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Content of reduced glutathione form as a biomarker of oxidative stress in spinach plants growing in soil contaminated with zinc

Arasimowicz M., Wiśniowska-Kielian B., Niemiec M.

Journal of Ecological Engineering

2014

Vol. 15, nr 1

61--66

The aim of the paper is the assessment of the efficiency of anti-oxidative system in spinach plants growing in the substratum polluted with zinc. The assessment was conducted on the basis of changes of reduced glutathione (GSH) form concentration in the plant aboveground organs. Spinach plants, ‘Matador’ c.v., were cultivated in soils contaminated with zinc in two pot experiments conducted in 2010 and 2011. The experimental substratum was light, slightly soil with granulometric composition of sandy silt loam. Zinc in the acetate form, was supplied to the soil in four doses corresponded to this metal critical concentrations in soil with 0, I, II and III degrees of pollution with this element according to IUNG classification. Simultaneously, the control with natural Zn content in soil was maintained. Zn concentrations in spinach ranged from 412.8 to 1722 mg kg-1 d.m. and increased with growing degree of substratum pollution with this element. Over the course of the vegetation period the content of Zn in plants was generally greater. GSH content in spinach grown in both years of experiments fluctuated from 31.70 to 238 μg g-1 f.m. The biggest content of this compound in spinach was stated in the initial phase of plants growth. The plants tolerated only the first two Zn doses supplied to the substratum. Spinach growing in the objects where zinc additions to the soil equalled II and III degree of substratum pollution died shortly after germination. The plants from these objects in the initial growth phase contained significantly less GSH than spinach from the objects with two first degrees (0 and I) of substratum pollution with zinc or from the control. The content of reduced glutathione form in spinach is a good biomarker of oxidative stress caused by zinc presence in plants. Synthesis of a bigger amount of GSH conditions spinach plant resistance to over the norm zinc content in soil. The efficiency of antioxidative system in spinach is bigger in the initial phase of this plant growth.

Convenient Method for Determination of Biomarkers of Oxidative Stress Developed in Organism by Environmental Factors

Rutkowski M., Grzegorczyk K.

Chemia i Inżynieria Ekologiczna

2002

Vol. 9, nr 9

1021-1032

A convenient method for environmental diseases diagnostics is described using determination of oxidative stress biomarker - the ratio of reduced to oxidized vitamin C concentrations in blood plasma, based on the use of a phosphotungstate reagent. It makes blue dye (maximum absorption 700 Dm) in reaction with reduced form of this vitamin present in tested sample, which allows spectrophotometrically to measure its concentration (I). Present here, also oxidized form of vitamin C undergoes reduction - the whole vitamin in the sample obtains reduced form, then determined as previously (II). Difference of the data (II-I) gives the concentration of vitamin C oxidized form. Conversion of oxidized vitamin C present in samples to the reduced form depends on used reducing agent and on reduction time: for dithiothreitol it was complete during 20 minutes. The method detection limit is 5 x 10-7 mol. dm-J. Results of clinical analyses in diseases with environmental background obtained with this method were consistent with data from HPLC method obtained from literature.

Opisano dogodną dla diagnostyki chorób środowiskowych metodę oznaczania biomarkera stresu oksydacyjnego, jakim jest stosunek stężeń zredukowanej i utlenionej witaminy C w osoczu krwi, opartą na zastosowaniu odczynnika fosforowolframowego. Tworzy on błękitny barwnik (maksimum absorpcji 700 nm) w reakcji z obecną w badanej próbce formą zredukowaną tej witaminy, co pozwala spektrofotometrycznie oznaczyć jej stężenie (I). Współobecną tu formę utlenioną witaminy C poddaje się redukcji - cała ilość witaminy w próbce uzyskuje postać zredukowaną, oznaczaną jak uprzednio (11). Różnica wyników (11-1) daje stężenie formy utlenionej witaminy C. Konwersja zawartej w próbkach utlenionej witaminy C do postaci zredukowanej jest uzależniona od zastosowanego reduktora i od czasu redukcji: dla ditiotreitolu przy 20 min była całkowita. Badania kontrolne wykazały pełną poprawność parametrów analitycznych opracowanej metody, granica wykrywalności wynosi 5 x 10-7 mol. dm-3. Otrzymywane przy jej użyciu wyniki analiz klinicznych w chorobach o podłożu środowiskowym są zgodne z danymi z piśmiennictwa uzyskanymi za pomocą HPLC.