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, 18 (2), 332-6

Analysis of Tear Glucose Concentration With Electrospray Ionization Mass Spectrometry

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Analysis of Tear Glucose Concentration With Electrospray Ionization Mass Spectrometry

Christopher R Taormina et al. J Am Soc Mass Spectrom.

Abstract

We have developed a mass spectrometry-based method that allows one to accurately determine the glucose concentration of tear fluid. We used a 1 microL micro-capillary to collect tear fluid from the tear meniscus with minimal irritation of the eye. We analyzed the 1 muL volume of collected tear fluid with liquid-chromatography electrospray ionization mass spectrometry with the use of D-glucose-6,6-d2 as an internal standard. Repeated measurements and a recovery experiment on pooled, onion-induced tears showed that the analysis of the glucose in tears was precise (4% relative standard deviation) and provided 100% recovery. We found the tear glucose concentration of one fasting nondiabetic subject to be 13 to 51 microM while the onion-induced tear glucose concentration of a different nondiabetic subject to be 211 to 256 microM.

Figures

Figure 1
Figure 1
Eleven spectra of aqueous solutions of known glucose concentrations (0 – 200 μM) with intensities normalized to the internal standard D-glucose-6,6-d2 signal intensity at m/z 205. The linear regression of the experimental glucose concentration verses the known glucose concentration provided the best fit equation of y = 0.99 x − 0.78 (R2 = 0.98).
Figure 2
Figure 2
Duplicate samples of pooled, induced tears at four different concentrations of added glucose were analyzed. Error bars are plus/minus one standard deviation from the average value of each sample. Linear regression analysis gave an equation of y = 0.99 x + 256 (R2 = 0.99) and a recovery range of 94% to 111% of the glucose spike.

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