Thin-layer Chromatography Results Research Articles

Identifikasi kandungan fucoidan yang diekstrak dari Sargassum sp. yang diambil dari Pantai Pantai Sundak, Gunung Kidul

This study aims to identify the fucoidan content extracted from Sargassum sp. collected from Sundak Beach, Gunung Kidul. Fucoidan is a sulfated polysaccharide compound with various biological activities, including anticoagulant, antitumor, and anti-inflammatory activities. Sargassum sp. was chosen as the source of fucoidan due to its abundance in coastal waters and its pharmacological potential. The extraction method was carried out using a hot water extraction technique with acid treatment, followed by a purification process using dialysis. The extracted fucoidan content was then analyzed using spectrophotometry and Thin Layer Chromatography (TLC) to determine its concentration and purity. Fourier Transform Infrared (FTIR) spectroscopy was used to identify the functional groups of the compound. The results showed that fucoidan was successfully extracted from Sargassum sp. with a significant yield. The main component of the extracted fucoidan from Sargassum sp. in this study was fucose, as indicated by the TLC results. The Rf value of the fucoidan hydrolysate from Sargassum sp. and the standard fucoidan, which was 0.51, matched that of fucose, indicating that both samples contained fucose. This study concludes that Sargassum sp. from Sundak Beach, Gunung Kidul, is a potential source for high-quality fucoidan production.

Identifikasi kandungan fucoidan yang diekstrak dari Sargassum sp. yang diambil dari Pantai Pantai Sundak, Gunung Kidul

This study aims to identify the fucoidan content extracted from Sargassum sp. collected from Sundak Beach, Gunung Kidul. Fucoidan is a sulfated polysaccharide compound with various biological activities, including anticoagulant, antitumor, and anti-inflammatory activities. Sargassum sp. was chosen as the source of fucoidan due to its abundance in coastal waters and its pharmacological potential. The extraction method was carried out using a hot water extraction technique with acid treatment, followed by a purification process using dialysis. The extracted fucoidan content was then analyzed using spectrophotometry and Thin Layer Chromatography (TLC) to determine its concentration and purity. Fourier Transform Infrared (FTIR) spectroscopy was used to identify the functional groups of the compound. The results showed that fucoidan was successfully extracted from Sargassum sp. with a significant yield. The main component of the extracted fucoidan from Sargassum sp. in this study was fucose, as indicated by the TLC results. The Rf value of the fucoidan hydrolysate from Sargassum sp. and the standard fucoidan, which was 0.51, matched that of fucose, indicating that both samples contained fucose. This study concludes that Sargassum sp. from Sundak Beach, Gunung Kidul, is a potential source for high-quality fucoidan production.

Kromatografi Lapis Tipis (KLT): Pendekatan Pola Kromatogram Untuk Mengkonfirmasi Rhodamin B Pada Perona Pipi

Blusher is a decorative cosmetic whose purpose is to beautify the face and emphasize the shape of the cheekbones. Rhodamine B is a synthetic dye in the form of a crystalline powder which is prohibited from being used in cosmetics according to the regulation by Head of The Indonesian Food and Drug Administration No. 18 of 2015 concerning Technical Requirements for Cosmetic Materials, because they are dangerous if contact with the skin. The purpose of this study was to determine rhodamine B in blusher which is traded in Kandat Market, Kediri Regency. In this study, five samples were used, namely sample A, sample B, sample C, sample D, and sample E. The qualitative analysis method used was Thin Layer Chromatography (TLC). In this method, the eluent N-butanol:ethyl acetate:ammonia is used in the ratio (55: 20: 25). TLC results showed visually in pink color, then when viewed under UV light 254 nm was yellow fluorescence. From the five samples, the Rf value was 0.52; 0.56; 0.61; 0.52 and 0.57, while the average Rf of the standard solution of rhodamine was 2.1. So that from the five samples it can be seen that they do not contain rhodamine B dye.

Antioxidant and hepatoprotective effects of novel heteropolysaccharide isolated from Lobularia maritima onCCl4-induced liver injury in rats.

