RESUMEN
The main form of folate in human plasma is 5-methyltetrahydrofolate (5MTHF). The observation that folate in human serum is photosensitive supports the hypothesis that humans developed dark skin in high ultraviolet fluences areas in order to protect folate in the blood from UV radiation. However, folates alone are quite photostable. Therefore, in this study, we examined for the first time the photodegradation of 5MTHF in the presence of the endogenous photosensitizer uroporphyrin (Uro), which is sometimes present in low concentration in human serum, under UV and near-UV light exposure. We found strong indications that while 5MTHF alone is rather photostable, it is degraded quickly in the presence of Uro. Using deuterium oxide (D(2)O) as an enhancer of the lifetime of singlet oxygen and the singlet oxygen sensor green reagent (SOSG) as a scavenger of singlet oxygen, we have found that the photodegradation most likely proceeds via a type II photosensitization. Our results show that singlet oxygen is likely to be the main intermediate in the photodegradation of 5MTHF mediated by Uro. Our findings may be useful for further studies the evolution of human skin colours.
Asunto(s)
Fotólisis , Tetrahidrofolatos/efectos de la radiación , Uroporfirinas/farmacología , Humanos , Trastornos por Fotosensibilidad , Fármacos Fotosensibilizantes/farmacología , Oxígeno Singlete , Piel/química , Piel/efectos de la radiación , Tetrahidrofolatos/química , Rayos UltravioletaRESUMEN
Second-order multivariate calibration methods in combination with a continuous flow system, which allows for the continuous on-line irradiation of the analytes, have been employed for the determination of folic acid and its main metabolite 5-methyltetrahydrofolic acid in serum samples. An experimental central composite design, together with response surface methodology, has been used to find the optimum instrumental variables to perform the photochemical reaction. The time evolution of the emission spectra of the generated photoproducts, in the range 330-540 nm, after irradiation at 275 nm for 20 min, provided the three-way data set employed. On the basis of the differences on the kinetic rates of the photoreaction of both analytes, direct determination of the compounds in human plasma has been accomplished. The second-order methods assayed were parallel factor analysis (PARAFAC), self-weighted alternating trilinear decomposition (SWATLD), and unfolded partial least-squares (U-PLS), multidimensional partial least-squares (N-PLS), and bilinear least-squares (BLLS), all three in combination with the residual bilinearization procedure (RBL).
Asunto(s)
Ácido Fólico/sangre , Tetrahidrofolatos/sangre , Calibración , Ácido Fólico/efectos de la radiación , Humanos , Cinética , Análisis Multivariante , Fotoquímica , Valores de Referencia , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Espectrometría de Fluorescencia/métodos , Espectrofotometría Ultravioleta/métodos , Tetrahidrofolatos/efectos de la radiación , Factores de Tiempo , Rayos UltravioletaRESUMEN
Aeration of aqueous solutions of 5,10-methenyltetrahydrofolic acid (MTHF) during exposure to ultraviolet irradiation (lambda 300-390 nm, 240 W/m2, 30 min) slowed down photolysis in comparison with deaerated solutions. The rate of photolysis in the presence of oxygen depended on buffer composition. It did not exceed 6% of the starting amount of MTHF. Photolysis of MTHF included opening of the imidazoline ring, dehydrogenation of the tetrahydropterin portion, and elimination of the p-aminobenzoylglutamate moiety. 6,7-Dimethyltetrahydropterin was used as a model compound to show that protonation of the reduced pterin heterocycle increased its tolerance to oxidation, and UV irradiation did not accelerate this process. The stabilizing effect of protonation of the pterin portion and the presence of the positively charged imidazoline moiety are assumed to hamper MTHF oxidation and photolysis. It is assumed that these factors favored the choice of MTHF molecules as photosensors in light-sensitive proteins in the course of evolution.