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Simultaneous Determination of Glucose and Choline Based on the Intrinsic Fluorescence of the Enzymes

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Abstract

It has been possible to perform the simultaneous determination of choline and glucose using the intrinsic fluorescence of the corresponding enzyme as an analytical signal. This can be done in two ways. First, for low glucose and choline concentrations (about 0.55 mM and 0.75 µM respectively) two differentiated signals, without mutual interference, are obtained for both analytes in the same measurement. Second, when glucose and choline concentrations are higher, a new model has been designed which permits the concentrations to be accurately determined in samples containing from 0.55 mM to 3.75 mM glucose and from 0.75 µM to 11.0 µM choline; the method has been applied to simultaneous glucose and choline determinations in serum samples with good results. This method gives a better performance than multivariate calibration based on Partial Least Squares Regression. The methodology here shown could be also used for the simultaneous determination of other pairs of analytes.

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Acknowledgements

This work was supported by the DGES (Ministerio de Educación y Ciencia of Spain): Project CTQ 2005-05761.

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Authors and Affiliations

  1. GBA, INA, Analytical Chemistry Department, Faculty of Sciences, University of Zaragoza, Pedro Cerbuna 12, 50009, Zaragoza, Spain

    J. J. Romero, S. de Marcos & J. Galbán

  2. GBA, INA, Analytical Chemistry Department, Faculty of Veterinary, University of Zaragoza, Miguel Servet 177, 50013, Zaragoza, Spain

    I. Sanz-Vicente

  3. Applied Chemistry Department, Faculty of Chemistry, University of País Vasco, Apdo 1072, 20080, San Sebastian, Spain

    M. Ostra & C. Ubide

Authors
  1. I. Sanz-Vicente
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  2. J. J. Romero
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  3. S. de Marcos
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  4. M. Ostra
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  5. C. Ubide
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  6. J. Galbán
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Corresponding author

Correspondence to I. Sanz-Vicente.

Electronic supplementary material

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Table IS

Effect of the choline concentration on the glucose linear response range and calibration line. (DOC 29.0 KB)

Table IIS

Analytical results obtained for glucose and choline simultaneous determination in the best conditions found using PLS multivariate calibration. (DOC 27.5 KB)

Figure 1S

I t ,n representations for solution indicated in Table 1. [GOx] = 2.0 IU mL−1, [ChOx] = 1.0 IU mL−1 in a buffer solution phosphate pH 8. The numbers in the graphs correspond to the solution indicated in Table 1. (DOC 276 KB)

Figure 2S

Variation of the intrinsic fluorescence of a mixtures of GOx 2.0 IU mL−1 and ChOx 1.0 IU mL−1 in a buffer solution phosphate pH 8. Instrumental conditions as indicated in section 2.1: a 0.55 mM of glucose and 5.50 µM of Choline is added. b Sum of two registers: one of them is the fluorescence intensity variation to a sample containing 0.55 mM of glucose and another one is the fluorescence intensity variation to a sample containing 5.50 µM of choline. c Overlay of (a) and (b). (DOC 220 KB)

Figure 3S

Variation of the intrinsic fluorescence of a mixtures of GOx 2.0 IU mL−1 and ChOx 1.0 IU mL−1 in a buffer solution phosphate pH 8. Instrumental conditions as indicated in section 2.1: a 11.0 µM of Choline is added; b 2.25 mM of glucose is added; c 2.25 mM of glucose and 11.0 µM of choline is added. (DOC 26.5 KB)

Figure 4S

I t ,n representations obtained for a glucose calibration study. Experimental conditions: [GOx] = 2.0 IU mL−1 in a buffer solution phosphate pH 8. Glucose concentrations: a 0.55 mM; b 1.00 mM; c 1.50 mM M; d 2.25 mM; e 3.00 mM; f 3.75 mM. (DOC 38.0 KB)

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Sanz-Vicente, I., Romero, J.J., de Marcos, S. et al. Simultaneous Determination of Glucose and Choline Based on the Intrinsic Fluorescence of the Enzymes. J Fluoresc 19, 583–591 (2009). https://doi.org/10.1007/s10895-008-0448-5

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  • DOI: https://doi.org/10.1007/s10895-008-0448-5

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