The aim of the present study was to investigate the extraction and the characterization of a novel heteropolysaccharide from Tunisian halophyte Lobularia maritima (LmPS). We were also interested in its antioxidant, anti‐inflammatory, and hepatoprotective effects on carbon tetrachloride (CCl4)‐induced liver injury in rats. LmPS physicochemical properties were evaluated by thin‐layer chromatography (TLC), high‐performance liquid chromatography (HPLC), thermogravimetric analysis (TGA), and UV absorption. According to TLC and HPLC results, LmPS was a heteropolysaccharide composed of glucose, galactose, and xylose. Its molecular weight was 130.62 kDa. This heteropolysaccharide was characterized by a significant antioxidant potential and was efficient against oxidative stress and CCL4‐induced hepatotoxicity in rat Wistar models (n = 8) treated with a single dose of LmPS 250 mg/kg of body weight. This was evidenced by a significant increase in serum marker enzymes specially aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH). The cytokines released after stimulation of rats with LmPS showed high anti‐inflammatory profiles with an increased rate of interleukine‐10 (IL‐10) with 0.03 pg/mL compared to animals treated only with CCl4. On the contrary, we noticed a decrease of the other cytokines (tumor necrosis factor α: TNF‐α, interleukine‐6: IL‐6, transforming growth factor beta 1: TGF‐β1) with average concentration values of <0.2, 0.1, and 0.04 pg/mL, respectively. Besides, histopathological examinations revealed that CCl4 causes acute liver damage, characterized by extensive hepatocellular necrosis, vacuolization, and inflammatory cell infiltration, as well as DNA fragmentation. LmPS administration at a dose of 250 mg/kg resulted in a significant hepatoprotection, evidenced by a reduction of CCl4‐induced oxidative damage for all tested markers. These findings eagerly confirmed that LmPS was effective in the protection against CCl4‐induced hepatotoxicity and genotoxicity. It, therefore, suggested a potential therapeutic use of this polysaccharide as an alternative medicine for patients with acute liver diseases.

POTENTIAL OF ACTIVE COMPOUNDS LEAVES OF MANGO PARASITE (Dendrophthoe pentandra (L). Miq) AS ANTIFUNGAL

Infection is a disease that is easily found in tropical areas such as Indonesia. The causes of infection that are easily found include infections due to fungi. One of the fungi that causes many infections is the Candida fungus. Tropical drugs commonly used to treat cutaneous candidiasis include nystatin, clotrimazole, miconazole, ketoconazole and other azoles. However, antifungal drugs have limitations, such as severe side effects, poor penetration into certain tissues, and the emergence of resistance. To overcome the negative effects caused by these synthetic antifungal drugs, it is necessary to explore natural antifungal drugs. One of the plants that has the potential as a medicinal plant is the mango parasite plant (Dendrophthoe pentandra (L.) Miq) which is a parasitic plant that attaches to the host which has the potential as herbal medicine. Research on the potential of active compounds in the leaves of mango parasite (Dendrophthoe pentandra (L). Miq) as antifungals has been carried out from November 2020 to June 2021 at the Genetics and Biotechnology Laboratory, Biology Department, FMIPA Sriwijaya University. The purpose of this study was to obtain fractions that had antifungal activity, to obtain isolates that had antifungal activity, what active compounds were present in the leaves of the mango parasite as antifungal, and to determine the minimum inhibitory concentration of the fungus Candida albicans. Based on research that has been carried out with methanol, n-hexane and ethyl acetate extracts that are active as antifungals, namely n-hexane and ethyl acetate extracts with an n-hexane inhibition zone diameter of 12 mm while ethyl acetate is 10 mm. Then purification of the n-hexane and ethyl acetate fractions was carried out using gravity column chromatography and the n-hexane fraction obtained two pure compounds that were active as antifungals, namely isolate N-1 with a diameter of 12 mm and N-4 with a diameter of 10 mm. Two pure acetate fractions were obtained which were active as antifungals, namely isolates E-1 with a diameter of 10 mm and E 5 with a diameter of 12 mm. Furthermore, the results of the eluate were tested by MIC and thin layer chromatography (TLC). The MIC results on isolates N-1, N-4, and E-1 were able to inhibit at a concentration of 500 ppm and were classified as strong. Meanwhile, isolate E-5 was able to inhibit at a concentration of 1000 ppm which was classified as moderate. The TLC results showed color spots on isolate N-1 which were blue, namely terpenoids, while in isolate N-4 which was orange in color were alkaloids, while in isolate E-1, the color was phenolic yellow, while in isolate E-5, which was purple, it was group terpenoids. The results of the bioautography of isolate N-1 with R¦ value: 0.9, isolate N-4 with R¦: 0.6, isolate E-1 with R¦ value: 0.8, and isolate E-5 with R¦ value: 0,9.

