J. Nig. Soc. Phys. Sci. 4 (2022) 891 Journal of the Nigerian Society of Physical Sciences Levels of Zinc (Zn), Copper (Cu), Iron (Fe), and Cadmium (Cd) in Soil, Rice Stalk, and Oryza Sativa Grain in Ishiagu Rice Field, Ebonyi State, Nigeria; Human Health Risk D. N. Ajaha, E. Agboeze ID a,∗, J. N. Ihediohab, E. Chukwudi-Madua, C. C. Chimea aDepartment of Industrial chemistry, Enugu State University of Science and Technology, Enugu State, Nigeria bDepartment of pure and Industrial chemistry, University of Nigeria Nsukka, Enugu State, Nigeria Abstract Levels of heavy metals (Zn, Cu, Fe, Cd) were determined in soil, rice grain, and rice stalk from Federal College of Agriculture Ishiagu rice field, Ebonyi state, Nigeria. The dried samples were digested with a 1: 3 (HNO3: HCl) mixture and analyzed with atomic absorption spectrophotometer (AAS). The mean concentration of the metals in the soil before planting, soil after harvest, and rice grain were as follows: Zn (7.28, 11.33 and24.90); Cu (3.40, 4.64 and 4.14); Fe (803.04, 735.47 and 107.78); Cd (1.14, ND and ND) and were all within FEPA and FAO/WHO limits. The daily intake values for a 60 kg adult were Zn (0.04), Cu (0.01), and Fe (0.18) and were all below the recommended limits by Codex Alimentarius standards. The Target Hazard Quotient (THQ) for Zn, Cu, and Fe was less than one (1<), and the total hazard index was less than 1, indicating that the population will not be exposed to the potential health risk from these metals. However, the metal levels should be monitored to ensure they stay at harmless levels. DOI:10.46481/jnsps.2022.891 Keywords: Heavy metals, Rice, Soil, Health risk, Environmental pollution Article History : Received: 26 June 2022 Received in revised form: 28 July 2022 Accepted for publication: 14 August 2022 Published: 25 September 2022 c© 2022 The Author(s). Published by the Nigerian Society of Physical Sciences under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0). Further distribution of this work must maintain attribution to the author(s) and the published article’s title, journal citation, and DOI. Communicated by: E. A. Emile 1. Introduction Advancement and mechanization of urban growth have pro- moted socio-economic development in developing countries like Nigeria and the world at large. But besides all the positive ef- fects, they cause ”environmental pollution”. Soil contamination ∗Corresponding author tel. no: +234 9039239802 Email address: emmanuel.agboeze@gmail.com (E. Agboeze ID ) by heavy metals is a significant environmental concern world- wide, as it concerns human health and food security/quality [1- 5]. According to Yap et al. [5], the significant sources of heavy metals include anthropogenic environmental activities like min- ing, smelting processes, steel and iron industries, chemical in- dustries, agriculture, and domestic activities [4]. These sources outweigh natural sources like weathering of the parent material and volcanic eruptions [5]. Recently, concern has been raised about possible contamination of the crop (rice) by heavy met- als. The plants can absorb these heavy metals in the soil through 1 https://orcid.org/0000-0001-5337-117X https://orcid.org/0000-0001-5337-117X Ajah et al. / J. Nig. Soc. Phys. Sci. 4 (2022) 891 2 their roots, stems, or leaves and accumulate in their organs [6, 7]. In specific concentrations, some of these heavy metals are essential to plants, but in higher concentrations can become toxic. These metals are a health hazard to living organisms due to their persistence, non-biodegradable, and non-thermal degradable environmental characteristics [5]. The uptake of metals in excessive amounts may either cause harm to the plant or enter the food chain and accumulate when these plants are taken up. Metals accumulated in the human body through the food chain cause diseases like lung cancer and damage the cen- tral nervous system, kidney, and liver [8-11]. Heavy metals, which can pose severe hazards to humans and the environment, are increasingly being found in the envi- ronment due to the growth of mining, smelting, and other in- dustrial operations. The quality of the surrounding air, soil, and water bodies is affected by pollution from heavy metals like lead (Pb), arsenic (As), nickel (Ni), cadmium (Cd), copper (Cu), and zinc (Zn), which endangers the lives of both animals and humans through the food chain [12]. Evaluation of potential impacts on human health in contam- inated environmental media is part of assessing the risk to hu- man health [4]. Human exposure to contaminants, the type of contaminants, and the affected person’s susceptibility all deter- mine how they affect human health [4]. Health impacts may in- clude increased risk of cancer, high blood pressure, acute neuro- logical abnormalities in fetuses, organ dysfunction, respiratory issues, physical and mental illness, shortened life expectancy, and immune system deterioration [12]. This study is intended to determine the levels of Zinc, Cop- per, Iron, and Cadmium in soil, rice grain, and stalk obtained from a rice field in a local area in Nigeria (Federal College of Agriculture Ishiagu, Ebonyi state) through the determination of the physicochemical properties of the soil, evaluation of the level of contamination, and health risk assessment. 