Improved bioautographic assay on TLC layers for qualitative and quantitative estimation of xanthine oxidase inhibitors and superoxide scavengers

Kong, Yao; Li, Xiangkun; Zhang, Na; Yu, Miao; Feng, Hao; Wu, Tao; Chen, Zhihong

doi:10.1016/j.jpba.2017.11.077

Cited by 20 publications

(7 citation statements)

References 27 publications

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“…Similarly, Kong et al. (2018) separated and identified four kinds of active components (AG I‐IV) in A. membranaceus through TLC method, and then they were further extracted by bioautographic assay, suggesting the bioautography based on TLC separation was rapid, simple, and stable for screening and estimation of potential xanthine oxidase inhibitors and superoxide scavengers. Besides, Lee et al.…”

Section: Resultsmentioning

confidence: 99%

Characterization of astragaloside I‐IV based on the separation of HPTLC from Pleurotus ostreatus cultivated with Astragalus

Yana

Yang

et al. 2020

Journal of Food Science

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In this study, total saponins were extracted from Pleurotus ostreatus cultivated with Astragalus as one of organic culture substrates. High Performance Thin Layer Chromatography (HPTLC) assay showed total saponins could be separated effectively, and four kinds of spots were identified as AG I, AG II, AG III, and AG IV, respectively. FTIR spectra based on HPTLC separation assay showed the saponin characteristic groups including-OH, C-H, C=O, and the glycoside linkaged to sapogenin group CO -C, suggesting the four kinds of spots belonged to cycloartane-type triterpene saponins. The primary mass spectra of precursor ion (HPTLC-ESI-MS) assay further proved the main composition of four kinds of spots was AG I-IV, respectively. Physical properties, including the detection of specific rotation and melting point, revealed the separation of high-purity saponin monomer by HPTLC. HPTLC-dual wavelength spectrodensitometric method detection showed that content of astragaloside I-IV was ranged from 0.2 to 0.5 mg/g, and the total astragalosides contents attained to 1.397 mg/g, indicating P. ostreatus could bioaccumulate astragalosides from Astragalus. These results demonstrated the characterization of astragalosides based on the separation of HPTLC was effective, and supported to consider astragalosides-enriched P. ostreatus as functional edible fungus for food and medical applications.

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Section: Resultsmentioning

confidence: 99%

Characterization of astragaloside I‐IV based on the separation of HPTLC from Pleurotus ostreatus cultivated with Astragalus

Yana

Yang

et al. 2020

Journal of Food Science

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“…Factors such as temperature, pH, incubation time, enzyme and substrate concentrations can affect the enzymatic reaction catalysed by aromatase 13 . The range selected for investigating each factor was based on prior information about the assay procedure 27 .…”

Section: Methodsmentioning

confidence: 99%

“…The range selected for investigating each factor was based on prior information about the assay procedure 27 . The effect of different substrate concentrations (0.05–0.25 μM), enzyme concentrations (0.8–2.4 nM), pH range (4.5–10.5), temperature range (15–60°C) and incubation periods (10–60 min) were investigated using one‐factor‐at‐a‐time design 13 . The effects of temperature and incubation periods on G‐6‐P dehydrogenase were also studied.…”

Section: Methodsmentioning

confidence: 99%

“…HPTLC‐bioautography has been widely employed for testing many biological activities including enzyme inhibitory activities such as anti‐lipase, anti‐xanthine‐oxidase, 8 anti‐cholinesterase, 6 anti‐α‐glucosidase, 9 antifungal 10 and radical scavenging activity 11 . In addition, several studies of quantitative evaluation for biologically active components in crude drugs were established 12–15 …”

Section: Introductionmentioning

confidence: 99%

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Development of a validated HPTLC‐bioautographic method for evaluation of aromatase inhibitory activity of plant extracts and their constituents

