ESR Study of the Singlet Oxygen Quenching and Protective Activity of Trolox on the Photodecomposition of Riboflavin and Lumiflavin in Aqueous Buffer Solutions

ABSTRACT:  Singlet oxygen quenching activity of Trolox, a water-soluble derivative of tocopherol, was studied by electron spin resonance (ESR) spectroscopy in a buffer solution (pH 7.4) containing methylene blue (MB), 2,2,6,6-tetramethyl-4-piperidone (TMPD) after light illumination for 30 min. Trolox at the concentration of 125 μM quenched 89.1% singlet oxygen in the system. Trolox showed significantly higher singlet oxygen quenching activity than ascorbic acid in the buffer solution ( P  < 0.05). Riboflavin in phosphate buffer solutions was degraded very fast under fluorescent light illumination. The photodegradation rate of riboflavin at pH 8.5 was significantly higher than pHs 4.5 and 6.5 ( P  < 0.05). Lumiflavin was also degraded under the fluorescent light illumination, but its degradation rate was much lower than that of riboflavin under the same light intensity. Unlike riboflavin, the rate of lumiflavin photodegradation was the greatest at pH 4.5 and followed by pHs 6.5 and 8.5, in a decreasing order. Trolox greatly protected the photodegradation of riboflavin and lumiflavin. The protective activities of Trolox against the photodegradation of riboflavin and lumiflavin were also pH dependent. The treatments of 5 mM Trolox in the buffer solutions of pHs 8.5 and 6.5 exhibited 56.1% and 31.7% protection of riboflavin against degradation during 120 min light illumination, respectively. The treatments of Trolox at the concentrations of 1, 3, and 5 mM in the buffer of 6.5 exhibited 14.8%, 58.4%, and 81.4% protection of lumiflavin against degradation during 24 h light illumination, respectively.