2. Experimental 2.1. Area of Study See figure 1. 2.2. Sample Collection The land (Federal College of Agriculture, ishiagu, rice field) is about 6 hectares, i.e. (240×75m). The land was divided into 4parts (A, B, C, and D). 2.3. Soil Sample Collection Eight (8) composite soil samples were collected from four sec- tions of the rice farm before plantation. Each composite is made of 5 grab samples randomly collected from each area of the rice farm. In the same way, twelve (12) composite soil sam- ples were collected after the rice harvest. Four (4) grab samples were randomly collected from different parts of the land which has not undergone any agricultural practice (a fallow land) as the soil control. These soil samples were taken from 0-20cm depth from the surface of the ground with a clean machete, and the collected samples were placed in well-labeled polyethylene bags. 2.4. Rice Sample Collection Twelve (12) composite rice plant samples were collected from four different sections of the farm. Each composite consists of five (5) randomly selected rice plants concerning the soil al- ready sampled. The rice grains from each composite were sepa- rated from the stalk. The rice grains and stalks were each placed in polyethylene bags. 2.5. Sample Preparation 2.5.1. Soil Preparation The soil samples were air-dried, crushed, and passed through a 0.16mm sieve to remove gravel-sized materials and then ho- mogenized with a mortar and pestle. The samples were then stored in polyethylene bags with labels for analysis. 2.5.2. Rice Preparation The rice grain samples were refined to remove the husk using mortar and pestle and winnowed with a tray pan. The polished rice grains were then stored in polyethylene bags well labeled. The rice stems were finely chopped with the knife on a wooden platform and stored in labeled polyethylene bags. 2.5.3. Ash Two (2) ash samples were collected from 2 different abattoir sites and were mixed to get a homogenous sample. The ash was used to mix the rice seeds before broadcasting to scare birds away or prevent them from eating them. 2.5.4. Sample Size The overall sample for analysis became: 8 for soil before plant- ing + 12 for soil after harvest + 12 for rice grain +12 for rice stem +4 for soil control, and + 1 for ash, totaling 49 samples. 2.5.5. Digestion of Soil Samples, Rice Grain Samples, and Rice Stalk Samples 10g each of the soil and rice grain samples, 5g of the rice stalk were measured into different digestion flasks, and 40ml of Aqua- regia (Nitric acid and HCl in the ratio of 1:3) was added. The mixtures were then heated on a heating mantle to boil until the fume became clear and were allowed to cool. Each of the var- ious digested solutions was diluted with deionized water and filtered through a filter paper into 100ml volumetric flasks and were made up to the mark with deionized water. The solu- tions were finally kept in test tubes. Concentrations of Zn, Cu, Fe, and Cd were determined using the AA-7000 atomic spec- troscopy. Statistical analysis of the data obtained was done us- ing SPSS version 17 for windows and Palisade analysis. 2.6. Quality Assurance The precision of the analytical procedure was investigated by carrying out recovery experiments. This was done by determin- ing the metals’ concentration in duplicate spiked and unspiked rice samples. Spiking was done by adding 1 ML of 2ppm and 2 Ajah et al. / J. Nig. Soc. Phys. Sci. 4 (2022) 891 3 Figure 1. Area of study 4ppm metal solution to 2 g of samples, which were later sub- jected to the digestion procedure. % recovery = a − b × 100 c , (1) where a denotes concentration in the spiked sample, b repre- sents concentration in the unspiked sample and c denotes con- centration of the metal ion added. 2.6.1. Risk Assessment This process evaluates the potential effects of a contaminant on humans from doses received through one or more exposure pathways. The health risk from rice consumption was assessed using the target hazard quotient (THQ) and total hazard index (THI) [13]. The target hazard quotient (THQ) is the ratio of the determined dose of a pollutant to a reference dose level. In con- trast, the total hazard index (THI) evaluates the potential risk of adverse health effects from a mixture of chemical constituents in rice. If this ratio is less than 1, the exposed population is unlikely to experience noticeable adverse effects [14-16]. The following equation calculated the THQ and THI values for the metals: T HQ = EF × ED × IR × C R f D × BW × AT × 10−3 (2) T HI = T HQ1 + T HQ2 + · · · + T HQn (3) Where THQ is the target hazard quotient, EF is the exposure frequency (365 days /year), ED is the exposure duration (54 years), IR is the rice ingestion(kg/person/day), and C is the metal concentration in rice (mg/kg), RFD is the oral reference dose (mg/kg/day) and AT is the average time for non–carcinogens (365 days /year× ED). The oral reference dose for metals (mg/kg/day) were: Cu(0.040), Zn(0.300), Fe(0.700) [17]. The calculated THQ value is also shown in Table 2. It is less than 1, thus indicating that the Nigerian population has not been ex- posed to the potential health risk of dietary copper via rice con- sumption. 