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All Journal HAYATI Journal of Biosciences Jurnal Rekayasa Proses MEDIA MEDIKA INDONESIANA Indonesian Journal of Cancer Chemoprevention Indonesian Journal of Biotechnology Majalah Obat Tradisional INDONESIAN JOURNAL OF PHARMACY Jurnal Rekayasa Proses

Ratna Asmah Susidarti

1Department Of Pharmaceutical Chemistry, Faculty Of Pharmacy, Universitas Gadjah Mada, Yogyakarta 55281, Indonesia. 2Cancer Chemoprevention Research Center, Faculty Of Pharmacy, Universitas Gadjah Mada, Yogyakarta 55281, Indonesia.

Author-ID : 2420860

Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Earth & Planetary Sciences Education Health Professions Immunology & microbiology Materials Science & Nanotechnology Medicine & Pharmacology Public Health Social Sciences

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Antimigratory Activity of Brazilin-Containing Fraction from Caesalpinia sappan L. on MDAMB-231 Cells Sri Handayani; Ratna Asmah Susidarti; Puspa Dewi Narrij Lotulung; Akhmad Darmawan; Edy Meiyanto; Riris Istighfari Jenie
HAYATI Journal of Biosciences Vol. 27 No. 4 (2020): October 2020
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.27.4.266

Caesalpinia sappan is studied for several biological activities. The aim of this research is to determine the cytotoxic and antimigratory activities of Caesalpinia sappan active fraction in combination with cisplatin on human TNBC cells (MDA-MB-231). Caesalpinia sappan heartwood was extracted with methanol. Then, several fractions of the methanol extract were obtained by using a liquid-liquid extraction method followed by column chromatography. The cytotoxicity was determined using MTT assay. Synergistic effects were analyzed by calculating the combination index (CI). Migration was examined using wound-healing assay. Levels of MMP2 activity were determined with gelatin zymography assay. The results showed that most of the fractions included in this study exhibited cytotoxic effects against MDA-MB-231 cells, and C fraction demonstrated the highest cytotoxic activity of all fractions. The combination of C-cisplatin revealed a synergistic inhibitory effect on MDA-MB-231 cell growth (CI<1). Furthermore, C fraction, alone and in combination with cisplatin, inhibited migration of MDA-MB-231 and suppressed MMP2 activity. The C fraction isolated from Caesalpinia sappan increased the cytotoxic and antimigratory activities of cisplatin on MDA-MB-231 cells. Based on these findings, the potential of Caesalpinia sappan to act as a supportive agent in metastatic TNBC treatment with cisplatin warrants further exploration.

Brazilein in combination with cisplatin inhibit proliferation and migration on highly metastatic cancer cells, 4T1 Sri Handayani; Ratna Asmah Susidarti; Zalinar Udin; Edy Meiyanto; Riris Istighfari Jenie
Indonesian Journal of Biotechnology Vol 21, No 1 (2016)
Publisher : Universitas Gadjah Mada

Brazilein performs anticancer activities on several cancer cells and potentially inhibits metastasis. The aims of this study is to observe the synergistic cytotoxic and migration inhibitory effect of brazilein combined with cisplatin on 4T1 breast cancer cells. Under MTT assay, we found that brazilein revealed cytotoxic effect on 4T1 cells in a dosedependent manner (IC50=50 ± 0.3 µM). Combination of brazilein and cisplatin showed synergistic effect (CI=0.72). Flowcytometry analysis on the cell cycle progression showed that single treatment of 25 µM brazilein induced G2/Mphase accumulation, 12.5 µM cisplatin induced Sphase accumulation, while combination of brazilein and cisplatin induced Sphase and G2/Mphase accumulation. Combination of brazilein and cisplatin induced apoptosis higher than that of the single treatments. Based on wound healing assay, 12.5 µM brazilein and its combination with 6.25 µM cisplatin inhibited cells migration. Immunoblotting and gelatin zymography analysis showed that combination of brazilein and cisplatin inhibited the expression level of Rac1 and MMP9 proteins. Based on these results, we conclude that brazilein enhanced cytotoxic activity of cisplatin and inhibited migration on 4T1 cells and potentially can be developed as an enhancing cytotoxic and antimetastasis agent.

