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Abstrak :
Temulawak (Curcuma xanthorrhiza Roxb.) merupakan salah satu tanaman obat yang banyak digunakan di Indonesia, dan di Asia Tenggara temulawak dimanfaatkan sebagai bumbu masak dan obat. Aktivitas antimikroba temulawak diuji terhadap Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Porphyromonas gingivalis ATCC 33277, Staphylococcus aureus ATCC 25923, Bacillus cereus ATCC 11778, Streptococcus mutans Type F (MUI), dan Candida albicans ATCC 10231 dengan menggunakan broth dilution method. Ekstrak etanol 70% temulawak dapat menghambat pertumbuhan bakteri Gram positif S. aureus dan S. mutans pada konsentrasi 1,0-5,0% b/v, dan B. cereus pada konsentrasi 2,0-5,0% b/v. Minimum Inhibitory Concentration (MIC) ekstrak etanol 70% temulawak adalah 0,1% b/v untuk S. aureus dan S. mutan, sedangkan terhadap B. cereus adalah 2,0% b/v.

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Temulawak (Curcuma xanthorrhiza Roxb.) is one of popular medicinal plant in Indonesia, has been used as spices and medicinal purposes in South-East Asian countries. Antimicrobial activity of temulawak was tested toward Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Porphyromonas gingivalis ATCC 33277, Staphylococcus aureus ATCC 25923, Bacillus cereus ATCC 11778, Streptococcus mutans Type F (MUI), and Candida albicans ATCC 10231. Antimicrobial assay was carried out by using broth dilution method. The ethanol 70% extract of temulawak inhibited the growth of Gram positive bacteria S. aureus and S. mutans at concentration of 1,0-5,0% w/v, while B. cereus at concentration 2,0-5,0% w/v. The Minimum Inhibitory Concentration (MIC) of ethanol 70% extract against S. aureus and S. mutans were 0,1% w/v, while against B. cereus were 2,0% w/v.

Abstrak :
Curcuma xanthorrhiza Roxb. adalah tanaman asli Indonesia yang memiliki kandungan xantorizol dan kurkuminoid, terutama pada bagian rimpangnya. Ekstraksi rimpang temulawak menggunakan pelarut cairan ionik (IL) dengan metode ultrasound assisted extraction (UAE). IL merupakan salah satu pelarut alternatif yang polaritasnya dapat dirancang bergantung pada kation dan anionnya dan UAE merupakan salah satu metode ekstraksi modern dengan keuntungan berupa konsumsi jumlah pelarut yang lebih rendah. Tujuan dari penelitian ini ialah mengoptimasi ekstraksi, mendapatkan kondisi optimum ekstraksi, dan membandingkan hasil ekstraksi rimpang temulawak C12MIMCl-UAE dengan maserasi-etanol 96%. Variabel bebas yang digunakan dalam kondisi optimasi ialah konsentrasi IL, waktu ekstraksi, dan rasio serbuk-pelarut. Ketiga variabel tersebut dilakukan perancangan menggunakan metode Response Surface Methodology-Box Behnken Design (RSM-BBD) dan dihasilkan 17 runs kondisi optimasi. Hasil optimum dari ke-17 runs tersebut terdapat pada run ke-5 dengan kondisi ekstraksi berupa konsentrasi IL 0,12 M, waktu ekstraksi selama 20 menit, dan rasio serbuk-pelarut 1:30 g/mL. Perolehan kadar xantorizol dan kurkuminoid terhadap serbuk pada kondisi optimum IL-UAE dengan penambahan Na2SO4 dan diklorometana ialah 43,121 mg/g dan 10,576 mg/g dengan rendemen sebesar 29,99%. Hasil tersebut lebih tinggi dibandingkan dengan perolehan kadar xantorizol dan kurkuminoid terhadap serbuk dengan metode ekstraksi maserasi-etanol 96%, yaitu 9,256 mg/g dan 2,163 mg/g dengan rendemen sebesar 8,92%. ......Curcuma xanthorrhiza Roxb. (javanese turmeric), is an Indonesia native plant that contains xanthorrhizol and curcuminoid, especially in its rhizome. Extraction of javanese turmeric rhizomes has been done using ionic liquid (IL) as a solvent by ultrasound assisted extraction (UAE) method. IL is one of the alternative solvents whose polarity can be designed according to its cation and anion and UAE is one of the modern extraction methods with the advantage of consuming a lower amount of solvent. The aim of this study is to optimize extraction, obtain optimum extraction conditions, and to compare the extraction results of javanese turmeric rhizome using C12MIMCl-UAE and