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ISOLASI DAN IDENTIFIKASI SENYAWA ANTIOKSIDAN DARI BATANG PAKIS (Alsophila glauca J.Sm) DENGAN METODE DPPH (2,2-DIFENIL-1-PIKRILHIDRAZIL) Sri Wahdaningsih; Subagus Wahyuono; Erna Prawita Setyowati
Majalah Obat Tradisional Vol 18, No 1 (2013)
Publisher : Faculty of Pharmacy, Universitas Gadjah Mada

Oxidative stress induced by the radicals have been known to affect the occurrence of various degenerative disease. In order to search natural antioxidant compounds, it had been studied the isolation and identification of antioxidant compounds in fern stems (Alsophila glauca J.Sm) using DPPH method (1,1-diphenyl-2-picrylhydrazyl). Fern stem extraction was done by maceration with wasbenzen and metanol. Extracts was obtained by evaporating the wasbenzen solvent with a rotavapor. Material was re-macerated with methanol after wasbenzen evaporation. The same way was done to obtain the metanol extract. These extracts were tested for antioxidant activity using DPPH method with TLC. Active extract was partitioned in 80% methanol. Both 80% methanol soluble and insoluble extract were tested their antioxidant activity using DPPH method (TLC). Active extract was fractionated in vacuum liquid chromatography using a mobile phase with different polarity gradient and different concentrations (wasbenzen:chloroform). Active fraction was isolated by preparative TLC method and purity of obtained compounds was tested by TLC. Antioxidant activity of obtained isolates was tested by DPPH method using spectrophotometry. Compounds known to had antioxidant activity as radical catcher with IC50 178.4µg/mL.

PENETAPAN KADAR ALKALOID DARI EKSTRAK ETANOLIK BUNGA KEMBANG SEPATU (Hibiscus rosa-sinensis L.) Mimiek Murrukmihadi; Subagus Wahyuono; Marchaban Marchaban; Sudibyo Martono
Majalah Obat Tradisional Vol 18, No 2 (2013)
Publisher : Faculty of Pharmacy, Universitas Gadjah Mada

Kembang sepatu flower (Hibiscus rosa-sinensis L.) was fractionally used as expectorant. Based on Bioassay Guided fractionation, an active fraction was separated, and the fraction was identified is Alkaloid was the major compound based on TLC analysis. Viscosity value measured by viscometer was used as a Bioassay model of expectorant activity in vitro and asetyl cysteine was used as positive control. Alkaloid content determination of the ethanolic extract was measured by TLC-Densitometric compared with standard curve of isolated alkaloid as the selected marker (Y=12.1360X+2901.4474). The alkaloid content in the ethanolic extract was determined as 2.35 ± 0.67 %.

AKTIVITAS PENANGKAP RADIKAL BEBAS DARI BATANG PAKIS (Alsophila glauca J. Sm) Sri Wahdaningsih; Erna Prawita Setyowati; Subagus Wahyuono
Majalah Obat Tradisional Vol 16, No 3 (2011)
Publisher : Faculty of Pharmacy, Universitas Gadjah Mada

Free radical induced oxidative stress that influences the occurrence of various degenerative diseases such as cancer, coronary heart disease and premature aging. In the case that body's antioxidant defense system does not have excessive antioxidants, additional natural antioxidant via food or other nutrients intake is needed. Stems of ferns are known to contain phenolic compounds that are known to have antioxidant activity. A study has been carried out to determine antioxidant potential of stems of ferns of western Borneo origin compared to quercetin as positive control. Initially, material was macerated gradually with wasbenzen followed by methanol. The extract obtained was filtered and evaporated to dryness to give dried wasbenzen extract and methanol extract respectively. Antioxidant properties of extracts were performed by DPPH on TLC. Active (wasbenzen) extract was then partitioned with 80% methanol to give soluble and insoluble fractions. Active (insoluble) fraction was further fractionated by vacuum liquid chromatography using a gradient mobile phase with increasing polarity (wasbezen: chloroform). Fractions obtained were tested antioxidant activity, and isolation was done by preparative TLC. The purity of isolated antioxidant compound was confirmed by TLC, and antioxidant potential was measured spectroscopically with DPPH method. This compound displayed antioxidant activity, but its value was lower than that of quercetin used as a positive control (IC50 178.4 v / s 2.17 microg / mL)

