Removal of added nitrate in cotton burr compost, mulch compost, and peat: mechanisms and potential use for groundwater nitrate remediation

We conducted batch tests on the nature of removal of added nitrate in cotton burr compost, mulch compost, and sphagnum peat that may be potentially used in a permeable reactive barrier (PRB) for groundwater nitrate remediation. A rigorous steam autoclaving protocol (121 degrees C for 2h each day for three consecutive days) for the cotton burr compost and autoclaving of all labware and the nitrate working solutions resulted in drastically different results compared to the non-autoclaved treatment. In the non-autoclaved cotton burr compost, added nitrate at 20 mg N l(-1) decreased rapidly and was not detected after 3d; whereas, the autoclaved cotton burr compost showed persistent nitrate above 15.5 mg N l(-1) even after 10d, which is comparable with nitrate concentrations above 17.6 mg N l(-1) in a treatment using NaN(3) at 1000 mg l(-1). Dewaxed cotton burr compost showed decreased nitrate reduction compared to the pristine cotton burr compost. No nitrate reduction was detected in the dewaxed sphagnum peat. It is concluded that nitrate removal in the organic media is controlled by microbiologically mediated processes. The use of readily available cotton burr and mulch composts may offer a cost-effective method of nitrate removal from contaminated groundwater.     

纳米铁系双金属-微生物体系去除地下水NO-3-N研究

采用不同液相还原法制备纳米Fe0、Fe/Ni和Fe/Cu粒子,将其与反硝化细菌混合应用于地下水NO3--N去除研究。考察3种体系对NO3--N去除速率的影响,并对其脱氮产物及RNA水平上纳米铁系双金属对反硝化细菌的毒性效应进行了分析和讨论。结果表明,9 d内纳米Fe0体系可完全将NO3--N去除,过程中伴随NO2--N先升高后降低的生成趋势,NH 4+-N生成52%;纳米Fe/Ni体系脱氮速率最快,6 d内可将NO 3--N完全去除,几乎未检测到NO 2--N的生成,而NH 4+-N的转化率高达69%;纳米Fe/Cu体系7 d内可将NO3--N去除完全,NH4+-N的生成率降低,仅39%,但是出现33%NO2--N积累。从反应前后反硝化细菌总RNA浓度变化看,3种纳米粒子对反硝化细菌的毒性大小为纳米Fe/Ni﹥纳米Fe/Cu﹥纳米Fe0。     

固态碳源去除地下水硝酸盐的模拟实验

选取了5种研究较少的固体材料,棉花、丝瓜络、甘蔗渣、可降解餐盒、木屑作为去除地下水硝酸盐的外加碳源。在锥形瓶中进行反硝化对比实验,研究了不同固态碳源下NO3--N、NO2--N、NH4+-N及pH的变化情况,分析了NO3--N及总氮的去除率。研究结果表明,反硝化过程中pH呈升高趋势,在6.9～8.5范围内浮动。可降解餐盒和丝瓜络相对于其他的固态碳源来说,对NO3--N和总氮有较高的去除率,但丝瓜络的总氮去除率明显低于可降解餐盒。可降解餐盒的硝酸盐去除率达到98.28%,总氮去除率达到93.48%。可降解餐盒能够有效地去除地下水硝酸盐,达到以废治废的效果,是经济有效的最佳固态碳源。     

Effects of macrophytes and external carbon sources on nitrate removal from groundwater in constructed wetlands

Several microcosm wetlands unplanted and planted with five macrophytes (Phragmites australis, Commelina communis, Penniserum purpureum, Ipomoea aquatica, and Pistia stratiotes) were employed to remove nitrate from groundwater at a concentration of 21-47 mg NO3-N/l. In the absence of external carbon, nitrate removal rates ranged from 0.63 to 1.26 g NO3-N/m2/day for planted wetlands. Planted wetlands exhibited significantly greater nitrate removal than unplanted wetlands (P<0.01), indicating that macrophytes are essential to efficient nitrate removal. Additionally, a wetland planted with Penniserum showed consistently higher nitrate removal than those planted with the other four macrophytes, suggesting that macrophytes present species-specific nitrate removal efficiency possibly depending on their ability to produce carbon for denitrification. Although adding external carbon to the influent improved nitrate removal, a significant fraction of the added carbon was lost via microbial oxidation in the wetlands. Planting a wetland with macrophytes with high productivity may be an economic way for removing nitrate from groundwater. According to the harvest result, 4-11% of nitrogen removed by the planted wetland was due to vegetation uptake, and 89-96% was due to denitrification.     