maceration-96% ethanol. The all variables used optimization conditions are IL concentration, extraction time, and powder-solvent ratio. The three variables were designed using the Response Surface Methodology-Box Behnken Design (RSM-BBD) method and produced 17 runs of optimization conditions. The optimum results of the 17 runs were found in the 5th run with extraction conditions: 0.12 M of IL concentrations, 20 minutes of extraction times, and 1:30 g/mL of powder-solvent ratios. The xanthorrhizol and curcuminoid levels in powder under optimum conditions of IL-UAE with the addition of Na2SO4 and dichloromethane were 43,12 mg/g and 10,58 mg/g with 29,99% of yield. The result were higher than those of maceration-96% ethanol extraction method, which could only extract 9,26 mg/g and 2,16 mg/g of xanthorrhizol and curcuminoid content with 8,92% of yield.

Abstrak :
Background: One of the major challenges in developing an antifungal against Candida albicans as a therapeutic strategy for oral candidiasis is its resistance towards antimicrobial agents. Currently, medicinal plants are continuously being developed as a therapeutic agent, including Javanese turmeric (Curcuma xanthorrhiza Roxb.), an Indonesian plant widely used as a traditional medicine. Its main active compound, xanthorrhizol, as well as the Javanese turmeric Ethanol Extract (EET) had been reported to have antifungal properties. However, extract quality as well as cultivation site of a medicinal plant may affect its’ effectivity as a therapeutic agent. Objective: To analyze the influence of Javanese turmeric cultivation site against its’ ethanolic extract effectivity in inhibiting C. albicans biofilms. Methods: 2 Javanese turmeric ethanolic extract were collected from different cultivation sites, which were Bogor, West Java and Malang, East Java. The extracts were tested for inhibitory activity against C. albicans biofilm using OD600, TPC, and MTT assay measurements. Results: There were concentration differences of xanthorrhizol content in between the two extracts. Despite so, both extracts showed insignificant MBIC50 differences at roughly 0,10 µg/mL, while their MBIC90 were measured to be constant at all biofilm development phases, at 0,30 µg/mL. Conclusion: The different cultivation site of Javanese turmeric in Java island does not influence the effectivity of JtET in inhibiting C. albicans biofilm. ......Background: One of the major challenges in developing an antifungal against Candida albicans as a therapeutic strategy for oral candidiasis, an opportunistic oral fungal infection, is its resistance towards antimicrobial agents. Currently, medicinal plants are being developed as a therapeutic agent, including Javanese turmeric (Curcuma xanthorrhiza Roxb.), an Indonesian plant widely used as a traditional medicine. Its main active compound, xanthorrhizol, is known to have antifungal properties. However, extract quality as well as cultivation site of a medicinal plant may affect its’ effectivity as a therapeutic agent. Up until now, there hasn’t been any datas that explain how these factors affect Javanese turmeric ethanolic extract as a C. albicans strategy. Objective: To analyze the effect of Javanese turmeric cultivation site against its’ ethanolic extract ability to inhibit C. albicans biofilms. Methods: 2 Javanese turmeric ethanolic extract were collected from different cultivation sites, which were Bogor, West Java and Malang, East Java. The extracts were tested for inhibitory activity against C. albicans biofilm using OD600, TPC, and MTT assay measurements. Results: There were concentration differences of xanthorrhizol between the extracts. Despite so, both extracts’ MBIC50 differences were insignificant at roughly 0,10 µg/mL, while their MBIC90 were measured to be constant at all biofilm development phases, at 0,30 µg/mL. Conclusion: There were no significant differences between the MBICs of both extracts against C. albicans biofilm. More research needs to be done to confirm the role of other factors such as storage time on their MBIC.