Isolation and Identification of New Pigment From Monascus purpureus

This research was aimed to isolate and identify the new pigment from the main pigments of Monascus purpureus, which derive from five different isolates and through with solid fermentation used IR-64 rice as substrate. The measurement of pigment was done by extracting t samples using methanol in the 14th day of fermentation process. The absorbance of main pigment was measured by a UV-Visible spectrophotometer at the wavelengths of 364, 365, 375, 400, 480 and 500 nm. The results showed that the absorbance of each pigments isolate varied within ranges from 0.219 to 0.559. Result followed of Thin Layer Chromatography (TLC) method with chloroform: acetone (9: 1). TLC results were followed by isolating IPB-B samples using Thin Layer Chromatography- Preparative (TLC-P), whereas the band obtained was identified by using Liquid Chromatography-Mass Spectrum (LC-MS) instrument. The isolated and identified IPB-B samples showed that there was found a new compound of Molecular Weight (MW), which was Monaphilol A with Molecular Weight (MW) of 384.47.

Quality and composition of Albendazole, Mebendazole and Praziquantel available in Burkina Faso, Côte d'Ivoire, Ghana and Tanzania.

BackgroundEven though the international combat against Neglected Tropical Diseases such as schistosomiasis or soil-transmitted helminthiases depends on reliable therapeutics, anthelminthic pharmacovigilance has been neglected on many national African drug markets. Therefore, quality and composition of Albendazole, Mebendazole and Praziquantel locally collected in Burkina Faso, Côte d’Ivoire, Ghana and Tanzania were analysed.MethodsSamples of 88 different batches were obtained from randomly selected facilities. Sampling took place in Northwest Tanzania, Western Burkina Faso, Southeast Côte d’Ivoire and Southwest Ghana. Visual examination of both packaging and samples was performed according to the WHO ‘Be Aware’ tool. Products were then screened with the GPHF Minilab, consisting of tests of mass uniformity, disintegration times and thin-layer chromatography (TLC). Confirmatory tests were performed according to international pharmacopoeiae, applying assays for dissolution profiles and high-performance liquid chromatography (HPLC).FindingsDespite minor irregularities, appearance of the products did not hint at falsified medicines. However, 19.6% of the brands collected in Ghana and Tanzania were not officially licensed for sale. Mass uniformity was confirmed in 53 out of 58 brands of tablets. 41 out of 56 products passed disintegration times; 10 out of the 15 failing products did not disintegrate at all. Evaluating TLC results, only 4 out of 83 batches narrowly missed specification limits, 18 batches slightly exceeded them. Not more than 46.3% (31 / 67) of the tablets assayed passed the respective pharmaceutical criteria for dissolution. HPLC findings confirmed TLC results despite shifted specification limits: 10 out of 83 tested batches contained less than 90%, none exceeded 110%.ConclusionIn the four study countries, no falsified anthelminthic medicine was encountered. The active pharmaceutical ingredient was not found to either exceed or fall below specification limits. Galenic characteristics however, especially dissolution profiles, revealed great deficits.

Phytochemical screening and antibacterial activity of essential oils of Cymbopogon citratus and Ocimum americanum : a comparative account

The present study highlights the extraction of essential oils from C. citratus and O. americanum using Microwave Hydrodiffusion and Gravity method (MHG) and Hydrodistillation Method (HD), the qualitative and quantitative screening along with determination of antibacterial potential. For the study, extraction of essential oils was performed using Microwave Hydrodiffusion and Gravity method and Hydrodistillation Method. Comparative study was performed to check the total flavonoid content (TFC), total phenolic content (TPC), biochemical test, thin layer chromatography (TLC) and determination of antibacterial activity of the essential oils. The results of the study depicted that the MHG method for extraction of essential oils was much better compared to extraction by HD method as supported by the biochemical tests, TFC, TPC and TLC results. However, the essential oils did not show antibacterial activity. More study related to the antimicrobial activity needs to be performed. Essential oils form an integral part of the pharmaceutical and healthcare industry hence, detailed study regarding the isolation and application of essential oils is required. The present study suggests the use of MHG method for extraction of essential oils.

