is a member of family Zingiberaceae. It is grows in tropical and subtropical Asia coutries, including Indonesia. Bangle is one of common name in Indonesia. It is used as spices and medicinal plants for the treatment of conditions such as inflammation, fever, headache, cough, rheumatism, carminative, and anthelmintic agent. The rhizome of Zingiber cassumunar Roxb. contain some active components such as phenylbutenoids which has anticancer and anti-inflammatory potency (Chung et al., 2009; Lee et al., 2007; Han et al., 2005) and immunostimulant (Chairul et al., 2009), phenylbutanoids (such as phlain I-IV) which has anti-inflammatory and antioxidant activity (Nakamura et al., 2009), fungicide/ antifungal (Tripathi et al., 2008; Jantan et al., 2003), cassumunin A-C (Masuda et al., 1993; Masuda and Jitoe, 1994; Nagano et al., 1997) and cassumunarin A-C (Nakatani and Kikuzaki, 2001), complex curcuminoids which has antioxidant activity, and also essential oils which has antimicrobial (Pithayanukul et al., 2007), and anthelmintic (Sukandar et al., 1997). Rhizome of Z. cassumunar Roxb. yield about 0.95% essential oil (Bhuiyan et al., 2008) which contains 4-terpeniol and sabinene as its main components. Phenolic compounds are some of active phytochemical. Phenolic compounds can reduce free radical species and can be used as natural antioxidant agent. Considering of phenolic compounds in Z. cassumunar Roxb. we determined antioxidant activity and total phenolic compound of its rhizome in this study. MATERIALS AND METHOD Materials Rhizomes of Zingiber cassumunar Roxb. were collected from Manoko, Lembang, Indonesia. The specimens were identified at ANTIOXIDANT ACTIVITY AND TOTAL PHENOLIC CONTENT OF BANGLE (Zingiber cassumunar Roxb.) RHIZOME Aktivitas Antioksidan dan Kandungan Fenol Total Rimpang Bangle (Zingiber cassumunar Roxb.) the Bandungense herbarium, School of Life Sciences and Technology, Bandung Institute of Technology. Rhizomes was dried in oven with temperature at 40 C. Dried rhizomes was extracted by macerate method used ethanol solvent for 3x24 hours. Extract was concentrated by rotary evaporated. rendemen extract was 13,98%. Methods Determination of free radical scavenging activity The free radical scavenging activity of extract and ascorbic acid as positive control was determined using the stable radical DPPH (1,1 diphenyl 2 picrylhydrazyl) (Seal, 2011). Aliquots (0.2 ml) of the tested sample were placed in test tubes and 3.8 ml of freshly prepared DPPH solution (60 ppm) in methanol was added in each test tube and mixed. 30 min later, the absorbance was measured at 515 nm (λ max of DPPH solution). The capability to scavenge the DPPH radical was calculated using the following equation: DPPH scavenged (%) = [(Ac As)/Ac] x 100 Where Ac is the absorbance of the control (DPPH solution) and As is the absorbance in presence of the sample of the extracts. The antioxidant activity of the extract was expressed as IC50. The IC50 value was defined as the concentration that inhibits the formation of DPPH radicals by 50%. Each value was determined from regression equation. Determination of total phenolic content Total phenolic were determined by Folin Ciocalteu reagent (Pourmorad et al., 2006). A dilute extract (5 ml) or gallic acid (standard phenolic compound) was mixed with Folin Ciocalteu reagent (5 ml, 1:10 diluted with distilled water) and aqueous Na2CO3 (4 ml, 1 M). The mixtures were allowed to stand for 15 min and the total phenolic content were determined by colorimetry at 765 nm. The standard curve was prepared using 100, 150, 200, 250, 300, and 350 μg/ml solutions of gallic acid in Volume 7, No.2, Desember

3 Lia Marliani, Winasih Rahmawati, Anju Sinurat methanol : water (50:50, v/v). Total phenol values are expressed in terms of gallic acid equivalent (mg/g of dry mass), which is a common reference compound. Absorbance of this reaction product, complex compound Molybdenum blue was increased due concentration increasing. The Calibration curve of standard solution is shown in figure 1. RESULTS AND DISCUSSION DPPH used a substrate to determine antioxidant activity based on the reduction of DPPH solution (purple coloured) on methanol in the presence of hydrogen donating antioxidants and form non radical DPPH-H (yellow coloured). The scavenging activity of Z. cassumunar Roxb. rhizome extract increased due increasing concentration of the extract. DPPH scavenged (%) of extract and ascorbic acid are shown in Table 1 and Table 2 Table 1. DPPH scavenged (%) of extract Concentration (μg/ml) DPPH scavenged (%) IC50 (μg/ml) Table 2. DPPH scavenged (%) of Ascorbic acid Concentration (μg/ml) DPPH scavenged (%) IC50 (μg/ml) That result showed that Z. cassumunar Roxb. rhizome has moderate antioxidant activity (IC μg/ml) but not as good as ascorbic acid as positive control (IC μg/ml). Total phenolic content were measured with using Folin-Ciocalteu reagent. The principle of this method is reduction ability of phenol functional group at alkali solution. The reduction of fosfotungstat-fosfomolibdenum complex (Folin-Ciocalteu reagent) by phenol ion will change its color to be dark blue. Volume 7, No.2, Desember