Abstrak :
Latar Belakang: Temulawak (Curcuma Xanthorrhiza Roxb.) yang mengandung zat aktif Xanthorrhizol merupakan tanaman asli Indonesia yang telah diketahui memiliki efek anti bakteri dan anti C. albicans. Ekstrak etanol temulawak mengandung turunan alkohol sehingga berpotensi menurunkan pH dan dapat memicu demineralisasi email. Dalam penelitian ini digunakan sediaan obat tetes mikroemulsi dengan kandungan 15% ekstrak etanol temulawak. Tujuan: Menguji adverse effect (efek yang tidak diinginkan) dari paparan obat tetes ekstrak etanol temulawak terhadap kekerasan mikro permukaan email gigi. Metode: 28 gigi premolar paska ekstraksi tanpa karies dan kerusakan struktural dipisahkan menjadi 4 kelompok yang akan direndam dalam Obat tetes ekstrak etanol temulawak , kelompok kontrol positif Obat kumur komersial tipe 1 dan Obat kumur komersial tipe 2 dan kelompok kontrol negatif Akuades. Paparan dilakukan selama 1 menit sesuai dengan kelompok bahan paparan, dibilas, lalu direndam selama 10 menit dalam akuades pada suhu 37oC yang dilakukan selama 42 siklus untuk simulasi pemakaian 2 minggu dan dilakukan 21 siklus tambahan untuk simulasi pemakaian 3 minggu. Pengukuran kekerasan mikro dilakukan dengan dengan menggunakan Shimadzu HMV-G – Micro Vickers Hardness Tester sebelum paparan, setelah simulasi pemakaian 2 minggu dan setelah simulasi pemakaian 3 minggu. Data dianalisis dengan Repeated ANOVA dan One-way ANOVA denganuji Post Hoc Tamehane T2. Hasil: Perendaman dalam Obat tetes ekstrak etanol temulawak selama 2 minggu dan 3 minggu menyebabkan penurunan kekerasan mikro yang berbeda bermakna dibandingkan nilai kekerasan mikro permukaan email awal (p<0,001). Pada simulasi pemakaian 3 minggu, rata-rata dibandingkan dengan kontrol negatif tidak memiliki perbedaan bermakna (p 0.065). Kesimpulan: Penurunan kekerasan mikro permukaan email setelah paparan Obat tetes temulawak 3 kali 1 menit dalam sehari selama 3 minggu masih dalam batas aman ......Introduction: Temulawak (Curcuma Xanthorrhiza Roxb.) is a native plant to the Indonesian Archipelago containing the active compound – Xanthorrhizol. Xanthorrhizol and ethanolic extract of temulawak have previously been studied to have anti-C. albicans activities. Ethanolic extract of temulawak contains alcohol derivative which have a potential to lower pH level and trigger enamel demineralisation. This study uses an oromucosal drops containing Curcuma xanthorrhiza ethanolic extract to examine its characteristic stability and it’s safety towards enamel surface Objective: This study is done to analyze the effect of oromucosal drops containing Curcuma xanthorrhiza ethanolic extract exposure on enamel surface micro-hardness. Method: 28 extracted premolars without caries or any other structural damages is used and grouped into different exposure groups, the of oromucosal drops containing Curcuma xanthorrhiza ethanolic extract, Obat kumur komersial tipe 1 (LO), and Obat kumur komersial tipe 2 (LFB) as positive control, and Distilled water as negative control. Exposure is done for 1 minute following the exposure group, then for another 10 minutes in distilled water at temperature 37oC. The cycle is done 21 times for exposure simulation of 2 weeks use and 42 times for exposure simulation of 3-weeks use. Data obtained before exposure, after simulation of 2-weeks use, and 3-weeks used are statistically analyzed with Repeated ANOVA and One-way ANOVA with Post Hoc Tamehane T2. Result: A decrease in enamel surface microhardness following exposure to oromucosal drops containing Curcuma xanthorrhiza ethanolic extract for 2 weeks and 3 weeks were found with significant difference compared to baseline number (p <0,001). After 3 weeks exposure, the mean deacreased of enamel surface hardness was not found significantly diffrenct than the negative control (p 0.065). Conclusion: exposure to oromucosal drops containing Curcuma xanthorrhiza ethanolic extract 3 times a day, 1 minute long for 3 weeks of exposure was still within normal limit.

