TLC Chromatogram Research Articles

Kinetic optimization of angkak – red ginger extraction and its impact on antioxidant activity

A mixture of angkak and red ginger is a Chinese herbal concoction with potential as an antioxidant. This herbal combination can be extracted using the kinetic hot maceration method by optimizing the stirring speed to be more efficient. Previous studies have shown that using 400, 600, and 800 rpm stirring speeds provides the best IC50 and yield. Based on these problems, this research aims to determine the effect of increasing stirring speed on IC50 and % yield in the extraction of angkak and red ginger, as well as prove the benefits of this herbal combination through the IC50 value. The method used is extracting a mixture of angkak and red ginger at kinetic variations of 400, 600, and 800 rpm using a magnetic hotplate stirrer at a temperature of 60ºC for 2 hours; determination of % yield; determining the chromatogram profile using TLC; determination of antioxidant activity using DPPH; as well as data analysis of percentage yield and IC50 using SPSS. Analysis using SPSS shows that the stirring speed significantly affects the % yield, where increasing the moving speed above 400 rpm results in a decrease in the % yield. However, the moving speed does not affect the IC50. The TLC chromatogram profile shows the presence of 6-gingerol compounds in angkak and red and yellow pigment compounds in red ginger, which have potential as antioxidants. The optimization results in this study obtained optimal % yield and IC50 at 400 rpm with average values, respectively, namely 15.933 ± 3.4771% and 103.76 ± 10.032 ppm, and the combination of angkak and red ginger ingredients can work synergistically to increase yield and antioxidant.

Screening and Molecular Characterization of Antibacterial Secondary Metabolite Producing Actinomycetes from Soils of Eastern Mountain Regions of Nepal

Antibacterial secondary metabolite is a bioactive compound like antibiotic that can be considered a substance, produced by one microorganism, which in low concentration inhibits the growth of other microorganisms. Actinomycetes, slow-growing gram-positive bacteria, are the major sources of bioactive compounds. This study aimed to screen and identify antibacterial secondary metabolite-producing actinomycetes by sequencing the 16S rRNA method (molecular identification) from the soils of the mountain region of eastern Nepal. Starch casein agar (SCA) medium was used for the isolation of actinomycetes which were confirmed by primary screening and secondary screening. Identification of presumptive genera was done based on macroscopic, microscopic, and biochemical characteristics and confirmed by sequencing their 16S rRNA genes. The antibacterial compound was produced by culturing the potential isolate in starch casein broth. Using organic solvents such as ethyl acetate, n-butanol, chloroform, dichloromethane, and methanol, the chemical was recovered from the fermented broth. TLC performed the antibacterial substance characterization. Only 9 (13.6%) of the 66 actinomycetes isolates showed antibacterial activity against test microorganisms. Only one of the nine isolates, M3, had antibacterial activity in primary screening against gram-positive (Staphylococcus aureus) and gram-negative (Escherichia coli, Salmonella Typhi, Shigella spp., and Pseudomonas aeruginosa) test bacteria. M3 was chosen for secondary screening due to its strong antibacterial activity. The minimum inhibitory concentration (MIC) of crude antibacterial substances was found to be 2.5 mg/mL against test organisms. According to the TLC chromatogram, the isolate produced only one compound with an Rf value of 0.81, completely distinct from the spot formed by gentamicin (standard), which had an Rf value of 0.89. The isolates were considered Streptomyces spp., a distinct taxonomic group based on characterization by macroscopic, microscopic, biochemical, physiological, and molecular techniques. This study concluded that Mountain regions are the reservoir of antibiotic- producing actinomycetes. Streptomyces is the most common genus.