Dawood

Shawky

Hammoda

et al. 2021

Phytochemical Analysis

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Introduction Aromatase is a CYP450 enzyme that catalyses the conversion of androgens into oestrogens, where the decrease in the production of oestrogens aided by aromatase inhibitors is considered a target in post‐menopausal breast cancer therapy. TLC‐bioautography is a technique employed for combining chromatographic separations on TLC plates with bioassays. This is the first report to evaluate aromatase inhibitory activity using this technique. Objectives The aim of this study is to develop and validate a new TLC‐bioautographic method for determination of aromatase inhibitory activity in 14 plant extracts. Two quantitation methods, the peak area method and reciprocal iso‐inhibition volume (RIV) method, were compared and investigated to attain reliable results. Factors affecting the enzymatic reaction (temperature, pH, enzyme and substrate concentrations … etc.) were also investigated to attain the optimum parameters. Methodology TLC assisted by digital image processing was implemented for quantitative estimation of the aromatase inhibition of 14 plant extracts using chrysin as positive control. The fluorometric substrate dibenzyl fluorescein (DBF) was utilised for the assay, where inhibitory compounds were visualised as dark spots against a blue fluorescent background. Two software programs, Sorbfil® videodensitometer (in the peak area method) and ImageJ® (in the RIV method), were thoroughly validated using the International Council on Harmonisation (ICH) guideline and used for quantitation. Results The RIV method showed superiority over the peak area method in the quantitation results of the tracks with non‐homogenous background with %RSD values of 0.98 and 1.49 compared with 2.86 and 3.58, respectively. Further, the methods allow the comparison of the activity of different unknown inhibitory compounds without the need for a reference or a positive control. Conclusion Using the TLC‐bioautographic method by image processing combined with the RIV quantitation method, simultaneous separation and quantitation of aromatase inhibitory components could be applied to estimate the relative activity of various plant extracts.

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“…The adsorbent must be inert with the substances to be analyzed and not act as a catalyst in decomposition reactions. The TLC has been used only for the qualitative determination of prebiotics [ 13 ].…”

Section: Introductionmentioning

confidence: 99%

A simple quantitative method using TLC-image analysis to determine fructooligosaccharides (FOS) in food samples

MÁRQUEZ¹,

CERVANTES²,

MARTINEZ³

et al. 2022

Turkish Journal of Chemistry

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The thin-layer chromatography technique (TLC) is a simple and inexpensive analysis commonly used to identify qualitatively the presence of carbohydrates in food samples such as mono- di and oligosaccharides particularly. TLC assay could be improved using image processing software for the semiquantitative determination of this type of compound. In the present work, TLC-image analysis with Silica Gel 60 TLC plates was used for the semiquantitative determination of 6 standards of carbohydrates (glucose, fructose, sucrose, 1-kestose, nystose, and fructofuranosylnystose). Subsequently, the areas of the spots of each compound were determined by digitizing in a conventional office scanner. Then, the segmentation of the images is carried out using software for image processing. The calibration curves were plotted in the Excel software using the average of the areas of the pigmentations obtained in pixels. In this study, the technique of thin-layer chromatography was also used to quantitatively determine the presence of carbohydrates in food samples such as honey, garlic, and onion. Values of determination coefficient (R2) greater than 0.97 in all the calibration curves were obtained. This technique could be useful for detecting carbohydrates (monosaccharides, disaccharides, and oligosaccharides) in analytical assays and food samples without needing specialized analytical equipment. In this work, it was possible to determine the concentration of carbohydrates in samples of garlic and onion that showed the presence of prebiotic carbohydrates in addition to sucrose, glucose, and fructose.

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Improved bioautographic assay on TLC layers for qualitative and quantitative estimation of xanthine oxidase inhibitors and superoxide scavengers

Cited by 20 publications

References 27 publications

Characterization of astragaloside I‐IV based on the separation of HPTLC from Pleurotus ostreatus cultivated with Astragalus

Characterization of astragaloside I‐IV based on the separation of HPTLC from Pleurotus ostreatus cultivated with Astragalus

Development of a validated HPTLC‐bioautographic method for evaluation of aromatase inhibitory activity of plant extracts and their constituents

A simple quantitative method using TLC-image analysis to determine fructooligosaccharides (FOS) in food samples

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