Concurrent nitrate and Fe(III) reduction during anaerobic biodegradation of phenols in a sandstone aquifer

The biodegradation of phenols (5, 60, 600 mg l⁻¹) under anaerobic conditions (nitrate enriched and unamended) was studied in laboratory microcosms with sandstone material and groundwater from within an anaerobic ammonium plume in an aquifer. The aqueous phase was sampled and analyzed for phenols and selected redox sensitive parameters on a regular basis. An experiment with sandstone material from specific depth intervals from a vertical profile across the ammonium plume was also conducted. The miniature microcosms used in this experiment were sacrificed for sampling for phenols and selected redox sensitive parameters at the end of the experiment. The sandstone material was characterized with respect to oxidation and reduction potential and Fe(II) and Fe(III) speciation prior to use for all microcosms and at the end of the experiments for selected microcosms.The redox conditions in the anaerobic microcosms were mixed nitrate and Fe(III) reducing. Nitrate and Fe(III) were apparently the dominant electron acceptors at high and low nitrate concentrations, respectively. When biomass growth is taken into account, nitrate and Fe(III) reduction constituted sufficient electron acceptor capacity for the mineralization of the phenols observed to be degraded even at an initial phenols concentration of 60 mg l⁻¹ (high) in an unamended microcosm, whereas nitrate reduction alone is unlikely to have provided sufficient electron acceptor capacity for the observed degradation of the phenols in the unamended microcosm.For microcosm systems, with solid aquifer materials, dissolution of organic substances from the solid material may occur. A quantitative determination of the speciation (mineral types and quantity) of electron acceptors associated with the solids, at levels relevant for degradation of specific organic compounds in aquifers, cannot always be obtained. Hence, complete mass balances of electron acceptor consumption for specific organic compounds degradation are difficult to confine. For aquifer materials with low initial Fe(II) content, Fe(II) determinations on solids and in aqueous phase samples may provide valuable information on Fe(III) reduction. However, in microcosms with natural sediments and where electron acceptors are associated with the sediments, complete mass-balances for substrates and electron acceptors are not likely to be obtained.     

Hydraulic constraints on the performance of a groundwater denitrification wall for nitrate removal from shallow groundwater

Denitrification walls are a practical approach for decreasing non-point source pollution of surface waters. They are constructed by digging a trench perpendicular to groundwater flow and mixing the aquifer material with organic matter, such as sawdust, which acts as a carbon source to stimulate denitrification. For efficient functioning, walls need to be permeable to groundwater flow. We examined the functioning of a denitrification wall constructed in an aquifer consisting of coarse sands. Wells were monitored for changes in nitrate concentration as groundwater passed through the wall and soil samples were taken to measure microbial parameters inside the wall. Nitrate concentrations upstream of the wall ranged from 21 to 39 g N m(-3), in the wall from 0 to 2 g N m(-3) and downstream from 19 to 44 g N m(-3). An initial groundwater flow investigation using a salt tracer dilution technique showed that the flow through the wall was less than 4% of the flow occurring in the aquifer. Natural gradient tracer tests using bromide and Rhodamine-WT confirmed groundwater bypass under the wall. Hydraulic conductivity of 0.48 m day(-1) was measured inside the wall, whereas the surrounding aquifer had a hydraulic conductivity of 65.4 m day(-1). This indicated that during construction of the wall, hydraulic conductivity of the aquifer had been greatly reduced, so that most of the groundwater flowed under rather than through the wall. Denitrification rates measured in the center of the wall ranged from 0.020 to 0.13 g N m(-3) day(-1), which did not account for the rates of nitrate removal (0.16-0.29 g N m(-3) day(-1)) calculated from monitoring of groundwater nitrate concentrations. This suggested that the rate of denitrification was greater at the upstream face of the wall than in its center where it was limited by low nitrate concentrations. While denitrification walls can be an inexpensive tool for removing nitrate from groundwater, they may not be suitable in aquifers with coarse textured subsoils where simple inexpensive construction techniques result in major decreases in hydraulic conductivity.     