Identification and characterization of major bioactive compounds from Andrographis paniculata (Burm. f.) extracts showed multi-biomedical applications

The Andrographis paniculata recognized as most valuable medicinal plant in folk medicine. Hence, this research was designed to evaluate antibacterial potential of petroleum ether (PE) and methanol (ME) extracts of A. paniculata against skin infection causing bacterial pathogens such as Staphylococcus aureus, Streptococcus pyogenes, Klebsiella pneumoniae, Enterobacter aerogenes, Proteus vulgaris, and Propionibacterium acnes. Also assessed the antidiabetic (α-glucosidase and α-amylase inhibition assay), antioxidant, and photoprotective potential of PE and ME extract analyses. The major bioactive compounds were identified and characterized through UV, FTIR, 1H-NMR and 13C-NMR spectra analyses. The ME extract contain more number of phytochemicals (alkaloids, flavonoids, saponins, terpenoids, glycoside, protein, and phytosterol) than PE extract. The antibacterial activity result also revealed that the ME (as dose dependent) extract showed better activity at 250 mg mL−1 as in the following order: P. acnes (6–29 mm) > K. pneumoniae (3–28 mm) > S. aureus (3–27 mm) > P. vulgaris (3–26 mm) > S. pyogenes (2–25 mm) > E. aerogenes (1–23 mm). PE: E. aerogenes (3–20 mm) > P. vulgaris (2–19 mm) > P. acnes (3–18 mm) > K. pneumoniae (3–17 mm) > S. aureus (2–16 mm) > S. pyogenes (0–11 mm). The MIC value of ME extract was found as 100–150 mg mL−1 and it was better than PE extract. Similarly, the ME also possesses dose based α-glucosidase inhibition activity as up to 85% at 250 mg mL−1 concentration. The fluorescence spectra analysis method also stated that the ME extract possess photoprotective bioactive agent. The ME fractions (F01 and F02) obtained from TLC and column chromatogram were identified as 3-O-β-d-glucosyl-14- deoxyandrographiside and 14-deoxyandrographolide respectively through UV, FTIR, 1H-NMR and 13C-NMR spectra analyses. Such compounds may be responsible for significant antibacterial activity against pathogenic bacteria causing skin infections, excellent antidiabetic activity, as well as photoprotective potential.

Antioxidant properties of Carallia brachiata stem bark and leaf

Carallia brachiata (Lour.) Merr. (Rhizophoraceae) had been used to treat various human disoders. This study reported antioxidant activity of C. brachiata of stem bark and leaf in various solvents. Each sample was macerated in various solvent separately. By using DPPH, ABTS, and FRAP assays, antioxidant activity was determined. The TLC method was created using dichloromethane and ethyl acetate mobile phase. The methanol extract of stem bark contained the highest TPC value (262.73 ± 1.58 mg GAE/ g extract), while the methanol extract of leaf contained the highest TFC value (45.46 ± 0.32 mg QE/ g extract). The total phenolic content of stem bark extract was higher than that of leaf extract, but the flavonoid content of the leaf extract was marginally higher. The methanol extract exhibited the highest antioxidant activity for DPPH (13.06±0.08 μg/mL), ABTS (7.19±0.08 μg/mL), and FRAP assay (155.19±0.34 μM Fe2+/g). In this research, stem bark has greater antioxidant activity than the leaf. In addition, the results of TLC profile were detailed. The TLC chromatogram indicates that C. brachiata stem bark contains more components than leaf. This study indicates that C. brachiata has a rich source of prospective antioxidants.

SEPARATION OF ANTIOXIDANT COMPOUNDS FROM THE STEM BARK EXTRACTS OF HAEMATOSTAPHIS BARTERI HOOK F. (ANACARDIACEAE) USING CHROMATOGRAPHIC TECHNIQUES

