This review (with 106 references) mainly deals with the analytical applications of flavin-adenine dinucleotide (FAD) fluorescence. In the first section, the spectroscopic properties of this compound are reviewed at the light of his different acid–base, oxidation and structural forms; the chemical and spectroscopic properties of flavin mononucleotide (FMN) and other flavins will be also briefly discussed. The second section discusses how the properties of FAD fluorescence changes in flavoenzymes (FvEs), again considering the different chemical and structural forms; the glucose oxidase (GOx) and the choline oxidase (ChOx) cases will be commented. Since almost certainly the most reported analytical application of FAD fluorescence is as an auto-indicator in enzymatic methods catalysed by FvE oxidoreductases, it is important to know how the concentrations of the different forms of FAD changes along the reaction and, consequently, the fluorescence and the analytical signals. An approach to do this will be presented in section 3. The fourth part of the paper compiles the analytical applications which have been reported until now based in these fluorescence properties. Finally, some suggestions about tentative future research are also given.
In the past twelve years, digital image colorimetry (DIC) on smartphones has acquired great importance as an alternative to the most common analytical techniques. This analysis method is based on fast, low-cost, and easily-accessible technology, which can provide quantitative information about an analyte through the color changes of a digital image. Despite the fact that DIC is very widespread, it is not exempt from a series of problems that are not fully resolved yet, such as variability of the measurements between smartphones, image format in which color information is stored, power distribution of the illuminant used for the measurements, among others. This article proposes a methodology for the standardization and correction of these problems using self-developed software, together with the use of a 3D printed light box. This methodology is applied to three different colorimetric analyses using different types and brands of smartphones, proving that comparable measurements between devices can be achieved. As color can be related to many target analytes, establishing this measurement methodology can lead to new control analysis applicable to diverse sectors such as alimentary, industrial, agrarian, or sanitary.
Tyramine oxidase (TAO), peroxidase (HRP), and Amplex Red (AR) have been immobilized on cellulose to obtain disposable biosensors for the determination of histamine. During the enzymatic reaction, AR is oxidized and a pink spot is obtained. Using a smartphone and measuring the G (green) color coordinate, histamine can be determined in the presence of other biogenic amines (putrescine and cadaverine) in concentrations ranging from 2·10−5 M to 5·10−4 M with a 7.5·10−6 M limit of detection (LoD). Despite tyramine interference, experimental conditions are provided which allow rapid and simple histamine and simultaneous histamine/tyramine (semi)quantitative determination in mixtures. Finally, tyramine and histamine were determined in a tuna extract with good results (compared to the reference HPLC–MS method). The methodology can also be applied in solution allowing histamine (and simultaneous histamine/tyramine) determination with a lower LoD (1.8·10−7 M) and a similar selectivity. Graphical abstract