ISOLASI DAN IDENTIFIKASI SENYAWA ANTIOKSIDAN DAUN KESEMEK (Diospyros kaki Thunb.) DENGAN METODEDPPH (2,2-DIFENIL-1-PIKRILHIDRAZIL) Isnindar Isnindar; Subagus Wahyuono; Erna Prawita Setyowati
Majalah Obat Tradisional Vol 16, No 3 (2011)
Publisher : Faculty of Pharmacy, Universitas Gadjah Mada

Antioxidant is a substance which in small concentrations can significantly inhibit or prevent the oxidation of the substrate. One plant is efficacious as an antioxidant is the persimmon (Diospyros kakiThunb.) where is cultivated widely in East Asia, Spain and Indonesia. In the search for natural antioxidant compounds, have been studied isolation and identification of antioxidant compound of persimmon leaves (DiospyroskakiThunb.) using DPPH (2,2-diphenyl-1-pikrilhidrazil) method. The extraction of persimmon leaves was carried out by maceration method with wasbenzen. Extracts obtained by evaporating the solvent wasbenzenon a rotary evaporator and were then re-macerated with methanol. Antioxidant activity of extracts was evaluated with DPPH 0.2% method. Active extract was partitioned with chloroform, methanol, distilled water and tested for antioxidant activity by DPPH 0.2% method. The active compound was purified byPTLC and the purified active isolates was then confirmed by TLC. The antioxidant activity (IC50) active isolates were analyzed using spectrophotometry. Analysisof active isolates resulted the antioxidant activity (IC50) of 100.00 ug / ml. UV-Vis spectrum of the active isolates showed absorption at λmax 285 and 401 nm. Infrared spectra (KBR) showed absorption at 3446 cm-1 (OH), 2926 cm-1 (CHaliphatic), 1456 cm-1 (CH2), 1384 cm-1 (CH3), 1255 cm-1, and 1115 cm-1 (C-0-C), 1631 cm-1 (C=Caromatic). GC-MS spectra gave two peaks with retention time 11.408 minutes with a similarity index of 79.243% with the molecular ion (M•)+ appeared at m/z 178, 163, 147. These spectra are expected to be methyl eugenol. Furthermore, the peak with retention time 12.982 minutes with a similarity index of 20.757%, producing a mass spectrum with molecular ion peaks (M•)+appeared at m/z 192, 177, 161. These spectra are expected to be myristicin compound.

ISOLASI DAN IDENTIFIKASI SENYAWA ANTIBAKTERI DARI DAUN PETAI CINA (Leucaena leucocephala (Lam.) De Wit.) Ari Sartinah; Puji Astuti; Subagus Wahyuono
Majalah Obat Tradisional Vol 15, No 3 (2010)
Publisher : Faculty of Pharmacy, Universitas Gadjah Mada

Plants are important natural resources widely explored for maintaining health and treatment of diseases. One of them is Leucaena leucocephala leaf which is used as traditional medicine. Indonesian has used this leaf to treat wound and inflammation. This study was conducted to isolate and identify antibacterial compounds from L. leucocephala leaves. Dry powder of this leaf was extracted using washbenzen by Soxhletation method followed by methanol extraction. Both extracts were tested for antibacterial activity against Staphylococcus aureus ATCC 25923 dan E. coli 25922 and analysed by TLC. Positive extract to S. aureus was further separated by vacuum liquid chromatography using different combination of washbenzene and ethyl acetate followed by analysis using TLC. Each fraction was tested for antibacterial activity and analysed by TLC. The positive fraction was further separated by TLC-preparative, assigned as p1a, p2a, p3a. The active compound (p2a) was tested for its purity using TLC with various mobile phase. Its structure was identified using combination data of UV-Vis and IR spectrophotometry, GC-MS and NMR. Based on UV-Vis data, this compound showed the absence of chromophoric group. IR data showed adsorption at 3409,4 cm-1 (OH), 2928,2 cm-1(CHaliphatic), 2854,3 cm-1(CHaliphatic), 1575,5 cm-1 (C=C), 1416,4 cm-1 (CH2), 1385,0 cm-1(CH3), 1258,3 cm-1 and 1082,3 cm-1 (C-O). GC-MS data showed molecule ion at m/z 482 (M + H+)and fragmented ion at m/z 427 (M + H+). 1H-NMR (CDCl3) spectrum showed resonance at δ 0,8; 1,4; 1,6; 2,0; 2,3; 3,6; 4,2 and 5,4 pp, suggesting that this compound is lupeol.