In the search for plants with antioxidant properties, Haematostaphis barteri Hook f. (Anacardiaceae) was found to be a plant often used in Nigeria and other nations for the management of cancer, anemia, haemorrhoid and many other ailments. The ethyl acetate and acetone extracts of the stem bark in previous research by the same author were assayed for phenolic compounds and the phenolic compounds were found to possess antioxidant properties when TLC chromatograms were stained with iodine vapour and several spots were revealed. The ethyl acetate and acetone fractions were further chromatographed on TLC with several solvent systems and the mixtures from toluene, ethyl acetate and formic acid (TEF) in the ratio of 4/2.5/3.5 and 3/5/2 and n-butanol, acetic acid and water (BAW) in the ratio of 12/4/6 gave better resolutions. When chromatograms were obtained and developed with a solution of p-anisaldehyde in concentrated sulphuric acid, different spots were visualizedwith different colours signifying the presence of different bioactive metabolites. The Presence of antioxidant compounds was determined by bioutography TLC chromatograms were stained with a solution of 0.2% DPPH (2, 2-Diphenyl-1-picrylhydrazyl) solution in methanol and dried with heat gun and the appearance of yellow spots on purple background signifies the presence of antioxidants. A selected spot with an Rf value of 0.31 was monitored and isolated from column chromatography. The purified bioactive metabolite was crystallized, weighed and saved for further analysis.

Qualitative and Quantitative Estimation of Total Phenols in Narcissus tazettaL. Bulbs

Narcissus tazetta, a member of the Amaryllidaceae family, is known to be rich in bioactive metabolites such as alkaloids, phenolics and flavonoids, which have been found in nearly every species in this family. N. tazetta cultivated in Iraq, had not previously been studied for its active components; thus, the current study used phytochemical screening and phenolic compounds estimation, both qualitatively and quantitatively. Results showed that the plant alcoholic extract was rich in alkaloids, polyphenols and flavonoids besides tannins, polysaccharides and saponins. Qualitatively; TLC and HPLC chromatogram for total phenolics and flavonoids compounds revealed the presence of Gallic acid (GA), Caffeic acid (CA), Paracumarin acid (P-c), Chlorogenic acid (Ch), Catechin (Ca), Hydroqunolinon (H) with more abundant content in Vanillic and Salycilic acids as phenolic compounds; beside, Rutin (R), Quercetin (Q), Apegenin (Ap), Luetolin (L), and Kaempferol (K) as flavonoids. Dried bulbs of N. tazetta contain total phenolic compounds reached 90 mg/g and total flavonoids in a quantity of 0.47 mg/g. Plant bulb analyzed by colorimetric reaction assay.

Diversity and Distribution of Potential Biosurfactant Producing Bacillus Sp MN 243657, GC-MS Analysis and its Antimicrobial Study

Biosurfactants are microbially produced surface-active compounds. They are amphiphilic molecules with hydrophilic and hydrophobic regions. The demand for biosurfactants has been exponentially growing as they are nontoxic and biodegradable. They have different applications in several industrial sectors. The objective of this study was to isolate and characterize the native bacteria which produce biosurfactants from oil contaminated soil of different places in Kerala and Tamil Nadu, India. The soil samples were collected from petrol pumps and workshops where the soil is contaminated with petrol, diesel and oil. The bacteria were isolated from contaminated soil samples and confirmed as Bacillus sp.The cultures were screened for biosurfactant production by different screening techniques such as blood hemolysis, oil spreading assay, emulsification ability assay, bacterial adherence to hydrocarbons activity. The potential biosurfactant producing culture was selected and identified using molecular techniques and submitted to NCBI Gene Bank (MN 243657 – Bacillus sp). The selected bacterial culture was used for biosurfactant production and these were characterized by UV, TLC, FTIR and GC -MS analysis. The derived biosurfactant's Rf value was 0.68 as determined by a TLC chromatogram. In a UV-visible spectroscopy study, the isolated biosurfactant displayed a highest peak at 415 nm. According to FTIR analysis, the isolated biosurfactant displayed an intense peak at 3340 cm -1. The large peaks of the biosurfactant were observed at various retention times of 12.75, 10.22, 4.98, and 3.87, respectively, after GC-MS analysis. Antibacterial and antifungal activity of the biosurfactant was identified against pathogenic bacteria such as P.aeruginosa, E. coli, K. pneumoniae, S. aureus and fungi Aspergillus niger, Aspergillus terreus, Aspergillus flavus.