Numerical simulation of a fine-grained denitrification layer for removing septic system nitrate from shallow groundwater

One of the most common methods to dispose of domestic wastewater involves the release of septic effluent from drains located in the unsaturated zone. Nitrogen from such systems is currently of concern because of nitrate contamination of drinking water supplies and eutrophication of coastal waters. It has been proposed that adding labile carbon sources to septic distribution fields could enhance heterotrophic denitrification and thus reduce nitrate concentrations in shallow groundwater. In this study, a numerical model which solves for variably saturated flow and reactive transport of multiple species is employed to investigate the performance of a drain field design that incorporates a fine-grained denitrification layer. The hydrogeological scenario simulated is an unconfined sand aquifer. The model results suggest that the denitrification layer, supplemented with labile organic carbon, may be an effective means to eliminate nitrogen loading to shallow groundwater. It is also shown that in noncalcareous aquifers, the denitrification reaction may provide sufficient buffering capacity to maintain near neutral pH conditions beneath and down gradient of the drain field. Leaching of excess dissolved organic carbon (DOC) from the denitrification layer is problematic, and causes an anaerobic plume to develop in simulations where the water table is less than 5-6 m below ground surface; this anaerobic plume may lead to other down gradient changes in groundwater quality. A drain field and denitrification layer of smaller dimensions is shown to be just as effective for reducing nitrate, but has the benefit of reducing the excess DOC leached from the layer. This configuration will minimize the impact of wastewater disposal in areas where the water table is as shallow as 3.5 m.     

复合介质渗透反应格栅去除地下水中的氨氮

针对受低浓度氨氮污染的地下水,实验筛选组合了不同的反应介质,利用串联的多介质填充柱模拟渗透反应格栅,通过物理吸附及生物硝化-反硝化作用来实现氮的去除。结果表明,在进水氨氮浓度为10 mg/L、流速为0.5 m/d的条件下,模拟柱对氨氮的去除率达到98%以上,且不会出现亚硝酸盐及硝酸盐浓度的升高。水体经过释氧柱后溶解氧由2mg/L升高至10 mg/L以上,表明释氧材料可提供硝化细菌所需的好氧环境。好氧柱中填充易于生物挂膜的生物陶粒及对氨氮有较强吸附能力的沸石,二者联用通过生物硝化-物理吸附协同作用实现对氨氮的去除,其中生物作用实现的氨氮去除量占总去除量的50%左右。后续厌氧反应柱填充海绵铁除氧并利用松树皮颗粒作为碳源,创造反硝化菌生长条件,硝酸盐氮浓度可由10 mg/L降低至5 mg/L以下,实现对好氧反应阶段所产生的硝酸盐的去除,避免了地下水的二次污染。     

Retained nitrate, hydrated sulfates, and carbonaceous mass in federal reference method fine particulate matter for six eastern U.S. cities

Material balance of fine particulate matter (PM2.5) measured with the Federal Reference Method (FRM) is developed for one rural and five urban locations in the eastern half of the United States using routine Speciation Trends Network (STN) and FRM chemical measurements and thermodynamic models. The Aerosol Inorganics Model is used to estimate retained particle bound water, and an ammonium nitrate evaporation model is used to estimate nitrate concentrations retained on the Teflon-membrane filter of the FRM. To address large uncertainties in carbonaceous mass calculated from STN carbon measurements, retained carbonaceous mass is derived by material balance between PM2.5 FRM mass and estimates of its non-carbon constituents. The resulting sulfate, adjusted nitrate, derived water, inferred carbonaceous material balance approach (SANDWICH) is compared with reconstructed fine mass (RCFM) using the Interagency Monitoring of Protected Visual Environments monitoring program equation. For this study, the SANDWICH method resulted in approximately 21-27% higher sulfate mass and approximately 24-85% lower nitrate mass. The combined mass associated with sulfates and nitrates, however, are well within +/- 10% of the proportion derived using the more traditional RCFM method. The discrepancies between SANDWICH and measurement-derived carbonaceous mass vary from -21% to +56% on an annual basis and are attributed in part to urban-rural source influences and uncertainties in estimating FRM-retained carbonaceous mass.     

