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Pengaruh Perbedaan Metode Ekstraksi Metabolit Sekunder Streptomyces sp. GMR22 terhadap Toksisitas pada Sel BHK-21 Mentari, Diani; Naima, Mirtani; Wulansari, Riska; Widada, Jaka; Nuringtyas, Tri Rini; Wibawa, Tri; Wijayanti, Nastiti
Pharmacon: Jurnal Farmasi Indonesia Vol 16, No 1 (2019)
Publisher : Universitas Muhammadiyah Surakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.23917/pharmacon.v16i1.8032

Streptomyces sp. GMR22 is local isolate from Wanagama 1 Forest in Yogyakarta. They have the potential to be developed to produce active compounds because have PKS and NRPS genes.The active compounds from isolation are strongly influenced by various factors, one of them is extraction techniques. Effect difference of extraction technique will be affected by the quality of secondary metabolites produced.The purpose of this study was to compare the cytotoxicity effects of secondary metabolites of Streptomyces sp. GMR22 which have extracted with different stages from previous studies. The extraction technique was carried out by multilevel separatory funnel extraction methods, which was first extracted using non-polar solvent (n-hexane) and then extracted using semi-polar solvent (ethyl acetate). This research is important because in previous studies (separatory funnel only extracted using ethyl acetate) with the use of the lowest concentration in the dengue virus antiviral test (further test) caused 100% of deaths in BHK-21 cells.This study indicate that multilevel extraction result in lower CC50 value than previous studies. There are 49.160 µl/ml (n-hexane extract) and 284.56 µl/ml (ethyl acetate extract) while water extract is 464,38 µl/ml. FTIR compound analysis show that the three extracts produced have different spectrum patterns, especially in the n-hexane and ethyl acetate extract. Value of CC50 is not too high, it is expected that the secondary metabolites contained in the extracts can be used for further analysis such as antiviral testing because it is safe for normal host cells such as BHK-21 cells

Optimasi Metode 1H-NMR Profiling pada Rimpang Kunyit (Curcuma domestica) Dwiseptianti, Caroline; Susanto, Febri Adi; Purwestri, Yekti Asih; Nuringtyas, Tri Rini
Bioeksperimen: Jurnal Penelitian Biologi Vol 5, No 2: September 2019
Publisher : Universitas Muhammadiyah Surakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.23917/bioeksperimen.v5i2.9238

The use of medicinal plants is increasing due to the lack of side effects caused and the number of bioactive compounds that cannot be represented by synthetic chemical synthesis compounds. However, the management and use of natural medicines for the main handling of diseases are often hampered by the quality of the ingredients which are low and unstable. The standardized quality control system of OAI (Indonesian Natural Medicine) is the main key to improve clinical assurance and safety of the use of herbal medicines in Indonesia. One of the medicinal plants known to the public is Curcuma longa L. (turmeric). The main active components contained in turmeric are curcumin, demetoksikurkumin, bis-demetoksikurkumin, and ar-turmeron. Information about the quality of turmeric is needed in its use as a raw material for drugs so we need an analytical technique that is able to identify the diversity of metabolite profiles of active compounds. In this research, an optimization method is used to improve efficiency in the extraction of turmeric rhizome metabolites so that the best solvent concentration is known for the analysis of fingerprinting secondary metabolites with 1H-NMR 500 MHz spectroscopy in turmeric rhizomes. The results were analyzed with MNOVA software and chemical shift obtained compared with the reference. From the results obtained a concentration of methanol-d4 (CD3OD) 100% able to extract curcumin better than other solvents. The solvent is able to extract saccharide (sugar) compounds in the form of sucrose, amino acids and fatty acids in the form of methionine, glutamine, acetate, and glycero phospho choline.