2.7. Data Analysis Data obtained were analyzed with SPSS 17 for windows. Cor- relation coefficient analysis was carried out to establish the cor- 3 Ajah et al. / J. Nig. Soc. Phys. Sci. 4 (2022) 891 4 relation pattern of various metal pairs in soil and rice grain sam- ples. The student’s t-test was carried out to determine any sig- nificant difference in the metal concentration in the soil before planting and after harvest at p<0.05. 3. Results and Discussion Table 1 shows recoveries ranging from 84% to 128% with a mean of 96% and precision of 5.7, a value lower than 10% indicating high accuracy. 3.1. Heavy Metals Concentration in Soil and Rice Grain 3.1.1. Copper in Soil The concentrations of Copper (Table 2) in the soil samples from the control portion ranged from 1.67 mg/kg - 3.33 mg/kg. The lowest value (1.67 mg/kg) was recorded at Control2, while the highest (3.33 mg/kg) was at Control4. These concentrations gave a mean ± SD of 2.22 ± 0 .75 (Table 3). As in Table 2, the concentration of copper in the ”soil before planting” ranged between 2.39 mg/kg - 5.18 mg/kg. The lowest value (2.39 mg/kg) was recorded at D2 and the highest (5.18 mg/kg) at A1. These gave a Mean ± SD of 3.40 ± 0.91 (Table 3). Also, the concentrations of copper (Table 2) in ”soil after harvest” ranged from 3.70 mg/kg to 6.07mg/kg. A3 had the lowest value (3.70mg/kg), and B2 had the highest value (6.07 mg/kg). The mean ± SD was 4.64 ± 0.69 (Table 3). It compared the mean concentrations of (”soil before plant- ing” and ”soil after harvest”) (3.40 mg/kg and 4.64 mg/kg, re- spectively) with the mean concentration of the ”soil control” (2.22 mg/kg). It can be seen that the soil was slightly contam- inated with copper. This contamination could be attributed to the use of pesticides or herbicides. This possibility was sup- ported by the fact that most pesticides used in agricultural soils were based on compounds containing Cu, Hg, Mn, Pb, or Zn [18]. The results compared with other researchers; Zhuang reported 502 mg/kg as the mean concentration of copper in agricultural soils around the Daboshan mine in Guandong, China. Also, Song recorded 8.41-148.73 mg/kg as the mean concentration of copper in agricultural soils of Suxian County, South China. This variation of results might be due to the copper concentra- tions in irrigation water and other agronomic practices in the respective areas [16, 19]. However, the mean concentrations of Cu in (soil before planting (3.40) and soil after harvest (4.64) is below the permissible limits; 1. 150 mg/kg as with Chinese Environmental Quality Stan- dards (1995) for soils [9]. 2. (70-80) mg/kg as with FEPA (1991) guidelines for heavy metals in soil. The low concentrations recorded in this study may be attributed to the continuous removal of cop- per by rice grown in the field. Therefore, the concentra- tion of Copper in Soil may not harm mice and humans when used for rice production [3]. 3.1.2. Copper in Rice The concentration of copper in “Rice stalk” as in Table 2 above ranged between 0.74mg/kg-11.06mg/kg with A1& B3 having the lowest value (0.74 mg/kg) and A3having the highest value(11.06mg/kg). These concentrations gave a mean ± SD of 2.46 ± 2.95, as in Table 3. As in Table 2 above, the concentration of copper in ”rice grain” ranged between 1.84mg/kg-14.81mg/kg, with A2 having the lowest value (1.84 mg/kg) and C3 having the highest value (14.81 mg/kg). The concentrations gave a mean ± SD value of 4.14 ± 3.92as seen in Table 3. In Tanzania, Machiwa reported the mean concentration of Cu in rice collected from different locations to be 3.7mg/kg. Also, in Guandong, China, Zhuang reported the mean concentration of Copper in Rice as 6.34 mg/kg [16, 20]. However, the mean Cu concentrations of the rice samples in this study are within China’s maximum permissible limits (10 mg/kg) [21]. They are also within the FAO/WHO recommended limits (20 mg/kg) for copper in rice grains, which means that the rice is suitable for consumption [21]. 3.1.3. Relationships Between Copper Concentration in the Soil Before Planting and Soil After Harvest Statistical analysis with student T-test indicates a significant difference in the metal (Copper) concentration of the soil before planting and after harvest at p=0.05 with a calculated value of 3.27. Table 3 shows the results of the student T-test for metal concentrations in the ”soil before planting and soil after har- vest.” 3.1.4. Zinc in Soil The Zinc concentration in the soil samples collected from the ”control portion” as in Table 2 ranged between 3.73 mg/kg - 8.43 mg/kg. The lowest value (3.73 mg/kg) was recorded in control2, while the highest (8.43 mg/kg) was in control2. These concentrations gave a mean ± SD of 5.26 ± 2.2, as shown in Table 3. Also, the concentration of Zinc in the “soil before planting” (Table 2) ranged between 2.75mg/kg - 13.17mg/kg with the lowest value (2.75mg/kg) at A1 and the highest value (13.17mg/kg) at A2. These gave a mean ± SD of 7.28 ± 3.90 (Table 3). As in Table 2, the Zinc concentration in ”soil after harvest” ranged between 6.88mg/kg-15.15mg/kg. The lowest value was C3(6.88 mg/kg), while the highest was D3(15.15 mg/kg). The mean ± SD is 11.33 ± 2.51, as in Table 3. Comparing the mean concentration of zinc in the ”soil before planting and soil after harvest” (7.28 mg/kg and 11.33 mg/kg respectively) with that of the soil control (5.26 mg/kg) as seen in Table 2, It can be seen that their values are more than that in the ”soil control” suggesting that the soil is contaminated with zinc. The contam- ination could be attributed to pesticides and herbicides on the field, as zinc is one of the constituents of most pesticides [18]. Also, we compared the results with those reported by researchers like; 1. Zhuang reported that the Zn concentrations of the agri- cultural soils around the Daboshan mine in Guangdong, 4 Ajah et al. / J. Nig. Soc. Phys. Sci. 4 (2022) 891 5 China is 498 mg/kg [14]. 