Assessment of Heavy Metals Contamination in Spinacia oleracea L. and Coriandrum sativum L. Irrigated with Wastewater: Implications for Food Safety and Environmental Health

Assessment of Heavy Metals Contamination in Spinacia oleracea L. and Coriandrum sativum L. 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Irrigated with Wastewater: Implications for Food Safety and Environmental Health Nida Mehboob, Shams ur Rehman, Maqsood Ahmed, Khalid F. Almutairi, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6150572/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Vegetables are vital for human nutrition but can accumulate heavy metals, posing risks to public health and the environment, particularly in regions using wastewater for irrigation. This study, conducted in Gujranwala, Pakistan, evaluated heavy metal contamination in wastewater, soil, Spinacia oleracea L., and Coriandrum sativum L. using Atomic Absorption Spectrometry (AAS). Results revealed significant levels of lead (0.2255 mg/L), cobalt (0.0721 mg/L), chromium (0.1173 mg/L), and cadmium (0.0232 mg/L) in wastewater. Spinach and coriander samples exhibited heavy metal concentrations exceeding World Health Organization (WHO) limits, including chromium (11.313 mg/kg), lead (0.541 mg/kg), and cadmium (0.331 mg/kg). Soil samples also showed high cadmium levels. The findings underscore the urgent need for sustainable irrigation practices and land management to mitigate heavy metals bioaccumulation and safeguard food safety. Plant Physiology and Morphology Heavy metals wastewater vegetable spectrometry contamination Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 1. Introduction Vegetables are grown on small scale as compared to other major commercial crops, however, their productivity is dependent on the fertile soil, balance nutrient and availability of good-quality water. Pakistan's vegetable farming area has been steadily growing over time, and due to water constraints, wastewater is now widely used in agriculture 1 . Continuous use of this contaminated water can lead to heavy metal contamination of vegetables and groundwater, which can become contaminated with these metals if used for extended periods 2 . In the largest towns of Pakistan's, the availability of treated water is negligible which is only 1%, while the other 99% is directly applied to crop growth and dumped into various bodies of water 3 . Among the commonly cultivated vegetables, Spinach, a seasonal edible food plant, is the most nutrient-dense vegetable globally, with China producing 27.52 million tons 4 . However, In Pakistan, spinach is consumed raw or cooked and is valued for its rich phytoconstituents, vitamin and mineral content. Pakistan imports more than 4500 kg of spinach annually from Belgium and 2000 kg from China, at a cost of almost 0.49 million Pakistani rupees and 0.17 million rupees, respectively 5 . The reliance on untreated wastewater for irrigation in Pakistan poses a critical threat to vegetable safety, particularly spinach, a nutrient-rich crop, highlighting the urgent need for sustainable water management to mitigate heavy metal contamination and ensure food security. The other commonly cultivated vegetables is Coriander, which is a tropical crop, is widely grown in Pakistan and is valued for its fresh leaves and seeds, which are rich in essential oils, vitamins, and nutrients 6 . The plant thrives in low temperatures and frost-free conditions, and its optimal growth range is 20 ºC to 25 ºC 7 . Due to a lack of freshwater, farmers in Pakistan are forced to irrigate their fields with sewage and industrial effluent, which contain a high concentration of dangerous metals and metallic compounds 8 . Coriander, a nutrient-rich tropical crop widely grown in Pakistan, faces contamination risks from wastewater irrigation, highlighting the urgent need for sustainable water management practices. Plants absorb metal from the soil and water, which later affect humans or cattle when they eat the plants. After heavy metals enter the food chain, it is difficult to take them out, leading to long-term adverse effects and health concerns for both humans and animals consuming contaminated vegetables 9 . Heavy metals causing long-term adverse effects and health concerns for both humans and animals consuming contaminated vegetables pose a serious risk to the environment and the health of the human beings 10 . Consuming contaminated vegetables over an extended period may cause serious health issues with the liver, kidneys, circulatory and neurological systems, and bones 11 . Vegetables may accumulate higher concentrations of heavy metals if they are cultivated near sources of pollution or in locations with hazardous metal contamination 12 . The toxicity level of these metals can be increased when the crops or vegetables grown through contaminated waste water 13 . Lead, a heavy metal, is concentrated in human bones and disrupts the normal maturation of erythroid components in the bone marrow, causing neurological and behavioral consequences in children 14 . Cadmium, the most mobile heavy metal in soil and plants, has the highest potential to spread through the food chain, damaging the kidney, liver, lung, and bones 15 . Cobalt exposure increases damage to fats in the liver and disrupts the liver's antioxidant defense system 16 . Another, important heavy metal Chromium, a necessary dietary vitamin, can be contaminated by consuming contaminated food, inhalation, and skin absorption 17 . Similarly, Copper poisoning is a serious concern, with most cases being unintentional 18 . To mitigate the risks associated with untreated wastewater for irrigation in Pakistan, measures such as wastewater treatment protocols, safer irrigation practices, and continuous monitoring of water quality are essential. Therefore, the current study was conducted for the assessment and quantification of heavy metals in wastewater irrigated Spinach ( Spinacia oleracea L. ) and Coriander ( Coriandrum sativum L. ). 