Chromogenic Assay Is More Efficient in Identifying α-Amylase Inhibitory Properties of Anthocyanin-Rich Samples When Compared to the 3,5-Dinitrosalicylic Acid (DNS) Assay
The inhibition of carbohydrate digestion by plant bioactive compounds is a potential dietary strategy to counteract type 2 diabetes. Indeed, inhibition of α-amylase, a key enzyme that carries out the bulk of starch digestion, has been demonstrated for a range of bioactive compounds including anthocy...
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MDPI AG
2023-09-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/28/17/6399 |
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| author | Sadia Zulfiqar Federica Blando Caroline Orfila Lisa J. Marshall Christine Boesch |
| author_facet | Sadia Zulfiqar Federica Blando Caroline Orfila Lisa J. Marshall Christine Boesch |
| author_sort | Sadia Zulfiqar |
| collection | DOAJ |
| description | The inhibition of carbohydrate digestion by plant bioactive compounds is a potential dietary strategy to counteract type 2 diabetes. Indeed, inhibition of α-amylase, a key enzyme that carries out the bulk of starch digestion, has been demonstrated for a range of bioactive compounds including anthocyanins; however, sample pigmentation often interferes with measurements, affecting colorimetric assay outcomes. Therefore, the present study compared the performance of a direct chromogenic assay, using 2-chloro-4 nitrophenyl α-D-maltotrioside (CNPG3) as a substrate, with the commonly used 3,5-dinitrosalicylic acid (DNS) assay. The direct chromogenic assay demonstrated a 5–10-fold higher sensitivity to determine α-amylase inhibition in various samples, including acarbose as a reference, pure anthocyanins, and anthocyanin-rich samples. The IC<sub>50</sub> values of acarbose presented as 37.6 μg/mL and 3.72 μg/mL for the DNS assay and the direct chromogenic assay, respectively, whereas purified anthocyanins from blackcurrant showed IC<sub>50</sub> values of 227.4 µg/mL and 35.0 µg/mL. The direct chromogenic assay is easy to perform, fast, reproducible, and suitable for high-throughput screening of pigmented α-amylase inhibitors. |
| first_indexed | 2024-03-10T23:16:11Z |
| format | Article |
| id | doaj.art-31b70a48e921473db8aca0157a53d6d5 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-10T23:16:11Z |
| publishDate | 2023-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-31b70a48e921473db8aca0157a53d6d52023-11-19T08:35:26ZengMDPI AGMolecules1420-30492023-09-012817639910.3390/molecules28176399Chromogenic Assay Is More Efficient in Identifying α-Amylase Inhibitory Properties of Anthocyanin-Rich Samples When Compared to the 3,5-Dinitrosalicylic Acid (DNS) AssaySadia Zulfiqar0Federica Blando1Caroline Orfila2Lisa J. Marshall3Christine Boesch4School of Food Science and Nutrition, Faculty of Environment, University of Leeds, Leeds LS2 9JT, UKInstitute of Sciences of Food Production (ISPA), National Research Council (CNR), Via Prov.le Lecce-Monteroni, 73100 Lecce, ItalySchool of Food Science and Nutrition, Faculty of Environment, University of Leeds, Leeds LS2 9JT, UKSchool of Food Science and Nutrition, Faculty of Environment, University of Leeds, Leeds LS2 9JT, UKSchool of Food Science and Nutrition, Faculty of Environment, University of Leeds, Leeds LS2 9JT, UKThe inhibition of carbohydrate digestion by plant bioactive compounds is a potential dietary strategy to counteract type 2 diabetes. Indeed, inhibition of α-amylase, a key enzyme that carries out the bulk of starch digestion, has been demonstrated for a range of bioactive compounds including anthocyanins; however, sample pigmentation often interferes with measurements, affecting colorimetric assay outcomes. Therefore, the present study compared the performance of a direct chromogenic assay, using 2-chloro-4 nitrophenyl α-D-maltotrioside (CNPG3) as a substrate, with the commonly used 3,5-dinitrosalicylic acid (DNS) assay. The direct chromogenic assay demonstrated a 5–10-fold higher sensitivity to determine α-amylase inhibition in various samples, including acarbose as a reference, pure anthocyanins, and anthocyanin-rich samples. The IC<sub>50</sub> values of acarbose presented as 37.6 μg/mL and 3.72 μg/mL for the DNS assay and the direct chromogenic assay, respectively, whereas purified anthocyanins from blackcurrant showed IC<sub>50</sub> values of 227.4 µg/mL and 35.0 µg/mL. The direct chromogenic assay is easy to perform, fast, reproducible, and suitable for high-throughput screening of pigmented α-amylase inhibitors.https://www.mdpi.com/1420-3049/28/17/6399α-amylaseanthocyaninsdirect chromogenic assayDNS assaycarbohydrate digestion |
| spellingShingle | Sadia Zulfiqar Federica Blando Caroline Orfila Lisa J. Marshall Christine Boesch Chromogenic Assay Is More Efficient in Identifying α-Amylase Inhibitory Properties of Anthocyanin-Rich Samples When Compared to the 3,5-Dinitrosalicylic Acid (DNS) Assay Molecules α-amylase anthocyanins direct chromogenic assay DNS assay carbohydrate digestion |
| title | Chromogenic Assay Is More Efficient in Identifying α-Amylase Inhibitory Properties of Anthocyanin-Rich Samples When Compared to the 3,5-Dinitrosalicylic Acid (DNS) Assay |
| title_full | Chromogenic Assay Is More Efficient in Identifying α-Amylase Inhibitory Properties of Anthocyanin-Rich Samples When Compared to the 3,5-Dinitrosalicylic Acid (DNS) Assay |
| title_fullStr | Chromogenic Assay Is More Efficient in Identifying α-Amylase Inhibitory Properties of Anthocyanin-Rich Samples When Compared to the 3,5-Dinitrosalicylic Acid (DNS) Assay |
| title_full_unstemmed | Chromogenic Assay Is More Efficient in Identifying α-Amylase Inhibitory Properties of Anthocyanin-Rich Samples When Compared to the 3,5-Dinitrosalicylic Acid (DNS) Assay |
| title_short | Chromogenic Assay Is More Efficient in Identifying α-Amylase Inhibitory Properties of Anthocyanin-Rich Samples When Compared to the 3,5-Dinitrosalicylic Acid (DNS) Assay |
| title_sort | chromogenic assay is more efficient in identifying α amylase inhibitory properties of anthocyanin rich samples when compared to the 3 5 dinitrosalicylic acid dns assay |
| topic | α-amylase anthocyanins direct chromogenic assay DNS assay carbohydrate digestion |
| url | https://www.mdpi.com/1420-3049/28/17/6399 |
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