Development of a Fluorescence Assay for Rapid Detection of Alkaline Phosphatase Activity in Fresh Milk Based on Inner Filter Effect of Carbon Dots
This work aimed at developing a fluorescence assay for rapid and sensitive determination of alkaline phosphatase (ALP) activity in fresh milk using nitrogen doped carbon dots (N-CDs) as fluorescence probe. Green-emissive N-CDs were synthesized by using ethylenediamine as the nitrogen source and cate...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | zho |
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The editorial department of Science and Technology of Food Industry
2023-09-01
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| Series: | Shipin gongye ke-ji |
| Subjects: | |
| Online Access: | http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2022110075 |
| _version_ | 1827861250330066944 |
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| author | Baozhu JIA Zhenxi HE Xinru HUANG Zhijing QIU Linyuan GUO Yupei YUAN Biman WANG Lin LUO |
| author_facet | Baozhu JIA Zhenxi HE Xinru HUANG Zhijing QIU Linyuan GUO Yupei YUAN Biman WANG Lin LUO |
| author_sort | Baozhu JIA |
| collection | DOAJ |
| description | This work aimed at developing a fluorescence assay for rapid and sensitive determination of alkaline phosphatase (ALP) activity in fresh milk using nitrogen doped carbon dots (N-CDs) as fluorescence probe. Green-emissive N-CDs were synthesized by using ethylenediamine as the nitrogen source and catechol as the carbon source through a hydrothermal method. The obtained N-CDs were characterized by transmission electron microscopy (TEM), UV-vis spectroscopy, X-ray photoelectron spectroscopy (XPS), and Fourier transform infrared (FT-IR) spectra. Control experiments were used to verify the feasibility for constructing the fluorescent assay using the N-CDs. Single-factor experiments were conducted to optimize the pH of working buffer, the concentration of disodium p-nitrophenyl phosphate (PNPP) and enzymatic reaction time for ALP detection. The results indicated that the as-synthesized N-CDs were successfully prepared with all the characterization results being consistent with those in previous works. The optimal working conditions for this assay were as follows: Tris-HCl (20 mmol/L, pH10) as working buffer, 1 mmol/L of PNPP as enzymatic substrate, incubation for 50 min. Under the optimal conditions, it was found that fluorescence intensity of N-CDs linearly corelated with ALP concentration from 0.01 to 25 U/L with correlation coefficients of 0.995. The linear regression equation was Y=15397.05X+70344.46 with a limit of detection (LOD) of 0.001 U/L (S/N=3). Recoveries for two kinds of fresh milk were in the range from 100.1% to 107.2%, and the coefficients of variations were less than 14.3%, which indicated that the proposed method had good accuracy and reliability. This study provides an accurate and efficient method for the detection of ALP in fresh milk. |
| first_indexed | 2024-03-12T13:35:15Z |
| format | Article |
| id | doaj.art-3665604c131c42c488ef715da822db78 |
| institution | Directory Open Access Journal |
| issn | 1002-0306 |
| language | zho |
| last_indexed | 2024-03-12T13:35:15Z |
| publishDate | 2023-09-01 |
| publisher | The editorial department of Science and Technology of Food Industry |
| record_format | Article |
| series | Shipin gongye ke-ji |
| spelling | doaj.art-3665604c131c42c488ef715da822db782023-08-24T05:51:58ZzhoThe editorial department of Science and Technology of Food IndustryShipin gongye ke-ji1002-03062023-09-01441733434110.13386/j.issn1002-0306.20221100752022110075-17Development of a Fluorescence Assay for Rapid Detection of Alkaline Phosphatase Activity in Fresh Milk Based on Inner Filter Effect of Carbon DotsBaozhu JIA0Zhenxi HE1Xinru HUANG2Zhijing QIU3Linyuan GUO4Yupei YUAN5Biman WANG6Lin LUO7College of Biological and Food Engineering, Guangdong University of Education, Guangzhou 510303, ChinaGuangdong Provincial Key Laboratory of Food Quality and Safety, College of Food, South China Agricultural University, Guangzhou 510642, ChinaCollege of Biological and Food Engineering, Guangdong University of Education, Guangzhou 