New methods of nitrate removal from water

Nitrate contamination in groundwater resources originates mainly from the excessive use of fertilisers and uncontrolled land discharges of treated wastewater. This can cause potential health hazards to infants and pregnant women, thus limiting the direct use of the groundwater resources for the human consumption in several parts of the world, including India. The conventional processes used to eliminate nitrate from water are ion exchange, reverse osmosis and electro-dialysis. The utility of these processes has been limited due to their expensive operation and subsequent disposal problem of the generated nitrate waste brine. This paper presents a comprehensive account of the methods/techniques used for the removal of nitrate ion from water during the last 10 years with special reference to the biological denitrification and fate of the metals in decontamination processes.     

纳米铁-微生物耦合体系去除硝酸盐的影响因素研究

采用液相还原法制备出纳米铁粒子,并与自养反硝化细菌耦合,以解决单独使用生物反硝化和纳米铁还原法的不足。本实验在纳米铁-微生物耦合体系可以有效还原硝酸盐的基础上,研究了pH、温度和DO等环境因素对该耦合体系脱氮速率和产物的影响,以期通过优化参数达到最好的脱氮效果。结果表明,该体系在中性条件下能够快速将硝酸盐还原,随pH升高,氨氮比例无显著变化,均在40%左右,但还原速率有所下降;随温度的升高,氨氮比例有所上升,而反应速率明显升高,但该体系在5℃时仍能将硝酸盐完全去除;耦合体系中的DO过高或过低都会导致产物中氨氮比例的增加,0.4 mg/L左右为较适宜DO水平,但对硝酸盐还原速率的影响不大,当DO为0.8 mg/L时,硝酸盐仍可以在8 d内完全去除。因此,该耦合脱氮体系对pH、温度和DO的适应能力较强,有利于实际地下水的原位修复。     

Evaluation of heavy metal contents in co-composts of poultry manure with barley wastes or chestnut burr/leaf litter

The presence of heavy metals in composts is a main cause of adverse effects on animal and human health, transmitted through the food chain from the soil, groundwater and plants. In this study, the contents of Zn, Cd, Pb and Cu present in co-composts of poultry manure (liquid or solid) with a co-composting material (barley wastes or chestnut burr/leaf litter) were assessed. A compost of solid manure was used as control because a compost cannot be obtained from the liquid manure. The original solid poultry manure showed a Zn content of 2134+/-75 mg/kg, exceeding the current legal limit in Spain of 1100 mg/kg. In the solid poultry manure co-compost with chestnut burr/leaf litter and barley wastes, Zn content decreased to 813+/-25 mg/kg and 883+/-37 mg/kg, respectively. The contents in heavy metals (Zn, Cd, Pb and Cu) of the co-composts were under the maximum limit permitted under the Spanish legislation, excepting for the Zn level in liquid poultry manure co-composted with chestnut burr/leaf litter.     

Transport and reduction of nitrate in clayey till underneath forest and arable land

Transport and reduction of nitrate in a typically macroporous clayey till were examined at variable flow rate and nitrate flux. The experiments were carried out using saturated, large diameter (0.5 m), undisturbed soil columns (LUC), from a forest and nearby agricultural sites. Transport of nitrate was controlled by flow along the macropores (fractures and biopores) in the columns. Nitrate reduction (denitrification) determined under active flow mainly followed first order reactions with half-lives (t(1/2)) increasing with depth (1.5-3.5 m) from 7 to 35 days at the forest site and 1-7 h at the agricultural site. Nitrate reduction was likely due to microbial degradation of accumulated organic matter coupled with successive consumption of O2 and NO3- in the macropore water followed by reductive dissolution of Fe and Mn from minerals along the macropores. Concentrations of total organic carbon measured in soil samples were near identical at the two study sites and consequently not useful as indicator for the observed differences in nitrate reduction. Instead the high reduction rates at the agricultural site were positively correlated with elevated concentration of water-soluble organic carbon and nitrate-removing bacteria relative to the forest site. After high concentrations of water-soluble organic carbon in the columns from the agricultural site were leached they lost their elevated reduction rates, which, however, was successfully re-established by infiltration of new reactive organics represented by pesticides. Simulations using a calibrated discrete fracture matrix diffusion (DFMD) model could reasonably reproduce the denitrification and resulting flux of nitrate observed during variable flow rate from the columns.     