2. Materials and methods 2.1 Study area Gujranwala division, the Punjab Province of Pakistan is situated at (31°32′–32°33′ N and 73°11′–74°28′ E) with a total land area of 3,622 km 2 as shown in figure 1. Gujranwala has an economy based primarily on the use of pesticides and fertilizers, which are long-term agricultural practices used to boost crop productivity. The majority of land conversion in the study region is from agricultural land to residential housing societies 19 . The climatic conditions from June to September, the summertime temperature ranges from 27 to 47°C. There are 5-19°C of temperature variation during the winter, which spans from November to March 20 . 2.2 Collection of Samples The samples were collected from three different sites from district Gujranwala. The sampling process included the collection of spinach and coriander along with the soil from each of the three sites. 3000g of each vegetable, 100g soil sample and 100ml wastewater was collected from the same field by individual sampling. Materials such as organic trash, boulders, and gravels manually extracted from the soil and then collected in plastic bags. Water sample was cleaned with deionized water to get rid of any impurities, and was labelled 1-liter plastic bottle that has been soaked in 10% HNO 3 for a full day. Before being transported to the lab, it was kept at 4°C for additional tests, the water sample was placed on ice for further Atomic Absorption Spectroscopy (AAS) 2 . 2.3 Processing of Vegetables The vegetable samples were washed by removing the dirt or soil with tap water. The leaves were carefully rinsed under running water and distilled water was used to completely rinse the sample in order to get rid of any remaining cleaning solution and outside impurities. The samples were washed, and any surplus water was patted dry with fresh paper towels, and were dried under the shade at room temperature as shown in figure 2 and 3 13 . 2.4 Homogenization To ensure a representative sample for examination, the samples were homogenized once they have dried. To ensure that the heavy metals in the sample are distributed evenly, the leaves were ground into a fine powder using a blender as shown in figure 4. Between samples, the homogenization apparatus was carefully cleaned to prevent contamination 13 . 2.5 Sample digestion One gram of each oven-dried and ground vegetable part or soil was digested at 80°C in a 15 ml tri-acid mixture containing HNO 3 , H 2 SO 4 , and HClO 4 in a 4.5:1:1 ratio to obtain a clear solution. The solution was then filtered, and the resulting filtrate was adjusted to a volume of 50 ml by adding distilled water 2 . A 50 ml wastewater sample was digested at 80°C with 10 ml of HNO 3 concentration to produce a transparent solution. The clear solution then underwent additional filtering, and distilled water was added to the filtrate to bring its volume to 50 ml 22 . 2.6 Atomic Absorption Spectroscopy (AAS) for Heavy metals detection Heavy metals concentrations in vegetables, soil, and water was detected through Flame Atomic Absorption Spectrophotometer (FAAS) 2 . Flame Atomic absorption is a quantitative technique for analyzing heavy metals. This technique uses light with a certain wavelength that is released by an element radiation source from a sample to determine the heavy metals concentrations. Using standard solutions with known metal concentrations, the instrument creates a calibration curve from which the absorbance of the unknown sample is compared. Parts per million (ppm) is usual reports of the metal's concentration in the sample, and this comparison makes quantitative reporting possible 23 . The essential parameters required for conducting atomic absorption spectroscopy analysis of several heavy metals. Each element such as lead, chromium, cobalt, copper, and cadmium which was characterized by its specific wavelength for optimal detection. The slit width, which affects spectral resolution, was uniform for most elements at 0.7 nm, except for cobalt with a narrower slit of 0.2 nm. An acetylene flame was employed across all analyses to achieve the necessary high temperatures for sample atomization as shown in table 1. These parameters are crucial for ensuring accurate and consistent measurements of heavy metal concentrations in the samples analyzed using FAAS 24 . Table 1. Performance Parameters of Determining the Elements by FAAS. Heavy Metals Wavelength (nanometer) Slit (nanometer) Flame type Flow (liters per minute) Burner. Height (millimeters) Lead 283 0.7 Acetylene 2.0 7 Chromium 357 0.7 Acetylene 2.8 9 Cobalt 240 0.2 Acetylene 1.6 7 Copper 324 0.7 Acetylene 1.8 7 Cadmium 228 0.7 Acetylene 1.8 7 2.7 Statistical analysis A t-test statistical analysis was used in heavy metal detection analysis to assess if the mean concentration of a heavy metal in a sample differs significantly from a hypothesized value, such as regulatory limits, based on calculated t-values and p-values 25 . 