Using the HPTLC-bioluminescence bacteria assay for the determination of acute toxicities in marine sediments and its eligibility as a monitoring assessment tool

Logemann, Anna E; Schafberg, Michaela; Brockmeyer, Berit

doi:10.1016/j.chemosphere.2019.05.246

Cited by 9 publications

(3 citation statements)

References 43 publications

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“…A solution through a holistic orthogonal heart-cut 2D LC superhyphenation was sought. The A. fischeri bioassay − was selected because it is more universally detecting bioactive compounds based on our experience. Often this bioassay indicated the most bioactive compounds.…”

Section: Resultsmentioning

confidence: 99%

Orthogonal Hyphenation of Planar and Liquid Chromatography for Mass Spectrometry of Biomarkers out of the Bioassay Matrix (NP-HPTLC-UV/vis/FLD-Bioassay-RP/IEX-HPLC-UV/vis-ESI-MS)

Kirchert

Morlock

2020

Anal. Chem.

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Bioprofiling on the planar chromatogram with in situ biological/enzymatic assays is a powerful bioanalytical screening tool for the nontargeted detection of known and especially unknown/unidentified bioactive compounds directly in multicomponent mixtures (e.g., foods, spices, and botanicals). However, together with the bioactive zone, the adsorbed bioassay medium is eluted into the mass spectrometer (MS) and interfering with evaluation. Another sample track without bioassay has thus been handled in parallel. Hence, for a direct zone elution from the bioautogram, different setups were investigated to reduce the impact of the bioassay medium load. A biocompatible filter, orthogonal reversed-phase/cation-exchange columns (RP/IEX-HPLC), UV/vis detector, and a Rheodyne valve were installed between the zone eluting interface (after normal-phase high-performance thin-layer chromatography-multi-imaging-bioassay, NP-HPTLC-UV/vis/FLD-bioassay) and the MS. For the negative electrospray ionization mode (ESI–), an RP-18e-HPLC column and valve switch were exploited. After gradient optimization, the RP-column retarded the eluted polar compounds and split-off the salts of the bioassay medium in the first minutes. This reduced the bioassay load and separated analyte signals thereof. However, most bioassay medium mass signals were predominantly detectable in ESI+-MS. Here, the reduction of bioassay matrix signals was achieved by integrating a mixed-mode RP/IEX column. Finally, two different superhyphenations were successfully proven: NP-HPLC-UV/vis/FLD-bioassay-RP-HPLC-UV/vis-ESI–-MS with a valve switch and NP-HPLC-UV/vis/FLD-bioassay-RP/IEX-HPLC-UV/vis-ESI±-MS with or without it. Although the original bioprofiling (NP-HPTLC-UV/vis/FLD-bioassay) was prolonged from 3 to 13 min per sample, such superhyphenations covering chemistry/biology/mass spectrometry are considered as an efficient nontarget bioanalytical tool for fast evaluation of complex samples.

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Section: Resultsmentioning

confidence: 99%

Orthogonal Hyphenation of Planar and Liquid Chromatography for Mass Spectrometry of Biomarkers out of the Bioassay Matrix (NP-HPTLC-UV/vis/FLD-Bioassay-RP/IEX-HPLC-UV/vis-ESI-MS)

Kirchert

Morlock

2020

Anal. Chem.

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“…Apart from UV/Vis/FLD detections and the use of derivatization reagents, the compounds in the extracts were detected via the Gram-negative Aliivibrio fischeri bioassay (Figure 2C). This assay detects more universally any bioactive chemicals interfering with the energetic metabolism of the bacteria and is commonly used in environmental research [30]. The bioautogram revealed a similar 2, 0.4 g/mL in methanol, 0.5 µL/area) on HPTLC plates silica gel 60 F 254 with acetonitrile-water 4:1, v/v, plus 8 mg diphenylboric acid-2aminoethylester (Table S1 from Supplementary Materials, mobile phase 6) derivatized with diphenylamine aniline o-phosphoric acid reagent and documented at white light illumination.…”

