A 96-well-plate–based method for the estimation of alpha-amylase activity using miniaturises 3,5-DINITROSALICYLIC ACID (DNSA) colorimetric method
The DNSA assay has been widely employed for the in vitro detection and quantification of alpha-amylase inhibitory activity. However, the conventional method is associated with inconsistencies between protocols and requires a large volume of samples and other assay reagents that can compromise accura...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Penerbit Universiti Kebangsaan Malaysia
2022
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| Online Access: | http://journalarticle.ukm.my/21752/ http://journalarticle.ukm.my/21752/1/MAs%2012.pdf |
| _version_ | 1848815427985080320 |
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| author | Noor Syaffinaz Noor Mohamad Zin, Nur Ain Sabrina Azmi, Nursarah Syamimi Anuar, Syahirah Ain Shafie, Muhamammad Aiman Faris Maznan, Yong, Meng Goh Nurdiana Samsulrizal, |
| author_facet | Noor Syaffinaz Noor Mohamad Zin, Nur Ain Sabrina Azmi, Nursarah Syamimi Anuar, Syahirah Ain Shafie, Muhamammad Aiman Faris Maznan, Yong, Meng Goh Nurdiana Samsulrizal, |
| author_sort | Noor Syaffinaz Noor Mohamad Zin, |
| building | UKM Institutional Repository |
| collection | Online Access |
| description | The DNSA assay has been widely employed for the in vitro detection and quantification of alpha-amylase inhibitory activity. However, the conventional method is associated with inconsistencies between protocols and requires a large volume of samples and other assay reagents that can compromise accurate quantitation. Therefore, the study aimed to develop a reliable, simple, and rapid analytical method for determining α-amylase activity. The developed method was carried out in 96-well microplates with a total volume of 250 μL and a total assay time of 1 hr, including pre-incubation. The method was validated for linearity, the limit of detection (LOD), the limit of quantitation (LOQ), and precision. A higher coefficient of determination (R2) value was observed for the developed method as compared to the conventional method (0.9983 ± 0.0003 vs 0.9667 ± 0.0383). The coefficient of variation (CV%) of each data point was less than 15%, indicating excellent data precision. The optimum assay conditions were identified at 2 U/mL of enzyme solution and 5% (w/v) starch solution at 50 °C incubation temperature with an IC50 value of 0.026 ± 0.005 mg/mL. It is concluded that the developed method is practical, precise, and accurate for estimating α-amylase inhibitory activity and would provide reproducible results. |
| first_indexed | 2025-11-15T00:49:49Z |
| format | Article |
| id | oai:generic.eprints.org:21752 |
| institution | Universiti Kebangasaan Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T00:49:49Z |
| publishDate | 2022 |
| publisher | Penerbit Universiti Kebangsaan Malaysia |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | oai:generic.eprints.org:217522023-06-21T05:18:25Z http://journalarticle.ukm.my/21752/ A 96-well-plate–based method for the estimation of alpha-amylase activity using miniaturises 3,5-DINITROSALICYLIC ACID (DNSA) colorimetric method Noor Syaffinaz Noor Mohamad Zin, Nur Ain Sabrina Azmi, Nursarah Syamimi Anuar, Syahirah Ain Shafie, Muhamammad Aiman Faris Maznan, Yong, Meng Goh Nurdiana Samsulrizal, The DNSA assay has been widely employed for the in vitro detection and quantification of alpha-amylase inhibitory activity. However, the conventional method is associated with inconsistencies between protocols and requires a large volume of samples and other assay reagents that can compromise accurate quantitation. Therefore, the study aimed to develop a reliable, simple, and rapid analytical method for determining α-amylase activity. The developed method was carried out in 96-well microplates with a total volume of 250 μL and a total assay time of 1 hr, including pre-incubation. The method was validated for linearity, the limit of detection (LOD), the limit of quantitation (LOQ), and precision. A higher coefficient of determination (R2) value was observed for the developed method as compared to the conventional method (0.9983 ± 0.0003 vs 0.9667 ± 0.0383). The coefficient of variation (CV%) of each data point was less than 15%, indicating excellent data precision. The optimum assay conditions were identified at 2 U/mL of enzyme solution and 5% (w/v) starch solution at 50 °C incubation temperature with an IC50 value of 0.026 ± 0.005 mg/mL. It is concluded that the developed method is practical, precise, and accurate for estimating α-amylase inhibitory activity and would provide reproducible results. Penerbit Universiti Kebangsaan Malaysia 2022 Article PeerReviewed application/pdf en http://journalarticle.ukm.my/21752/1/MAs%2012.pdf Noor Syaffinaz Noor Mohamad Zin, and Nur Ain Sabrina Azmi, and Nursarah Syamimi Anuar, and Syahirah Ain Shafie, and Muhamammad Aiman Faris Maznan, and Yong, Meng Goh and Nurdiana Samsulrizal, (2022) A 96-well-plate–based method for the estimation of alpha-amylase activity using miniaturises 3,5-DINITROSALICYLIC ACID (DNSA) colorimetric method. Malaysian Applied Biology, 51 (4). pp. 95-102. ISSN 0126-8643 https://jms.mabjournal.com/index.php/mab/index |
| spellingShingle | Noor Syaffinaz Noor Mohamad Zin, Nur Ain Sabrina Azmi, Nursarah Syamimi Anuar, Syahirah Ain Shafie, Muhamammad Aiman Faris Maznan, Yong, Meng Goh Nurdiana Samsulrizal, A 96-well-plate–based method for the estimation of alpha-amylase activity using miniaturises 3,5-DINITROSALICYLIC ACID (DNSA) colorimetric method |
| title | A 96-well-plate–based method for the estimation of alpha-amylase activity using miniaturises 3,5-DINITROSALICYLIC ACID (DNSA) colorimetric method |
| title_full | A 96-well-plate–based method for the estimation of alpha-amylase activity using miniaturises 3,5-DINITROSALICYLIC ACID (DNSA) colorimetric method |
| title_fullStr | A 96-well-plate–based method for the estimation of alpha-amylase activity using miniaturises 3,5-DINITROSALICYLIC ACID (DNSA) colorimetric method |
| title_full_unstemmed | A 96-well-plate–based method for the estimation of alpha-amylase activity using miniaturises 3,5-DINITROSALICYLIC ACID (DNSA) colorimetric method |
| title_short | A 96-well-plate–based method for the estimation of alpha-amylase activity using miniaturises 3,5-DINITROSALICYLIC ACID (DNSA) colorimetric method |
| title_sort | 96-well-plate–based method for the estimation of alpha-amylase activity using miniaturises 3,5-dinitrosalicylic acid (dnsa) colorimetric method |
| url | http://journalarticle.ukm.my/21752/ http://journalarticle.ukm.my/21752/ http://journalarticle.ukm.my/21752/1/MAs%2012.pdf |