2.8 Bio-Concentration Factor In order to assess the heavy metal transfer capability and vegetable yield based on soil metal content, the bio-transfer factor Bio-Concentration Factor (BCF) of the samples was determined. The calculation was performed using the following formula, BCF is calculated as follows, BCF = Cv/Cs Where Cv represents the metal concentration found in the veggies, and Cs represents the metal concentration found in the soil 26 . Values greater than 1, BCF (Bioconcentration Factor) indicate higher BCR (Bioconcentration Ratio) that the plant is a potential heavy metal and vice versa. 3. Results 3.1 Detection of heavy metals The concentrations of Cd, Cr, Cu, Co, and Pb in spinach, coriander, soil, and wastewater samples were measured in mg/kg for solid samples (spinach, coriander, and soil) and mg/L for liquid samples (wastewater). Some vegetables, including coriander and spinach, are eaten in the form of leaves. Thus, the number of heavy metals in each leaf of a particular vegetable along with their soil samples were determined. The results represented in table 2 shows the concentrations of various heavy metals such as cobalt, copper, cadmium, chromium, and lead in spinach samples and their corresponding soil samples. Cobalt concentrations in the spinach samples varied from 0.2611 to 0.3412 mg/kg. The levels of copper, which range from 10.215 to 11.313 mg/kg, were noticeably higher. Much smaller amounts of cadmium, between 0.0131 and 0.0267 mg/kg, were recorded in the tested samples. Lead values range from 0.4353 to 0.54132 mg/kg, whereas chromium concentrations recorded in the range of 2.4147 to 2.6713 mg/kg. In contrast, the soil samples have greater cobalt contents, ranging from 0.4471 to 0.5132 mg/kg. The soil had substantially lower levels of copper, ranging from 3.0722 to 3.2013 mg/kg, than the spinach. However, the soil had much higher concentrations of cadmium, ranging from 0.361 to 0.4652 mg/kg, suggesting that a large amount of Cd is retained in the soil instead of being absorbed by the spinach. The amounts of chromium in the soil, which ranges from 2.4512 to 2.6306 mg/kg, were comparatively similar to those in spinach. The soil has greater lead (Pb) values than the spinach samples, ranging from 0.6214 to 0.7415 mg/kg. These results showed that the spinach absorbs certain metals more effectively (e.g., Cu) while other metals (e.g., Cd and Pb) tend to retained in higher concentrations by the soil. Table 2. Concentration of Heavy Metals Detected in Spinach and Soil Samples. Heavy Metals Samples Spinach 1 (mg/kg) Spinach 2 (mg/kg) Spinach 3 (mg/kg) Soil 1 mg/L Soil 2 mg/L Soil 3 mg/L Co 0.3412 0.2736 0.2611 0.4471 0.4541 0.5132 Cu 11.1740 10.215 11.313 3.0722 3.1312 3.2013 Cd 0.0267 0.0213 0.0131 0.4278 0.361 0.4652 Cr 2.4147 2.5631 2.6713 2.6306 2.4512 2.5713 Pb 0.4353 0.54132 0.4512 0.7415 0.6341 0.6214 The table 3 presented the concentrations of various heavy metals such as cobalt, copper, cadmium, chromium, and lead in three coriander samples and their corresponding soil samples. Cobalt values in the coriander samples ranged from 0.1649 to 0.2613 mg/kg. The levels of copper, which ranged from 6.1319 to 6.7688 mg/kg, were noticeably high. Significant concentrations of cadmium were recorded with in the coriander samples, ranging from 0.2146 to 0.3313 mg/kg. However, the range of chromium levels was quite small, which was in the range 0.4121 to 0.5131 mg/kg. Moreover, concentrations of lead in the samples was recorded in the range of 0.3712 to 0.5631 mg/kg. The same heavy metals were present in different amounts in the soil samples. The amounts of cobalt in the soil were similar to those in the coriander plants, ranging from 0.2131 to 0.3126 mg/kg. Interestingly, the soil had greater concentrations of copper, ranging from 6.3791 to 7.2134 mg/kg. The soil contained somewhat more cadmium than the coriander samples (0.3219–0.3513), suggesting that some cadmium was not properly absorbed by the plant and instead stayed in the soil. Similarly, the soil had greater quantities of chromium than the coriander, ranging from 0.5617 to 0.6301 mg/kg. Lead levels in the soil, which ranged from 0.5131 to 0.8721 mg/kg, were significantly higher than in the coriander samples as shown in figure 5. Table 3. Concentration of Heavy Metals Detected in Coriander and Soil Samples. Heavy Metals Samples Coriander 1 (mg/kg) Coriander 2 (mg/kg) Coriander 3 (mg/kg) Soil 1 mg/L Soil 2 mg/L Soil 3 mg/L Co 0.1649 0.2541 0.2613 0.2521 0.3126 0.2131 Cu 6.7688 6.2613 6.1319 7.1634 6.3791 7.2134 Cd 0.3173 0.2146 0.3313 0.3513 0.3412 0.3219 Cr 0.4302 0.5131 0.4121 0.5617 0.6301 0.6213 Pb 0.4374 0.3712 0.5631 0.7642 0.8721 0.5131 3.2 Heavy metals detection in Wastewater The results regarding heavy metal concentration in waste water samples are shown in figure 3.2. The amount of cobalt (Co) in the wastewater was 0.0721 mg/L, more above the 0.05 mg/L WHO acceptable limit. In small doses, cobalt (Co) is a necessary as trace element, but higher concentrations can be hazardous. The concentration of copper in the wastewater was 0.2183 mg/L, which was well below the WHO permissible limit of 2.0 mg/L. The wastewater sample had 0.0232 mg/L of cadmium (Cd), which was more than the 0.01 mg/L WHO acceptable limit. The amount of chromium (Cr) in the wastewater was 0.1173 mg/L, more than twice the 0.05 mg/L WHO acceptable level. The concentration in the wastewater indicated that there is a serious pollution issue that requires attention. The wastewater has 0.2255 mg/L of lead (Pb), more than twice the WHO recommended limit of 0.1 mg/L as shown in figure 6. 