A TLC bioautographic assay for the detection of nitrofurantoin resistance reversal compound

Shahverdi, Ahmad Reza; Abdolpour, Farid; Monsef-Esfahani, Hamid Reza; Farsam, Hassan

doi:10.1016/j.jchromb.2006.11.011

Cited by 40 publications

(29 citation statements)

References 7 publications

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“…Traditionally, bioautographic technique has used the growth inhibition of microorganisms to detect anti-microbial components of extracts chromatographed on a TLC layer. This methodology has been considered as the most efficacious assay for the detection of anti-microbial compounds (Shahverdi, 2007). Bio-autography localizes antimicrobial activity on a chromatogram using three approaches: (i) direct bio-autography, where the micro-organism grows directly on the thin-layer chromatographic (TLC) plate, (ii) contact bio-autography, where the antimicrobial compounds are transferred from the TLC plate to an inoculated agar plate through direct contact and (iii) agar overlay bio-autography, where a seeded agar medium is applied directly onto the TLC plate (Hamburger and Cordell, 1987;Rahalison et al, 1991).…”

Section: Thin-layer Chromatography (Tlc) and Bio-autographic Methodsmentioning

confidence: 99%

Extraction, Isolation And Characterization Of Bioactive Compounds From Plants’ Extracts

Sasidharan

Chen²,

Dharmaraj³

et al. 2010

Afr. J. Trad. Compl. Alt. Med.

852

746

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Section: Thin-layer Chromatography (Tlc) and Bio-autographic Methodsmentioning

confidence: 99%

Extraction, Isolation And Characterization Of Bioactive Compounds From Plants’ Extracts

Sasidharan

Chen²,

Dharmaraj³

et al. 2010

Afr. J. Trad. Compl. Alt. Med.

852

746

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“…This culture was sprayed onto a developed TLC plate and incubated overnight at 37°C in 100% relative humidity. After incubation, the plate was sprayed with a 2% (w/v) solution of triphenyl-tetrazolium chloride (TTC) and incubated for further 6 h. Inhibition zone was observed as clear area against a red-coloured background on the TLC plate [13, 14]. …”

Section: Methodsmentioning

confidence: 99%

Isolation and Partial Characterization of Bioactive Fucoxanthin fromHimanthalia elongataBrown Seaweed: A TLC-Based Approach

Rajauria¹,

Abu‐Ghannam²

2013

International Journal of Analytical Chemistry

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Seaweeds are important sources of carotenoids, and numerous studies have shown the beneficial effects of these pigments on human health. In the present study, Himanthalia elongata brown seaweed was extracted with a mixture of low polarity solvents, and the crude extract was separated using analytical thin-layer chromatography (TLC). The separated compounds were tested for their potential antioxidant capacity and antimicrobial activity against Listeria monocytogenes bacteria using TLC bioautography approach. For bio-autography, the coloured band on TLC chromatogram was visualized after spraying with DPPH and triphenyl-tetrazolium chloride reagents which screen antioxidant and antimicrobial compounds, respectively, and only one active compound was screened on the TLC plate. Preliminary identification of this active compound was done by comparing its colour and R f (retention factor) value with the authentic fucoxanthin standard. Further, the active compound was purified using preparative TLC. This purified compound showed a strong antioxidant (EC50: 14.8 ± 1.27 µg/mL) and antimicrobial (inhibition zone: 10.27 mm, 25 µg compound/disc) activities, which were examined by DPPH scavenging and agar disc-diffusion bioassay, respectively. The bioactivity shown by the purified compound was almost similar to the fucoxanthin standard. The characteristic UV-visible and FT-IR spectra of the purified active compound completely matched with the standard. Hence, the main active compound in H. elongata was identified as fucoxanthin.

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Section: Introductionmentioning

confidence: 99%

“…Bioautography is particularly important to avoid the time-consuming isolation of inactive compounds. TLC bioautographic methods combine chromatographic separation and in situ activity determination facilitating the localization and target-directed isolation of active constituents in a mixture (Shahverdi et al, 2007).A number of bioautographic assays have been developed, which can be divided into three groups (Rios et al, 1988). These include direct bioautography; where the microorganisms grow directly on thin-layer chromatography (TLC) plates, contact bioautography; where the antimicrobial compounds are transferred from the TLC plate to an inoculated agar plate through direct contact, and agar overlay or immersion bioautography; where a seeded agar medium is applied onto the TLC plate.…”

mentioning

confidence: 99%

Detection of antimicrobial compounds by bioautography of different extracts of leaves of selected South African tree species

Сулейман¹,

McGaw²,

Naidoo³

et al. 2010

Afr. J. Trad. Compl. Alt. Med.

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The hexane, acetone, dichloromethane and methanol extracts of Combretum vendae A.E. van Wyk (Combretaceae), Commiphora harveyi (Engl. ) test organisms included bacteria (Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus), and fungi (Aspergillus fumigatus, Candida albicans, Cryptococcus neoformans, Microsporum canis and Sporothrix schenckii).A simple bioautographic procedure, involving spraying suspensions of the bacteria or fungi on thin layer chromatography (TLC) plates developed in solvents of varying polarities was used to detect the number of antibacterial and antifungal compounds present in the extracts. All the extracts had antimicrobial activity against at least one of the test microorganisms. This activity was denoted by white spots against a red-purple background on the TLC plates after spraying with tetrazolium violet. Twenty seven TLC plates; 9 for each solvent system and 3 different solvent systems per organism were tested in the bioautographic procedure. Of the bacteria tested, S. aureus was inhibited by the most compounds separated on the TLC plates from all the tested plants. Similarly, growth of the fungus C. neoformans was also inhibited by many compounds present in the extracts. Loxostylis alata appeared to be the plant extract with the highest number of inhibition bands when compared with other plants tested against both bacteria and fungi. This species was selected for in depth further study.Key words: Bioautography; Medicinal plants; Antifungal; Antibacterial; Synergism IntroductionDespite the existence of conventional antimicrobial agents, resistant or multi-resistant strains of pathogenic microorganisms are continuously appearing, imposing the need for a thorough search for and development of new drugs (Silver and Bostian, 1993). Fungi and bacteria cause important human and animal diseases, especially in tropical and subtropical regions, and commonly occur in immunocompromised or immunodeficient patients. Over the last decade, there has been a renewed interest in plants; and the pharmaceutical industry considers plants as a viable option for the discovery of new leads (Soejarto, 1996). In fact, it is also estimated that natural products are implicated in the development of 44% of all new drugs, generally as leads for the preparation of semi-synthetic derivatives (Hostettmann et al., 2001).In an effort to discover new lead compounds, many research groups screen plant extracts to detect secondary metabolites with relevant biological activities. In this regard, several bioassays were developed for screening purposes (Hostettmann, 1991).Once the technique has been mastered, bioautography is a highly efficacious assay for the detection of antimicrobial compounds because it allows localization of activity even in a complex matrix, and therefore facilitates the target-directed isolation of the active constituents (Rahalison et al., 1991). Bioautography has enabled rapid progress for quick detection of new antimicrobial compounds from plants and othe...

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A TLC bioautographic assay for the detection of nitrofurantoin resistance reversal compound

Cited by 40 publications

References 7 publications

Extraction, Isolation And Characterization Of Bioactive Compounds From Plants’ Extracts

Extraction, Isolation And Characterization Of Bioactive Compounds From Plants’ Extracts

Isolation and Partial Characterization of Bioactive Fucoxanthin fromHimanthalia elongataBrown Seaweed: A TLC-Based Approach

Detection of antimicrobial compounds by bioautography of different extracts of leaves of selected South African tree species

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