期刊
TALANTA
卷 197, 期 -, 页码 105-112出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.talanta.2018.12.076
关键词
Flavin adenine dinucleotide; Flavin mononucleotide; Cancer cell; Fluorescence; Four-way calibration
资金
- National Natural Science Foundation of China [21175041, 21575039]
- Foundation for Innovative Research Groups of NSFC [21521063]
- Science and Technology Project of Guizhou Province [[2017]7293, [2017]5788]
- Natural Science Project of Department of Education of Guizhou Province [[2017] 116]
- Scientific Research Foundation for Introduced Talent of Guizhou University [201564, 201669]
The simultaneous quantitative analysis of intracellular metabolic coenzymes flavin adenine dinucleotide (FAD) and flavin mononucleotide (FMN) is of interest because they participate in many electron-transfer reactions of metabolism. But, the simultaneous quantitative analysis of FAD and FMN is hard to be achieved by traditional analytical methods. This paper proposes a novel strategy of intrinsic fluorescence coupled with four-way calibration method for simultaneous quantitative analysis of intracellular metabolic coenzymes FAD and FMN. Through mathematical separation, this proposed analytical method efficiently achieved the simultaneous quantitative analysis of metabolic coenzymes FAD and FMN in the cell, despite the fact that uncalibrated spectral interferents coexist in the system. The predicted concentrations of FAD and FMN in the cell are 217.0 +/- 6.9 and 155.0 +/- 1.7 pmol/10(6) cells respectively, which were validated by the approved liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. This analytical method with second-order advantage simply requires the cell solution to be diluted by a buffer, it could introduce an interesting analytical strategy for multianalyte direct quantitative analysis in complex biological systems. In addition, we explore the third-order advantage of four-way calibration by a comparative study based on this real fluorescence data. The comparisons indicate that a four-way calibration method can provide higher sensitivity and more resolving power than a three-way calibration method.
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