Rutin in buckwheat herbs grown at different UV-B radiation levels: comparison of two UV spectrophotometric and an HPLC method

Rutin is an antioxidant with many interesting pharmacological effects. It can also be found in buckwheat (Fagopyrum esculentum Moench). UV radiation stimulates the activity of enzymes of the phenylpropanoid pathway and there is some evidence that it influences the rutin content in plants. The aim of the present research was (1) to examine the influence of different levels of UV-B radiation on rutin content and (2) to compare the results obtained by three analytical methods. The plants were grown under three UV-B levels: reduced, ambient and enhanced, simulating 17% ozone depletion. Analyses were performed by HPLC and two spectrophotometric methods. In one, the absorbancies were measured at 420 nm with and without the addition of AlCl(3). In another method the concentration was calculated from absorbancies at 352.5 nm and 366.5 nm according to the Official Methods of Analysis of AOAC International. The highest amounts of rutin were found in flowers, followed by leaves and stems. A comparison of the different treatments revealed that the highest amounts of rutin were in plants grown under ambient radiation, followed by the plants cultivated under enhanced UV-B and then under reduced UV-B radiation. Treatments caused more effect on leaves than on flowers. Leaves developed under ambient light conditions contained 97% more rutin than leaves grown under reduced UV-B radiation. In flowers, the contents differed by 19% only. The results obtained using the three methods showed a good correlation, but the absolute differences were surprisingly high. The AOAC and the AlCl(3) methods gave, on average, 140% and 30% higher results than HPLC, respectively.