The study of ascorbic acid as a reducing agent for iron(III) has been investigated in order to obtain an alternative carcinogenic reducing agent, hydroxylamine, used in spectrophotometric standard method based on the formation of a red-orange complex of Fe(II)-o-phenanthroline. The study was optimised with regards to ascorbic acid concentration as well as pH solution. The results showed that ascorbic acid showed maximum capacity as reducing agent of iron(III) under concentration of 4.46.10 -4 M and pH solution of 1-4.Under these conditions, ascorbic acid reduced iron(III) proportionally and performed similarly to that of hydroxylamine. The method gave result to linear calibration over the range of 0.2-2 mg/L withhigh accuracy of 97 % and relative standard deviation of less than 2 %. This method was successfully applied to assay iron speciation in water samples.Keyword: Iron, speciation, ascorbic acid, o-phenanthroline, spectrophotometry INTRODUCTIONIron is the second most abundant metallic element in the earth's crust and is essential in the metabolism of plants and animals. If presented in excessive amounts; however, it forms oxyhydroxide precipitate that stains laundry and porcelain. As a result, the recommended limit for iron in domestic water supplies is 0.3 mg/L. 1 Determination of Fe(II) and Fe(III) is important in many natural sciences like geology, agriculture, biology or environmental protection, mainly on account of different bioavailability and metabolism of iron on its two oxidation states. Iron(II) is essential for good health, as it helps transport oxygen in the blood and storage of oxygen by means of haemoglobine and myoglobine.2 Iron deficiency may lead to low immunity and infectious diseases. If the human body takes in too much iron will cause excretory functional disturbance and likely have greater carcinogenic potential. Both Fe (II) and Fe(III) are important in the biosphere, serving as an active centre of a wide range of proteins such as oxidases, reduces and dehydrates.3 However the plants need a certain amount of iron(II) to survive. Iron helps them create chlorophyll and aids in several other chemical processes plants perform. However, too much iron can have a toxic effect on the plant, weakening and eventually killing it. It should be noted that plants only absorb ferrous iron particles from the soil, and that other forms of iron will not affect plants.

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This work presents a simple and innovative protocol employing a microfluidic paper-based analytical device (µPAD) for equipment-free determination of mercury. In this method, mercury (II) forms an ionic-association complex of tetraiodomercurate (II) ion (HgI42−(aq)) using a known excess amount of iodide. The residual iodide flows by capillary action into a second region of the paper where it is converted to iodine by pre-deposited iodate to liberate I2(g) under acidic condition. Iodine vapor diffuses across the spacer region of the µPAD to form a purple colored of tri-iodide starch complex in a detection zone located in a separate layer of the µPAD. The digital image of the complex is analyzed using ImageJ software. The method has a linear calibration range of 50–350 mg L−1 Hg with the detection limit of 20 mg L−1. The method was successfully applied to the determination of mercury in contaminated soil and water samples which the results agreed well with the ICP-MS method. Three soil samples were highly contaminated with mercury above the acceptable WHO limits (0.05 mg kg−1). To the best of our knowledge, this is the first colorimetric µPAD method that is applicable for soil samples including mercury contaminated soils from gold mining areas.

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