Design and Evaluation of Vaginal Douche Formulation for the Ethanolic Extract of Cabbage (Brassica oleracea var. Capitata alba) as Anti-Flour Albus

The purpose of this present study was to determine the most suitable vaginal douche formulations of the Brassica oleracea var. capitata alba ethanolic extract as anti-flour albus. Determination of the basic vaginal douche formula was carried out using different thickener substances, as follows: carbopol 980, esaflor HM22 and SMC-20 viscolam. The concentration of acnibio Ac as a preservative with no antifungal activity against Candida albicans, was determined. The B. oleracea extract with different concentration (7 & 8.75%w/v) were formulated in the suitable basic formula. All formulas were stored at room temperature for 56 d and periodically evaluated for the stability of organoleptic characteristics, pH, and density. The effect of formulation on the active substance, existence was analyzed using thin layer chromatography (TLC). The antifungal effectivity of the formulas were evaluated using the agar diffusion method and its antifungal strength was compared to Povidone iodine solution. Among basis formula, the formula with SMC-20 viscolam provided the best base solubility. As a result of preservative concentration test, it was found that acnibio Ac at a concentration of 0.0125% w/v did not provide antifungal activity against C. albicans. The evaluation results showed that all vaginal douche formulas with varying extract concentrations were stable. Based on TLC results, the formulation did not showed the loss of active substance in the extract. The formula with cabbage extract concentration of 8.75% w/v revealed the best results and provided a good strength as an herbal preparation in compared to povidone iodine, because it has a small comparative value of 1: 0.2576. Thus, the cabbage leaf ethanolic extract concentration of 8.75% w/v can be recommended as reliable and effective active agents to be formulated as a vaginal douche using the studied formula.

ANALYSIS OF ILLICIT DRUGS AND NICOTINE IN A BUCCAL TOBACCO BRAND MARKETED IN YEMEN

Objectives: The objective of the present study was to check the potential presence of illicit drugs and to quantify the amount of nicotine in a buccal tobacco brand that had been observed to be increasingly used by Yemeni youths, since 2014, causing narcosis resembling states among them.&#x0D; Methods: Thin-layer chromatography (TLC) described by the United Nations Office on Drugs and Crime (UNODC) was used to screen illicit drugs in the tested brand. The illicit drugs investigated included opiates, heroin, amphetamines, and cocaine. The TLC results were confirmed as recommended by the UNODC using color chemical tests. Identification and quantification of nicotine in the brand was carried out using an appropriate high-performance liquid chromatography (HPLC) system.&#x0D; Results: No illicit drug was found in the tested tobacco brand. On the other hand, it was found that the amount of nicotine in just a single dose (sachet) of the buccal brand was 17.67±0.901 mg, which was 3.53-fold greater than usual buccal dose of nicotine (5 mg).&#x0D; Conclusion: With the exception of cannabis, opioids, and hallucinogens that were not investigated in this study due to technical obstacles, other major illicit narcotic drugs are not found in the brand. The brand contains high amount of nicotine/sachet. However, knowing that the user may use more than one sachet of the brands a day, there is a great potential of nicotine overdosing due to intake of the brand. This may cause a narcosis resembling state called “Nesbitt’s paradox,” characterized by reducing neuronal activity of the user.

Antioxidant activity of Indonesian water spinach and land spinach (Ipomoea aquatica): A comparative study

Ipomoea aquatica is Convolvulaceae plant that is widely consumed in Southeast Asia as a vegetable. The plant contains various bioactive components, e.g. phenols and flavonoids. Water spinach and land spinach are two varieties of Ipomoea aquatica in Indonesia. Both extracts have been analyzed for their free radical scavenging activity using DPPH (2,2-diphenyl-1-picrylhidrazyl). Total phenol was measured using Folin Ciocalteu and the total flavonoid was measured using AlCl3. Both extracts also have been analyzed by TLC (Thin layer chromatography) using DPPH and AlCl3 spray reagents. Water spinach has higher antioxidant activity than land spinach and there was a correlation between antioxidant activity and total phenol/flavonoid content. Based on TLC results, it was supposed that the compound responsible for antioxidant activities in water spinach and land spinach was flavonoid.

