Article

Automated high-throughput Vibrio fischeri assay for (eco)toxicity screening: application to ionic liquids.

REQUIMTE, Departamento de Química, Faculdade de Farmácia, Universidade do Porto, Rua Jorge Viterbo Ferreira, 228, 4050-313 Porto, Portugal.
Ecotoxicology and Environmental Safety (Impact Factor: 2.2). 03/2012; 80:97-102. DOI: 10.1016/j.ecoenv.2012.02.013
Source: PubMed

ABSTRACT An automated high-throughput Vibrio fischeri assay was developed and further applied to the evaluation of ionic liquids (ILs) (eco)toxicity. The assay was based on the reduction of bacterial bioluminescence in the presence of test compounds and the results were presented as EC(50). The assays were performed with eight commercially available ILs with distinct cationic head groups, alkyl side chains and anions. EC(50) values between 6.5 and 691.9 mmol L(-1) were obtained for the tested ILs, being hmim [Cl] the most toxic and bmim [Cl] the less toxic ones, confirming the influence of the different structural elements. Moreover, all the tested ILs exhibited a (eco)toxicity lower than Cu(II), used as a positive control during the optimization and analysis steps. The automated assay assured the precise control of the contact time between V. fischeri and test compound by means of a simple protocol that guaranteed adequate aspiration and handling of the solutions as well as the precise implementation of a computer controlled stop period. Furthermore, a significant reduction of the assay costs was achieved through automation mainly by a drastic reduction of the volume of bacterial suspension and test compound. The methodology was validated by comparison with a microplate assay; it was stated that the results, obtained after a 3min contact time, changed proportionally relatively to Cu(II) in both assays. This confirmed the applicability of the methodology as an (eco)toxicity screening assay, with reduction of time and increase of robustness and repeatability (n=10; rsd<1.1%). It is expected that due to its simplicity and reduced cost the developed assay can be integrated in the early stage of development of new compounds as a rapid screening test.

1 Bookmark
 · 
103 Views
  • [Show abstract] [Hide abstract]
    ABSTRACT: Due to the merits of being time-saving, cost effective and simple operation, the luminescent bacteria toxicity assay (LBTA) has been widely used for environmental pollution monitoring. Based on numerous studies since 2007, this critical review aims to give an overview on the mechanisms, developments and applications of LBTA. Firstly, based on the introduction of the mechanisms of LBTA, this review shows the interferences from the characteristics of testing samples (such as inorganic nutrients, color, turbidity) and summarizes the improvements on pretreatment method, test methods and test systems in recent years. Regarding the factors that affect the toxicity prediction of single chemicals, the correlation between the toxicity index expressed as median effective concentration (EC50) and characters (such as Kow, the alkyl chain length, the anion and the cation) of known chemicals, especially the emerging ionic liquids (ILs), were given an in-depth discussion. The models for predicting the joint effect of mixtures to luminescent bacteria were also presented. For the factors that affect the toxicity of actual waters, the correlation of toxicity of actual samples to luminescent bacteria and their conventional indexes were discussed. Comparing the sensitivity of the LBTA with other bioassays could indicate the feasibility of the LBTA applied on specific samples. The summary on the application of LBTA to environmental samples has been made to find the future research direction.
    Science of The Total Environment 08/2013; 468-469C:1-11. · 3.26 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Covering: 2008 to 2012Since the last comprehensive review by Otto Sticher on natural product isolation in NPR (O. Sticher, Nat. Prod. Rep., 2008, 25, 517), a plethora of new reports on isolation of secondary compounds from higher plants, marine organisms and microorganisms has been published. Although methods described earlier like the liquid-solid chromatographic techniques (VLC, FC, MPLC, HPLC) or partition chromatographic methods are still the major tools for isolating pure compounds, some developments like hydrophilic interaction chromatography (HILIC) have not been fully covered in previous reviews. Furthermore, examples of using different preparative solid-phase extraction (SPE) columns including molecular imprinting technology have been included. Special attention is given to chiral stationary phases in isolation of natural products. Methods for proper identification of plant material, problems of post-harvest changes in plant material, extraction methods including application of ionic liquids, de-replication procedures during natural product isolation are further issues to be discussed by the review. Selected work published between 2008 and mid-2012 is covered.
    Natural Product Reports 02/2013; · 10.18 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: The toxicity of 16 pharmaceutical active ionic liquids (IL-APIs) was evaluated by automated approaches based on sequential injection analysis (SIA). The implemented bioassays were centered on the inhibition of human carboxylesterase 2 and Vibrio fischeri, in the presence of the tested compounds. The inhibitory effects were quantified by calculating the inhibitor concentration required to cause 50% of inhibition (EC50). The EC50 values demonstrated that the cetylpyridinium group was one of the most toxic cations and that the imidazolium group was the less toxic. The obtained results provide important information about the safety of the studied IL-APIs and their possible use as pharmaceutical drugs. The developed automated SIA methodologies are robust screening bioassays, and can be used as a generic tools to identify the (eco)toxicity of the structural elements of ILs, contributing to a sustainable development of drugs.
    Journal of hazardous materials 12/2013; 265C:133-141. · 4.14 Impact Factor