2. Machiwa reported the mean zinc concentration as 65.46mg/kg in the agricultural soils of Lake Victoria basin, Tanzania [20]. 3. Kibassa reported the average range concentration of zinc in agricultural soils collected from six sites in Daressalan to be 33.18 mg/kg [22]. The mean concentrations gotten from the different soil por- tions (soil before planting and soil after harvest, 7.28 mg/kg and 11.33 mg/kg, respectively) in this study did not exceed that recorded by the researchers mentioned earlier and also are far below the maximum permissible limits (300-400 mg/kg) for zinc in soils by (FEPA) and (300 mg/kg) as with Grade ii En- vironmental Quality standards for agricultural soils in China. Therefore, the soil may not be harmful to rice production for consumption by human beings. 3.1.5. Zinc in Rice The concentration of zinc in ”Rice stalks” (Table 2) ranged between 15.14 mg/kg and 69.03 mg/kg, with D2 having the lowest value (15.14 mg/kg) and A1 having the highest value (69.03mg/kg). These concentrations gave a mean ± SD of 45.14 ± 13.73, as shown in Table 3. Also, the concentration of zinc in ”rice grain” ranged between 14.78 mg/kg-32.89mg/kg, as shown in Table 2, with D1 having the lowest value (14.78 mg/kg) and B2 has the highest value (32.89 mg/kg). The mean ± SD value was 24.90 ± 6.06, as shown in Table 3. It compared the mean concentration of zinc in rice grains (24.90 mg/kg) in this study with those reported by [20] (21.7 ug/g) and that reported by [23] (21.5 ug/g). It was found that they are still within the range, although most of the concentrations of zinc in rice individually in this study fall within these limits ex- cept for a few which are a bit higher in the rice grains in areas A2, B2, B3, and C1(31.37,32.89,30.81, and 29.48 mg/kg respec- tively). The concentrations in the rice stalk were relatively high. This could be attributed to the ash from the abattoir (rubber tire), which was used to mix the rice grains before broadcast- ing on the field to avoid /scare birds from eating the broadcasted rice. This ash contains zinc, which was made highly available to the rice stalk then, followed by the little the rice grain absorbed. However, the mean concentrations are still within the Chinese maximum permissible limits for zinc in rice (50 mg/kg) and the FAO/WHO (2002) recommended limits for zinc in rice grains. Therefore, rice is suitable for consumption [17]. 3.1.6. Relationships Between Zinc Concentrations in the Soil Before Planting and Soil After Harvest The statistical analysis with the student T-test indicated a signif- icant difference in the soil’s metal (Zinc) concentration before and after planting at p=0.05 with a calculated value of 2.60. Table 3 shows the results of the student T-test for metal concen- tration in the soil before and after planting. 3.1.7. Iron in Soil The concentration of iron in ”soil control” ranged between 805.26 mg/kg - 823.69 mg/kg, as shown in Table 2, with control1 hav- ing the lowest value (805.26) and control4having the highest value (823.69 mg/kg) and a Mean ± SD of 814.40 ± 7.52 as in Table 3. As in Table 2, the iron concentrations in the ”soil before plant- ing” ranged between 764.29 mg/kg - 845.59 mg/kg. D2 had the lowest value (764.29 mg/kg) while A1 had the highest value (845.59mg/kg), and the Mean ± SD value was 803.04 ± 26 as in Table 3. Also, the iron concentrations in ”soil after harvest” ranged be- tween 611.14-838.81 mg/kg, as in Table 2. The lowest value (611.14 mg/kg) was recorded at C2 and the highest (838.81 mg/kg) at D2. The Mean ± SD value was 735.4 ± 73.20, as shown in Table 3. Comparing the mean concentration of iron in (soil before planting and soil after planting) (803.04 mg/kg and 735.47 mg/kg, respectively) with soil control (814.40mg/kg). These values are lower than the soil control value meaning the soil is not contaminated with iron. However, the mean concen- trations in soils (before planting (803.04mg/kg) and after har- vest (735.47 mg/kg) are still within the maximum permissible limits (7000-550000) as with Chine’s grade ii Environmental quality standards [24]. 3.1.8. Iron in Rice The iron concentrations in ”Rice stalk” ranged between 56.61mg/kg-300.12mg/kg, as shown in Table 2. The lowest value (56.61 mg/kg) was recorded at C2, and the highest 300.12 mg/kg value was recorded at B2. The Mean ± SD value was 150.02 ± 85.20as shown in Table 3. The iron concentrations in ”rice grain,” as shown in Table 2, ranged between 0.66mg/kg-392 mg/kg. The lowest value (0.66 mg/kg) was recorded at B3 and the highest value (392.35 mg/kg) at D2. The Mean ± SD value was 107.78 ± 138.15, as in Table 3. 