Kinetics Study of Paracetamol Production from Para-Aminophenol and Acetic Anhydride Rifki Wahyu Kurnianto; Muhammad Fahrurrozi; Hilda Ismail; Raden Rara Endang Lukitaningsih; Indah Tri Nugraha; Pudjono Pudjono; Rochmadi Rochmadi; Ari Sudarmanto; Ratna Asmah Susidarti
Jurnal Rekayasa Proses Vol 15, No 1 (2021)
Publisher : Departemen Teknik Kimia Fakultas Teknik Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jrekpros.64551

In the last decade, Indonesia intensifies the efforts to reduce pharmaceutical imports. One of the initiatives is establishing a paracetamol production facility to start operating in 2024. Kinetics study is needed as a basis to design the paracetamol reactor. This study investigated the optimal temperature, reactant mole ratio, and agitation speed in the reactor for paracetamol production. In this study, aqueous solution of para-aminophenol was reacted with acetic anhydride. The mole ratio of para-aminophenol to acetic anhydride was varied to 1:1, 1:1.2, 1:1.5, and 1:2 while the temperature was varied to 80 °C, 90 °C, and 110 °C. However, due to uncontrolled heat of the reaction and limitation of the mixture’s boiling point, the actual reaction temperatures were 86 °C, 90 °C, and 108 °C. In addition, the agitation speed of 250 RPM and 350 RPM were also studied. Thin layer chromatography (TLC) and densitometry were used to determine the concentration of paracetamol in the reacting mixture. The optimum temperature, reactant mole ratio, and agitation speed in this study were 108 °C, 1:1.5, and 350 RPM, respectively. In addition, a reaction performed under those operating parameters gave the reaction rate constant of 1.95 L mol-1 min-1.Keywords: acetic anhydride; kinetics; para-aminophenol; paracetamol; pharmaceutical industry A B S T R A KDalam sepuluh tahun terakhir ini, Indonesia bertekad mengurangi impor bahan baku farmasi. Salah satu upaya yang dilakukan adalah membangun fasilitas produksi parasetamol yang akan mulai beroperasi pada tahun 2024. Studi kinetika diperlukan sebagai dasar perancangan reaktor parasetamol. Oleh karena itu, penelitian ini mengkaji kondisi operasi optimal pada reaksi produksi parasetamol yang akan dibutuhkan sebagai dasar perancangan pabrik. Pada percobaan ini, para-aminofenol direaksikan dengan anhidrida asetat dengan media air. Rasio mol para-aminofenol terhadap asetat anhidrida divariasikan 1:1 1:1,2, 1:1,5, dan 1:2 sedangkan temperatur divariasikan 80 °C, 90 °C, dan 110 °C. Akan tetapi, karena panas reaksi yang tidak dikontrol dan batasan berupa titik didih dari campuran reaksi, temperatur aktual reaksi menjadi 86 °C, 90 °C, dan 108 °C. Selain itu, kecepatan putaran pengadukan juga divariasikan pada angka 250 RPM dan 350 RPM. Kromatografi lapis tipis (KLT) dan densitometri digunakan untuk menentukan konsentrasi parasetamol dalam campuran reaksi. Temperatur, rasio mol reaktan, dan kecepatan putaran pengadukan yang optimum pada penelitian ini masing-masing adalah 110 °C, 1:1,5, dan 350 RPM. Selain itu, reaksi yang dilakukan dengan kondisi operasi tersebut menghasilkan konstanta laju reaksi 1,95 L mol-1 menit-1.Kata kunci: anhidrida asetat, industri farmasi, kinetika, para-aminofenol, parasetamol

Metode RPTLC dan Optimasi Fase Gerak Dalam Penetapan Harga Rm Sebagai Salah Satu Parameter Lipofilisitas Dalam Rancangan Obat Gunardi Gunardi; Ratna Asmah S.; Bambang Tri Purwanto; Edy Sulistyowati; Siti Musinah
MEDIA MEDIKA INDONESIANA 2009:MMI VOLUME 43 ISSUE 5 YEAR 2009
Publisher : MEDIA MEDIKA INDONESIANA

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (229.154 KB)