Comparison of the Determination of Some Antihypertensive Drugs in Clinical Human Plasma Samples by Solvent Front Position Extraction and Precipitation Modes.

The determination of the selected antihypertensive drugs in human plasma samples with the novel solvent front position extraction (SFPE) technique is presented. The SFPE procedure combined with LC-MS/MS analysis was used for the first time to prepare a clinical sample containing the drugs mentioned above from different therapeutic groups. The effectiveness of our approach was compared with the precipitation method. The latter technique is usually used to prepare biological samples in routine laboratories. During the experiments, the substances of interest and the internal standard were separated from other matrix components using a prototype horizontal chamber for thin-layer chromatography/high-performance thin-layer chromatography (TLC/HPTLC) with a moving pipette powered by a 3D mechanism, which distributed the solvent on the adsorbent layer. Detection of the six antihypertensive drugs was performed by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) in multiple reaction monitoring (MRM) mode. Results obtained by SFPE were very satisfactory (linearity R2 ≥ 0.981; %RSD ≤ 6%; LOD and LOQ were in the range of 0.06-9.78 ng/mL and 0.17-29.64 ng/mL, respectively). The recovery was in the range of 79.88-120.36%. Intra-day and inter-day precision had a percentage coefficient of variation (CV) in the range of 1.10-9.74%. The procedure is simple and highly effective. It includes the automation of TLC chromatogram development, which significantly reduced the number of manual operations performed, the time of sample preparation and solvent consumption.

Antioxidant activity from the endemic aquatic Pothos tener Wall lives in Bantimurung waterfalls

Bantimurung is located in the early tertiary isolated small land area in Sulawesi, and this is the factor that caused the endemism of its flora and fauna. Pothos tener Wall is an excellent aquatic plant used as a bioindicator in rain tropical forests and its lives under the Bantimurung waterfalls. Aquatic plant biodiversity has a great potential for bringing about new pharmaceutical inventions derived from freshwater aquatic flora. The genus Pothos is quite popular as a medicinal plant in India and Srilanka similarly P. scandens and P.chinensis as an anti-inflammatory, anticancer, cytotoxic potential, anti-oxidant, antibacterial, anthelmintic potential, antipyretic, anti-diabetic, and bronchodilator. However, the research regarding P.tenner is quite lacking. The study aimed to analyze the antioxidant activity from three extracts (methanol, ethyl acetate, and n-hexane) of P.tenner as an endemic plant from Indonesia. The TLC chromatogram pattern of three extracts had identified the free radical scavenging after being sprayed with a 0,2% DPPH spraying agent. The DPPH method with minor modification was adopted in three extracts of P.tener to examine the antioxidant activity. The IC50 value of quercetin (standard), n-hexane, ethyl acetate, and methanol extracts were 7.4 ppm, 156,26 ppm, 196,25 ppm, and 163,78 ppm, respectively. In general, the potential antioxidant activity of three extracts was n-hexane>methanol>ethyl acetate. To conclude, the n-hexane extract of P.tener has the strongest antioxidant activity among others.

SKRINING FITOKIMIA DAN PROFIL KLT DARI FRAKSI N-HEKSANA DAN ETIL ASETAT PADA KULIT Pithecellobium jiringa (Jack) Prain

Jengkol (Pithecellobium jiringa (Jack) Prain) is a plant that is very often used as a popular processed food ingredient. Based on previous research that the seeds and rind of jengkol have a hypoglycemic effect and reduce glucose levels, and other ingredients are alkaloids, saponins, quinone and flavonoids. This study aims to determine the content of secondary metabolite compounds and the TLC profile contained in the rind of jengkol at fractionation of n-hexane and ethyl acetate. The methods are screening phytochemical, column chromatography, and the TLC. The phytochemical test showed that the rind of jengkol contained terpenoid, flavonoid and saponin compounds. Fraction A was shown spot bluish-green with Rf value 0,64 by the TLC test. The result of the TLC test was indicated to- saponin compound which is characterized by the presence of foam. The result of ethyl acetate fractionation was shown as light blue on The TLC chromatogram with an Rf value of 0,47 which is characterized flavonoid test.