Chitosan Nanoparticles as a Carrier for Indigofera intricata Plant Extract: Preparation, Characterization and Anticancer Activity

Background: Cancer is one of the major causes of the death and affects people of all ages throughout the world. The drugs that are currently available to treat cancer have many side effects. Hence, there is considerable scientific interest in the continuing discovery of new anticancer agents from natural sources. The aim of this study was to prepare and characterize nanoparticles combining Indigofera intricata crude alcoholic extract and chitosan and to evaluate the anticancer cell proliferative activity for both extract and nanoparticles. Methods: Dried alcoholic extract was prepared and characterized for its phenolic and flavonoid contents. Chitosan extract nanoparticles was prepared by ionic gelation method and characterized by thin layer chromatography (TLC), Fourier-transform infrared spectroscopy (FTIR), particle size and zeta-potential analysis. The anticancer cell proliferative activities of both plant extract and nanoparticles at different concentrations were evaluated using breast cancer cell line (MCF 7). Results: The alcoholic extract showed high contents from both phenolic and flavonoid constituents (15 % and 22 % respectively). The interaction of polyphenolic compounds of the extract with chitosan was confirmed by the TLC and FTIR results. The particle size and zeta-potential of nanoparticles found to be 400.6nm ± 101.8 nm and +42.1 mV ± 9.27 mV respectively. The plant extract showed the lowest cell viability of 45.21% ± 4.8% at the highest dose (250 mg) tested in this investigation. Almost 500-fold reduction (from 250 mg to 0.5 mg) in the extract concentration required to achieve same anticancer cell proliferative activity when formulated as nanoparticles. Also 2.5 mg extract containing nanoparticles showed similar anticancer cell proliferative activity as 5 mg 5-FU. Conclusion: Our results revealed that traditional medicinal plants could be an excellent source of natural anticancer agents and the chitosan-extract nanoparticles is a promising formulation strategy to enhance their clinical effectiveness.

Biocolorants Production by Pigment-Producing Bacteria Isolated from Soil

Background: The use of synthetic organic colors has been acknowledged for many years as the most reliable and economical method of restoring some of the food’s original shade to the processed products. However, from the health safety point of view, they are not accepted by consumers because they produce skin allergies, less stable and also produce highly toxic wastes that pose a threat to the environment.&#x0D; The Aim of the Study: The aim was to isolate and identify pigment-producing bacteria from soil and to study various growth parameters for their pigment production.&#x0D; Materials and Methods: Soil samples were collected from different site within Sokoto State metropolis and were screened on nutrient agar for isolation of pigment-production bacteria. The isolated pigment-producing bacteria were subjected morphological, biochemical and molecular characterization. The phylogenetic analyses of bacterial isolates were carried out using Molecular Evolutionary Genetics Analysis (MEGA 6 software). Ethanol, methanol and chloroform were used for pigments extraction and the extracted pigments were characterized using Ultraviolet-Visible spectroscopy, Fourier Transformed Infrared (FTIR) spectroscopy and thin-layer chromatography (TLC). The effects of growth medium, pH, temperature, incubation time, shaking and static conditions on pigments production was determined. The stability of the pigments was tested toward pH and temperature.&#x0D; Results: Three (3) isolates that showed purple, orange and blue-green pigment were selected for pigment productions. The isolates were identified as Chromobacterium violaceum, Pseudomonas aeruginosa and Salinococcus roseus. The optimization studies revealed that the Chromobacterium violaceum produced highest purple pigment in nutrient broth at pH 8 for 96 hours of incubation at 35°C under shaking condition while Pseudomonas aeruginosa produced green pigment in nutrient broth at pH 7, 72 hours of incubation at 37°C under shaking condition and Salinococcus roseus produced highest orange pigment on nutrient broth at pH 7, after 96 hours of incubation at 40°C under shaking condition. The characteristics of the pigments corresponded to that violacein, pyocyanin and zeaxanthin based on their FTIR, UV-visible spectroscopy and TLC results. It was found that all the pigments showed good stability at the temperatures of 200°C and fairly stable at lower pH (2).&#x0D; Conclusion: It therefore concluded that the soil could be the source for isolating pigment-producing bacteria that would offer various industrial applications such textile industries.

