<b>Application of ascorbic acid 2-phosphate as a new voltammetric substrate for alkaline phosphatase determination in human serum</b>
An electrochemical assay of the enzyme alkaline phosphatase (ALP) using ascorbic acid 2-phosphate (AAP) as a new voltammetric substrate has been described in this paper. In the alkaline buffer solution the ALP enzymatic hydrolysis product of AAP was ascorbic acid (AA), which was an electro-active su...
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| Format: | Article |
| Language: | English |
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Chemical Society of Ethiopia
2005-12-01
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| Series: | Bulletin of the Chemical Society of Ethiopia |
| Subjects: | |
| Online Access: | http://www.ajol.info/index.php/bcse/article/view/21122 |
| _version_ | 1819139712655294464 |
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| author | Wei Sun Kui Jiao |
| author_facet | Wei Sun Kui Jiao |
| author_sort | Wei Sun |
| collection | DOAJ |
| description | An electrochemical assay of the enzyme alkaline phosphatase (ALP) using ascorbic acid 2-phosphate (AAP) as a new voltammetric substrate has been described in this paper. In the alkaline buffer solution the ALP enzymatic hydrolysis product of AAP was ascorbic acid (AA), which was an electro-active substance and had a sensitive differential pulse voltammetric (DPV) oxidative response on glassy carbon electrode (GCE) at +380 mV (versus Ag/AgCl), so the activity of ALP could be monitored voltammetrically of the oxidative peak current of AA. The electrochemical behaviours of AA were carefully studied and the AA standard solution could be measured by DPV method in the linear range from 10.0 to 1000.0 μmol/L with the detection limit of 8.0 μmol/L. The optimal conditions for ALP enzymatic reaction and the voltammetric detection were optimized. Under the optimal conditions the calibration curve for ALP assay exhibited a linear range from 0.4 to 2000.0 U/L with a detection limit of 0.3 U/L. This proposed method was further applied to determine the ALP content in healthy human serum and the results were in good agreement with the traditional <i>p</i>-nitrophenyl phosphate spectrophotometric method. The kinetic constants of enzymatic reaction were also investigated with the apparent kinetic constant K<sub>m</sub> as 2.77 mmol/L and the maximum velocity V<sub>max</sub> as 0.33 mol/min. |
| first_indexed | 2024-12-22T11:27:01Z |
| format | Article |
| id | doaj.art-fc4421204fd34acdb1654a4561ec107d |
| institution | Directory Open Access Journal |
| issn | 1011-3924 1726-801X |
| language | English |
| last_indexed | 2024-12-22T11:27:01Z |
| publishDate | 2005-12-01 |
| publisher | Chemical Society of Ethiopia |
| record_format | Article |
| series | Bulletin of the Chemical Society of Ethiopia |
| spelling | doaj.art-fc4421204fd34acdb1654a4561ec107d2022-12-21T18:27:43ZengChemical Society of EthiopiaBulletin of the Chemical Society of Ethiopia1011-39241726-801X2005-12-01192163173<b>Application of ascorbic acid 2-phosphate as a new voltammetric substrate for alkaline phosphatase determination in human serum</b>Wei SunKui JiaoAn electrochemical assay of the enzyme alkaline phosphatase (ALP) using ascorbic acid 2-phosphate (AAP) as a new voltammetric substrate has been described in this paper. In the alkaline buffer solution the ALP enzymatic hydrolysis product of AAP was ascorbic acid (AA), which was an electro-active substance and had a sensitive differential pulse voltammetric (DPV) oxidative response on glassy carbon electrode (GCE) at +380 mV (versus Ag/AgCl), so the activity of ALP could be monitored voltammetrically of the oxidative peak current of AA. The electrochemical behaviours of AA were carefully studied and the AA standard solution could be measured by DPV method in the linear range from 10.0 to 1000.0 μmol/L with the detection limit of 8.0 μmol/L. The optimal conditions for ALP enzymatic reaction and the voltammetric detection were optimized. Under the optimal conditions the calibration curve for ALP assay exhibited a linear range from 0.4 to 2000.0 U/L with a detection limit of 0.3 U/L. This proposed method was further applied to determine the ALP content in healthy human serum and the results were in good agreement with the traditional <i>p</i>-nitrophenyl phosphate spectrophotometric method. The kinetic constants of enzymatic reaction were also investigated with the apparent kinetic constant K<sub>m</sub> as 2.77 mmol/L and the maximum velocity V<sub>max</sub> as 0.33 mol/min.http://www.ajol.info/index.php/bcse/article/view/21122Ascorbic acid 2-phosphateAlkaline phosphataseVoltammetryAscorbic acidEnzymatic assay |
| spellingShingle | Wei Sun Kui Jiao <b>Application of ascorbic acid 2-phosphate as a new voltammetric substrate for alkaline phosphatase determination in human serum</b> Bulletin of the Chemical Society of Ethiopia Ascorbic acid 2-phosphate Alkaline phosphatase Voltammetry Ascorbic acid Enzymatic assay |
| title | <b>Application of ascorbic acid 2-phosphate as a new voltammetric substrate for alkaline phosphatase determination in human serum</b> |
| title_full | <b>Application of ascorbic acid 2-phosphate as a new voltammetric substrate for alkaline phosphatase determination in human serum</b> |
| title_fullStr | <b>Application of ascorbic acid 2-phosphate as a new voltammetric substrate for alkaline phosphatase determination in human serum</b> |
| title_full_unstemmed | <b>Application of ascorbic acid 2-phosphate as a new voltammetric substrate for alkaline phosphatase determination in human serum</b> |
| title_short | <b>Application of ascorbic acid 2-phosphate as a new voltammetric substrate for alkaline phosphatase determination in human serum</b> |
| title_sort | b application of ascorbic acid 2 phosphate as a new voltammetric substrate for alkaline phosphatase determination in human serum b |
| topic | Ascorbic acid 2-phosphate Alkaline phosphatase Voltammetry Ascorbic acid Enzymatic assay |
| url | http://www.ajol.info/index.php/bcse/article/view/21122 |
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