3.1.9. Relationships Between the Iron Concentration in the Soil (Before Planting) and Soil (After Harvest) The statistical analysis with the student T-test indicated a signif- icant difference in the soil’s metal (Iron) concentration before and after planting at p=0.05 with a calculated value of 2.92. Table 3 shows the results of the student T-test for Metal con- centrations in the soil before and after planting. 3.1.10. Cadmium in Soil Cadmium concentrations were detected in only one portion; in the ”soil before planting” area A1, with a value (1.14 mg/kg), as shown in Table 2. 3.1.11. Cadmium in Rice The concentrations of Cadmium in ”rice stalk” were scantily detected at A1, B1, B2, C3 & D1 with the range (0.02mg/kg-2.03mg/kg) as shown in Table 2. The lowest value (0.02 mg/kg) was recorded at B2 and the highest value at D1. The mean ±SD was 0.53±0.85, as shown in Table 3. The mean 5 Ajah et al. / J. Nig. Soc. Phys. Sci. 4 (2022) 891 6 cadmium concentration in rice stalk was higher than the Codex standards (1993-1995) (0.1mg/kg) for Cadmium in Rice stalk. This high value could be attributed to the ash (from the abattoir (rubber tire) that was used to mix the rice seeds before broad- casting on the rice field [25]. However, none of these concen- trations were made available to the rice grain, which leaves the conclusion that the rice grain is free from Cadmium contamina- tion and is, therefore, fit for consumption. 3.1.12. Zinc Concentrations in Soil Zinc concentration in the ”soil before planting” ranged between 2.75mg/kg and 13.17, with a mean of 7.28. Zinc concentra- tion in ”soil after harvest” ranged between 6.88mg/kg and 15.15 mg/kg, with a mean of 11.33. The mean concentrations of zinc from the different soil portions (soil before planting and soil af- ter harvest) in this study are below the 33.18 mg/kg and 65.46 mg/kg reported by kibassa and Machiwa as average range con- centration of zinc in agricultural soils of Daressalan and lake victoria basin, Tanzania respectively [20, 22]. Moreover, the mean zinc concentration in the soil did not exceed the maxi- mum permissible limits (300-400 mg/kg) for zinc in soils by FEPA and (300 mg/kg) as with Grade ii Environmental Quality standards for agricultural soils in China [24]. 3.1.13. Copper Concentration in Soil Copper concentration in ”soil before planting” ranged between 2.39mg/kg and 5.18 mg/kg, with a mean of 3.40. The Con- centrations of copper in the soil after harvest ranged between 3.70 mg/kg and 6.07 mg/kg, with a mean of 4.64. These mean concentrations are below the permissible limits of 150 mg/kg as with Chinese Environmental quality standards for soil and 70-80 mg/kg as with EPA guidelines for heavy metals in soil. The low concentrations are in line with 8.41-148.73mg/kg as reported by Song as the mean concentration of copper in agri- cultural soils of Suxian County, South China [19]. 3.1.14. Iron Concentration in Soil Iron concentration in ”soil before planting” ranged between 764.29 mg/kg and 845.59 mg/kg, with a mean of 803.04. Iron concentrations in ”soil after harvest” ranged between 611.14 and 838.81 mg/kg with a mean of 735.4. However, the mean concentrations are still within the maximum permissible limits (7000-550000) as with Chinese grade ii Environmental quality standards [24]. 3.1.15. Heavy Metals in Rice Grain Zinc in Rice Grain Zinc concentration in rice grain ranged between 14.78 mg/kg and 32.89 mg/kg, with a mean of 24.90. This mean value is within the 21.5g/g reported by Herawati and the 21.7g/g re- ported by Machiwa as the mean concentration of zinc in rice collected from different locations in Tanzania [23, 20]. How- ever, the zinc mean concentration in this study is below the 50mg/kg recommended limits by FAO/WHO (2002) for zinc in rice grains. Hence it is worth noting that the rice may not pose any kind of danger to human health as a result of Zinc contamination [26]. Copper in Rice Grain The concentration of copper in rice grain ranged between 1.84 mg/kg-14.81mg/kg with a mean of 4.14. This mean value is not far from the 3.7 mg/kg reported as a copper concentration in rice grain collected from different locations In Tanzania by Machiwa and the 6.34 mg/kg reported as the mean concentra- tion of copper in rice from Guangdong, China, by Zhuang [16]. However, the mean Copper concentration of the rice grain sam- ples in this study is within the maximum permissible limits,10 mg/kg by China and 20 mg/kg by FAO/WHO (2002), which shows that the rice is good for consumption. [20, 14]. Iron in Rice Grain Iron concentrations in ”rice grain” ranged from 0.66mg/kg-392mg/kg with a mean of 107.78. Cadmium in Rice Grain Cadmium was not detected in ”rice grain” for all the rice grain samples. Correlation Matrix Correlation is a statistical technique that shows whether and how strongly pairs of variables are related. Table 4 presents the correlation matrix for the metals in the soil before plant- ing. A strong positive and significant correlation was observed between copper and iron with an R-value of 0.787. The strong positive correlation suggests the similar origin of the metal pairs, probably from agrochemicals used on the farm. Liu reported strong correlations among Cu, Ni, and Cr in the soil around an electroplating plant and have implied that the metals have the same pollution sources [27, 28]. A weak positive correla- tion was observed between zinc and iron (r=0.345). However, a weak negative correlation (r=-0.030) was observed between copper and zinc, showing that an increase in the concentration of one metal results in a decrease in the concentration. Table 5 presents the correlation matrix for the metals in the soil after harvest. Weak positive correlations were observed be- tween Cu-Fe, Cu-Zn, and Fe-Zn pairs. Table 6 presents the correlation matrix for the metals in the rice grain. A weak pos- itive correlation was observed between Cu-Zn, showing that an increase in the concentration of one metal results in a decrease in the concentration of another. 