3.3 Descriptive statistical analysis of heavy metals in spinach samples Descriptive data of spinach samples are shown in figure 7. Including the concentrations of heavy metals and comparing them with World Health Organization (WHO) limits. The average (mean) concentration was 0.29 mg/kg for cobalt. The World Health Organization (WHO) has set a cobalt limit of 0.1 mg/kg in specific dietary and environmental situations in an effort to protect human health. The average content of copper in spinach was determined to be 10.9 mg/kg. These amounts of copper are far below the 73.3 mg/kg WHO guideline. With an average of 0.02 mg/kg, the amounts of cadmium in spinach are below the WHO limit, which is 0.1 mg/kg. Spinach has an average chromium content of 2.54 mg/kg. The average content of chromium is marginally higher than the 2.3 mg/kg WHO limit. The mean concentration of lead was 0.47 mg/kg. These levels are higher than the 0.3 mg/kg lead limit set by the World Health Organization (WHO). 3.4 Descriptive statistical analysis of heavy metals in spinach soil samples Descriptive data of spinach soil samples are shown in figure 8. Including the concentrations of heavy metals and comparing them with World Health Organization (WHO) limits. The average level of cobalt (Co) in the soil used to grow spinach was 0.47 mg/kg. The cobalt (Co) contents in the soil were substantially lower than the WHO standard of 30 mg/kg for cobalt in soil. The average content of copper (Cu) in the spinach soil was 3.13 mg/kg. The copper (Cu) levels in soil were much below the 36 mg/kg WHO standard for copper in soil. This shows that there is no copper contamination in the soil, and the amounts of copper in spinach are safe for consumption. With an average value of 0.41 mg/kg, cadmium (Cd) levels in the soil range from 0.36 mg/kg to 0.46 mg/kg. These levels were higher above the WHO threshold of 0.31 mg/kg. The average amount of chromium (Cr) in the soil was 2.55 mg/kg. The amounts of chromium in soil were far lower than the WHO limit, which was 68 mg/kg. This indicates that chromium (Cr) levels in spinach are mostly a result of the plant's natural absorption and that chromium contamination of the soil is unlikely. The average lead (Pb) concentration in the soil was 0.66 mg/kg. The lead levels in soil were well below the 85 mg/kg WHO standard for lead in soil. This implies that there was no lead contamination in the soil, and the slightly higher lead levels in spinach could be the result of other causes such irrigation water. 3.5 Descriptive statistical analysis of heavy metals in coriander samples Descriptive data of coriander samples are shown in figure 9. Including the concentrations of heavy metals and comparing them with World Health Organization (WHO) limits. Coriander had cobalt (Co) values on average 0.22 mg/kg. The average amount of cobalt in coriander surpasses the safety threshold of 0.1 mg/kg set by the WHO for cobalt. The average level of chromium (Cr) in coriander was 0.45 mg/kg. These amounts of copper (Cu) were far below the 73.3 mg/kg WHO guideline. The average level of copper (Cu) in coriander was 6.38 mg/kg, with a range of 0.56 to 0.51 mg/kg. These amounts of copper (Cu) are far below the 2.3 mg/kg WHO guideline. With an average of 0.28 mg/kg, the average level of cadmium (Cd) in coriander exceeds the safety threshold of 0.1 mg/kg as shown in table 4.8, which is the limit set by the WHO. The average lead (Pb) content in coriander was 0.45 mg/kg. The average level of lead in coriander surpasses the 0.3 mg/kg WHO standard for lead. 3.6 Descriptive statistical analysis of heavy metals in coriander soil samples Descriptive data of coriander soil samples are shown in figure 10. It showed the concentrations of heavy metals and comparing them with World Health Organization (WHO) limits. The average value of cobalt (Co) in the soil used to grow coriander was 0.25 mg/kg. The cobalt contents in the soil were substantially lower than the WHO standard of 30 mg/kg for cobalt in soil. The average content of copper (Cu) in the coriander soil was 6.91 mg/kg. However, the copper levels in soil were much lower to 36 mg/kg of standard for copper in soil. Moreover, the average amount of cadmium (Cd) in the soil was 0.33 mg/kg. The WHO has set a limit of 0.31 mg/kg of cadmium. Similarly, the levels of cadmium in the soil are somewhat higher than this threshold. The average contents of chromium (Cr) in the soil was 0.60 mg/kg. The amounts of chromium in soil were far lower than the WHO limit, which is 68 mg/kg. The average lead (Pb) concentration in the soil was 0.71 mg/kg. The lead levels in soil were well below the 85 mg/kg WHO standard for lead in soil. 3.7 One sample T-test statistical analysis of spinach samples T-test showed the t value, df and P value of each heavy metal in spinach as shown in table 4. In spinach samples, Co, Cu, Cd and Pb showed significant differences as their P values were less than 0.05, which means that concentration of these heavy metals were different than WHO standards. while Cr showed insignificant results with p-value 0.07, which means that the concentrations of Cr were within limit of WHO standards. Table 4. T-Test Results for Heavy Metal Concentrations in Spinach samples. Heavy Metals T-value Df P-value Co 7.7158 2 0.01639* Cu -180.78 2 0.0000306* Cd -20.142 2 0.02456* Cr 3.3575 2 0.07843 Pb 5.3307 2 0.03344* (*) indicates significant differences (p-value < 0.05), while others are non-significant. 