PRODUCTION OF MALTOOLIGOSACCHARIDES FROM HUTAN JATI VARIETY CULTIVAR TACCA (Tacca leontopetaloides) STARCH

This research aimed to extract and characterize the physicochemical properties of starch from Tacca tuber, to determine the optimum conditions for enzymatic hydrolysis to produce maltooligosaccharides, and to analyze the character of these maltooligosaccharides. The analysis was conducted by calculating the amount of reducing sugar, total sugar, and the degree of polymerization, and by using the TLC (Thin Layer Chromatography) and HPLC (High-Performance Liquid Chromatography) analyses. The Hutan Jati variety cultivar of Tacca was selected from three Tacca variety cultivars (Hutan Jati, Pulau Katang and Gunung Batur) to produce maltooligosaccharides by enzymatic hydrolysis of crude Brevibacterium sp. α-amylase. The optimum conditions for the enzymatic hydrolysis of Hutan Jati variety cultivar Tacca starch for the production of maltooligosaccharides were obtained at a substrate concentration of 3% (w/v) and a ratio of 1:5 enzyme and substrate at 6 hours incubation time. From 250 mL of fresh hydrolysate, some 34.49 grams of powder maltooligosaccharide were produced. The TLC and HPLC results showed a similar yield of both the liquid and powder maltooligosaccharides with maltose, maltotriose, and maltotetraose as the main products. Considering its physicochemical characteristics and the product of its maltooligosaccharides, the starch from the tuber of Hutan Jati variety cultivar Tacca possessed strong potential for the future production of maltooligosaccharides particularly,  maltotriose and maltotetraose, in food industries.

Molecular identification of aflatoxigenic Aspergillus species in feedstuff samples.

Background and Purpose:Aflatoxins are naturally produced by some species of Aspergillus, such as A. flavus and A. parasiticus. Aflatoxins reportedly have carcinogenic effects on human, poultry, and livestock, and therefore could be linked to severe human illnesses. Aflatoxin biosynthesis pathway involves different clustered genes, including structural, regular, and unassigned genes. The present study was conducted to detect aflR, aflP, and aflD as three important genes contributing to aflatoxin B1 production cycle in Aspergillus species isolated from the feedstuffs of animal husbandry.Materials and Methods:This study was conducted on 25 isolates of A. flavus, A. parasiticus, A. nomius, and A. nidulans, isolated from animal feedstuff as a test group. The test group was compared with two standard strains (i.e., A. flavus and A. parasiticus) as aflatoxigenic reference organisms and negative controls (i.e., A. fumigatus, A. fusarium, and A. penicillium) in terms of the presence of aflR, aflP, and aflD genes using polymerase chain reaction (PCR). The determination of the toxigenicity and aflatoxin production of isolated Aspergillus species was accomplished using thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC).Results: The results obtained by the amplification of the selected genes by PCR method for the detection of aflatoxigenic Asprgillus species were significantly correlated with TLC and HPLC results. Accordingly, all samples, having positive results for aflatoxin B1 production in TLC and HPLC, were able to show the amplification of three target genes. However, 4 cases out of 6 (66%) non-aflatoxigenic isolates were positive for three or two genes.Conclusion: Based on the findings, the molecular detection of aflatoxin biosynthesis genes (i.e., aflP, aflD, and aflR) could be considered as a quick and reliable method for the detection of aflatoxigenic Aspergillus. Furthermore, this method could be useful in planning and implementing strategies targeted toward improving the safety of human or animal food.

Two factor at a time approach by response surface methodology to aggrandize the Bacillus subtilis KLP2015 surfactin lipopeptide to use as antifungal agent