Characterisation of indole alkaloids and phenolic acids from wild mushroom Tropicoporus linteus and its chemical profiles compared with other Sanghuang mushrooms

Two new indole alkaloids, 1-methylindole-7-methoxy-3-carboxaldehyde (2) and 7-methoxyindole-3-carboxaldehyde (4), together with 7-methoxyindole-3-carboxylic acid methyl ester (1 ) and 1-methylindole-3-carbaldehyde (3) were isolated from the fruiting bodies of wild Sanghuang mushroom Tropicoporus linteus (TL663). TLC, 1H-NMR and LC-MS chemical profiles of this mushroom compared with other three genera of wild Sanghuang mushroom extracts were investigated. The TLC, 1H-NMR and LC-MS profiles of TL663 and Sanghuangporus sanghuang (SS664) were similar and significantly different from other mushrooms. These two samples indicated the same TLC chromatograms by showing prominent bands of 1 − 4 when observed under UV 254 nm and having sharp aldehyde proton signals of 3-carboxaldehyde indole type in 1H-NMR spectra. From LC-MS analyses, peaks of isolated compounds 1 − 4 and indole-3-carboxaldehyde (5) in TL663 extract and peaks of protocatechuic acid (6), caffeic aldehyde (7), caffeic acid (8) and 3,4-dihydroxybenzalacetone (9) phenolic acids in TL663 fraction were identified.

Comparative phytochemicals screening and estimation of constituents present in the leaves of Decalepis species through HPLC

Background and objectivesThe variation in the active phytoconstituents in different plants causes changes in their biological properties. The present study was aimed with the comparative phytochemicals and estimation of the constituent present in the leaves of Decalepis species using high performance liquid chromatography (HPLC). Materials and MethodsThe leaves of Decalepis hemiltonii Wight & Arn. Decalepis arayalpathra(J. Joseph and V. Chandras.) Venter. and Decalepis nervosa(Wight &Arn.) Venter.were selected and methanol solvent was used for the preparation of the extracts. Various proximate analysis viz. moisture content, ash values, crude fibers, extractive values were estimated. Thereafter, series of basic chemical tests for the methanol leaves extracts followed by total phenolics, and total flavonoids content were also carried out. Furthermore, TLC chromatogram and RP-HPLC study was carried out for all the plant extracts to find out the presence of phenolic compounds and compared the estimated amount present in all the leaves extracts. ResultsThe outcome of the study revealed that all the three species varied with the yield and the phytochemical screening showed varied concentration of alkaloids, glycosides, flavonoids, proteins, steroids, phenols, and tannins. Among the three species Decalepis hemiltonii(DH) showed presence of higher percentage of moisture (8.6%), crude fibers (8.32%) and total ash content (4.95%) than others. Further, presence of high concentration of phytochemicals was showed by Decalepis arayalpathra (DA) and the higher yield in methanol solvent than other two species. The same plant sample showed presence of higher amount of gallic acid (0.86 mg/100 g) than other two species. Total alkaloids, total phenolic and total flavonoids were higher in DA leaves flowed by Decalepis nervosa (DN) and DH leaves samples. InterpretationOverall the results indicated that DA leaves extracts was better in terms of yield, presence of phytoconstituents, presence of total alkaloids, total flavonoids, and total phenolics and presence of gallic acid followed by DN and DH leaves.