3.8 One sample T-test statistical analysis of spinach soil samples T-test showed the t value, df and p value of each heavy metal in spinach soil samples as shown in table 5. In spinach soil samples, all heavy metals like Co, Cu, Cd, Cr and Pb showed significant differences as their p values were less than 0.05, which means that concentration of these heavy metals were different than WHO standards. Table 5. T-Test Results for Heavy Metal Concentrations in Spinach soil samples. Heavy Metals T-value Df P-value Co -1194.1 2 0.00000070148* Cu -72.715 2 0.0001891* Cd -73.258 2 0.0001863* Cr -879.99 2 0.000001291* Pb -2213.9 2 0.0000000204* (*) indicates significant differences (p-value < 0.05), while others are non-significant. 3.9 One sample T-test statistical analysis of coriander samples In coriander samples, Cu and Cr showed significant differences as their p values were less than 0.05, which means that concentration of these heavy metals were different than WHO standards. While Co, Cd and Pb showed insignificant results with p values above than 0.05 as shown in table 6, which indicated the concentrations of these heavy metals were within limit of WHO standards as shown in . Table 6. T-Test Results for Heavy Metal Concentrations in Coriander samples. Heavy Metals T-value Df P-value Co 4.0888 2 0.05493 Cu -344.28 2 0.000008437* Cd -0.60525 2 0.6065 Cr -59.443 2 0.0002829* Pb 2.7939 2 0.1078 (*) indicates significant differences (p-value < 0.05), while others are non-significant. 3.10 One sample T-test statistical analysis of coriander soil samples In coriander’s soil samples, Co, Cu, Cr and Pb showed significant differences as their p values were less than 0.05, which means that concentration of these heavy metals were different than WHO standards. while Cd showed insignificant results with p value 0.08 as shown in table 7, which indicated the concentrations of Cd were within limit of WHO standards. Table 7. T-Test Results for Heavy Metal Concentrations in Coriander soil samples. Heavy Metals T-value Df P-value Co -1027.5 2 0.00000009473* Cu -101.65 2 0.00008628* Cd 3.265 2 0.08251 Cr -3137 2 0.000000001016* Pb -792.53 2 0.0000001592* (*) indicates significant differences (p-value < 0.05), while others are non-significant. 3.11 Bio-concentration Factor (BCF) of heavy metals from soil to vegetables The average bio-concentration factor values for each heavy metal (Co, Cd, Cr, Cu, Pb) in spinach and coriander samples are shown in table 8. Spinach has an average cobalt concentration of 0.291967 mg/kg. The average cobalt concentration in the soil used to grow spinach was 0.471467 mg/kg. For cobalt (Co) in spinach, the bio-concentration ratio (BCR) is 0.619273. This value suggests that spinach had a moderate soil-based cobalt accumulation. Coriander had an average cobalt content of 0.226767 mg/kg. The average content of cobalt (Co) in the soil used to grow coriander was 0.259267 mg/kg. However, in coriander, the BCR for cobalt (Co) has been determined as 0.874646. This indicates that coriander is somewhat more capable than spinach of absorbing cobalt from the soil. Spinach has an average copper concentration of 10.900667 mg/kg. The average concentration of copper (Cu) in the soil used to grow spinach was 3.1349 mg/kg. It is determined that spinach had a higher BCR of 3.477198 for copper. This suggests that spinach is an extremely effective way for the plant to absorb copper (Cu) from the soil. Coriander has an average copper (Cu) content of 6.387333 mg/kg. The average content of copper (Cu) in the soil used to cultivate coriander was 6.918633 mg/kg. In coriander, the BCR for copper (Cu) was determined to be 0.923207. This indicates that spinach is somewhat more capable than coriander of absorbing cobalt (Co) from the soil. Cadmium (Cd) concentration in spinach was 0.020367 mg/kg on average. The average level of cadmium (Cd) in the soil used for the growth of spinach was 0.418 mg/kg. The BCR for spinach's cadmium content is 0.048724. This suggests that spinach absorbs less cadmium (Cd) from the soil. Cadmium (Cd) content in coriander is 0.287733 mg/kg on average. The average quantity of cadmium (Cd) exhibited by the soil for growing coriander is 0.338133 mg/kg. It is determined that the BCR for cadmium in coriander is 0.850946 which indicates that coriander is somewhat more capable than spinach of absorbing cadmium from the soil. Spinach has an average chromium (Cr) concentration of 2.5497 mg/kg. Likewise, the mean concentration values of chromium (Cr) in the soil used to grow spinach was 2.551033 mg/kg. For spinach, the BCR for chromium is 0.999477. This suggests that spinach takes up a moderate amount of chromium (Cr) from the soil. Coriander had an average chromium concentration of 0.4518 mg/kg. The average amount of chromium (Cr) in the soil used to cultivate coriander was 0.604367 mg/kg. It is calculated that the BCR for chromium (Cr) in coriander was 0.747559 which implies that coriander absorbs chromium (Cr) from the soil in a moderate amount which indicated that spinach is slightly more capable than coriander of absorbing chromium from the soil. Spinach has an average lead concentration of 0.47594 mg/kg. The average lead (Pb) concentration in the soil used to grow spinach was 0.665667 mg/kg. The calculated BCR for lead (Pb) in spinach was 0.714982. This suggests that spinach absorbs lead (Pb) from the soil to a moderate extent. Coriander has an average lead (Pb) content of 0.457233 mg/kg. Lead (Pb) concentrations in