scholarly journals Extraction and Isolation of Antioxidant-antibacterial Compounds From Lactobacillus casei Strain K1C by Thin-layer Chromatography

2021 ◽  
Vol In Press (In Press) ◽  
Author(s):  
Zahra Pourramezan ◽  
Mana Oloomi ◽  
Rouha Kasra Kermanshahi ◽  
Hassan Rezadoost

Background: Nowadays, searching for natural bioactive compounds with potential use in food industries is a major issue. Because of simple purification, natural compounds from microbial sources attract more attention. These encompass antioxidant and antibacterial materials derived from probiotics. Methods: In this study, Lactobacillus strains were isolated from kefir specimens. The antioxidant and antibacterial activity of the methanol extract of the supernatants was determined using 2, 2-diphenyl-picyril hydrazil (DPPH) and minimum inhibitory concentration (MIC) methods, respectively. In order to increase the antioxidant properties, a minimum medium fermented aerobically was used. Results: Antibacterial activity of Lactobacillus supernatant increased against E. coli ATCC 11303 in case of minimum medium (25.32 mg/mL) compared to MRS broth (32 mg/mL); however, aerobic condition decreased antibacterial production (65.44 mg/mL). After fractionation by thin-layer chromatography (TLC), this value reached the highest level (500 µg/mL). Production analysis at different times showed that maximum antibacterial activity was obtained in the middle of the logarithmic growth phase until the beginning of the stationary growth phase. The antioxidant traits increased significantly in minimum culture media and anaerobic condition (492.1 ± 0.25 µg/mL) compared to the similar condition in MRS broth (880.96 ± 0.05 µg/mL). The highest antioxidant production was observed in the stationary growth phase of the aerobically fermented minimum medium (266.82 ± 0.17 µg/mL). Conclusions: The findings of this study showed that the best antibacterial and antioxidant-producing isolate, L. casei strain K1C (accession no.: KU954559), could be useful as a natural preservative in food industries.

2010 ◽  
Vol 7 (1) ◽  
pp. 88-92 ◽  
Author(s):  
Pipin T. Kurniawati ◽  
H. Soetjipto ◽  
Leenawati Limantara

Research on Bixa orellana L. have been done to isolate, identify and determine bixin percentage, the antioxidant and antibacterial activities of bixin from B. orellana seed.  Isolation and identification of bixin was done by thin layer chromatography (TLC), column chromatography, chemical test of bixin and UV-Vis double beam spectroscopy. Percentage of bixin was calculated by JECFA method, the antioxidant activity was determined by DPPH (1-1 diphynilpicrylhidrazil) method while antibacterial activity was analyzed by the use of agar diffusion method. Thin layer chromatography (TLC) for the crude extract contained 5 spot, where spot 5th was bixin. Bixa orellana has 75±3% of bixin. Antioxidant activity of bixin had IC50 548.5±20.0 ppm. Whereas the antibacterial activity of bixin against the Escherichia coli and Staphylococus aureus could be classified as weak inhibition category at 500-750 μg and medium inhibition category at 1500 μg.   Keywords: Bixa orellana L., bixin, antioxidant, antibacteria


Molekul ◽  
2007 ◽  
Vol 2 (1) ◽  
pp. 1
Author(s):  
Purwantiningsih Sugita

Antibacterial compound from methanol fraction of methylene dichloride extract of pelawan bark had been studied. Pelawan bark was extracted using ethanol 95% and CH2Cl2-water (1:1), respectively. Extract of CH2Cl2 was partitioned using hexane-methanol 90% (1:1). Afterwards, methanol fraction was analyzed by thin layer chromatography methods to find the best eluent for column chromatography. The best eluent came from mixture of acetone-chloroform-methanol (0,6:4:0,4). The fractionation yielded 16 fractions with the first (M1) and the third (M5) fractions became the one spotted fractions. M1 and M5 fractions were examined for their antibacterial activity using paper diffusion methods by determining minimum inhibitory concentration value. M1 fraction showed greater activity than M5 fraction with minimum inhibitory concentration value of 2,7857 mg/mL. The result of phytochemistry test showed that both fractions contain terpenoid.