Abstrak :
Latar Belakang: Ekstrak etanol temulawak (Curcuma xanthorrhiza Roxb.) telah terbukti secara in vitro memiliki khasiat sebagai anti Candida albicans (C.albicans). Dalam upaya pengembangan tanaman obat tersebut sebagai obat herbal terstandar anti C.albicans, ekstrak etanol temulawak telah diformulasikan menjadi obat tetes oromukosa. Temulawak mengandung kurkumin yang merupakan senyawa polifenolik berwarna kuning yang dapat menyebabkan diskolorasi gigi. Tujuan: Mengetahui pengaruh paparan obat tetes ekstrak etanol temulawak terhadap warna email gigi. Metode: Gigi premolar tanpa karies dan defek struktural dicelupkan dalam obat tetes ekstrak etanol temulawak, CHX 0,2%, dan akuades selama 1 menit kemudian dibilas dan direndam dalam akuades selama 10 menit pada suhu 37oC. Tahapan dilakukan sebanyak 42 siklus (simulasi penggunaan 2 minggu) dan 63 siklus (simulasi penggunaan 3 minggu). Analisis warna dilakukan menggunakan colorimeter pada 3 tahap waktu yaitu sebelum paparan, setelah paparan, dan setelah penyikatan gigi. Nilai yang didapatkan berupa ΔE yang menunjukkan selisih nilai pengukuran warna email sebelum dan setelah paparan obat serta sebelum dan setelah penyikatan. Hasil: Pada tahap waktu T1-T3 simulasi penggunaan 2 minggu dan 3 minggu, nilai ΔE>3.3 pada ketiga kelompok sehingga terlihat adanya perubahan warna yang signifikan antara warna gigi awal dan setelah penyikatan gigi. Terdapat perubahan warna gigi yang signifikan setelah dilakukan penyikatan dengan pasta gigi. Kesimpulan: Obat tetes ekstrak etanol temulawak mengakibatkan perubahan warna email gigi yang signifikan. Penyikatan gigi dapat mengurangi efek perubahan warna pada email gigi. ......Background: Javanese Turmeric (Curcuma xanthorrhiza Roxb.) ethanol extract is known to have antifungal properties against Candida albicans (C.albicans) based on in vitro studies. The next step in developing a standardised herbal medicine is by formulating Javanese Turmeric Ethanol Extract into oromucosal drops. Curcumin found in javanese turmeric is a yellowish polyphenolic compound that has the potential to cause staining on the enamel. Objective: This study is aimed to evaluate the effect Javanese Turmeric ethanol extraxt oromucosal drops on discoloration of the dental enamel. Method: Premolars with no caries and structural defects are immersed in the Javanese Turmeric ethanol extract oromucosal drops, a 0,2% CHX mouthwash, and distilled water for 1 minute. After rinsing, they are then immersed in distilled water for 10 minutes at 37oC. The method mentioned is repeated for 42 cycles (2-week simulation) and 63 cycles (3-week simulation). Color assessment is done using a colorimeter at three different time points: before immersion, after immersion, and after brushing. Results will be shown as ΔE which is the color difference of enamel before and after immersion, as well as before and after toothbrushing. Result: At time point T1-T3 for the 2-week and 3-week simulation, the ΔE score is greater than 3.3 on all three groups indicating a significant color difference before immersion and after toothbrushing. A significant color difference is observed after toothbrushing with toothpaste. Conclusion: Javanese Turmeric ethanol extract oromucosal drops cause a significant tooth discoloration. Brushing had significant effect on removal of induced stains.

