Rapid autographic method for detection of enzymatic browning inhibitors based on enzyme immobilization
This work describes a TLC-coupled bioautographic assay suitable for the separation and detection of apple polyphenol oxidase (PPO) inhibitors from natural extracts. PPO was immobilised in agar containing L-DOPA as substrate and 3-methyl-2-benzothiazolinone hydrazone hydrochloride (MBTH) to enhance c...
| Main Authors: | , , , |
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| Format: | Artículo |
| Language: | Inglés |
| Published: |
2018
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| Subjects: | |
| Online Access: | http://hdl.handle.net/20.500.12123/2935 https://www.sciencedirect.com/science/article/pii/S0308814618311646 https://doi.org/10.1016/j.foodchem.2018.07.025 |
| _version_ | 1855483148039618560 |
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| author | Micheloni, Oscar B. Farroni, Abel Eduardo Garcia, Paula Furlan, Ricardo Luis Eugenio |
| author_browse | Farroni, Abel Eduardo Furlan, Ricardo Luis Eugenio Garcia, Paula Micheloni, Oscar B. |
| author_facet | Micheloni, Oscar B. Farroni, Abel Eduardo Garcia, Paula Furlan, Ricardo Luis Eugenio |
| author_sort | Micheloni, Oscar B. |
| collection | INTA Digital |
| description | This work describes a TLC-coupled bioautographic assay suitable for the separation and detection of apple polyphenol oxidase (PPO) inhibitors from natural extracts. PPO was immobilised in agar containing L-DOPA as substrate and 3-methyl-2-benzothiazolinone hydrazone hydrochloride (MBTH) to enhance colour development. The inhibition was detected as white spots on reddish background. Minimum amount of PPO inhibitors detected was 0.0125 μg of 4-hexylresorcinol, 0.025 μg of ascorbic acid, 0.5 μg of cysteine and 1 μg of kojic acid. The assay was compatible with normal and reverse phase TLC systems and allows detecting compounds that directly had action on the enzyme as well as agents that could convert quinones back to their reduced form. The chromatographic run evidenced the different nature of enzymatic browning inhibitory compounds from garlic and onion extracts. Using natural enzymes will provide a fast and cheap alternative for target specific exploration of natural enzymatic inhibitors. |
| format | Artículo |
| id | INTA2935 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2018 |
| publishDateRange | 2018 |
| publishDateSort | 2018 |
| record_format | dspace |
| spelling | INTA29352024-03-11T10:11:05Z Rapid autographic method for detection of enzymatic browning inhibitors based on enzyme immobilization Micheloni, Oscar B. Farroni, Abel Eduardo Garcia, Paula Furlan, Ricardo Luis Eugenio Tecnología de Alimentos Food Technology Polifenoles Pardeamiento Enzimático Extractos Vegetales Polyphenols Enzymic Browning Plant Extracts This work describes a TLC-coupled bioautographic assay suitable for the separation and detection of apple polyphenol oxidase (PPO) inhibitors from natural extracts. PPO was immobilised in agar containing L-DOPA as substrate and 3-methyl-2-benzothiazolinone hydrazone hydrochloride (MBTH) to enhance colour development. The inhibition was detected as white spots on reddish background. Minimum amount of PPO inhibitors detected was 0.0125 μg of 4-hexylresorcinol, 0.025 μg of ascorbic acid, 0.5 μg of cysteine and 1 μg of kojic acid. The assay was compatible with normal and reverse phase TLC systems and allows detecting compounds that directly had action on the enzyme as well as agents that could convert quinones back to their reduced form. The chromatographic run evidenced the different nature of enzymatic browning inhibitory compounds from garlic and onion extracts. Using natural enzymes will provide a fast and cheap alternative for target specific exploration of natural enzymatic inhibitors. EEA Pergamino Fil: Micheloni, Oscar B. Universidad Nacional del Noroeste de la Provincia de Buenos Aires (UNNOBA). Departamento de Ciencias Básicas; Argentina. Fil: Farroni, Abel E. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino. Laboratorio de Calidad de Alimento, Suelos y Agua; Argentina. Fil: García, Paula. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina. Fil: Furlan, Ricardo L.E.. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina. 2018-07-31T18:27:30Z 2018-07-31T18:27:30Z 2018-07 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/2935 https://www.sciencedirect.com/science/article/pii/S0308814618311646 0308-8146 https://doi.org/10.1016/j.foodchem.2018.07.025 eng info:eu-repo/semantics/restrictedAccess application/pdf Food chemistry. 269 : 638-643. (2018) |
| spellingShingle | Tecnología de Alimentos Food Technology Polifenoles Pardeamiento Enzimático Extractos Vegetales Polyphenols Enzymic Browning Plant Extracts Micheloni, Oscar B. Farroni, Abel Eduardo Garcia, Paula Furlan, Ricardo Luis Eugenio Rapid autographic method for detection of enzymatic browning inhibitors based on enzyme immobilization |
| title | Rapid autographic method for detection of enzymatic browning inhibitors based on enzyme immobilization |
| title_full | Rapid autographic method for detection of enzymatic browning inhibitors based on enzyme immobilization |
| title_fullStr | Rapid autographic method for detection of enzymatic browning inhibitors based on enzyme immobilization |
| title_full_unstemmed | Rapid autographic method for detection of enzymatic browning inhibitors based on enzyme immobilization |
| title_short | Rapid autographic method for detection of enzymatic browning inhibitors based on enzyme immobilization |
| title_sort | rapid autographic method for detection of enzymatic browning inhibitors based on enzyme immobilization |
| topic | Tecnología de Alimentos Food Technology Polifenoles Pardeamiento Enzimático Extractos Vegetales Polyphenols Enzymic Browning Plant Extracts |
| url | http://hdl.handle.net/20.500.12123/2935 https://www.sciencedirect.com/science/article/pii/S0308814618311646 https://doi.org/10.1016/j.foodchem.2018.07.025 |
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