4 ANTIOXIDANT ACTIVITY AND TOTAL PHENOLIC CONTENT OF BANGLE (Zingiber cassumunar Roxb.) RHIZOME Aktivitas Antioksidan dan Kandungan Fenol Total Rimpang Bangle (Zingiber cassumunar Roxb.) Figure 1. Calibration curve of Gallic acid (GA) as standard solution. Table 3. Total phenolic content of extract _ Total phenolic content No A C C (µg/ml) (µg/ml) g GAE/100 g g GAE/100 g extract crude drug (dry) _ X±SD ± ± n = 3 This result showed that total phenolic content of Z. cassumunar Roxb. rhizome was 2.17± g GAE/100 g extract or 0.30 g GAE/ 100 g crude drugs. Moderate antioxidant activity of Z. cassumunar Roxb. rhizome can be related with its total phenolic content. Phenolic compounds are one of active antioxidant compound. Their hydroxyl group can reduced free radical species. CONCLUSION Zingiber cassumunar Roxb. rhizome has moderate antioxidant activity (IC μg/ ml). Total phenolic content of Z. cassumunar Roxb. rhizome was 2.17± g GAE/100 g extract or 0.30 g GAE/ 100 g crude drugs. Volume 7, No.2, Desember

5 Lia Marliani, Winasih Rahmawati, Anju Sinurat REFERENCES Bhuiyan MNI., Chowdhury JU., Begum J Volatile constituents of essential oils isolated from leaf and rhizome of Zingiber cassumunar Roxb. Bangladesh J Pharmacol, 3: Chairul P. and Chairul SM Phagocytosis Effectivity Test of Phenylbutenoid Compounds Isolated from Bangle (Zingiber cassumunar Roxb.) Rhizome. Biodiversitas, 10(1): Chung SY., Han AR., Sung MK., Jung HJ., Nam JW., Seo Ek., Lee HJ Potent Modulation of P-glycoprotein Activity by Naturally Occurring Phenylbutenoids from Zingiber cassumunar. Phytother. Res., 23: Han AR., Kim MS., Jeong YH., Lee SK., Seo EK Cyclooxygenase-2 Inhibitory Phenylbutenoids from the Rhizomes of Zingiber cassumunar. Chem. Pharm. Bull., 53(11): Jantan I., Mohd Yassin MS., Chin CB., Chen LL., Sim NL Antifungal Activity of the Essential Oils of Nine Zingiberaceae Species. Pharmaceutical Biology, 41(5): Lee JW., Min HY., Han AR., Chung HJ., Park EJ., Park HJ., Hong JY., Seo EK., Lee SK Growth Inhibition and Induction of G1 Phase Cell Cycle Arrest in Human Lung Cancer Cells by a Phenylbutenoid Dimer Isolated from Zingiber cassumunar, Biol. Pharm. Bull., 30(8): Masuda T., Jitoe A., Nakatani N Structure of Cassumunin A, B, and C, New Potent Antioxidants from Zingiber cassumunar. Chemistry Letters, Masuda T. and Jitoe A Antioxidative and Antiinflammatory Compounds from Tropical Gingers: Isolation, Structure Determination, and Activities of Cassumunins A, B, and C, New Complex Curcuminoids from Zingiber cassumunar. J. Agric. Food Chem., 42: Nagano T, Oyama Y, Kajita N, Chikahisa L, Na kata M, Okazaki E, Masuda T New Curcuminoids Isolated from Zingiber cassumunar Protect Cells Suffering from Oxidative Stress: A Flow-Cytometric Study Using Rat Thymocytes and H202, Jpn. J. Pharmacol., Nakatani N. and Kikuzaki H Antioxidants in Ginger Family In: Quality Management of Nutraceuticals, Ho, C., et al., ACS Symposium Series, American Chemical Society: Washington, DC Nakamura S., Iwami J., Matsuda H., Wakayama H., Pongpiriyadacha Y., Yoshikawa M Structures of New Phenylbutanoids and Nitric Oxide Production Inhibitors from the Rhizomes of Zingiber cassumunar. Chem. Pharm. Bull., 57(11): Pithayanukul P., Tubprasert J., Wuthi- Udomlert M In Vitro Antimicrobial Activity of Zingiber cassumunar (Plai) Oil and a 5% Plai Oil Gel. Phytother. Res., 21: Pourmorad F., Hosseinimehr SJ., Shahabimajd N Antioxidant activity, phenol and flavonoid contents of some selected Iranian medicinal plants. African Journal of Biotechnology, 5(11): Sukandar EY., Suganda AG, Kristiana AS Efek Anthelmintika Zingiber zerumbet, Zingiber cassumunar, dan Curcuma xanthorrhiza terhadap Cacing Ascaris summ. Majalah Farmasi Indonesia, 8(1): Seal T Evaluation of Antioxidant Activity of Some Wild Edible Fruits of Meghalaya State In India. Int J Pharm Pharm Sci, 3(4): Tripathi P., Dubey NK., Shukla AK Use of some essential oils as post-harvest botanical fungicides in the Volume 7, No.2, Desember

6 ANTIOXIDANT ACTIVITY AND TOTAL PHENOLIC CONTENT OF BANGLE (Zingiber cassumunar Roxb.) RHIZOME Aktivitas Antioksidan dan Kandungan Fenol Total Rimpang Bangle (Zingiber cassumunar Roxb.) management of grey mould of grapes caused by Botrytis cinerea, World J Microbiol Biotechnol., 24: Volume 7, No.2, Desember