A Gram’s positive, rod shaped and endospores-forming ‘bacterial isolate KLP15′ was identified as Bacillus subtilis (Accession number KT459335) by the biochemical tests and 16S rRNA based sequencing. Lipopeptide(s) production from B. subtilis KLP2015 increased from 200.0 to 547.0 mg/l after the optimization of physico-chemical parameters like choice of nitrogen source, fermentation time, fermentation temperature and broth pH by ‘One Factor at a Time’ (OFAT) approach. Further, the yield of lipopeptide(s) increased to 985 mg/l from an earlier value of 547 mg/l when a “Two Factors at a Time (TFAT) approach using Response Surface Methodology (RSM) was attempted. The extracted crude lipopeptide fraction from the cell-free fermentation broth of B. subtilis KLP2015 was purified and characterized by High Performance Liquid Chromatography (HPLC), Thin Layer Chromatography (TLC), UV-Visible Spectrophotometry (UV-vis), Fourier Transform Infrared (FTIR) and MALDI-TOF studies. HPLC and TLC results confirmed the purity of the lipopeptide preparation and its Surfactin-type nature. The FTIR and MALDI-TOF spectra of the purified lipopeptide were found to be similar to that of an authentic Surfactin molecule. The purified lipopeptide showed potent 75.1% and 41.9% antifungal activity against Mucor sp. and Aspergillus niger, respectively on the basis of zone of inhibition of growth of these fungal strains. Moreover, the purified lipopeptide preparation of B. subtilis KLP2015 was most effective against the growth of Mucor sp. as reflected by the recorded MIC values of 6.25 µg/ml while MIC of lipopeptide for the A. niger was 12.5 µg/ml.

ANTIBACTERIAL ACTIVITY OF DRACONTOMELON DAO EXTRACTS ON METHICILLIN-RESISTANT S. AUREUS (MRSA) AND E. COLI MULTIPLE DRUG RESISTANCE (MDR).

Background:Staphylococcus aureus, methicillin-resistant and Escherichia coli, multidrug-resistant included in the list of antibiotic-resistant priority pathogens from WHO. As multidrug-resistant bacteria problem is increasing, it is necessary to probe new sources for identifying antimicrobial compounds. Medicinal plants represent a rich source of antimicrobial agents. One of the potential plants for further examined as antibacterial is Dracontomelon dao (Blanco) Merr. & Rolfe. The present study designed to find the antibacterial activity of D. dao stem bark extracts on Methicillin-resistant S. aureus (MRSA) and E. coli Multiple Drug Resistance (MDR), followed by determined secondary metabolites with antibacterial activity and determined the value of MIC (minimum inhibitory concentration) and MBC (minimum bactericidal concentration).Materials and Methods:D. dao stem bark extracted using 60% ethanol. Disc diffusion test methods used to find the antibacterial activity, following by microdilution methods to find the value of MIC and MBC. Secondary metabolites with antibacterial activity determined by bioautography using TLC (thin layer chromatography) methods.Results:D. dao stem bark extracts are sensitive to MSSA, MRSA and E.coli MDR bacteria. The inhibition zone is 16.0 mm in MSSA, 11.7 mm in MRSA and 10.7 mm in E. coli MDR. The entire MBC/MIC ratios for MSSA, MRSA and E.coli MDR is lower than 4. The ratio showed bactericidal effects of D. dao stem bark extracts. In TLC results, colorless bands found to be secondary metabolites with antibacterial activity.Conclusion:D. dao stem bark extracts are potential to develop as antibacterial agent especially against MRSA and E. coli MDR strain.

ANTIBACTERIAL ACTIVITY OF ETHANOLIC EXTRACT AND INFUSE OF Bauhinia variegata LEAVES AGAINST Streptococcus pyogenes

Bauhinia variegata is a plant that has many virtues and widely grown in Indonesia. Yet the virtues and uses are still limited as the shade and ornamental plant. The purpose of this study is to determine the antibacterial ethanol extract activity and the infuse of Bauhinia Variegata leaves toward Streptococcus pyogenes and to determine the classes of compounds that have antibacterial activity. Bauhinia variegata leaves are extracted by maceration and infundation with 96% ethanol and distilled water. The test method of antibacterial activity is used disc difusson Kirby Bauer methods. The Analyses of the compounds which are contained in the Bauhinia Variegata leaves are done by TLC (Thin Layer Chromatography) and test tubes. The results showed that ethanol extract and infuse of Bauhinia Variegata leaves has no antibacterial activity toward Streptococcus pyogenes. TLC results and test tubes showed that the ethanol extract of Bauhinia Variegata leaves contain tannins, saponins, terpenoids, and alkaloids. While, the infuse of Bauhinia variegata leaves are saponins and alkaloids. Keywords: Bauhinia variegata, antibacterial, Streptococcus pyogenes, maceration, infundation.