RPTLC and optimizing mobile phase methods in Rm value determination as one of lipophylicity parameters in drug designBackground: The early process to successfully achieve its target is drug penetration or absorption. Of the three physicochemical parameters i.e, lipophyllicity, electronic and steric parameters, the lipophyllicity parameter is the most reponsible in drug absorption process. The research was aimed to determine retention modified (Rm) value of barbituric acid compound derivatives using RPTLC metod and mobile phase optimizing.Methods: This study was conducted on barbituric acid compound derivatives by using reverse phase thin layer chromatography (RPTLC). Silica Gel GF 254 that had been submerget in the mixture of liquid paraffin and petroleum eter (95:5) was used as a stationary phase. The mixture of polar to non polar solvent was used as mobile phase.Results: Research showed that in this method the most optimum of mobile phase was indicated by methanol and acetic acid mixture in the ratio of (1:9). The resulted Rm values of 5,5-diethylbarbituric acid, 5,5-diallylbarbituric acid, 5-allyl-5-isopropylbarbituric acid, 5-allyl-5-isobuthylbarbituric acid, 5-etil-5-(1-methylbutyl) barbituric acid, 5-(1-cyclohexene-1-yl)-1,5-dimethylbarbituric acid and 5-ethyl-5-phenylbarbituric acid were as follow 0.116; 0.144; 0.162; 0.221; 0.262; 0.187 and 0.199.Conclusions: The most optimum mobile phase in this method was the mixed solvents that had lower polarity, i.e, the mix of methanol and acetic acid in the ratio of (1:9). The H1 , H2 and H3 substituens in barbituric acid nuclei showed, the longer carbon chain, the higher the Rm values, howover the existing of double bond in such substituents will decrease the Rm value.Keywords: RPTLC, mobile phase, Rm value ABSTRAK Latar belakang: Proses awal keberhasilan obat dalam mencapai target adalah penetrasi atau absorpsi. Parameter lipofilisitas paling bertanggung jawab terhadap proses absorpsi obat dibanding parameter elektronik dan stearik. Tujuan penelitian ini adalah penggunaan metode RPTLC dan optimasi fase gerak dalam penentuan harga retention modified (Rm) senyawa turunan asam barbiturat.Metode: Penelitian dilakukan terhadap senyawa turunan asam barbiturat, dengan metoda kromatografi lapis tipis fase terbalik. Digunakan fase diam silika gel GF 254 yang telah dibacem dengan campuran parafin cair dan petroleum eter dengan perbandingan (95:5). Fase gerak yang digunakan dipilih campuran pelarut dari yang sangat polar sampai yang kurang polar.Hasil: Fase gerak yang paling optimum digunakan dalam metode ini adalah campuran metanol dan asam asetat dengan perbandingan (1:9). Harga Rm yang diperoleh secara berturutan, asam 5,5-dietilbarbiturat, asam 5,5-dialilbarbiturat, asam 5-alil-5- isopropilbarbiturat, asam 5-alil-5-isobutilbarbiturat, asam 5-etil-5-(1-metilbutil) barbiturat, asam 5-(1-sikloheksen-1-il) 1,5- dimetilbarbiturat dan asam 5-etil-5-fenilbarbiturat adalah: 0,116; 0,144; 0,162; 0,221; 0,262; 0,187 dan 0,199.Simpulan: Fase gerak yang paling optimum dalam metoda ini adalah bukan fase berair, tetapi berupa campuran pelarut yang mempunyai polaritas rendah, yakni campuran metanol dan asam asetat dengan perbandingan (1:9). Substituen atom H1 , H2 dan H3 pada inti asam barbiturat, menunjukkan makin panjang rantai karbon, makin tinggi harga Rm-nya. Adanya ikatan rangkap pada substituen menurunkan harga Rmnya.

PENGARUH MEDIA PADA PERTUMBUHAN FUNGI ENDOFIT IP-2 DAN PRODUKSI METABOLIT AKTIF INHIBITOR POLIMERISASI HEM Indah Purwantini; Wahyono Wahyono; Mustofa Mustofa; Ratna Asmah Susidarti
Majalah Obat Tradisional Vol 20, No 1 (2015)
Publisher : Faculty of Pharmacy, Universitas Gadjah Mada

Fungi endofit IP-2 adalah fungi endofit yang diisolasi dari tanaman Artemisia annua L. Fungi ini diketahui mempunyai aktivitas sebagai inhibitor polimerisasi hem. Dalam penelitian ini akan diteliti mengenai media yang sesuai untuk pertumbuhan maupun pembentukan metabolit aktifnya serta menentukan waktu yang sesuai untuk pemanenan metabolit aktifnya. Fungi endofit difermentasi dalam 4 jenis media selama 14 hari. Setiap hari dilakukan pengambilan sampel yang akan digunakan untuk mengetahui bobot sel keringnya yang selanjutnya digunakan untuk analisis pertumbuhannya serta digunakan untuk uji aktivitas inhibitor polimerisasi. Uji dilakukan dengan menggunakan metoda yang dikembangkan oleh Bassilico dkk (1998). Data bobot sel kering dan prosentase penghambatan inbihitor polimerisasi dianalisis sehingga diperoleh kurva pertumbuhan dan kurva produksi metabolitnya yang akan digunakan untuk menentukan waktu yang tepat untuk memanen metabolit aktifnya. Hasil penelitian menunjukkan bahwa dalam media PDB fungi endofit IP-2 menunjukkan pertumbuhan yang paling baik dan mampu menghasilkan metabolit aktif paling tinggi dibandingkan 3 media lainnya. Dalam media PDB, pada hari ke-7 terlihat jumlah metabolit aktif yang dihasilkan paling banyak sehingga waktu panen metabolit yang tepat adalah pada hari ke-7.