Abstrak :
Latar belakang: Salah satu faktor virulensi C. albicans adalah pembentukan biofilm yang dapat meningkatkan resistensi terhadap agen antijamur. Temulawak merupakan tanaman obat khas Indonesia yang diketahui memiliki efek antijamur karena mengandung zat aktif xanthorrhizol. Tujuan: Mengetahui potensi penggunaan ekstrak etanol temulawak dalam mengeradikasi biofilm C. albicans isolat klinis. Metode: Pemaparan ekstrak etanol temulawak kepada biofilm C. albicans selama 1 jam pada berbagai fase pembentukan biofilm. MTT assay digunakan untuk mengukur persentase eradikasi biofilm. Hasil: Ekstrak etanol temulawak memiliki nilai KHM dan KBM 15 terhadap C. albicans isolat klinis planktonik. Nilai Konsentrasi Eradikasi Biofilm Minimal KEBM50 ekstrak etanol temulawak terhadap biofilm C. albicans isolat klinis pada fase awal, fase menengah, dan fase maturasi adalah 25 , 15 , dan 15. Kesimpulan: Ekstrak etanol temulawak mampu mengeradikasi biofilm C. albicans isolat klinis. ...... Background: An ability to form biofilm is one of the C. albicans rsquo s virulence factor that increase resistance towards antifungal agents. Java turmeric is an Indonesian medicinal plant which reported to have antifungal effects due to its active component, xanthorrhizol. Objective: To measure in vitro potential use of Java turmeric ethanol extract in eradicating C. albicans clinical isolate biofilm. Method: One hour exposure of Java turmeric ethanol extract to C. albicans biofilm formation phases. MTT assay is used to test the percentage of biofilm eradication. Result: The Minimum Inhibitory Concentration MIC and Minimum Fungicidal Concentration MFC of Java turmeric ethanol extract towards planktonic C. albicans was 15. The Minimum Biofilm Eradication Concentration MBEC50 in the early phase was 25, intermediate phase 15 and maturation phase 15. Conclusion: Java turmeric ethanol extract is effective in eradicating clinical isolate of C. albicans biofilm.

Abstrak :
Latar Belakang: Temulawak (Curcuma xanthorrhiza Roxb.) merupakan salah satu tanaman obat unggul Indonesia yang memiliki potensi untuk menghambat pembentukan biofilm C. albicans. Faktor virulensi yang dapat menyebabkan C. albicans menjadi fungi patogen diantaranya adalah pembentukan biofilm dan sekresi enzim hidrolitik. Fosfolipase merupakan salah satu enzim hidrolitik yang dapat merusak membran sel inang. Tujuan: Menganalisis aktivitas fosfolipase pada biofilm C. albicans ATCC 10231 fase awal, menengah, dan maturasi yang terhambat ekstrak etanol temulawak (Curcuma xanthorrhiza Roxb.). Metode: Nilai Kadar Hambat Biofilm Minimal (KHBM50) C. albicans ditentukan dengan uji MTT-assay. Ekstrak etanol temulawak dengan konsentrasi sesuai KHBM50 dipaparkan pada biofilm fase awal, menengah, dan maturase. Kontrol negative tidak dipaparkan apapun, kontrol positif dipaparkan Nystatin 100.000 IU. Aktivitas fosfolipase biofilm C. albicans dianalisis dengan mengukur proporsi antara diameterzona presipitasi dengan diameter koloni C. albicans pada medium Egg Yolk Agar (EYA). Hasil: Nilai KHBM50 ekstrak etanol temulawak terhadap biofilm C. albicans ATCC 10231 pada fase awal, fase menengah, dan fase maturasi berturut-turut adalah 25%, 30%, dan 35%. Pada kontrol positif, aktivitas fosfolipase biofilm C. albicans fase awal, fase menengah, dan fase maturasi bernilai 1. Aktivitas fosfolipase biofilm C. albicansfase awal, fase menengah, dan fase maturasi yang terhambat ekstrak etanol temulawak berturut-turut 0.84, 0.80, dan 0.83. Pada kontrol negatif, aktivitas enzim fosfolipase biofilm C. albicans fase awal, fase menengah, dan fase maturasi berturut-turut 0.59, 0.57, dan 0.57. Kesimpulan: Terdapat kecenderungan penurunan aktivitas enzim fosfolipase pada biofilm C. albicans yang terhambat > 50% ekstrak etanol temulawak.