Secondary metabolite profiling of four host plants leaves of wild silk moth Attacus atlas L. Lisna Hidayati; Tri Rini Nuringtyas
Indonesian Journal of Biotechnology Vol 21, No 2 (2016)
Publisher : Universitas Gadjah Mada

Secondary metabolites may affect insect herbivores’ host plant preferences. Attacus atlas L. larvae are known have a wider variety of host plants compared with other members of the Attacus genus. This research compared the metabolic profiles of four A. atlas host plants: keben (Barringtonia asiatica (L.) Kurz), dadap (Erythrina lithosperma Miq.), gempol (Nauclea orientalis L.), and soursop (Annona muricata L.). Leaves were collected from Sawit Sari Research Station, Yogyakarta. Terpenoid was extracted by macerating the leaves in ethyl acetate and subjecting them to GC-MS analysis, while alkaloid, tannin, and flavonoid were extracted through percolation. Total alkaloids, tannins, and flavonoids were measured using spectrophotometric analysis. Multivariate data analysis using PAST ver. 3.0 was performed on the GC-MS data. Based on the PCA scatter plot of the GC-MS data, keben leaves were clustered separately from the other three leaves by PC1. Dadap and gempol leaves were clustered together due to the phytol content while caryophyllene was detected only in soursop leaves. Neophytadiene was detected in all of the leaves, suggesting that this terpenoid may serve as a signal to locate the host plants. Keben leaves contained the lowest alkaloids and highest tannins and flavonoids compared with the other leaves. These secondary metabolites may determine the host plant suitability for culturing the A. atlas.

NMR metabolite comparison of local pigmented rice in Yogyakarta Dio N. Wijaya; Febri Adi Susanto; Yekti Asih Purwestri; Dyah Ismoyowati; Tri Rini Nuringtyas
Indonesian Journal of Biotechnology Vol 22, No 2 (2017)
Publisher : Universitas Gadjah Mada

Pigmented rice may have a black or red color due to higher anthocyanin content in its grain. A natural antioxidant, many studies on anthocyanin have reported its positive effects on human health. This fact has spurred the development of pigmented rice as a functional food. This study aimed to compare the metabolite profiles of black and red rice. Three black rice cultivars, namely Melik, Pari Ireng, and Cempo Ireng Sleman, and two red rice cultivars, Inpari 24 and RC 204, were used. After husk removal, grain samples were ground in liquid nitrogen and dried with a freeze dryer. The dried samples were extracted using 50% MeOD4 (in a D2O phosphate buffer pH 6 containing 0.01% TSP as an internal standard). Metabolomic analysis was performed using 500 MHz NMR followed by multivariate data analysis. An orthogonal partial least squares-discriminant analysis (OPLS-DA) model ađer PCA was constructed to discriminate between the five different cultivars. The resulting OPLS-DA score plot revealed a clear separation between black rice and red rice. The metabolites that could influence the separation of red rice and black rice were valine, threonine, alanine, glutamate, galactinol, β-glucose, α-glucose, raffinose, and fumaric acid.

Plant growth‐promoting activity of endophytic bacteria from sweet sorghum (Sorghum bicolor (L.) Moench) Charlie Ester de Fretes; Donny Widianto; Yekti Asih Purwestri; Tri Rini Nuringtyas
Indonesian Journal of Biotechnology Vol 26, No 4 (2021)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/ijbiotech.64893

Application of high levels of chemical fertilizers for optimal growth of sweet sorghum causes environmental degradation. Plant growth‐promoting bacteria have biotechnological importance because they can improve the growth and health of important agronomic plants. This study aimed to isolate, characterize, and identify endophytic bacteria associated with sweet sorghum (cv. KCS105), and also to study the inoculation effects of selected isolates on sorghum growth. In this study, 35 isolates were evaluated for their ability to support plant growth. The results showed that seven isolates were diazotrophic, six were capable of dissolving phosphate, six produced IAA and could detect ACC‐deaminase activity, and three inhibited the growth of pathogenic fungi. Nine isolates exhibiting mechanisms for promoting plant growth from the Alphaproteobacteria (Devosia), Firmicutes (Bacillus, Paenibacillus, Staphylococcus), and Actinobacteria (Microbacterium, Brachybacterium) phyla were identified. In addition, the Paenibacillus sp. BB7, Bacillus sp. PIB1B, and Bacillus sp. PLB1B isolates showed increasing effects on plant growth in greenhouse tests. Endophytic bacterial isolates which display plant growth‐promoting features can potentially be employed as biofertilizer agents. They may also address environmental damage problems resulting from the use of chemical fertilizers and pesticides.