3.1.16. Dietary Intake of Heavy Metals and Potential Health Risks Via Rice Consumption Table 7 shows the metal concentrations (mg/kg) in rice, provisional maximum tolerable daily intake (PMTDI) (mg/kg BW) with estimated daily intake (EDI)(mg/kg BW/day), and target hazard quotient (THQ) data for a 60 kg adult. The intake of heavy metals was estimated by multiplication of daily con- sumption rate with metal content in rice divided by the body- weight EDI = MC ×IR BW (4) Where; Mc= Concentration of the heavy metal in contaminated rice, IR= ingestion rate or average rice consumption in the study region, and BW= Bodyweight. 6 Ajah et al. / J. Nig. Soc. Phys. Sci. 4 (2022) 891 7 Table 1: Recoveries and precision (%) of metals Cu, Zn, Fe, and Cd from spiked soil, ash, rice grain, and stalk samples after digestion Element Sample Spike (g/mL) Conc in unspiked sample (g/mL) Conc in spiked sample (g/mL) Recovered conc. (g/m L) Recovery (%) Precision ZINC Soil before plant- ing A 2ppm 4ppm 2.000 2.000 3.8182 5.9827 1.8182 3.9827 92 100 6.3 Soil before plant- ing B 2ppm 4ppm 0.2251 0.2251 1.9134 3.8009 1.6883 3.5758 84 89 4.1 Control 1 2ppm 4ppm 0.3723 0.3723 2.1126 3.9307 1.7403 3.5584 87 89 2.5 Control 2 2ppm 4ppm 0.2078 0.2078 2.1039 4.0260 1.8961 3.8182 95 95 0 Ash 2ppm 4ppm 5.3506 5.3506 7.4632 9.2987 2.1126 3.9481 106 99 4.8 Soil after harvest A 2ppm 4ppm 0.7100 0.7100 2.4242 4.5801 1.7142 3.8701 86 97 8.5 Soil after harvest B 2ppm 4ppm 0.6494 0.6494 2.4242 4.4675 1.7748 3.8181 89 95 4.6 Grain 1 2ppm 4ppm 0.9524 0.9524 2.7965 4.6667 1.8441 3.7143 92 93 1.1 Grain 2 2ppm 4ppm 0.7013 0.7013 2.5108 4.5628 1.8095 3.8615 90 97 5.3 Stalk 1 2ppm 4ppm 2.6753 2.6753 4.4242 6.554 1.7489 3.8787 87 97 3.5 Stalk 2 2ppm 4ppm 1.7489 1.7489 3.6190 5.6190 1.8701 3.8701 94 97 2.3 IRON Soil before plant- ing A 2ppm 4ppm 111.2664 111.2664 113.4672 115.4236 2.2008 4.1572 110 103 4.7 Soil before plant- ing B 2ppm 4ppm 128.2620 128.2620 129.9738 132.5546 1.7118 4.524 86 113 14.3 Control 1 2ppm 4ppm 132.7860 132.7860 134.2533 137.5546 1.4673 4..7686 72 119 33.9 Control 2 2ppm 4ppm 128.5066 128.5066 131.0742 131.9301 2.5676 3.4235 128 86 27.8 Ash 2ppm 4ppm 106.8646 106.8646 108.5764 110.2882 1.7118 3.4236 86 86 0 Soil after harvest A 2ppm 4ppm 134.7424 134.7424 136.8210 138.5328 2.0786 3.7904 104 95 6.4 Soil after harvest B 2ppm 4ppm 132.4192 132.4192 134.1310 136.0873 1.7118 3.6681 86 92 4.7 Grain 1 2ppm 4ppm 1.9563 1.9563 3.6681 5.9913 1.7118 4.035 86 101 11.3 Grain 2 2ppm 4ppm 1.2227 1.2227 3.1790 5.5022 1.9563 4.2795 99 107 5.5 Stalk 1 2ppm 4ppm 6.6026 6.6026 8.6812 10.5153 2.0786 3.9127 104 98 4.2 Stalk 2 2ppm 4ppm 10.0262 10.0262 11.9825 14.0611 1.9563 4.0349 99 101 1.4 COPPER Soil before Planting A 2ppm 4ppm 0.0323 0.0323 2.0693 4.0416 2.037 4.0093 102 100 1.4 Soil before plant- ing B 2ppm 4ppm 0.0000 0.0000 1.9723 3.9769 1.9723 3.9769 99 99 0 7 Ajah et al. / J. Nig. Soc. Phys. Sci. 4 (2022) 891 8 Control 1 2ppm 4ppm 0.0000 0.0000 1.8430 3.9769 1.8430 3.9769 92 99 5.2 Control 2 2ppm 4ppm 0.0647 0.0647 1.9400 3.8799 1.8753 3.8152 94 95 1.1 Ash 2ppm 4ppm 5.2702 5.2702 7.3395 9.2148 2.0693 3.9446 103 99 2.8 Soil after harvest A 2ppm 4ppm 0.2587 0.2587 2.0370 3.8476 1.7783 3.5889 89 90 1.1 Soil after harvest B 2ppm 4ppm 0.2587 0.2587 2.2633 4.0739 2.0046 3.8152 100 95 3.7 Grain 1 2ppm 4ppm 0.0000 0.0000 1.7460 4.1709 1.7460 4.1709 87 104 12.5 Grain 2 2ppm 4ppm 0.0647 0.0647 2.0693 3.9446 2.0046 3.8799 100 97 2.2 Stalk 1 2ppm 4ppm 0.0647 0.0647 1.9400 3.8476 1.8753 3.7829 94 95 1.1 Stalk 2 2ppm 4ppm 0.0970 0.0970 1.8753 3.7182 1.7783 3.6212 89 91 X=96 1.6 X=5.7 Table 2: Metals concentrations (mg/kg) Sample Copper Zinc Iron Cadmium Soil control 1 1.85 3.81 814.32 - 2 1.67 3.73 814.31 - 3 2.04 5.06 805.26 - 4 3.33 8.43 823.69 - Soil before planting A1 5.18 2.75 845.59 1.14 A2 4.05 13.17 821..89 - B1 3.32 9.11 812.46 - B2 3.30 7.48 803.78 - C1 3.68 6.98 785.79 - C2 2.59 3.99 774.81 - D1 2.68 11.68 815.74 - D2 2.39 3.09 764.29 - Ash 1 156.81 42.07 848.28 - Soil after harvest A1 5.55 13.74 789.75 3-.68 A2 4.25 12.51 761.50 - A3 3.70 12.16 761.35 - B1 4.25 9.49 746.86 - B2 6.07 9.00 759.38 - B3 4.04 8.29 736.97 - C1 4.25 10.21 703.58 - C2 4.26 12.96 611.14 - C3 4.60 6.88 659.74 - D1 5.32 13.11 625.99 - D2 4.80 12.41 838.81 - D3 4.59 15.15 830.55 - Rice stalk A1 0.74 69.03 61.86 0.20 A2 5.15 46.61 86.51 - A3 11.06 58.33 144.68 - 8 Ajah et al. / J. Nig. Soc. Phys. Sci. 4 (2022) 891 9 B1 1.48 33.16 280.15 0.16 B2 1.85 39.05 300.12 0.02 B3 0.74 41.26 185.20 - C1 1.48 46.10 197.25 - C2 1.85 41.01 56.61 - C3 1.11 41.74 60.60 0.22 D1 1.47 51.36 126.97 2.03 D2 1.11 15.44 213.29 - D3 1.48 58.62 86.95 - Rice grain A1 2.03 23.06 15.13 - A2 1.84 31.37 10.49 - A3 2.03 28.46 61.16 - B1 1.85 19.88 2.63 - B2 2.03 32.89 213.71 - B3 2.04 30.81 0.66 - C1 7.92 29.48 22.92 - C2 8.86 24.62 16.42 - C3 14.81 23.78 22.37 - D1 2.21 14.78 289.63 - D2 2.04 24.59 392.35 - D3 2.03 15.07 245.83 - Table 3. Mean ± SD of metal concentrations (mg/kg) Sample Copper Zinc Iron Cadmium Soil control 2.22 ± 0.75 5.26 ± 2.2 814.40 ± 7.52 - Soil b/4 planting 3.40 ± 0.91 7.28 ± 3.90 803.04 ± 26.76 1.14 Ash 156.81 ± 156.81 42.07 ± 42.07 848.28 ±848.28 - Soil after harvest 4.64 ± 0.69 11.33 ± 2.51 735.47 ± 73.20 - Rice stalk 2.46 ± 2.95 45.14 ± 13.73 150.02 ± 85.20 0.53±0.85 Rice grain 4.14 ± 3.92 24.90 ± 6.06 107.78 ± 138.15 - Table 4. Pearson’s correlations between different metals in the soil before planting Cu Zn Fe Cu 1 Zn -0.030 1 - Fe 0.787 0.345 1 Table 5. Pearson’s correlations between different metals in the soil after harvest Cu Zn Fe Cu 1 - - Zn 0.071 1 - Fe 0.079 0.274 1 Table 6. Pearson’s correlations between different metals in rice grain Cu Zn Fe Cu 1 - - Zn 0.031 1 - Fe -0.338 -0.373 1 9 Ajah et al. / J. Nig. Soc. Phys. Sci. 4 (2022) 891 10 Table 7. Metal concentrations (mg/kg) in rice, PMTDI (mg/kg /person/day) with EDI (mg/kg bw/day) and THQ data for a 60 kgadult METAL MEAN ± SD PMTDI EDI THQ Copper 4.14±3.92 0.05-0.5 0.01 0.00017 Zinc 24.90±6.06 0.3-1 0.04 0.00014 Iron 107.78±138.15 0.8 0.18 0.00026 Cadmium - - - Provisional maximum tolerable daily intake (PMTDI) by JECFA According to the international rice research institute (2001), the average Nigerian consumes 24.8kg of rice per year, equivalent to 0.1kg per person/day. The daily intakes for a 60 kg adult were compared to the provisional maximum tolerable daily in- takes as stipulated by Joint FAO/WHO Expert Committee Food Additive (JECFA). It was found that the EDIS of the metals falls within the range of the safe values stipulated by JECFA [29]. The target hazard quotient (THQ) of heavy metals from rice consumption is in decreasing order: Fe>Cu>Zn and are all less than 1 indicating there will be no health risk. The total hazard index (THI) for rice consumption for a 60-kg adult is 0.00057, which is less than 1. Thus, the consumption of rice from this field will show no adverse effects from the metals. 4. Conclusion This research demonstrated that Zn, Cu, and Fe concentrations in soil and rice grains did not exceed the threshold set by several international organizations. The preliminary maximum toler- ated daily intake (PMTDI) established by JECFA was less than the metals’ estimated daily intake (EDI). The total hazard index was also less than one, indicating no potential health risk asso- ciated with the intake of rice in this field. The metal’s target hazard quotient (THQ) was less than one. The result from the physicochemical analysis of the soil showed that the soil is a type suitable for rice production. However, we recommend that the field be monitored con- tinuously to ensure that the metals stay at harmless levels. Acknowledgment The authors appreciate the editor and the anonymous re- viewers for their valuable comments towards the improvement of this article. References [1] Y. Jin, L. Wang, Y. Song, J. Zhu, M. Qin, L. Wu & D. Hou, “Integrated life cycle assessment for sustainable remediation of contaminated agricultural soil in China”, Environmental Science & Technology 55 (2021) 12032. [2] C. C. Onoyima, F. G. Okibe, E. Ogah & Y. A. Dallatu, “Heavy metal pol- lution and ecological risk assessment in the sediments of River Kaduna, Nigeria”, Journal of Research in Forestry, Wildlife and Environment 2 (2021) 205. [3] M. Qin, J. Gong, G. Zeng, B. Song, W. Cao, M. Shen & Z. Chen, “The role of microplastics in altering arsenic fractionation and microbial com- munity structures in arsenic-contaminated riverine sediments”, Journal of Hazardous Materials 12 (2022). [4] B. Hikon, G. G. Yebpella, L. Jafiya & S. Ayuba, “Preliminary Investiga- tion of Microplastic as a Vector for Heavy Metals in Bye-ma Salt Mine, Wukari, Nigeria”, Journal of the Nigerian Society of Physical Sciences 3 (2021) 25. [5] D. W. Yap, J. Adezrian, J. Khairiah, B. S. Ismail & R. Ahmad-Mahir, “The uptake of heavy metals by paddy plants (Oryza sativa) in Kota Marudu, Sabah, Malaysia”, Am Eurasian J Agric Environ Sci 6 (2009) 16. [6] S. Cheng, “Heavy metals in plants and phytoremediation”, Environmental Science and Pollution Research 33 (2003) 335. [7] C. S. Romero-Oliva, V. Contardo-Jara, T. Block & S. Pflugmacher, “Accumulation of microcystin congeners in different aquatic plants and crops–A case study from lake Amatitlán, Guatemala”, Ecotoxicology and environmental safety 1 (2014) 335. [8] N. Coen, C. Mothersill, M. Kadhim & E. G. Wright, “Heavy metals of rel- evance to human health induce genomic instability. The Journal of Pathol- ogy”, A Journal of the Pathological Society of Great Britain and Ireland 195 (2001) 293. [9] F. Zaccaria, S. C. van der Lubbe, C. Nieuwland, T. A. Hamlin & C. Fon- seca Guerra, “How divalent cations interact with the internal channel site of guanine quadruplexes”, ChemPhysChem 22 (2021) 2286. [10] M. Jiang, H. R. Chen, S. S. Li, R. Liang, J. H. Liu, Y. Yang & X. J. Huang, “The selective capture of Pb2+ in rice phloem sap using glutathione- functionalized gold nanoparticles/multi-walled carbon nanotubes: en- hancing anti-interference electrochemical detection”, Environmental Sci- ence: Nano 5 (2018). [11] D. D. Bwede, R. A. Wuana, G. EGAH, A. U. Itodo, E. Ogah, E. A. Yerima & A. I. Ibrahim, “Characterization and Evaluation of Human Health Risk of Heavy Metals in Tin Mine Tailings in Selected Area of Plateau State, Nigeria”, Journal of the Nigerian Society of Physical Sciences 3 (2021) 406. [12] K. O. Sodeinde, S. O. Olusanya, D. U. Momodu, V. F. Enogheghase & O. S. Lawal, “Waste glass: An excellent adsorbent for crystal violet dye, Pb2+ and Cd2+ heavy metals ions decontamination from wastewater”, Journal of the Nigerian Society of Physical Sciences 3 (2021) 414. [13] M. N. Amirah, A. S. Afiza, W. I. W. Faizal, M. H. Nurliyana & S. Laili, “Human health risk assessment of metal contamination through consump- tion of fish”, J Environ Pollut Hum Health. 