AKTIVITAS ANTIINFLAMASI EKSTRAK METANOLIK BUAH MANGGA KASTURI (Mangifera casturi) MELALUI PENGHAMBATAN MIGRASI LEUKOSIT PADA MENCIT YANG DIINDUKSI THIOGLIKOLAT Nanang Fakhrudin; Peni Susilowati Putri; Sutomo Sutomo; Subagus Wahyuono
Majalah Obat Tradisional Vol 18, No 3 (2013)
Publisher : Faculty of Pharmacy, Universitas Gadjah Mada

Kasturi (Mangifera casturi) is a plant originally from Kalimantan, mainly in South Kalimantan. Although there is no report regarding the usage of the fruit in traditional healings, current research demonstrate that methanolic extract of Mangifera casturi fruit demonstrated high antioxidant activity. As oxidant plays crucial role in inflammatory related diseases, we investigate the antiinflammatory effect of Mangifera casturi fruit methanolic extract. To assess the antiinflammatory activity, we used thioglycollate-induced leukocyte migration in mice. Male balb/c mice were pretreated (i.p) with Mangifera casturi fruit methanolic extract followed by thioglikolat (i.p) to induce leukocyte migration. After 4,5 hours, the mice were sacrificed and the number of leukocytes were counted using hemocytometer under light microscope. Our experiment indicates that Mangifera casturi fruit methanolic extract significantly exhibited antiinflammatory activity by inhibiting the migration of leukocytes induced by thioglycollate. However, the activity of the extracts was lower than the positive control, indomethacin. Further purification was required to obtain the active compound with the activity comparable to indomethacin. Thin layer chromatography (TLC) analysis demonstrates that the methanolic extract of Mangifera casturi fruit contains terpenoid and phenolic compounds.

ISOLATION AND IDENTIFICATION OF FREE RADICALS SCAVENGER FROM Daucus carota L LEAVES Sri Sudewi; Subagus Wahyuono; Puji Astuti
Majalah Obat Tradisional Vol 19, No 3 (2014)
Publisher : Faculty of Pharmacy, Universitas Gadjah Mada

The presence of free radicals produced inside the cells is thought to cause a range of diseases such as degenerative diseases. Antioxidant properties of carrot root (Daucus carota L.) have been reported elsewhere to inhibit the formation of these free radicals, but the compounds responsible for antioxidant activity inside its leaves have not yet determined. In Indonesia, Daucus carota L. leaves are regarded as waste product after harvesting the root. This study was conducted to isolate and identify the compound having antioxidant activity from the leaves of D.carota. n-Hexane, ethyl acetate, and methanol extracts were examined for free radical scavenger activity by using DPPH method. The n-hexane extract showing free radical scavenger activity was solid-solvent partitioned, fractionated using vacuum liquid chromatography and tested for free radical scavenger activities. The active compound was isolated by using preparative thin layer chromatography and its purity was determined by TLC with three mobile phase systems and two dimensional TLC. Based on spectroscopy UV-Vis, IR as well as 1H-NMR (CDC13) data, the active compound was suggested to be xanthophyll derivate, lutein. The compound at concentration of 0.616, 1.025, and 2.05 ppm had antioxidant activities of 0.94 ± 0.05; 18.53 ± 0.15; and 49.07 ± 0.86, respectively.

OPTIMASI FORMULASI SIRUP FRAKSI TIDAK LARUT ETIL ASETAT YANG MENGANDUNG ALKALOID DARI BUNGA KEMBANG SEPATU (Hibiscus rosa-sinensis L.) Mimiek Murrukmihadi; Subagus Wahyuono; Marchaban Marchaban; Sudibyo Martono
Majalah Obat Tradisional Vol 16, No 2 (2011)
Publisher : Faculty of Pharmacy, Universitas Gadjah Mada