Section: Selection Of the Extraction Solvent For Effect-directed Profilingmentioning

confidence: 99%

Section: Selection Of the Extraction Solvent For Effect-directed Profilingmentioning

confidence: 99%

Puree and Juice of Thai Mango and Pineapple Analyzed by High-Performance Thin-Layer Chromatography Hyphenated with Effect-Directed Assays

2021

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The requirements for analytical tools are changing due to the global production chain, the increasing cases of adulteration, and the growing trend towards consumption of plant-based food products worldwide. The assessment of bioactivity of natural foods is currently not a quality criterion, and a paradigm shift is postulated. A non-targeted effect-directed profiling by high-performance thin-layer chromatography hyphenated with five different effect-directed assays was developed exemplarily for the puree and juice products of mango Mangifera indica L. (Anacardiaceae) and pineapple Ananas comosus (L.) Merr. (Bromeliaceae). Several bioactive compounds were detected in each sample. The additional bioactivity information obtained through effect-directed profiles improves, expands and modernizes product control. Non-target effect-directed profiling adds a new perspective to previous target analysis results that can be used not only to ensure health claims based on bioactive compounds, but also to detect unknown bioactive compounds coming from contamination or residues or changes caused by food processing.

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Progress, applications, and challenges in high-throughput effect-directed analysis for toxicity driver identification — is it time for HT-EDA?

Alvarez-Mora,

Arturi,

Béen

et al. 2024

Anal Bioanal Chem

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The rapid increase in the production and global use of chemicals and their mixtures has raised concerns about their potential impact on human and environmental health. With advances in analytical techniques, in particular, high-resolution mass spectrometry (HRMS), thousands of compounds and transformation products with potential adverse effects can now be detected in environmental samples. However, identifying and prioritizing the toxicity drivers among these compounds remain a significant challenge. Effect-directed analysis (EDA) emerged as an important tool to address this challenge, combining biotesting, sample fractionation, and chemical analysis to unravel toxicity drivers in complex mixtures. Traditional EDA workflows are labor-intensive and time-consuming, hindering large-scale applications. The concept of high-throughput (HT) EDA has recently gained traction as a means of accelerating these workflows. Key features of HT-EDA include the combination of microfractionation and downscaled bioassays, automation of sample preparation and biotesting, and efficient data processing workflows supported by novel computational tools. In addition to microplate-based fractionation, high-performance thin-layer chromatography (HPTLC) offers an interesting alternative to HPLC in HT-EDA. This review provides an updated perspective on the state-of-the-art in HT-EDA, and novel methods/tools that can be incorporated into HT-EDA workflows. It also discusses recent studies on HT-EDA, HT bioassays, and computational prioritization tools, along with considerations regarding HPTLC. By identifying current gaps in HT-EDA and proposing new approaches to overcome them, this review aims to bring HT-EDA a step closer to monitoring applications. Graphical Abstract

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Using the HPTLC-bioluminescence bacteria assay for the determination of acute toxicities in marine sediments and its eligibility as a monitoring assessment tool

Cited by 9 publications

References 43 publications

Orthogonal Hyphenation of Planar and Liquid Chromatography for Mass Spectrometry of Biomarkers out of the Bioassay Matrix (NP-HPTLC-UV/vis/FLD-Bioassay-RP/IEX-HPLC-UV/vis-ESI-MS)

Orthogonal Hyphenation of Planar and Liquid Chromatography for Mass Spectrometry of Biomarkers out of the Bioassay Matrix (NP-HPTLC-UV/vis/FLD-Bioassay-RP/IEX-HPLC-UV/vis-ESI-MS)

Puree and Juice of Thai Mango and Pineapple Analyzed by High-Performance Thin-Layer Chromatography Hyphenated with Effect-Directed Assays

Progress, applications, and challenges in high-throughput effect-directed analysis for toxicity driver identification — is it time for HT-EDA?

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