1 (2013) 1. [14] H. Zhang, F. Zhang, J. Song, M. L. Tan & V. C. Johnson, ”Pollutant source, ecological and human health risks assessment of heavy metals in soils from coal mining areas in Xinjiang, China”, Environmental Re- search 202 (2021) 111702. [15] F. Chen, L. Saqlain, J. Ma, Z. I. Khan, K. Ahmad, A. Ashfaq & Y. Yang, “Evaluation of potential ecological risk and prediction of zinc accumula- tion and its transfer in soil plants and ruminants: public health implica- tions”, Environmental Science and Pollution Research 29 (2022) 3386. [16] P. Zhuang, B. Zou, N. Y. Li & Z. A. Li, “Heavy metal contamination in soils and food crops around Dabaoshan mine in Guangdong, China: implication for human health”, Environmental Geochemistry and Health 31 (2009) 707. [17] F. A. O. Joint, WHO working group report on drafting guidelines for the evaluation of probiotics in food, London, Ontario, Canada (2002). [18] R. A. Wuana &. F. E. Okieimen, “Heavy metals in contaminated soils: a review of sources, chemistry, risks and best available strategies for reme- diation”, International Scholarly Research Notices (2011) 1. [19] D. Song, D. Zhuang, D. Jiang, J. Fu & Q. Wang, “Integrated health risk assessment of heavy metals in Suxian County, South China”, International Journal of Environmental Research and Public Health 12 (2015) 7100. [20] J. F. Machiwa, “Heavy metal levels in paddy soils and rice (Oryza sativa (L)) from wetlands of Lake Victoria Basin, Tanzania”, Tanzania Journal of Science (2010) 36. [21] S. Binda, C. Hill, E. Johansen, D. Obis, B. Pot, M. E. Sanders & A. C. Ouwehand, “Criteria to qualify microorganisms as “probiotic” in foods and dietary supplements”, Front Microbiol. 11 (2020). [22] D. Kibassa, A. A. Kimaro & R. S. Shemdoe, “Heavy metals concentra- tions in selected areas used for urban agriculture in Dar es Salaam, Tan- zania”, Scientific Research and Essays 827 (2013) 1296. [23] N. Herawati, S. Suzuki, K. Hayashi, I. F. Rivai & H. Koyama, “Cadmium, copper, and zinc levels in rice and soil of Japan, Indonesia, and China by soil type”, Bulletin of Environmental Contamination and Toxicology 64 (2000) 33. [24] M. Wang, B. Markert, W. Chen, C. Peng & Z. Ouyang, “Identification of 10 Ajah et al. / J. Nig. Soc. Phys. Sci. 4 (2022) 891 11 heavy metal pollutants using multivariate analysis and effects of land uses on their accumulation in urban soils in Beijing, China”, Environmental Monitoring and Assessment 184 (2021) 5889. [25] M. W. Fiers, G. A. Kleter, H. Nijland, A. A. Peijnenburg, J. P. Nap & R. C. Van Ham, “AllermatchTM, a webtool for the prediction of potential aller- genicity according to current FAO/WHO Codex alimentarius guidelines”, BMC Bioinformatics 5 (2004) 1. [26] M. Arabameri, M. Mohammadi Moghadam, L. Monjazeb Marvdashti, S. M. Mehdinia, A. Abdolshahi & A. Dezianian, “Pesticide residues in pistachio nut: a human risk assessment study”, International Journal of Environmental Analytical Chemistry (2020) 1. [27] Y. Jin, L. Wang, Y. Song, J. Zhu, M. Qin, L. Wu & D. Hou, “Integrated life cycle assessment for sustainable remediation of contaminated agricultural soil in China”, Environmental Science & Technology 55 (2021) 12032. [28] H. Liu, A. Probst & B. Liao, “Metal contamination of soils and crops affected by the Chenzhou lead/zinc mine spill (Hunan, China)”, Science of the Total Environment 339 (2005) 153. [29] W. H. Organization, Safety evaluation of certain food additives: prepared by the eighty-ninth meeting of the Joint FAO/WHO Expert Committee on Food Additives (JECFA), (2022). 11 Introduction Experimental Area of Study Sample Collection Soil Sample Collection Rice Sample Collection Sample Preparation Soil Preparation Rice Preparation Ash Sample Size Digestion of Soil Samples, Rice Grain Samples, and Rice Stalk Samples Quality Assurance Risk Assessment Data Analysis Results and Discussion Heavy Metals Concentration in Soil and Rice Grain Copper in Soil Copper in Rice Relationships Between Copper Concentration in the Soil Before Planting and Soil After Harvest Zinc in Soil Zinc in Rice Relationships Between Zinc Concentrations in the Soil Before Planting and Soil After Harvest Iron in Soil Iron in Rice Relationships Between the Iron Concentration in the Soil (Before Planting) and Soil (After Harvest) Cadmium in Soil Cadmium in Rice Zinc Concentrations in Soil Copper Concentration in Soil Iron Concentration in Soil Heavy Metals in Rice Grain Dietary Intake of Heavy Metals and Potential Health Risks Via Rice Consumption Conclusion