the coriander-growing soil average 0.716467 mg/kg. Coriander's BCR for lead (Pb) was determined to be 0.638178. This implies that coriander, like spinach, accumulates lead (Pb) from the soil in a moderate amount indicated its more capability than coriander in absorbing lead from the soil. Table 8. Bio-concentration Ratio for each heavy metal from soil to vegetables Heavy metals Average conc. in Spinach (mg/kg) Average conc. in soil (mg/kg) BCR Soil to Spinach Average conc. in Coriander (mg/kg) Average conc. in soil (mg/kg) BCR Soil to Coriander Co 0.2919 0.4714 0.6192 0.2267 0.2592 0.8746 Cu 10.900 3.1349 3.4771 6.3873 6.918 0.9232 Cd 0.0203 0.4180 0.0487 0.2877 0.3381 0.8509 Cr 2.5497 2.5510 0.99947 0.45180 0.6043 0.7475 Pb 0.4759 0.6656 0.71498 0.4572 0.7164 0.6381 4. Discussion Pakistan is considered as a developing country with numerous serious problems that have a negative impact on people's health. One of the main issues facing by the human beings in the current situation is the lack of fresh water availability for humans and cultivation of crops and vegetables, which eventually affects the nation's agriculture sector by making it difficult to meet the fundamental needs of agriculturally produced food to feed the expanding population. In order to get rid of this problem, farmers are growing vegetables in peri-urban regions by mindlessly using untreated sewage or industrial water (Jabeen et al., 2020). In this study, a survey was conducted in 2023 and sites were selected where vegetables have been irrigated by wastewater of industry and sewage. Later on wastewater has been collected from the field at Gujranwala for the analysis of heavy metal concentration in waste water and compared with WHO limits. Our findings on assessment and quantification showed serious concerns about cobalt (Co), cadmium (Cd), chromium (Cr), and lead (Pb) levels over the WHO permitted limits by the examination of heavy metal concentrations in the wastewater. Both lead and chromium can cause serious health problems, when their levels are more than double the recommended limits and decisive action is essential in order to reduce these contaminants to assure the environmental and public health's safety. Other researchers in Pakistan additionally investigated higher levels of heavy metals in wastewater and discovered abnormal quantities. Jabeen et al., (2020), assessed higher contents of Manganese and Nickel in wastewater in Faisalabad, Pakistan. Similarly, Rehman et al., (2019), reported higher quantities of heavy metals such as Cd and Manganese in wastewater in district Sahiwal, Pakistan, where wastewater has been applied for irrigation. The analysis of heavy metal concentrations in spinach, coriander, and their respective soils reveals several significant findings. The quantification of heavy metals in the spinach samples varied, with lead, chromium, and cobalt concentrations above WHO recommendations. The cobalt levels were higher above the WHO limit of 0.1 mg/kg, ranging from 0.26 to 0.34 mg/kg on average. The average chromium concentration was 2.54 mg/kg, which is marginally more than the 2.3 mg/kg of WHO standard. With a mean of 0.47 mg/kg, lead levels in spinach ranged from 0.43 to 0.54 mg/kg, exceeding the WHO recommendation of 0.3 mg/kg. Our findings were supported by Kalhoro et al., (2023), who reported higher level of six heavy metals such as Cr, Ni, Cd, Pb, As and Hg in cabbage, cauliflower, turnip and radish. Similarly, Iqbal et al., (2020), found higher levels of heavy metals including Fe, Pb and Cr in sponge gourd in Pakistan. In contrast, levels of cadmium and copper in spinach's were within acceptable limits. The values of cadmium were between 0.01 and 0.02 mg/kg, which were below the WHO limit of 0.1 mg/kg, and copper varied from 10.2 to 11.3 mg/kg, both significantly below the WHO recommendation of 73.3 mg/kg. Lead, copper, chromium, and cobalt levels in the spinach soil were all much below WHO recommendations, suggesting that plant uptake of the metals rather than soil contamination is most likely to be responsible for the increased levels in spinach. The WHO standard of 0.31 mg/kg for cadmium is exceeded by the average of 0.41 mg/kg in the soil, indicating possible pollution and the need for soil remediation to reduce health concerns. Mahmood et al., (2020), investigated higher level of heavy metals such as Fe, Mg, Na, Ca and K in soil due to waste water irrigation.. Leblebici et al., (2020), also assessed higher level of heavy metals such as Zn and Mn in soil where waste water was applied. Aftab et al., (2023), found higher levels of heavy metals like Cr and Co in crops irrigated with waste water. Lead, cadmium, and cobalt levels in coriander samples exceeded the WHO recommendations. The mean cobalt content was 0.22 mg/kg, exceeding the WHO threshold of 0.1 mg/kg. Our investigations indicated Lead concentrations which varied from 0.37 to 0.56 mg/kg, with an average of 0.45 mg/kg, exceeding the WHO limit of 0.3 mg/kg. Moreover, cadmium levels averaged 0.28 mg/kg, exceeding the WHO threshold of 0.1 mg/kg. Our findings are in agreement with Ugulu et al., (2023), who investigated higher level of heavy metals of cadmium, cobalt, chromium, copper, lead in coriander when irrigated with waste water. Similarly, Hussain et al., (2022), showed higher levels of Cd in coriander while lower level of Mn accumulation as compared to WHO limits. On the other hand, coriander has safe copper levels of 6.38 mg/kg, below the 73.3 mg/kg WHO