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Background: Javanese turmeric (Curcuma xanthorrhiza Roxb.) is one of medical plant from Indonesia that has potency to inhibit biofilm formation of C. albicans. Biofilm formation and hydrolyticenzymes are two among manyvirulence factors of C. albicans. Phospholipaseisone of hydrolyticenzymesthat could degrade the hostcell membrane. Objective: To observe the activities ofphospholipase in early phase, intermediate phase, and maturation phase of biofilm C. albicans ATCC 10231 that has been inhibited by Javanese turmeric ethanolic extract. Method: MTT-assay wasused to measure the minimum biofilm inhibitory concentration (MBIC50) of C. albicans ATCC 10231in three phases of C. albicans biofilm. Those concentrations were used to observe phospholipase activities of biofilm in the relevant phases. The negative control were not exposed to anything, while the positive control were exposed to Nystatin 100.000 IU. Phospholipase activities were determined bymeasuring the proportion of precipitation zone diameter and C. albicans colony diameter onan egg yolk-agar medium. Results: The MBIC50of Javanese turmeric ethanolic extract towards formation of C. albicans biofilm ATCC 10231 in early phase, intermediate phase, and maturation phase were 25%, 30%, and 35%, respectively. Phospholipase activities value in early phase, intermediate phase, and maturation phase of C. albicans biofilm exposed by Nystatin were 1. Phospholipase activities value in early phase, intermediate phase, and maturation phase of C. albicans biofilms exposed by Javanese turmeric ethanolic extract were 0.84, 0.80, and 0.83, respectively. Phospholipase activities value in early phase, intermediate phase, and maturation phase of unexposed C. albicans biofilm were 0.59, 0.57, and 0.57, respectively. Conclusion: There istendency of decreased phospholipase activity in early phase, intermediate phase, and maturation phase of biofilm C. albicans that has been inhibited by Javanese turmericethanolic extract.

Abstrak :
Pendahuluan: Salah satu faktor virulensi utama yang mempengaruhi perubahan Candida albicans(C. albicans) dari flora komensal menjadi patogen adalah pembentukan biofilm. Pada biofilm fase maturasi, C. albicansmenjadi lebih resisten terhadap agen antifungal. Telah dibuktikan efek inhibisi ekstrak etanol temulawak (Curcuma xanthorrhiza Roxb.) terhadap pertumbuhan biofilm C. albicans. Tujuan: Menganalisis gambaran TEM sel C. albicans ATCC 10231pada biofilm fase maturasi yang terinhibisi ekstrak etanol temulawak (EET). Metode: Penelitian ini dibagi dua kelompok yaitu kelompok perlakuan dan tanpa perlakuan. Sebanyak 100μL suspensi C. albicans ATCC 10231dalam 96-well-plate diinkubasi selama 1.5 jam pada 370C,kemudian diaspirasi dan dibilas menggunakan PBS. Pada kelompok perlakuan dipapar 100μLEET dengan konsentrasi KHBM50(35%), sedangkan kelompok tanpa perlakuan tidak dipapar EET. Kedua kelompok di inkubasi kembali hingga 48 jam, kemudian dipindahkan ke Eppendorf tube untuk difiksasi dalam 2.5% glutaraldehyde dan dilakukan pemeriksaan TEM. Kontrol positif dipapar nystatin oral suspension. Hasil: Pemeriksaan TEM pada kelompok perlakuan, sel C. albicansATCC 10231 pada biofilm fase maturasi yang terpapar ekstrak etanol temulawak terlihat perubahan gambaran ultrastruktur yang berupa perubahan bentuk sel, penebalan dinding sel, pembesaran vakuola, dan iregularitas sitoplasma berikut organel-organel di dalamnya seperti disorganisasi pada nukleus, mitokondria, dan retikulum endoplasma. Sedangkan pada kelompok tanpa perlakuan menunjukan gambaran sel C. albicans ATCC 10231 normal. Kesimpulan: Pemeriksaan TEM dapat menunjukkan perubahan sel C. albicans ATCC 10231 pada biofilm fase maturasi yang terinhibisi ekstrak etanol temulawak. ...... Introduction: One of main virulence factor that influence the alteration of Candida albicans (C. albicans) from commensal flora into pathogenic flora is biofilm formation. At the maturation phase, C. albicans ismore resistant to antifungal agent. It has been proven that there is an