510303, ChinaCollege of Biological and Food Engineering, Guangdong University of Education, Guangzhou 510303, ChinaCollege of Biological and Food Engineering, Guangdong University of Education, Guangzhou 510303, ChinaCollege of Biological and Food Engineering, Guangdong University of Education, Guangzhou 510303, ChinaGuangdong Provincial Key Laboratory of Food Quality and Safety, College of Food, South China Agricultural University, Guangzhou 510642, ChinaGuangdong Provincial Key Laboratory of Food Quality and Safety, College of Food, South China Agricultural University, Guangzhou 510642, ChinaThis work aimed at developing a fluorescence assay for rapid and sensitive determination of alkaline phosphatase (ALP) activity in fresh milk using nitrogen doped carbon dots (N-CDs) as fluorescence probe. Green-emissive N-CDs were synthesized by using ethylenediamine as the nitrogen source and catechol as the carbon source through a hydrothermal method. The obtained N-CDs were characterized by transmission electron microscopy (TEM), UV-vis spectroscopy, X-ray photoelectron spectroscopy (XPS), and Fourier transform infrared (FT-IR) spectra. Control experiments were used to verify the feasibility for constructing the fluorescent assay using the N-CDs. Single-factor experiments were conducted to optimize the pH of working buffer, the concentration of disodium p-nitrophenyl phosphate (PNPP) and enzymatic reaction time for ALP detection. The results indicated that the as-synthesized N-CDs were successfully prepared with all the characterization results being consistent with those in previous works. The optimal working conditions for this assay were as follows: Tris-HCl (20 mmol/L, pH10) as working buffer, 1 mmol/L of PNPP as enzymatic substrate, incubation for 50 min. Under the optimal conditions, it was found that fluorescence intensity of N-CDs linearly corelated with ALP concentration from 0.01 to 25 U/L with correlation coefficients of 0.995. The linear regression equation was Y=15397.05X+70344.46 with a limit of detection (LOD) of 0.001 U/L (S/N=3). Recoveries for two kinds of fresh milk were in the range from 100.1% to 107.2%, and the coefficients of variations were less than 14.3%, which indicated that the proposed method had good accuracy and reliability. This study provides an accurate and efficient method for the detection of ALP in fresh milk.http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2022110075nitrogen-doped carbon dots (n-cds)fluorescent probeinner filter effect (ife)alkaline phosphatasefresh milk |
| spellingShingle | Baozhu JIA Zhenxi HE Xinru HUANG Zhijing QIU Linyuan GUO Yupei YUAN Biman WANG Lin LUO Development of a Fluorescence Assay for Rapid Detection of Alkaline Phosphatase Activity in Fresh Milk Based on Inner Filter Effect of Carbon Dots Shipin gongye ke-ji nitrogen-doped carbon dots (n-cds) fluorescent probe inner filter effect (ife) alkaline phosphatase fresh milk |
| title | Development of a Fluorescence Assay for Rapid Detection of Alkaline Phosphatase Activity in Fresh Milk Based on Inner Filter Effect of Carbon Dots |
| title_full | Development of a Fluorescence Assay for Rapid Detection of Alkaline Phosphatase Activity in Fresh Milk Based on Inner Filter Effect of Carbon Dots |
| title_fullStr | Development of a Fluorescence Assay for Rapid Detection of Alkaline Phosphatase Activity in Fresh Milk Based on Inner Filter Effect of Carbon Dots |
| title_full_unstemmed | Development of a Fluorescence Assay for Rapid Detection of Alkaline Phosphatase Activity in Fresh Milk Based on Inner Filter Effect of Carbon Dots |
| title_short | Development of a Fluorescence Assay for Rapid Detection of Alkaline Phosphatase Activity in Fresh Milk Based on Inner Filter Effect of Carbon Dots |
| title_sort | development of a fluorescence assay for rapid detection of alkaline phosphatase activity in fresh milk based on inner filter effect of carbon dots |
| topic | nitrogen-doped carbon dots (n-cds) fluorescent probe inner filter effect (ife) alkaline phosphatase fresh milk |
| url | http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2022110075 |
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