2014 ◽  
Vol 2014 ◽  
pp. 1-7 ◽  
Author(s):  
Nesa Jafari ◽  
Reza Behroozi ◽  
Davoud Farajzadeh ◽  
Mohammad Farsi ◽  
Kambiz Akbari-Noghabi

Staphylococcus aureusis a Gram-positive bacterium that causes many harmful and life-threatening diseases. Some strains of this bacterium are resistant to available antibiotics. This study was designed to evaluate the ability of indigenous actinomycetes to produce antibacterial compounds againstS. aureusand characterize the structure of the resultant antibacterial compounds. Therefore, a slightly modified agar well diffusion method was used to determine the antibacterial activity of actinomycete isolates against the test microorganisms. The bacterial extracts with antibacterial activity were fractionated by silica gel and G-25 sephadex column chromatography. Also, the active fractions were analyzed by thin layer chromatography. Finally, the partial structure of the resultant antibacterial compound was characterized by Fourier transform infrared spectroscopy. One of the isolates, which had a broad spectrum and high antibacterial activity, was designated asPseudonocardiasp. JB05, based on the results of biochemical and 16S rDNA gene sequence analysis. Minimum inhibitory concentration for this bacterium was 40 AU mL−1againstS. aureus. The antibacterial activity of this bacterium was stable after autoclaving, 10% SDS, boiling, and proteinase K. Thin layer chromatography, using anthrone reagent, showed the presence of carbohydrates in the purified antibacterial compound. Finally, FT-IR spectrum of the active compound illustrated hydroxyl groups, hydrocarbon skeleton, and double bond of polygenic compounds in its structure. To the best of our knowledge, this is the first report describing the efficient antibacterial activity by a local strain ofPseudonocardia.The results presented in this work, although at the initial stage in bioactive product characterization, will possibly contribute toward thePseudonocardiascale-up for the production and identification of the antibacterial compounds.


2019 ◽  
pp. 1-11
Author(s):  
R. Abdulaziz ◽  
M. H. Usman ◽  
U. B. Ibrahim ◽  
B. M. Tambari ◽  
A. Nafiu ◽  
...  

The aim of study was to evaluate the antibacterial activity of Cochlospermum tinctorium against ten (10) strains of antibiotic resistant food-borne pathogens Staphylococcus aureus and Listeria monocytogene. Ten (10) strains of antibiotic resistant food-borne pathogens Staphylococcus aureus and Listeria monocytogene procured from Microbiology Research Laboratory Usman Danfodiyo University Sokoto. The roots of Cochlospermum tinctorium were collected from the rock side in Dambu Gomo, Zuru Local Government Area of Kebbi State, Nigeria. The roots were washed, air-dried and milled to powder using mortal and pestle and sieved to obtained fine powder. Maceration was used for extraction using methanol as solvent. The antibacterial activity of the plant was determined on Mueller Hinton agar using agar well diffusion method. Minimum concentration (MIC) and minimum inhibitory concentration (MBC) of plant extract was also determined. Thin layer chromatography and column chromatography was employed for separation and fraction of different compounds in the plant extract. The fractions were screened for antibacterial activity and active fractions having high antibacterial activity were subjected Gas Chromatography Mass Spectoscopy (GC-MS) analysis. The result of methanol extraction yield 5.17% extracts. The methanol extract of Cochlospermum tinctorium was effective in inhibiting the isolates at high concentration of 10 mg/mL. The results thin layer chromatography revealed four spots with Rf values 0.02, 0.37, 0.44 and 0.80 respectively. The GC-MS analysis of the active methanol extract of Cochlospermum tinctorium root powder revealed the existence of major peaks 1-(+)-Ascorbic acid 2,6-dihexadecanoate (R.T: 13.666), Diethyl phthalate (R.T: 10.440), Undecyl acetate (R.T: 10.007), 3-tetradecanone (R.T: 9.793), 3-hexadecanone (R.T: 12.427). It therefore concluded that the root of Cochlospermum tinctorium has immense potential to be used in the area of pharmacology as it possess antimicrobial activity against the antibiotic resistant food-borne pathogens, thus could be exploited as alternative antimicrobial drugs.