inhibition effect of Javanese turmeric (Curcuma xanthorrhiza Roxb) on the growth of C. albicans biofilm. Objective: To analyze the TEM image of C. albicans cell on the maturation phase of biofilm inhibit by Javanese turmeric ethanol extract. Method: This research divided into two groups, there were treated group and untreated group. A 100μLC. albicans ATCC 10231 suspension on a 96-well-plate incubated for 1.5 hour at 370C, and then aspirated and washed by phosphate buffer saline (PBS). The treated group was exposed to 100μL Javanese turmeric ethanol extract as MBIC50 concentration (35%), while the untreated group was not exposed to Javanese turmeric ethanol extract. Both groups were incubated until 48 h, and then moved into the Eppendorf tube and fixed with glutaraldehyde 2.5% to examine using TEM. The positive control was exposed to nystatin oral suspension. Result: Compared to the negative control, C. albicans cell on biofilm maturation phase treated by Javanese turmeric extract ethanol extract shows the alteration on the its ultra structure. The alteration showed in shape, the thickness of cell wall, the enlargement of vacuole, and irregularity of cytoplasm include the organelles in it such as disorganization on nucleus, cytoplasm, and endoplasmic reticulum. Conclusion: TEM examination showed the alteration of C. albicans ATCC 10231 cell on maturation phase biofilm inhibited by Javanese turmeric ethanol extract.

Abstrak :
Rimpang temulawak (Curcuma xanthorrhiza Roxb.) mengandung senyawa kurkuminoid dan xantorizol yang memiliki banyak manfaat untuk kesehatan. Kedua senyawa tersebut dapat diekstraksi dengan pelarut Natural Deep Eutectic Solvent (NADES) dengan metode UAE. Oksidasi yang terjadi secara alami ekstrak dapat mempengaruhi stabilitas ekstrak cair sehingga senyawa kurkuminoid dan xantorizol yang dapat mengalami degradasi. Penelitian ini menganalisis pengaruh penambahan BHA dan suhu penyimpanan terhadap stabilitas ekstrak cair NADES temulawak. Ekstrak NADES rimpang temulawak ditambahkan BHA hingga mengandung BHA sebanyak 30 ppm, 40 ppm, dan 50 ppm kemudian disimpan di tiga suhu penyimpanan berbeda, yaitu 30°C ± 2°C, 5°C ± 3°C, dan -20°C ± 5°C selama 54 hari. uji stabilitas yang dilakukan adalah uji stabilitas fisik (organoleptik dan homogenitas) dan uji stabilitas kimia (pH, penetapan kadar kurkuminoid dan xantorizol). Hasil penelitian menunjukkan bahwa ekstrak yang ditambahkan BHA memiliki stabilitas yang lebih baik daripada ekstrak tanpa penambahan BHA. Ekstrak cair NADES rimpang temulawak yang mengandung 50 ppm dengan suhu penyimpanan -20°C ± 5°C menunjukkan kestabilan fisik dan kimia yang lebih baik daripada ekstrak cair NADES rimpang temulawak tanpa BHA. Berdasarkan hasil uji One-way ANOVA terdapat pengaruh yang signifikan dari BHA dan suhu terhadap kadar kurkuminoid dan xantorizol dengan nilai p < 0,05. ......Javanese turmeric (Curcuma xanthorrhiza Roxb.) contain curcuminoid and xanthorrhizol which have many benefit for health. Both compounds can be exctracted with Natural Deep Eutectic Solvent (NADES) by UAE method. Oxidation which occur naturally in liquid extract can effect the stability of compounds so that curcuminoid and xanthorrhizol can undergo degradation.  This study analyzed the effect of adding BHA and storage temperature on stability of liquid extract of javanese turmeric. Liquid extracts are added with BHA with different concentration (30 ppm, 40 ppm, and 50 ppm) and stored at three different temperature (30°C ± 2°C, 5°C ± 3°C, dan -20°C ± 5°C) for 54 days. Stability test carried on this study include physical stability test (organoleptic and homogeneity) and chemical stability test (pH, determination of curcuminoid and xanthorrhizol). The result showed that the addition of BHA can improve the stability of liquid extract. Liquid extract with 50 ppm BHA with a storage temperature at -20°C ± 5°C showed better physical and chemical stability. Based on One-way ANOVA test, there is a significant influence of BHA and storage temperature on curcuminoids and xanthorrhizol with p value is p<0,05.