Optimasi Metode 1H-NMR Profiling pada Rimpang Kunyit (Curcuma domestica) Caroline Dwiseptianti; Febri Adi Susanto; Yekti Asih Purwestri; Tri Rini Nuringtyas
Bioeksperimen: Jurnal Penelitian Biologi Vol 5, No 2: September 2019
Publisher : Universitas Muhammadiyah Surakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.23917/bioeksperimen.v5i2.9238

The use of medicinal plants is increasing due to the lack of side effects caused and the number of bioactive compounds that cannot be represented by synthetic chemical synthesis compounds. However, the management and use of natural medicines for the main handling of diseases are often hampered by the quality of the ingredients which are low and unstable. The standardized quality control system of OAI (Indonesian Natural Medicine) is the main key to improve clinical assurance and safety of the use of herbal medicines in Indonesia. One of the medicinal plants known to the public is Curcuma longa L. (turmeric). The main active components contained in turmeric are curcumin, demetoksikurkumin, bis-demetoksikurkumin, and ar-turmeron. Information about the quality of turmeric is needed in its use as a raw material for drugs so we need an analytical technique that is able to identify the diversity of metabolite profiles of active compounds. In this research, an optimization method is used to improve efficiency in the extraction of turmeric rhizome metabolites so that the best solvent concentration is known for the analysis of fingerprinting secondary metabolites with 1H-NMR 500 MHz spectroscopy in turmeric rhizomes. The results were analyzed with MNOVA software and chemical shift obtained compared with the reference. From the results obtained a concentration of methanol-d4 (CD3OD) 100% able to extract curcumin better than other solvents. The solvent is able to extract saccharide (sugar) compounds in the form of sucrose, amino acids and fatty acids in the form of methionine, glutamine, acetate, and glycero phospho choline.

Pengaruh Perbedaan Metode Ekstraksi Metabolit Sekunder Streptomyces sp. GMR22 terhadap Toksisitas pada Sel BHK-21 Diani Mentari; Mirtani Naima; Riska Wulansari; Jaka Widada; Tri Rini Nuringtyas; Tri Wibawa; Nastiti Wijayanti
Pharmacon: Jurnal Farmasi Indonesia Vol 16, No 1 (2019)
Publisher : Universitas Muhammadiyah Surakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.23917/pharmacon.v16i1.8032

Streptomyces sp. GMR22 is local isolate from Wanagama 1 Forest in Yogyakarta. They have the potential to be developed to produce active compounds because have PKS and NRPS genes.The active compounds from isolation are strongly influenced by various factors, one of them is extraction techniques. Effect difference of extraction technique will be affected by the quality of secondary metabolites produced.The purpose of this study was to compare the cytotoxicity effects of secondary metabolites of Streptomyces sp. GMR22 which have extracted with different stages from previous studies. The extraction technique was carried out by multilevel separatory funnel extraction methods, which was first extracted using non-polar solvent (n-hexane) and then extracted using semi-polar solvent (ethyl acetate). This research is important because in previous studies (separatory funnel only extracted using ethyl acetate) with the use of the lowest concentration in the dengue virus antiviral test (further test) caused 100% of deaths in BHK-21 cells.This study indicate that multilevel extraction result in lower CC50 value than previous studies. There are 49.160 µl/ml (n-hexane extract) and 284.56 µl/ml (ethyl acetate extract) while water extract is 464,38 µl/ml. FTIR compound analysis show that the three extracts produced have different spectrum patterns, especially in the n-hexane and ethyl acetate extract. Value of CC50 is not too high, it is expected that the secondary metabolites contained in the extracts can be used for further analysis such as antiviral testing because it is safe for normal host cells such as BHK-21 cells

1H NMR Fingerprinting of Medicinal Herbs Contain Chemical Drug Material Allopurinol Adita Yuniati Puspitasari; Harno Dwi Pranowo; Respati Tri Swasono; Tri Rini Nuringtyas
Majalah Obat Tradisional Vol 23, No 3 (2018)
Publisher : Faculty of Pharmacy, Universitas Gadjah Mada