Author(s):  
Sumathy Rengarajan ◽  
Vijayalakshmi Melanathuru ◽  
Deecaraman Munuswamy ◽  
Sankaranarayanan Sundaram ◽  
Saravanan Thiruverkadu Selvaraj

ABSTRACTObjective: The present study was to evaluate the in vitro antibacterial activity, and thin-layer chromatography (TLC) studies from the petals of fourdifferent Indian medicinal plants (Punica granatum, Hibiscus rosa-sinensis, Cassia auriculata, and Moringa oleifera).Methods: The phytochemical screening of the methanol extract of petals of four different Indian medicinal plants was performed using standardprocedures. The antimicrobial activity was tested against various test organisms using the agar disc diffusion method.Results: The preliminary phytochemical screening for petals of four different medicinal plants revealed the presence of flavonoids, alkaloids, tannins,and saponins. From the above study, the results indicated that the methanol extract of M. oleifera petals showed the highest antimicrobial activityagainst Staphylococcus aureus and Bacillus subtilis with zone of inhibition 17.93 and 23.40, respectively, at the concentration of 20 µl/ml and alsoshowed the maximum inhibitory activity at the highest concentration (20 µl/ml) than the lowest concentration (5 µl/ml) against Gram-negativebacteria such as Escherichia coli, Proteus vulgaris, Pseudomonas aeruginosa, and Gram-positive B. subtilis and S. aureus. TLC studies of methanolextracts of petals of Indian medicinal plants revealed the presence of different phytoconstituents as evidenced by separated compounds with differentRf values.Conclusion: The results obtained in the present study indicate that the petals of four different Indian medicinal plants showed the highest antibacterialactivity and can be used as an antibacterial agent against bacterial diseases.Keywords: Phytochemicals, Antibacterial activity, Thin-layer chromatography.


2019 ◽  
Vol 7 (22) ◽  
pp. 3882-3886
Author(s):  
Yessi Febriani ◽  
Vriezka Mierza ◽  
Novi Putri Handayani ◽  
Surismayanti Surismayanti ◽  
Ibrenaita Ginting

BACKGROUND: Infection is the most common diseases in developing country, including Indonesia. Bacteria that often causes infection is Escherichia coli and Staphylococcus aureus. One of the traditional plants that can be used as an antibacterial is lidah mertua. AIM: The purpose of this study was to find out the profile of chemical compounds by thin layer chromatography method and determine the antibacterial activity of Lidah Mertua leaves by in vitro. METHODS: This research conducted an experimentally using non-polar, semipolar, and polar as solvents to get extract against E. coli and S. aureus as bacterial testing. The antibacterial activity using agar diffusion method to get minimum inhibitory concentration (MIC). RESULTS: The result of the research on thin layer chromatography showed that the compounds contained in the Lidah Mertua leaves were polifenol, steroids and alkaloids. The data obtained were tabulated and analysed descriptively. The antibacterial activity show that n-hexane extract does not provide inhibitory activity. MIC value show that aethyl acetate extract of lidah mertua leaves inhibited the growth of E. coli and S. aureus at concentration 50 mg/mL and 25 mg/mL with diameters of inhibition zone is 8.50 mm and 8.20 mm and methanol extract of lidah mertua leaves inhibited the growth of E. coli and S. aureus at concentration 12.5 mg/mL and 25 mg/mL with diameters of inhibition zone is 8.46 mm and 8.32 mm. CONCLUSION: The profile of chemical compounds by thin layer chromatography method showed that the compounds contained in the Lidah Mertua leaves were polifenol, steroids and alkaloids. The antibacterial activity show that n-hexane extract does not provide inhibitory activity, but aethyl acetate extract of lidah mertua leaves inhibited the growth of E. coli and S. aureus.