Abstrak :
Latar Belakang: Temulawak (Curcuma xanthorrhiza Roxb.), sebagai salah satu tanaman obat unggulan Indonesia, dilaporkan memiliki efek eradikasi terhadap biofilm Candida albicans (C. albicans). Studi analisis Scanning Electron Microscopy (SEM) melaporkan penurunan densitas biofilm serta perubahan morfologi sel C. albicans yang terpapar Ekstrak Etanol Temulawak (EET). Namun bagaimana perubahan ultrastruktur sel C. albicans pada biofilm fase maturasi yang tereradikasi EET belum diketahui. Tujuan: Menganalisis gambaran ultrastruktur sel C. albicans ATCC 10231 pada biofilm fase maturasi setelah paparan EET. Metode: Biofilm C. albicans diinkubasi selama 48 jam agar mencapai fase maturasi, kemudian kelompok perlakuan dipaparkan EET dengan konsentrasi sesuai kadar eradikasi biofilm minimal (KEBM) yaitu 30%, kelompok kontrol positif diberikan paparan nystatin 100000 IU/ml, dan kelompok kontrol negatif tidak diberikan perlakuan. Transmission Electron Microscopy (TEM) dipakai untuk melihat perubahan ultrastruktur sel C. albicans pada biofilm. Hasil: Perubahan gambaran ultrastruktur sel C. albicans pada biofilm fase maturasi yang dipaparkan EET meliputi perubahan bentuk sel, perubahan ketebalan dinding sel, serta kerusakan organel. Kesimpulan: Analisis TEM menunjukkan perubahan sel Candida albicans ATCC 10231 pada sebagian sel biofilm fase maturasi yang tereradikasi EET. ......Background: Javanese turmeric (Curcuma xanthorrhiza Roxb.), as one of Indonesia’s eminent medicinal plant, had been reported for having eradication effect towards Candida albicans biofilm. SEM studies have shown that the exposure of javanese turmeric ethanol extract decreases the biofilm density and made changes in C. albicans cells morphology. However, the changes in cellular ultrastructure of C. albicans in biofilm at maturation phase which has been eradicated by javanese turmeric extract is not yet known. Objective: Analyzing the image of cellular ultrastructure of C. albicans ATCC 10231 biofilm at maturation phase after exposed to Javanese turmeric ethanol extract using Transmission Electron Microscopy. Method: C. albicans biofilm was incubated for 48 hours to achieve maturation phase, then the treatment group was exposed to 30% javanese turmeric ethanol extract according to the minimum biofilm eradication concentration, the positive control group was exposed to nystatin 100000 IU/ml, and the negative control group wasn’t exposed to anything. TEM was used to observe the cellular ultrastructure changes of the C. albicans biofilm. Result: The changes observed in the image of cellular ultrastructure of C. albicans biofilm treatment group were different cellular shape, different cell wall thicknesses, and damaged organelles. Conclusion: TEM analysis showed the changes occurred in the image of some cells in C. albicans biofilm at maturation phase which has been eradicated by javanese turmeric ethanol extract.