ANALISIS SILDENAFIL SITRAT DALAM JAMU KUAT DI KECAMATAN BOGOR BARAT DAN TANAH SAREAL DENGAN MENGGUNAKAN KROMATOGRAFI CAIR SPEKTROMETRI MASSA

Herbal medicineisthe treatment of choicebecause ofmerit in maintainingthe health ofthe peopleof the human body. Strongherbal medicineisherbal medicinethat isefficacious asstaminaandvitality ofman, but in its developmentpuritypowerfulherbsare addedsildenafilcitratebyparties who are notresponsible. This study was adescriptiveanalysis ofexperimentaldesign. Analysis of sildenafilcitrate compounds using Thin Layer Chromatography (TLC) and Liquid Chromatography Mass Spectrometry (LC-MS) were sampled in two districts, namely sub-district, Bogor Barat and Tanah Sareal thus obtained 32 samples, of which 18 samples of Bogor Barat coded A and 14 samples from Tanah Sareal coded B. TLC results were eluted at Silica gel 60 F254 with eluent methanol:chloroform (4:1), there were 5 samples showed close Rf value standard sildenafil citrate is 0.83, the sample code A4 value of Rf 0.84, A13 Rf value of 0.84, A17 Rf value of 0.83, B11 Rf value of 0.83, and B14 Rf value of 0.84. Results LC-MS where separation using column Phenomenex Synergi Fusion RP-100A, the pumping system with a mobile phase gradient of 0.2 M ammonium acetate pH 7 in water and 0.2 M ammonium acetate pH 7 in acetonitrile, was obtained the samples for A4, A13, A17, B11 dan B14 amounted to 198.01; 1734.14; 226.00; 156.77; 443.87 μg/kg

Degradation of Fat by a Bioaugmentation Product Comprising of Bacillus spp. Before and After the Addition of a Pseudomonas sp

A bioaugmentation product, BFL, comprising strains of the genus Bacillus, is evaluated for its ability to degrade fat in laboratory‐scale experiments. Addition of a Pseudomonas putida strain CP1 to the commercial mixed population (BFL‐CP1) is tested for optimization of fat degradation. Experiments are carried out in aerobic batch culture, at 30 °C and 150 rpm for 13 days incubation. A minimal medium (MM) and an enriched nutrient medium (ENM) are investigated supplemented with 1% (w/v) butter. Fat removal is determined gravimetrically and the lipid content is analyzed using thin layer chromatography (TLC) and gas chromatography (GC). No degradation of butter by the product is recorded after 13 days of incubation, while up to 97% degradation is observed by BFL‐CP1. All the Bacillus isolates produced lipase but not the Pseudomonas putida. TLC and GC results suggested that while the Bacillus spp. hydrolyzed the fat to fatty acids and glycerol, complete metabolism of the breakdown products only took place in the presence of the Pseudomonad sp. A citrate buffer is used to investigate fat removal by BFL‐CP1in low and stable pH using citrated minimal buffer. Similar fat removal is observed. The use of citrated minimal buffer caused flocculation of the mixed culture, a phenomenon desirable for fats, oils, and grease (FOGs) degradation in grease traps.Practical Applications: The bioaugmentation product, BFL, comprising strains of the genus Bacillus, only promoted high fat removal in a few days incubation after the addition of a Pseudomonas putida strain CP1. Analysis of the remaining fat suggested a cooperative activity between the Bacillus spp., which hydrolyzed the fat to fatty acids and glycerol, and the Pseudomonas putida CP1, which assimilated the released fatty acids. Formation of flocs were observed when the inoculum was tested under different environmental conditions with low pH. This phenomenon is desirable and along with the high degradative ability of the that new inoculum, it showed good potential for use in the treatment of FOG in grease traps.Fats, oils, and greases (FOGs) are generated in high amounts from food facilities presenting potential blockages and wastewater management problems. FOG may be intercepted at source using grease traps and may be treated biologically in situ using bioaugmentation, an environmentally desirable approach that involves the introduction of suitable microorganisms. In this study, combination of Gram‐positive and Gram‐negative bacteria successfully cooperated and degraded the FOG breaking down the lipids to fatty acids and glycerol by the activity of lipases, and also assimilating the produced fatty acids to carbon dioxide and water through the β‐oxidation process.