Phytochemical Analysis, Estimation of Quercetin, and in Vitro Anti-Diabetic Potential of Stevia Leaves Samples Procured from Two Geographical Origins

The current study used RP-HPLC to compare phytochemicals and estimate the bioactive constituents found in Stevia rebaudiana Bert. (SRB) leaves collected from two different geographical sources. SRB leaves were collected from Bangalore, Karnataka, India, and Reduit, Mauritius. Extracts were prepared using ethanol and aqueous solvents. Proximate analysis was used to evaluate moisture content, ash values, crude fibers, and extractive values. Following that, preliminary phytochemical screening was done on both ethanol leaves extracts, and subsequently total flavonoid content was determined. In addition, TLC chromatograms and RP-HPLC studies were performed on both plant extracts to determine the presence of flavonoid components in both leaves extracts, followed by in vitro anti-diabetic activity was performed with alpha amylase and alpha glucosidase enzymes against Acarbose as standard. Results revealed that both the extracts from two different geographical sources varied significantly with the yield, content of chemicals, and presence of quercetin (flavonoid) content when estimated through the RP-HPLC standardized method. Glycosides, flavonoids, proteins, steroids, saponins, terpenoids, and phenols were found in various concentrations during phytochemical screening. Among both zones, the ethanol leaves extract of SRB taken from Mauritius had a greater content of phytochemicals and a higher yield than other extracts due to the soil nature. The Mauritius sample had greater total flavonoid levels as well as more quercetin (0.92 ± 0.011) than the other extracts. Following that, ethanol extract inhibited enzymes (alpha amylase, alpha glucosidases) more than aqueous extract, and this inhibition was dose dependent. Among them, the Mauritius ethanol sample showed higher anti-diabetic efficacy than the Indian sample, but this difference was not significant. Overall, SRB ethanol leaves extracts outperformed other leaves extracts in terms of yield, phytoconstituents, and total flavonoids. Overall, both SRB samples had high quercetin levels and possessed anti-diabetic potential, but they were greater in the Mauritius sample, demonstrating that plant traits are influenced by geographic location.

Uji Sitotoksik Ekstrak Alkaloid Daun Belimbing Wuluh (Averrhoa bilimbi L.) terhadap Larva Udang (Artemia salina)

Research on cytotoxic test of alkaloids extract of belimbing wuluh leaves has been conducted. The objective of this research are to determine the yield, toxicity, TLC chromatogram pattern, and compounds in alkaloids extract of belimbing wuluh leaves with GC-MS. Alkaloid extraction process uses acid base method, the phytochemical tests was carried out with reagents Dragendorff, Wagner and Meyer. Toxicity test of alkaloids extract of belimbing wuluh leaves with BSLT method on A. salina larvae to determine LC50 value. Alkaloids extract of belimbing wuluh leaves were analyzed by TLC and GC-MS. The yield of alkaloids extract of belimbing wuluh leaves obtained was 10, 13 %. The results of phytochemical tests were carried out which showed positive containing alkaloids. LC50 value of the alkaloids extract of belimbing wuluh leaves indicated that is very toxic. The LC50value obtained is 9.170 ppm. Analysis using TLC with eluent of n-hexane: ethyl acetate (8:2) showed 5 stains. GC-MS analysis showed are alkaloid compound with an odd molecular weight.

Total Flavonoids Content of Polar Extracts of Cayratia trifolia Leaves

Cayratia trifolia leaves contain flavonoids compound that can dissolve well in polar solvents. This study aimed to evaluate the total flavonoids content of polar extracts which extracted sequentially from C. trifolia leaves. The dried C. trifolia leaves were macerated continually using n-hexane, ethyl acetate, and ethanolic solvent. The total flavonoids content was determined using spectrophotometric methods with a AlCl3 reagent. TLC chromatogram of the extracts were evaluate using chloroform: aceton: formic acid (10:2:1) as a mobile phase and silica gel F254 as a stationary phase. The citroborate and ammonia vapour were used as detection spray reagents. The highest total flavonoids content (27.95±0.62 mgQE/g) were found in 70% ethanol extract of C. trifolia leaves compared to ethyl acetate extract (17.98 ± 0.89 mgQE/g). The ethanolic extract is found six compound spots, while the ethyl acetate extract found four compound spots which were thought to be a flavonoid group. Thus, it can be concluded that the different polar extracting solvents are able to attract flavonoid compounds with different levels. The flavonoids in C. trifolia extract need to be separated to determine the types.