recommendation. However, Lead, chromium, copper, and cobalt concentrations in the coriander soil were all found to be substantially below the corresponding WHO standards, indicating no evidence of pollution. Whereas, the average level of cadmium in the coriander soil was 0.33 mg/kg, which is somewhat higher than the WHO guideline of 0.31 mg/kg. Aftab et al., (2023), found lower levels of heavy metals like Ni, Cu, Pb, and Mn concentrations which were in line with WHO standards in coriander. To determine the statistical significance differences, the t-value, degrees of freedom (df), and associated p-value for each heavy metal were computed. A p-value less than the conventional significance level (typically 0.05) suggests strong evidence against the null hypothesis, leading to its rejection. The results for chromium (Cr) were non-significant (p = 0.07843), meaning that the amount of the heavy metal in spinach was within WHO-acceptable limits. The quantities of heavy metals in relation to WHO guidelines were revealed by the one-sample t-test studies performed on coriander, spinach, and soil samples. Significant variations were found in the amounts of copper and chromium in coriander samples, suggesting levels exceeding WHO guidelines, but acceptable ranges were found for cobalt, cadmium, and lead. Ali et al., (2021) demonstrated a significant difference in soil samples for Pb, Cr, Cd, Ni, Zn and Cu at p ≤ 0.001, for Mn at p ≤ 0.05 while no significant difference was observed for Fe respectively. Also exhibited the highest mean value for Pb, Cr, Cd and Zn in vegetables. Hussain & Qureshi, (2020), showed results indicate significant differences that iron (Fe) was highest in lettuce followed by spinach, and Zn and Cr were second and third element in the vegetables. Eggplant and radish showed non-significant differences the lowest concentrations of heavy metals. Ahmed et al., (2022), showed similar results by assessing higher accumulation of heavy metals like chromium and cadmium as compared to WHO limits, in green leafy vegetables along with their soils. Similarly, Shahid et al., (2019), showed similar results which indicated significant differences in Cu and Zn in cucumber and chili. The heavy metal Bioconcentration Ratios (BCRs) for soil, spinach, and coriander show distinct patterns of accumulation that emphasize the bioindicators' importance in contaminated agricultural settings. Our research findings showed that Spinach exhibits a high BCR and effective copper accumulation, suggesting that it could be a sensitive bioindicator of soil copper contamination. A study conducted by Gupta et al., (2022), assessed significant accumulation of heavy metals like Zn and Mn in vegetables, whereas, spinach has less ability to carry lead, cadmium, and cobalt, but its uptake of chromium is balanced. Likewise, coriander shows moderate cobalt, copper, and cadmium accumulation efficiency which suggested that in contaminated soils, coriander can act as a reliable bioindicator for these heavy metals. in comparison, spinach, accumulates chromium and lead less efficiently, suggesting that it is not as effective as spinach at monitoring these specific elements. These research findings were supported by Oguh & Obiwulu, (2020), who reported low to moderate BCR values for heavy metals like As, Pb, Cd and Hg from soil to vegetables. Ali et al., (2021), also demonstrated similar higher BCR results, the transfer factor for Cr, Pb, Zn, Mn, Ni, Cd and Cu was greater than 0.5 due to contamination caused by discharges and industrial effluents. Understanding these accumulation patterns is crucial for developing effective strategies for monitoring and managing heavy metal contamination to ensure food safety and protect environmental health Additional research is aimed on more vegetables to detect heavy metals concerning with the health hazardous issue on human beings and healthier generation. 5. Conclusion This study concludes that there is a potential risk of heavy metal contamination in vegetables as a result of wastewater irrigation. This study evaluated the presence of heavy metals (Co, Cu, Cd, Cr, Pb) in wastewater, soil, and vegetable samples spinach and coriander. The atomic absorption spectrophotometry, revealed the levels of Pb, Co, Cr, and Cd in wastewater were alarmingly high. Spinach and coriander samples also exhibited heavy metal concentrations exceeding WHO permissible limits, with significant levels of Co, Cr, and Pb detected. Similarly, the one-sample t-test analysis on spinach and coriander samples revealed significant deviations in cobalt, copper, cadmium, and lead concentrations from WHO standards. However, the bioaccumulation factor (BCR) indicated higher Cu accumulation compared to Cr, Cd, Pb, and Co in the vegetables. These findings underscore the critical issue of heavy metal toxicity in urban areas due to wastewater irrigation. Therefore, the study suggested the urgent need for sustainable land and water resource management practices to mitigate heavy metal contamination for human health and food safety. Declarations Credit authorship contribution statement NM, and SR: Experimentation, Data analysis and manuscript writing; MA, and LW: Data analysis and Reviewing and editing; AM: S upervision; MA, and AA: Visualization; AM, KFA, and MA: Conceptualization and resources, AM, AES, and MMS: Analysis, reviewing, revising and editing; All authors have read and approved the final manuscript. Acknowledgment Authors are thankful to all the contributors who work and helped during the whole experiment. Cooperation of Assistant Professor Abdul Mateen and Director Dr. Maqsood Ahmed is greatly acknowledged for the supervision, conceiving of the manuscript and financial assistance. The authors extend their appreciation to Researchers Supporting Project number (RSPD2025R561), King Saud University, Riyadh, Saudi Arabia. Competing interests The authors declare no competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Consent for publication All authors have been agreed to the publication of this manuscript. Funding: Not applicable. The funding information will be provided if required by the journal. The authors extend their appreciation to Researchers Supporting Project number (RSPD2025R561), King Saud University, Riyadh, Saudi Arabia. 