2010 ◽  
Vol 29 (1) ◽  
pp. 30-38 ◽  
Author(s):  
J.N. Eloff

Several Leonotis species are used widely for medicinal purposes in Africa. There have been drastic changes in the taxonomic treatment of Leonotis species during the past decade. Two species, L. dysophylla and L. microphylla occurring in Pretoria have been considered as varieties of the same species and as different species by different authors. Because Leonotis species are used widely as medicinal plants inter alia against bacterial infections, we decided to compare the chemical composition and antibacterial activity of four plants from each of two populations of the species. The chemical composition of acetone extracts of finely ground leaves was determined by thin layer chromatography followed by spraying with vanillin-sulphuric acid. There were hardly any differences between plants from the same population. There were major differences between the two species in the composition of pigments separated by thin layer chromatography and for compounds visualized with the vanillin-sulphuric acid spray reagent. This supported the viewpoint that the two species should not be considered as varieties. The major differences found in chemical composition indicate that chemical parameters may play an important role in resolving taxonomic differences. Because such a small quantity of material is needed, it may be feasible to analyze one or two leaves obtained from herbarium sheets as an additional taxonomic parameter. The antibacterial activity of the acetone extracts was determined using a two-fold serial dilution microplate method with tetrazolium violet as indicator of growth. The specific strains of the four most important nosocomial bacterial pathogens suggested by the United States National Committee for Clinical Laboratory Standards were used: Staphylococcus aureus (American Type Culture Collection 29213), Pseudomonas aeruginosa (ATCC 27853), Escherichia coli (ATCC 25922) and Enterococcus faecalis (ATCC 21212). The minimum inhibitory activity of the crude extract of L. microphylla was 58, 33, 113 and 15 µg/ml against the four pathogens respectively. The MIC of the L. dysophylla extracts were 110, 95, 113 and 63 µg/ml respectively. If the total activity was calculated by dividing the quantity in mg extracted from one gram of each species with the MIC, the following values were obtained against the respective bacteria using the L. microphylla extract: 700, 1400, 381 and 2100 ml/g. The values for the L. dysophylla extract were 381, 420, 420 and 700 ml/g. This means that if one gram of dried leaves of L. microphylla were extracted with acetone it could be diluted to 2100 ml and it would still kill E. faecalis. These results not only prove the possible use of chemical and biological activity as taxonomic markers, but also the potential value of L. microphylla acetone extracts in treating infections with P. aeruginosa and E. faecalis. The activity of the crude extract against P. aeruginosa was as good as or better than the activity of ampicillin, gentamicin, nitrofurantoin, trimethroptin or sulfi soxazole. The activity against E. faecalis was as good as or better than ampicillin, gentamicin, nitrofurantoin or sulfi soxazole. If the acetone leaf extract of L. microphylla is stable and not toxic there is a good possibility of developing a commercially useful antibacterial product from it. The results indicate the taxonomic value of chemical parameters and biological activity and support the view that the two taxa should be considered as different species.


Planta Medica ◽  
2015 ◽  
Vol 81 (16) ◽  
Author(s):  
G Horváth ◽  
K Ács ◽  
W Jesionek ◽  
I Choma ◽  
A Böszörményi ◽  
...  

2021 ◽  
Vol 20 (66) ◽  
pp. 4-14
Author(s):  
A Yegorova ◽  
◽  
Yu Loskutova ◽  
G Voitiuk ◽  
A Maltsev ◽  
...  

Currently, high-performance thin-layer chromatography (HPTLC) is widely used to control product quality in the pharmaceutical and food industries. In the manufacture of food products, the most important requirement is the control of the content of various additives (preservatives, dyes, antioxidants). For the first time, a method for determining curcumin by high-performance thin-layer chromatography in bouillon cubes "Gallina Blanca" was proposed. Detection was carried out by densitometric scanning using CAMAG equipment when measuring absorbance at a wavelength 265 nm. The method is based on determining the peak area of curcumin in the chromatogram depending on its concentration. Curcumin content is determined by the calibration curve. The developed method was validated by the following tests: specificity, linearity, accuracy and detection limit. The calibration curve is linear in the range of curcumin concentrations from 120 to 520 ng/spot, the detection limit is 65 ng/spot. The specificity of the method is based on the ability to unambiguously evaluate the analyte in the presence of other components and is confirmed by using an external standard. The spots on the chromatograms of the test solution and the reference solution coincide in the value of Rf, which confirms the specificity of the method. Accuracy was evaluated according to the results of the analysis of various samples. The requirement for statistical insignificance of systematic error is fulfilled. The proposed method is express, characterized by satisfactory metrological characteristics, ease of implementation. Key words: high-performance thin-layer chromatography, curcumin, validation.


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