Colorimetric detection of salivary α‑amylase using maltose as a noncompetitive inhibitor for polysaccharide cleavage
This paper describes an approach for colorimetric detection of salivary α-amylase, one of the potential biomarkers of autonomic nervous system (ANS) activity, for enabling assessment of fatigue. The ability of α-amylase to cleave α-bonds of polysaccharides is utilized for developing a colorimetric a...
| Main Authors: | , , |
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| Format: | Journal Article |
| Language: | English |
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2020
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| Online Access: | https://hdl.handle.net/10356/143474 |
| _version_ | 1826130478215725056 |
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| author | Tsyrulneva, Iuna Alagappan, Palaniappan Liedberg, Bo |
| author2 | School of Materials Science and Engineering |
| author_facet | School of Materials Science and Engineering Tsyrulneva, Iuna Alagappan, Palaniappan Liedberg, Bo |
| author_sort | Tsyrulneva, Iuna |
| collection | NTU |
| description | This paper describes an approach for colorimetric detection of salivary α-amylase, one of the potential biomarkers of autonomic nervous system (ANS) activity, for enabling assessment of fatigue. The ability of α-amylase to cleave α-bonds of polysaccharides is utilized for developing a colorimetric assay. In the proposed approach, 2-chloro-4-nitrophenyl-α-d-maltotrioside as substrate releases a colored byproduct upon cleavage by salivary α-amylase. Introduction of maltose as a noncompetitive inhibitor yields desirable linear responses in the physiologically relevant concentration range (20–500 μg/mL) with a limit of detection (LOD) of 8 μg/mL (in aqueous solution). The concentrations of substrate and noncompetitive inhibitor are subsequently optimized for colorimetric detection of salivary α-amylase. A facile paper-based “strip” assay is proposed for analysis of human saliva samples with marginal interference from saliva components. The proposed assay is rapid, specific, and easy-to-implement for colorimetric detection of salivary α-amylase between 20 and 500 μg/mL. Complementary RGB (red, green, blue components) analysis offers quantitative detection with a LOD of 11 μg/mL. The two assay formats are benchmarked against the Phadebas test, a state of the art method for spectrophotometric detection of α-amylase. The reported paper-based methodology possesses a high potential for estimation of altered ANS responses toward stressors that possibly could find applications in assessment of fatigue and for monitoring onset of fatigue. |
| first_indexed | 2024-10-01T07:57:07Z |
| format | Journal Article |
| id | ntu-10356/143474 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T07:57:07Z |
| publishDate | 2020 |
| record_format | dspace |
| spelling | ntu-10356/1434742023-03-04T17:21:51Z Colorimetric detection of salivary α‑amylase using maltose as a noncompetitive inhibitor for polysaccharide cleavage Tsyrulneva, Iuna Alagappan, Palaniappan Liedberg, Bo School of Materials Science and Engineering School of Mechanical and Aerospace Engineering Institute for Sports Research Center for Biomimetic Sensor Science Engineering::Mechanical engineering α-Amylase Paper-based Strip This paper describes an approach for colorimetric detection of salivary α-amylase, one of the potential biomarkers of autonomic nervous system (ANS) activity, for enabling assessment of fatigue. The ability of α-amylase to cleave α-bonds of polysaccharides is utilized for developing a colorimetric assay. In the proposed approach, 2-chloro-4-nitrophenyl-α-d-maltotrioside as substrate releases a colored byproduct upon cleavage by salivary α-amylase. Introduction of maltose as a noncompetitive inhibitor yields desirable linear responses in the physiologically relevant concentration range (20–500 μg/mL) with a limit of detection (LOD) of 8 μg/mL (in aqueous solution). The concentrations of substrate and noncompetitive inhibitor are subsequently optimized for colorimetric detection of salivary α-amylase. A facile paper-based “strip” assay is proposed for analysis of human saliva samples with marginal interference from saliva components. The proposed assay is rapid, specific, and easy-to-implement for colorimetric detection of salivary α-amylase between 20 and 500 μg/mL. Complementary RGB (red, green, blue components) analysis offers quantitative detection with a LOD of 11 μg/mL. The two assay formats are benchmarked against the Phadebas test, a state of the art method for spectrophotometric detection of α-amylase. The reported paper-based methodology possesses a high potential for estimation of altered ANS responses toward stressors that possibly could find applications in assessment of fatigue and for monitoring onset of fatigue. Nanyang Technological University Accepted version This work is supported by Provost’s office and Institute for Sports Research (ISR), School of Mechanical and Aerospace Engineering, Nanyang Technological University, Singapore. 2020-09-03T06:52:49Z 2020-09-03T06:52:49Z 2019 Journal Article Tsyrulneva, I., Alagappan, P., & Liedberg, B. (2019). Colorimetric detection of salivary α‑amylase using maltose as a noncompetitive inhibitor for polysaccharide cleavage. ACS Sensors, 4(4), 865-873. doi:10.1021/acssensors.8b01343 2379-3694 https://hdl.handle.net/10356/143474 10.1021/acssensors.8b01343 30895774 2-s2.0-85064353988 4 4 865 873 en ACS Sensors This document is the Accepted Manuscript version of a Published Work that appeared in final form in ACS Sensors, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://doi.org/10.1021/acssensors.8b01343 application/pdf |
| spellingShingle | Engineering::Mechanical engineering α-Amylase Paper-based Strip Tsyrulneva, Iuna Alagappan, Palaniappan Liedberg, Bo Colorimetric detection of salivary α‑amylase using maltose as a noncompetitive inhibitor for polysaccharide cleavage |
| title | Colorimetric detection of salivary α‑amylase using maltose as a noncompetitive inhibitor for polysaccharide cleavage |
| title_full | Colorimetric detection of salivary α‑amylase using maltose as a noncompetitive inhibitor for polysaccharide cleavage |
| title_fullStr | Colorimetric detection of salivary α‑amylase using maltose as a noncompetitive inhibitor for polysaccharide cleavage |
| title_full_unstemmed | Colorimetric detection of salivary α‑amylase using maltose as a noncompetitive inhibitor for polysaccharide cleavage |
| title_short | Colorimetric detection of salivary α‑amylase using maltose as a noncompetitive inhibitor for polysaccharide cleavage |
| title_sort | colorimetric detection of salivary α amylase using maltose as a noncompetitive inhibitor for polysaccharide cleavage |
| topic | Engineering::Mechanical engineering α-Amylase Paper-based Strip |
| url | https://hdl.handle.net/10356/143474 |
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