Combination of high-performance thin-layer chromatography with a specific bioassay - A tool for effect-directed analysis

Spira, Denise; Reifferscheid, Georg; Buchinger, Sebastian

doi:10.1556/jpc.26.2013.5.2

Cited by 30 publications

(22 citation statements)

References 10 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…S-5), which required continuous calibration. Such a deviation was also evident for EE2 in literature [31]. In Figure S-5, E2 showed less deviation if compared to EE2, which confirmed analogous results showing R 2 0.96 for E2 versus R 2 0.81 for EE2 [32].…”

Section: Hptlc-pyes Dose-response Curves Lod Loq and Working Rangesupporting

confidence: 83%

“…It has to be noted that yeast cells in a very good condition showed LODs down to 250 fg/zone in the experiments on RP-18 W phases, however, a very good yeast cell colony was not regularly caught. LODs reported for chromatographed zones in literature were in the same range, however, the LODs of 1.3 pg/zone for E2 [30] and 0.5 pg/zone for EE2 [30,31] were calculated based on extrapolation according to the calibration curve method, whereas our LOD values were experimentally proven via the S/N ratio (Fig. 2).…”

Section: Hptlc-pyes Dose-response Curves Lod Loq and Working Rangementioning

confidence: 62%

“…As water has a very high solvent strength on silica gel plates, a strong zone diffusion (large, cloudy-shaped diffuse zones) was observed after several hours of aqueous plate incubation [29][30][31][32]. Although it was possible to detect individual standard substances with the so-called planar YES (pYES), standard mixtures could not be quantified due to the strong diffusion effects.…”

Section: Substantial Improvement Generating Sharp-bounded Zonesmentioning

confidence: 99%

“…For all YES assays reported in literature [28][29][30][31][32], pH 7 was applied for the enzyme-substrate reaction in the lac-Z buffer. However, the pH value of the MUG substrate solution had to be adapted to the lipophilic plate surface and was investigated in the range of pH 6-12.…”

Section: Optimization Of Individual Pyes Steps and Plate Ph Adjustmentmentioning

confidence: 99%

“…L-YES was initially applied on a planar plate (without chromatography), but broad spots were obtained after several hours of plate incubation with the aqueous yeast cell suspension [29]. Although the transfer of L-YES to the TLC/HPTLC field has been demonstrated [30][31][32], the clear substance assignment and reliable quantification of substance mixtures always failed due to arising zone diffusion. In the present study, a substantial improvement was demonstrated resulting in sharp-bounded zones even after several hours of incubation with aqueous bioassay media.…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Sharp-bounded zones link to the effect in planar chromatography-bioassay-mass spectrometry

Klingelhöfer

Morlock

2014

Journal of Chromatography A

View full text Add to dashboard Cite

Section: Hptlc-pyes Dose-response Curves Lod Loq and Working Rangesupporting

confidence: 83%

Section: Hptlc-pyes Dose-response Curves Lod Loq and Working Rangementioning

confidence: 62%

Section: Substantial Improvement Generating Sharp-bounded Zonesmentioning

confidence: 99%

Section: Optimization Of Individual Pyes Steps and Plate Ph Adjustmentmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Sharp-bounded zones link to the effect in planar chromatography-bioassay-mass spectrometry

Klingelhöfer

Morlock

2014

Journal of Chromatography A

View full text Add to dashboard Cite

Sample preparation for planar chromatography

Poole

2023

J of Separation Science

View full text Add to dashboard Cite

High‐performance thin‐layer chromatography has favorable properties for high‐throughput separations with a high matrix tolerance. Sample preparation, however, is sometimes required to control specific matrix interferences and to enhance the detectability of target compounds. Trends in contemporary applications have shifted from absorbance and fluorescence detection to methods employing bioassays and mass spectrometry. Traditional methods (shake‐flask, heat at reflux, Soxhlet, and hydrodistillation) are being challenged by automated instrumental approaches (ultrasound‐assisted and microwave‐assisted solvent extraction, pressurized liquid extraction, and supercritical fluid extraction) and the quick, easy cheap, efficient, rugged, and safe extraction method for faster and streamlined sample processing. Liquid‐liquid extraction remains the most widely used approach for sample clean‐up with increasing competition from solid‐phase extraction. On‐layer sample, clean‐up by planar solid‐phase extraction is increasingly used for complex samples and in combination with heart‐cut multimodal systems. The automated spray‐on sample applicator, the elution head interface, biological detection of target and non‐target compounds, and straightforward mass spectrometric detection are highlighted as the main factors directing current interest toward faster and simpler sample workflows, analysis of more complex samples, and the determination of minor contaminants requiring high concentration factors.

show abstract

Coupling of In Vitro Bioassays with Planar Chromatography in Effect-Directed Analysis

Weiß

Egetenmeyer

Schulz

2016

In Vitro Environmental Toxicology - Concepts, Application and Assessment

View full text Add to dashboard Cite

Modern analytical test methods increasingly detect anthropogenic organic substances and their transformation products in water samples and in the environment. The presence of these compounds might pose a risk to the aquatic environment. To determine a possible (eco)toxicological risk, aquatic samples are tested using various bioassays, including sub-organismic assays such as the luminescent bacteria inhibition test, the acetylcholinesterase inhibition test, and the umu-test. The effect-directed analysis (EDA) combines physicochemical separation methods with biological (in vitro) tests. High-performance thin-layer chromatography (HPTLC) has proved to be particularly well suited for the separation of organic compounds and the subsequent analysis of effects by the application of the biotests directly on the surface of the HPTLC plate. The advantage of using HPTLC in comparison to high-performance liquid chromatography (HPLC) for EDA is that the solvent which is used as a mobile phase during chromatography is completely evaporated after the separation and therefore can no longer influence the applied bioassays.A prioritization during the complex identification process can be achieved when observed effects are associated with the separated zones in HPTLC. This increases the probability of identifying the substance responsible for an adverse effect from the multitude of organic trace substances in environmental samples. Furthermore, by comparing the pattern of biological effects of a separated sample, it is possible to track and assess changes in biological activity over time, over space, or in the course of a process, even without identifying the substance. HPTLC has already been coupled with various bioassays.Because HPTLC is a very flexible system, various detection techniques can be used and combined. In addition to the UV/Vis absorption and fluorescence measurements, TLC can also be coupled with a mass spectrometer (MS) for compound identification. In addition, detection of functional groups by means of derivatization reagents can support this identification. It is also possible to combine derivatization and HPLC-MS.Two case studies are used to illustrate the significance of HPTLC-EDA in investigating water quality: Study on a wastewater treatment plant Possible influence of an artificial turf surface on ground water.

show abstract

Combination of high-performance thin-layer chromatography with a specific bioassay - A tool for effect-directed analysis

Cited by 30 publications

References 10 publications

Sharp-bounded zones link to the effect in planar chromatography-bioassay-mass spectrometry

Sharp-bounded zones link to the effect in planar chromatography-bioassay-mass spectrometry

Sample preparation for planar chromatography

Coupling of In Vitro Bioassays with Planar Chromatography in Effect-Directed Analysis

Contact Info

Product

Resources

About