Microbial reduction of nitrate in the presence of nanoscale zero-valent iron

Microbial reduction of nitrate in the presence of nanoscale zero-valent iron (NZVI) was evaluated to assess the feasibility of employing NZVI in the biological nitrate treatment. Nitrate was completely reduced within 3 d in a nanoscale Fe(0)-cell reactor, while only 50% of the nitrate was abiotically reduced over 7 d at 25 °C. The removal rate of nitrate in the integrated NZVI-cell system was unaffected by the presence of high amounts of sulfate. Efficient removal of nitrate by Fe(II)-supported anaerobic culture in 14 d indicated that Fe(II), which is produced during anaerobic iron corrosion in the Fe(0)-cell system, might act as an electron donor for nitrate. Unlike abiotic reduction, microbial reduction of nitrate was not significantly affected by low temperature conditions. This study demonstrated the potential applicability of employing NZVI iron as a source of electrons for biological nitrate reduction. Use of NZVI for microbial nitrate reduction can obviate the disadvantages associated with traditional biological denitrification, that relies on the use of organic substrates or explosive hydrogen gas, and maintain the advantages offered by nano-particle technology such as higher surface reactivity and functionality in suspensions.     

Characteristics of an aerobic denitrifier that utilizes ammonium and nitrate simultaneously under the oligotrophic niche

Introduction

An aerobic denitrifier was isolated from the Hua-Jia-Chi pond in China and identified as Pseudomonas mendocina 3-7 (Genbank No. HQ285879). This isolated strain could express periplasmic nitrate reductase which is essential for aerobic denitrification occurred when the dissolved oxygen (DO) level maintains at 3?C10?mg?L^?1.

Methods

To determine whether the ability of isolated strain is exhibited in the bioremediation of polluted drinking source water, the heterotrophic nitrification and aerobic denitrification characteristics of P. mendocina 3-7 under different cultural conditions such as oxygen level, nitrate and organic concentrations were studied from the nitrogenous balance in the paper.

Results and conclusions

By measuring the nitrogen balance in all experiments under different culture conditions, the removal of total organic carbon and ammonium was positively correlated with total nitrogen removal, especially under high substrate level. With substrate concentration decreasing, ammonium and nitrate removal occurred separately, and ammonium was completely utilized first under low substrate concentration. Compared to that under high substrate level, the specific growth rate of P. mendocina 3-7 was not low under the low substrate level and the pollutant removal efficiencies remained high, which implies the stronger nitrogen removal and acclimatization capacities of the strain in oligotrophic niches.     

Accumulation of nitrite in denitrifying barriers when phosphate is limiting

Permeable in situ denitrifying barriers can remove nitrate from groundwater. Barriers may be constructed by filling an excavated area with a porous mixture of sand, fine gravel, and substrate or by the injection of a nonaqueous phase substrate into an aquifer. The substrate stimulates the development of a denitrifying microbial community by providing an electron donor. The objective of this study was to determine the ability of denitrifying barriers to function under low-phosphate conditions. Sand columns injected with a soybean oil emulsion were used as laboratory models of denitrifying barriers. When a natural groundwater containing 17 mg l(-1) nitrate-N and 0.009 mg l(-1) phosphate-P was pumped through the columns, only a small amount of nitrate was removed from the water and, in some effluent fractions, 52% to 88% of the influent nitrate had converted to nitrite. Nitrite also accumulated when the phosphate concentration of the groundwater was increased to 0.040 or 0.080 mg l(-1) phosphate-P. Only when a 0.160 mg l(-1) phosphate-P supplement was added to the groundwater was there a loss of nitrate without a large accumulation of nitrite. The addition of solid calcium phosphate or rock phosphate to the sand columns was found to provide adequate phosphate for denitrification in short-term studies. These studies point out the need to ensure that adequate phosphate is present in denitrifying barriers especially when such barriers are used beneath phosphate-binding soils.     

不同阳离子对Fe~0还原硝酸盐的影响

由于水中硝酸盐污染的普遍性、难去除性和对人体健康的潜在危害性而引起人们的广泛关注。通过批实验,考察了不同阳离子(Fe2+、Fe3+和Cu2+)对Fe0还原硝酸盐的影响。结果表明,由于加入阳离子可直接或间接地增加溶液中的Fe2+而都能促进硝酸盐的还原,作用顺序为Fe3+Fe2+Cu2+;Fe2+对硝酸盐的还原具有重要作用,并随着反应的进行,转化为铁氧化物附着在铁表面而降低铁的活性;硝酸盐还原的主要产物为氨氮,亚硝酸盐只在反应初期有少量积累,尤其是加Cu2+的体系中,但随后都很快降低;在所有体系中,检测到的三氮(NO3--N、NO2--N和NH4+-N)之和只占理论总氮的51.5%~82.6%;动力学分析表明,硝酸盐的还原在不加阳离子的体系中更符合一级反应,而加了阳离子的处理更符合Lo-gistic模型。本研究结果阐明了Fe2+对Fe0还原硝酸盐的重要性。     