PENETAPAN KADAR DIOSGENIN DALAM EKSTRAK AIR Costus speciosus SECARA HPLC Hari Susanti; Subagus Wahyuono; Ratna Asmah Susidarti; Ika Puspita Sari
Majalah Obat Tradisional Vol 22, No 1 (2017)
Publisher : Faculty of Pharmacy, Universitas Gadjah Mada

Costus speciosus has been reported to have biological activity among antidiabetic, antihyperlipidemia, antioxidants. One of the chemical constituents that is responsible for some of the biological activity is diosgenin. This study aimed to determine diosgenin content of Costus speciosus water extract (CS). Water extract of CS was obtained with infundation method. The extract was dried using freeze dryer. The assay of diosgenin was performed by HPLC method. HPLC system used: LiChrospher® 100 RP-18 endcapped (5 µm) column length 25cm, colimn id 4 mm as stationary phase, acetonitrile-water (9: 1 v/v) as mobile phase, flow rate 2mL/min and UV detection at 205 nm. HPLC method developed was linear in the range 20-200 μg mL with R = 0.9999 ; slope = 6423.7; and intercept = -4280.5; LOD = 2.14 ug/mL and LOQ = 6.50 mg/mL; and recovery 100.63 %. Diosgenin levels in CS was 0.279 %. The developed HPLC method is relatively simple, rapid, sensitive, and accurate to determine the content of diosgenin in water extracts of Costus speciosus.

8-Methoxycapnolactone and stigmasterol From Micromelum minutum Ratna Asmah Susidarti; Marwadi Rahmani; Abdul Manaf Ali; M. Aspollah Sukari; Hazar B.M. Ismail; Julius Kulip; Peter G. Waterman
Indonesian Journal of Pharmacy Vol 18 No 2, 2007
Publisher : Faculty of Pharmacy Universitas Gadjah Mada, Yogyakarta, Skip Utara, 55281, Indonesia

The coumarin, 8-methoxycapnolactone and stigmasterol were isolated from the leaves of Micromelum minutum (Rutaceae) which collected from Sepilok, Sabah, Malaysia and their structures were characterized by spectroscopic methods.Key words: Micromelum minutum; Coumarin; 8-Methoxycapnolactone, Stigmasterol

Isolation and identification of kaempferol from jangkang (Homalocladium platycladum (F. Muell) Bailey) leaves and its antibacterial activity Maulita Cut Nuria; Wahyono .; Ratna Asmah Susidarti
Indonesian Journal of Pharmacy Vol 22 No 1, 2011
Publisher : Faculty of Pharmacy Universitas Gadjah Mada, Yogyakarta, Skip Utara, 55281, Indonesia

Isolation and identification of kaempferol from jangkang (Homalocladium platycladum (F. Muell) Bailey) leaves and its activity against B. subtilis ATCC 9466 dan S. typhi ATCC 1408 has been done. The dried ground leaves (300.04 gram) were macerated with petroleum ether, chloroform and methanol respectively. The methanol extract was the most active extract against both bacteria. Further separations of this extract using preparative thin layer chromatography yielded flavonoid glycoside. Its aglicon was identified as kaempferol based on its spectroscopic data (MS, IR, and UV-Vis) and UV spectrum (densitometry). This compound shows iradical zone of 6.62 mm (Bacillus subtilis) and 6.27 mm (Salmonella typhi) at concentration of 200 µg/disk.Key words : kaempferol, jangkang, glycoside, aglicon

The Effect of Brazilin from Caesalpinia sappan on Cell Cycle and Modulation and Cell Senescence in T47D cells Riris Jenie; Sri Handayani; Ratna Asmah Susidarti; Edy Meiyanto
Indonesian Journal of Pharmacy Vol 31 No 2, 2020
Publisher : Faculty of Pharmacy Universitas Gadjah Mada, Yogyakarta, Skip Utara, 55281, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjpharm31iss2pp84

