Development of a Reliable Device for ‘Fluorokinetic’ Analysis Based on a Portable Diode Array MEMS Fluorimeter

A device was developed to study the evolution of fluorescence spectra as a function of time. A previously designed fluorimeter based on the diode array mini-spectrometer CM12880MA was used. The control and measurement were carried out by programming a SAM21D microcontroller. Considerations regarding the optimization of acquisition speed, memory, and computer interface have been analyzed and optimized. As a result, a very versatile device with great adaptability, reduced dimensions, portability, and a low budget (under EUR 500) has been built. The sensitivity, controlled by the integration time of the photodiodes, can be adjusted between 10 µs and 20 s, thus allowing sampling times ranging from 10 ms to more than 10 h. Under these conditions, chemical rate constants from 20 s−1 to 10−8 s−1 can be experimentally determined. It has a very wide operating range for the kinetic rate constant determination, over six orders of magnitude. As proof of the system performance, the oxidation reaction of Thiamine in a basic medium to form fluorescent Thiochrome has been employed. The evolution of the emission spectrum has been followed, and the decomposition rate constant has been measured at 2.1 × 10−3 s−1, a value which matches those values reported in the literature for this system. A Thiochrome calibration curve has also been performed, obtaining a detection limit of 13 nM, consistent with literature data. Additionally, the stability of Thiochrome has been tested, being the photo-decomposition rate constants 1.8 × 10−4 s−1 and 3.0 × 10−7 s−1, in the presence and absence of UV light (365 nm), respectively. Finally, experiments have been designed to obtain, in a single measurement, the values of both rate constants: the formation of Thiochrome from Thiamine and its photo-decomposition under UV light to a non-fluorescent product. The rate constant values obtained are in good agreement with those previously obtained through independent experiments under the same experimental conditions. These results show that, under these conditions, Thiochrome can be considered an unstable intermediate in a chemical reaction with successive stages.