Produksi dan Pemurnian Xilooligosakarida dari Xilan Tongkol Jagung menggunakan Xilanase Streptomyces P26B4 dan Khamir IP4

Xylooligosaccharides (XOS) are sugar oligomers from xylan that can be used as prebiotics to improve digestive tract health. Xylan can be extracted from corncobs which are a by-product of agriculture. The purpose of this study was to produce XOS through hydrolysis of corn cobs xylan using Streptomyces P26B4 xylanase. The products of hydrolysis also consisted of monomer xylose; for that xilooligosaccharides were purified using yeast IP4. The xylan hydrolysis products are quantitatively analyzed based on the value of reducing sugars and degree of polymerization (DP), strengthened qualitatively with TLC. Sugar component was analyzed after the addition of yeast by HPLC. P26B4 xylanase isolates had the highest activity on the 7th day incubation, pH 5,5 buffer citrate and temperature of 50°C. The lowest DP value of xylan hydrolysis was 2.49 at a concentration of 6%, and the 24th hour of incubation. TLC chromatograms showed that xylose and XOS products were produced. Purification of XOS at 6%, showed a decreasing in the area of xylose before and after receiving yeast respectively 1.87% and 1.41%.Therefore, yeast IP4 has the potential to consume xylose amnd purify the XOS. 
 Keywords: corncobs xylan, IP4 yeast, purification, Streptomyces P26B4 xylanase, xylooligosaccharides

Production of Bacterial Amylase and Evaluation for Starch Hydrolysis

In starch degrading industry, the first step to be performed is gelatinization of starch granules at elevated temperature. This process tends to form a viscose suspension, thus making mixing and pumping a huge challenge. Therefore, to lower the viscosity of starch paste thermostable enzymes are added for breaking down the polysaccharides into oligosaccharides by a process known as liquefaction. Therefore, the availability of these enzymes can be considered a prerequisite to start successful starch liquefaction and saccharification industry. The main objective of this study was, therefore, to produce efficient microbial amylolytic enzymes for starch hydrolysis. One bacterial isolate designated as BACC 2107 was isolated from locally collected samples. The enzyme has optimum activity at pH of 5.0 and 70°C. The addition of calcium in the reaction mixture was shown to stabilize the enzyme. The isolate BACC 2107 was able to grow under solid-state fermentation (SSF) using wheat bran as a solid substrate. Optimum enzyme production was recorded at 32°C, and at 120 h of incubation and at a moisture content of 75%. Addition of peptone to the SSF medium as an organic nitrogen supplement enhanced enzyme production. The enzyme was used to hydrolyze a 25% (w/v) of starch at 70°C and pH 5.0, and the resulting hydrolysate was analyzed on TLC plates. After 4 h hydrolysis, the TLC chromatogram showed the release of glucose, maltose, maltotriose, and other oligosaccharides, indicating that the enzyme is endo-acting alpha-amylase. The results of this study show that amylase from BACC 2107 has a good potential to be used in starch liquefaction and saccharification processes. The ability of the isolate to grow and produce the enzyme under SSF using cheap agro-industrial wastes could help to greatly reduce the production cost of the enzyme.