Ethanolic extract and brazilein-containing fraction of Caesalpinia sappan L., has been reported to inhibit cell proliferation in T47D (ER+ PR+/- cell, Luminal A subtype model). The Luminal A subtype is the most common subtype of breast cancer in Indonesian women. In this study, we explored the activity of the reduced form of brazilein, i.e. brazilin, in T47D cells proliferation and the mechanism that involved. The cytotoxicity activity of brazilin was observed using MTT assay. While the cell cycle modulation analysis was done by using flowcytometry, and the senescence assay was observed using S-A-β-galactosidase assay. The results showed that brazilin inhibited cell growth in a dose-dependent manner with an IC50 value of 50μM (or 14.3μg/mL). That was higher than a brazilein-containing fraction, which was reported previously by our group to have an IC50 value of 68μg/mL against the same cell. Cell cycle analysis showed that cells treated with brazilin were accumulated at the G2/M phase in a dose-dependent manner. Furthermore, cells treated with a combination of brazilin and doxorubicin was accumulated at the G2/M phase and sub G1 phase. Cells accumulation at sub G1 phase indicates that the cells undergo apoptosis. Our data of S-A-β-galactosidase assay showed that cells treated with 1/4IC50, 1/2IC50, and IC50 brazilin had lower senescent cells compared to the untreated cells. The morphology of cells treated with IC50 (50μM) brazilin changed. The cells shape became rounded, cells were shrinkage and detached from the well plate, indicating that cells may undergo apoptosis. These results suggested that brazilin was cytotoxic towards T47D cells and its combination with dox potentially induced apoptosis and decreased cell senescence. The ability of brazilin to decrease cell senescence provides new insight of utilization of C. sappan or its constituents, particularly brazilin, as anti-ageing.

Combination of Doxorubicin and Areca Ethanolic Extract Induces Apoptosis by Increasing Caspase-3 Level on Breast Cancer (T47D) Cells Fitria Rahmi; Edy Meiyanto; Ratna Asmah Susidarti
Indonesian Journal of Cancer Chemoprevention Vol 3, No 1 (2012)
Publisher : Indonesian Society for Cancer Chemoprevention

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjcanchemoprev3iss1pp339-344

Despite causing many side effects, doxorubicin (Dox) is still one of breast cancer drug of choice. Thus, combination of chemotherapy is developed in order to decrease doxorubicin regimen dose. The aim of this research is to examine the combination effect of doxorubicin (dox) and areca extract (AE) on T47D human breast cancer cells. The cytotoxic activity was determined using MTT assay. The combination index (CI) of the combination treatment was calculated to determine the effects (synergistic, additive or antagonistic). The combination application of dox (6-22nM) and AE (8-30µg/ml) on T47D cells showed synergistic (CI<0.9) or additive effect (CI=0.9-1.1). The effective combination of dox-AE was 6 nM - 8 µg/ml on CI<0.5. Apoptosis induction of AE solely and its combination with dox was then observed using double staining method. Moreover, expression of Bax and caspase-3 protein which mediated apoptosis, were observed using immunocytochemistry. Combination of AE and Dox increased expression of Caspase-3 but did not increase expression of Bax. This result showed AE increase the effectiveness of doxorubicin against T47D cells.Keywords: Breast cancer, doxorubicin, areca extract, T47D cells

Co-Authors Abdul Manaf Ali Akhmad Darmawan Andi Eko Wibowo Ari Sudarmanto Bambang Tri Purwanto Edy Meiyanto Edy Meiyanto Edy Meiyanto Edy Meiyanto Edy Sulistyowati Endang Lukitaningsih Fitria Rahmi Gunardi Gunardi Hari Susanti Hazar B.M. Ismail Hilda Ismail, Hilda Ika Puspita Sari Indah Purwantini Indah Tri Nugraha Jenie, Riris Istighfari Julius Kulip Kurnianto, Rifki Wahyu M. Aspollah Sukari Marwadi Rahmani Maulita Cut Nuria Muhammad Fahrurrozi Mustofa Mustofa Ni Putu Linda Laksmiani Nur Ismiyati Peter G. Waterman Pudjono Pudjono Puspa Dewi Narrij Lotulung Riris Jenie Rochmadi Rochmadi Sismindari Sismindari Siti Musinah Sri Handayani Sri Handayani Sri Handayani Subagus Wahyuono Ulfatul Husnaa Wahyono . Wahyono Wahyono Warsinah Warsinah Yuli Puspito Rini Zalinar Udin

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