Ethylacetate insoluble fraction containing alkaloids from the hibiscus flower (Hibiscus rosa-sinensis L.) with a concentration of 0.6% w/v has mucolityc activity which is equivalent to 0.1% acetylcysteine based on the capasity to decrease viscosity of the mucus. The use of hibiscus as a new cough medicine done traditionally. Syrup of ethylacetate insoluble fraction containing alkaloids was the appropiate liquid dosage form that has some advantages for examples easy to use and fast absorption of the active substance. The study aims to determine the optimum formulation syrup ethanolic fractions containing alkaloids from hibiscus flowers using the method of Simplex Lattice Design. Ethylacetate insoluble fraction containing alkaloids from the hibiscus flower is obtained by using the Vacuum Liquid Chromatography (VLC). Simplex Lattice Design Methods used to optimize the formulation syrup ethylacetate insoluble fraction containing alkaloids from hibiscus flower with seven formulas with variations in the amount of glycerin, sorbitol solution 70%, and mucilago 0.5% CMC-Na. Physical stability was analized to find higher total response value (R total) as parameter of the optimum formula by Simplex Lattice Design Methods with Design Expert ® software version 8.0.2. Optimum formula syrup ethylacetate insoluble fraction containing alkaloids from the hibiscus flower obtained with the proportion of 25.376% glycerine, 51.985% of 70% sorbitol solution, and 22.639% of 0.5% CMC Na mucilago. Data obtained physical characteristics test compared with prediction value using Design Expert ® software version 8.0.2. Physical characteristics of optimum syrup ad the zero week compared with physical characteristics on the forth week using paired t-test. The result showed that physical properties of syrup formula optimum didn’t different significantly for easy for pouring and tasted response except for viscosity and acidity. Ethylacetate fraction syrup containing alkaloids from hibiscus flower optimization results are less stable during the four weeks of storage.

PATCH EKSTRAK DAUN SIRIH (Piper betle L.): EVALUASI AKTIVITAS ANTIBAKTERI, PROFIL PELEPASAN EUGENOL dan TOLERANSI LOKAL Mufrod Mufrod; Suwaldi Suwaldi; Subagus Wahyuono
Majalah Obat Tradisional Vol 21, No 2 (2016)
Publisher : Faculty of Pharmacy, Universitas Gadjah Mada

Piper betle leaf in extract form is more effective than crude drug. Eugenol is a component in the extract that has antibacterial activity but irritate. Patch of piper betle leaf extract was used on the mucosa to make oral cavity hygiene. Antibacterial activity was influenced by the release of eugenol from the patch. Release enhancer substances (RES) such as glycerin, propylen glicol and tween 80 were added in patch formulation to increase the release of active substances. The aim of the research was to investigate the physicochemical properties, eugenol release profiles, and local tolerance test of the patch. Extract of piper betle leaf was made using infundation method. Patch was made according to the variation concentration of extract (1, 2 and 4%) and RES (glycerine, propylen glycol and tween 80) using chitosan as vehicle. Patch produced solvent casting method. Patch obtained was tested for swelling index, folding endurance, surface pH, antibacterial activity, release of eugenol, and local tolerance. Data obtained were analyzed descriptively. The result showed that the addition of RES did not affect the surface pH but increase the water absorption with in inconsistent way except patch with tween 80. The flexibility (folding endurance value) increased, and the highest amount of eugenol released was achieved by patch using propylen glicol. Patch with tween 80 and glycerin for all extract concentration and patch with 1% extract concentration using propylen glycol showed medium sensation (local tolerance), and patch with 2 and 4% extract using propylen glycol showed severe sensation.

PENENTUAN AKTIVITAS ANTIOKSIDAN DARI DAUN BUAS-BUAS (Premna serratifolia L.) MENGGUNAKAN METODE DPPH (2,2-diphenyl-1-picrylhydrazyl) Isnindar Isnindar; Subagus Wahyuono; Sitarina Widyarini; Yuswanto Yuswanto
Majalah Obat Tradisional Vol 21, No 3 (2016)
Publisher : Faculty of Pharmacy, Universitas Gadjah Mada

Antioxidant is a substance that in small concentrations can significantly inhibit or prevent oxidation on the substrate. Buas-buas (Premna serratifolia L.) is one of the plants that have antioxidant effect. The antioxidant activities of the wasbenzen and ethanol extracts of buas-buas leaves were determined by DPPH method using spectrophotometer UV-Vis. Data resulted from the analysis of antioxidant activity (IC50) is 532.24 μg/mL of wasbenzen extract, 24.40 μg/mL of ethanol extract, and 1.83 μg/mL of vitamin C.

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