Hydrochemical and multivariate statistical interpretations of spatial controls of nitrate concentrations in a shallow alluvial aquifer around oxbow lakes (Osong area, central Korea)

Hydrochemical and multivariate statistical interpretations of 16 physicochemical parameters of 45 groundwater samples from a riverside alluvial aquifer underneath an agricultural area in Osong, central Korea, were performed in this study to understand the spatial controls of nitrate concentrations in terms of biogeochemical processes occurring near oxbow lakes within a fluvial plain. Nitrate concentrations in groundwater showed a large variability from 0.1 to 190.6 mg/L (mean = 35.0 mg/L) with significantly lower values near oxbow lakes. The evaluation of hydrochemical data indicated that the groundwater chemistry (especially, degree of nitrate contamination) is mainly controlled by two competing processes: 1) agricultural contamination and 2) redox processes. In addition, results of factorial kriging, consisting of two steps (i.e., co-regionalization and factor analysis), reliably showed a spatial control of the concentrations of nitrate and other redox-sensitive species; in particular, significant denitrification was observed restrictedly near oxbow lakes. The results of this study indicate that sub-oxic conditions in an alluvial groundwater system are developed geologically and geochemically in and near oxbow lakes, which can effectively enhance the natural attenuation of nitrate before the groundwater discharges to nearby streams. This study also demonstrates the usefulness of multivariate statistical analysis in groundwater study as a supplementary tool for interpretation of complex hydrochemical data sets.     

Kinetics of reductive denitrification by nanoscale zero-valent iron

Zero-valent iron powder (Fe0) has been determined to be potentially useful for the removal of nitrate in the water environment. This research is aimed at subjecting the kinetics of denitrification by nanoscale Fe0 to an analysis of factors affecting the chemical denitrification of nitrate. Nanoscale iron particles with a diameter in the range of 1-100 nm, which are characterized by the large BET specific surface area to mass ratio (31.4 m2/g), removed mostly 50, 100, 200, and 400 mg/l of nitrate within a period of 30 min with little intermediates. Compared with microscale (75-150 microm) Fe0, end product is not ammonia but N2 gas. Kinetics analysis from batch studies revealed that the denitrification reaction with nanoscale Fe0 appeared to be a pseudo first-order with respect to substrate and the observed reaction rate constant (k(obs)) varied with iron content at a relatively low degree of application. The effects of mixing intensity (rpm) on the denitrification rate suggest that the denitrification appears to be coupled with oxidative dissolution of iron through a largely mass transport-limited surface reaction (<40 rpm).     

Nitrate reduction by fluoride green rust modified with copper

Nitrate reduction by the fluoride form of green rust modified with copper (GR-F(Cu)) was investigated using a batch reactor system. The extent of nitrate reduction was measured by measuring the increase in concentration of ammonia, which is the final product of nitrate reduction by GR. This approach was required, because nitrate could be removed from solution by ion exchange without reduction. The rate of ammonium production was investigated over the range of pH 7.8-11. The fastest reaction was achieved at pH 9 when GR was present at a concentration of 0.083M as Fe(II) and 1mM of Cu(II) was added. The rate at pH 9 was enhanced by a factor of 2.5 compared to that at pH 7.8 by comparing the time elapsed to transform all nitrate to ammonium. Kinetics of nitrate reduction by GR-F at pH 7.8 were affected by the concentration of Cu(II) added. The rate constants for ammonium production increased from 0.012 to 1.52h(-1) as Cu(II) additions increased from 0 to 2.5mM, but the reaction rate at 5mM was slightly decreased to 1.25h(-1). The mechanism of enhanced rates of nitrate reduction by addition of Cu(II) could not be fully determined in this study. However, XRD results showed that magnetite was produced in the reaction of Cu(II) and GR-F and SEM shows the production of nano-size particles which were not fully identified in this study. In addition, the concentration of Fe(II) in GR was observed to linearly decrease with concentration of Cu(II) added.