Aqueous extract of fresh leaves from Alternanthera brasiliana (L.) Kuntze: chemical evaluation and antimycobacterial and anticandidal activities

The focus of this study was to evaluate the presence and type of carbohydrates and phenolic molecules inaqueous extract (Aq-E) of Alternanthera brasiliana fresh leaves and its antioxidant and antimicrobial activities and cytotoxicity in vitro. The amount and types of the carbohydrates were measured by colorimetric and TLC methods. Phenolic compounds was detected by colorimetric assay and analysed by HPLC profiles. The antioxidant activity was evaluated by ABTS and Phosphomolybdenum methods. Antimicrobial activity was tested by microdilution method using microorganism models and antibiotic as positive control, the cytotoxicity in vitro was tested using Artemia salina. The results showed the presence of high amount of total sugars and uronic acids. TLC chromatograms showed mainly D-glucose, D-fructose, oligosaccharides and uronic acids in the Aq-E and a sugar alcohol in the methanolic extract. The colorimetric determination showed high concentration of phenolic compounds, which were visualised on the HPLC profiles, such as chemical markers of the Amaranthaceae family and several phenolic acids and flavonoids. The Aq-E demonstrated optimal antioxidant activities. The most important results were the excellent antibacterial and bactericidal activities against Mycobacterium megmatis (MIC = 15.6 µg/mL and MBC = 1000 µg/mL) and antimicrobial activity against Candida albicans (MIC = 31.2 µg/mL) and low cytotoxicity. Further possibilities for this plant extract will be to improve the pharmacological potential for developing new herbal medicines and possibly to study its association to allopathic antibiotics for prevention or treatment of infection diseases.

Spectral characterization of bioactive compounds from microalgae: N. Oculata and C. Vulgaris

Microalgae are unicellular eukaryotic organisms that thrive in a wide variety of habitats such as fresh, salt water and marine environment. Microalgae are considered to be a good source of fatty acids, pigments and various other bioactive metabolites. Hence, the current study attempts to identify bioactive metabolites from two microalgae species: Nannochloropsis Oculata and Chlorella Vulgaris. This diverse group of autotrophic organisms can rapidly produce large biomass per surface area. These phyto-compounds were qualitatively analysed using biochemical methods. Solvent extracts of fresh samples were subjected to preliminary identification by UV spectra and thin layer chromatography. Furthermore, GC-MS and HPLC analysis were conducted to identify and characterize pharmacologically potent molecules. Major Phytochemicals such as phenolic, flavonoids, pigments, protein, and carbohydrates were present in both the samples. This was primarily subjected to UV spectral analysis and TLC chromatogram. The chemical diversity of compounds was further confirmed using GC MS and HPLC.

Assessment of in vitro Antibacterial and Antifungal Activities of Leaf Extracts of Melia azedarach Linn

The present study was aimed to analyse in vitro, the antibacterial and antifungal activities of the leaf extracts of Melia azedarach. Leaves were extracted using different solvents such as ethanol, methanol and acetone by hot percolation method. Antibacterial and antifungal activity was measured using agar well diffusion assay and broth microdilution methods, respectively against different pathogenic bacteria and fungi. Thin-layer chromatography-directed bioautography was performed to determine the phytochemicals of the extracts and their antimicrobial activity. The gas chromatography-mass spectroscopy analysis was performed for the isolated active components exhibited antimicrobial activity. Among the various extracts tested, ethanol extract showed significant antimicrobial activity and exhibited zone of inhibition of 16 mm against Bacillus subtilis, 15 mm against Staphylococcus aureus, 14 mm against both Vibrio cholerae and Klebsiella pneumoniae, 12 mm against both Escherichia coli and Pseudomonas aeruginosa. The ethanol extract showed minimum inhibitory concentration of 250 μg/ml against both Aspergillus niger and Candida albicans. Phytocompounds in the ethanol extract were separated using TLC chromatogram and it showed seven spots. The bioautography of TLC chromatogram showed single band exhibited antibacterial activity and phytocompound from this band was isolated and identified as 5,6,7,8-tetrahydro-1,2,4-benzotriazine-3-amine using gas chromatography-mass spectroscopy analyses. The ethanol extract of leaf of Melia azedarach exhibited significant antimicrobial activity and the compound 5,6,7,8-tetrahydro-1,2,4-benzotriazine-3-amine might be responsible for antimicrobial activity.