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Pyridoxal 5′-phosphate assay based on lucigenin chemiluminescence

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Abstract

The authors describe the first chemiluminescence (CL) based method for determination of pyridoxal 5′-phosphate (PLP). PLP is found to generate intense CL with lucigenin higher than that of the conventional lucigenin-H2O2 system by a factor of about 9.0. This new finding is used to be in a detection method for PLP via flow injection analysis (FIA). Response is linear in the 50 nM to 200 μM PLP concentration range with a correlation coefficient of 0.998, and the detection limit (at an S/N of 3) is 6.9 nM. The assay is highly selective over various amino acids, vitamins, sugars, coenzymes and metal ions cofactors. It exhibits advantages over the commonly employed HPLC methods in that it is rapid, more economic, eco-friendly and high throughput FIA detection of PLP without the need for toxic derivatization reagents, organic solvents, and HPLC instrumentation. The method was successfully applied to the determination of PLP in (spiked) human blood samples with recoveries in the range from 96.2–101.6% with % RSD < 4.0. The new system is also employed to determine lucigenin in the linear range of 0.3 to 100.0 μM with a correlation coefficient of 0.994 and the limit of detection is 0.04 μM.

Schematic of the chemiluminescent assay for pyridoxal 5′-phosphate (PLP). Lucigenin−PLP demonstrates 9-fold stronger chemiluminescence intensity than the lucigenin−H2O2 system. The detection limit of PLP is 6.9 nM. The method can detect PLP in human serum with good recoveries.

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Acknowledgements

We appreciate the kind support from the National Natural Science Foundation of China (Nos. 21675148 and 21505128), the Ministry of Science and Technology of the People’s Re-public of China (No. 2016YFA0201300), and the Chinese Academy of Sciences (CAS)–the Academy of Sciences for the Developing World (TWAS) President’s Fellowship Program.

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Authors and Affiliations

  1. State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin, 130022, People’s Republic of China

    Mohamed Ibrahim Halawa, Muhammad Saqib, Wenyue Gao, Liming Qi, Wei Zhang & Guobao Xu

  2. University of Chinese Academy of Sciences, Beijing, 100049, People’s Republic of China

    Mohamed Ibrahim Halawa, Wenyue Gao & Liming Qi

  3. Department of Pharmaceutical Analytical Chemistry, Faculty of Pharmacy, University of Mansoura, Mansoura, 35516, Egypt

    Mohamed Ibrahim Halawa

  4. Sulaiman Bin Abdullah Aba Al-Khail – Centre for Interdisciplinary Research in Basic Science (SA-CIRBS), Faculty of Basic and Applied Sciences, International Islamic University, Islamabad, Pakistan

    Muhammad Saqib

  5. University of Science and Technology of China, Hefei, China

    Guobao Xu

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  1. Mohamed Ibrahim Halawa
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Correspondence to Wei Zhang or Guobao Xu.

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Halawa, M.I., Saqib, M., Gao, W. et al. Pyridoxal 5′-phosphate assay based on lucigenin chemiluminescence. Microchim Acta 185, 381 (2018). https://doi.org/10.1007/s00604-018-2887-2

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