A study to differentiate the pure medicinal herbs from the mix medicinal herbs with chemical drug material has been done. For this purpose, we conducted fingerprinting of commercial medicinal herbs and chemical drug material allopurinol using 1H-NMR followed with chemometrics analysis. Nine commercial traditional herbal medicines claimed for rheumatic were used as samples as well as allopurinol as the chemical drug standard. Extraction of samples was done by ultrasonicator for 15 min in methanol-d4 containing 0.01% TMSP as an internal standard. Each type of herbal medicine was prepared in three replicates. The phytochemical analysis was done by 500 MHz JEOL NMR. The chemometrics analysis was done using SIMCA software following the 1H NMR spectra processing with MNOVA software. All spectra showed no contamination with allopurinol. The specific signals of allopurinol at aromatic regions were confirmed not present when the spectra were stacked together. Hence, the result of OPLS-DA analysis convinced that the herbs were clearly separated the medicinal herbs into 3 classes. Jamu 1 is separated from others showed very high intensity of several signals which may indicate an addition of chemical medicines but not allopurinol. The clear separation of other two groups may corresponds to the similarity of ingredients. These results also showed that most of traditional medicines which produced by small industries, the traditional medicines contain no active pharmaceutical ingredients (allopurinol) indicating a high safety of Indonesia traditional medicines.

Immunomodulatory and Antioxidant Activity of Green Grass Jelly Leaf Extract (Cyclea barbata Miers.) In Vitro Rendi Mahadi; Mustafid Rasyiid; Krisnanda Surya Dharma; Lindia Anggraini; Rahma Nurdiyanti; Tri Rini Nuringtyas
Journal of Tropical Biodiversity and Biotechnology Vol 3, No 3 (2018): December
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jtbb.33441

Green grass jelly (Cyclea barbata Miers.) is known for its benefit to human health especially in supporting body’s immune system and wellness. This research aimed to determine immunomodulatory and antioxidant activity of green grass jelly leaf extracts in vitro. Old leaves were collected as sample then dried and ground to powder. The extraction was done with sohxletation using three different solvents, chloroform, ethyl acetate, and ethanol. The immunomodulatory activity was evaluated by treating the crude extracts at concentrations of 50, 100, and 500 mg/mL on macrophages of rat in vitro. Macrophage cells separated form peritoneal fluid used RPMI medium. Phagocytosis activity and phagocytosis capacity of macrophages were performed in vitro using latex beads that suspended in phosphate buffered saline (PBS). The antioxidant activity was measured by spectrophotometry technique with 1,1-diphenyl-2-picrylhydrazyl (DPPH) solution. All treatments were done three replicates. Detection of the bioactive groups of the extracts was done by Thin Layer Chromatography (TLC). The results showed that ethyl acetate extract has the highest phagocytosis activity followed by chloroform extract and ethanol extract, respectively. Optimum concentration was reached at 100 mg/mL of ethyl acetate extract. The ethyl acetate extract was also the highest antioxidant activity index 7.7 followed by both extracts of chloroform and ethanol similar index value of 6.25 and 6.3, respectively. The ethyl acetate extract has a high immunomodulatory activity and antioxidant activity which contained phenolics, flavonoids, tannins, and terpenoids.

Comparison of Metabolomics Expression in The Root and Leaf of Resistance and Susceptible Tomato against Root-Knot Nematode Enik Nurlaili Afifah; Rudi Hari Murti; Tri Rini Nuringtyas
AGRIVITA, Journal of Agricultural Science Vol 42, No 3 (2020)
Publisher : Faculty of Agriculture University of Brawijaya in collaboration with PERAGI

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.17503/agrivita.v42i3.2440

The resistant tomato against nematode produces various biochemical compounds associated with the defense mechanisms for nematode attack. These compounds allegedly expressed in all cells not only in the infected area. These metabolites are useful for plant development program to point out the candidate traits based on specific metabolites. This study aimed to compare expression of metabolomics as defense mechanism in root and leaf of the plant. Four cultivars consisted of the resistant [GM2 and F1 (a cross GM2 and Hawaii 7996)] and susceptible (Gondol Putih and Gondol Hijau) were used as plant materials. Fifty mg of freeze-dried of root and leaf were taken for assessing 1H-NMR (Nuclear Magnetic Resonance) for metabolomic analysis. Tomato roots resulted in 16 metabolites, while in the leaf detected 15, including amino acid, sugar, and aromatic compounds. Both root and leaf showed the same metabolites that play an important role in nematode-resistant mechanism, these metabolites were α- and β-glucose, and caffeic acid. These compounds had the same concentration within the root and leaf. It is implied that the metabolites associated with defense mechanism of tomato plants against nematode not only expressed locally in the infected area but also produced by not infected tissues throughout the plant.

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