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Human risk on heavy metal pollution and bioaccumulation factor in soil and some edible vegetables around active auto-mechanic workshop in Chanchaga Minna Niger state, Nigeria. Ann Ecol Environ Sci. 2020;4(1):12–22. Additional Declarations The authors declare no competing interests. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6150572","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":423705126,"identity":"d19c10ca-d316-4748-a245-a0fffbc49d8f","order_by":0,"name":"Nida Mehboob","email":"","orcid":"","institution":"Department of Biological Sciences, Faculty of Sciences, University of Sialkot, 51310, Pakistan","correspondingAuthor":false,"prefix":"","firstName":"Nida","middleName":"","lastName":"Mehboob","suffix":""},{"id":423705127,"identity":"11549a4d-2f1d-4e71-8abc-a8846cfcc198","order_by":1,"name":"Shams ur Rehman","email":"","orcid":"","institution":"State Key Laboratory of Wheat Improvement, Peking University Institute of Advanced Agricultural Sciences, Shandong Laboratory of Advanced Agriculture Sciences in Weifang, Weifang 261325, Shandong, China","correspondingAuthor":false,"prefix":"","firstName":"Shams","middleName":"ur","lastName":"Rehman","suffix":""},{"id":423705128,"identity":"2dc8384d-ba7a-49ff-bfb8-f5bb2061f618","order_by":2,"name":"Maqsood Ahmed","email":"","orcid":"","institution":"Department of Agriculture, Pest Warning and Quality Control of Pesticides, Sialkot, 51310, Pakistan","correspondingAuthor":false,"prefix":"","firstName":"Maqsood","middleName":"","lastName":"Ahmed","suffix":""},{"id":423705129,"identity":"7d61019e-3956-4b30-af18-1827a6c3d615","order_by":3,"name":"Khalid F. 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9","display":"","copyAsset":false,"role":"figure","size":26352,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eComparison of Heavy metals concentration of coriander samples with WHO limits.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"9.png","url":"https://assets-eu.researchsquare.com/files/rs-6150572/v1/e1945a99d6f38af78cead003.png"},{"id":77746834,"identity":"34b58b76-34d6-472e-8b5a-ee74e036e146","added_by":"auto","created_at":"2025-03-05 06:48:57","extension":"png","order_by":10,"title":"Figure 10","display":"","copyAsset":false,"role":"figure","size":28115,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eComparison of Heavy metals concentration of coriander soil samples with WHO limits.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"10.png","url":"https://assets-eu.researchsquare.com/files/rs-6150572/v1/4b0e8120e5205456bb0fd12f.png"},{"id":77749143,"identity":"bf676cf2-859d-403f-b634-58d087d49791","added_by":"auto","created_at":"2025-03-05 07:20:59","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":5603853,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6150572/v1/280a1096-6597-4ac5-817a-3b8724c31bbb.pdf"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"\u003cp\u003eAssessment of Heavy Metals Contamination in \u003cem\u003eSpinacia oleracea \u003c/em\u003eL. and \u003cem\u003eCoriandrum sativum \u003c/em\u003eL. Irrigated with Wastewater: Implications for Food Safety and Environmental Health\u003c/p\u003e","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eVegetables are grown on small scale as compared to other major commercial crops, however, their productivity is dependent on the fertile soil, balance nutrient and availability of good-quality water. Pakistan's vegetable farming area has been steadily growing over time, and due to water constraints, wastewater is now widely used in agriculture \u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e. Continuous use of this contaminated water can lead to heavy metal contamination of vegetables and groundwater, which can become contaminated with these metals if used for extended periods \u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e. In the largest towns of Pakistan's, the availability of treated water is negligible which is only 1%, while the other 99% is directly applied to crop growth and dumped into various bodies of water \u003csup\u003e\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u003c/sup\u003e. Among the commonly cultivated vegetables, Spinach, a seasonal edible food plant, is the most nutrient-dense vegetable globally, with China producing 27.52\u0026nbsp;million tons \u003csup\u003e\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u003c/sup\u003e. However, In Pakistan, spinach is consumed raw or cooked and is valued for its rich phytoconstituents, vitamin and mineral content. Pakistan imports more than 4500 kg of spinach annually from Belgium and 2000 kg from China, at a cost of almost 0.49\u0026nbsp;million Pakistani rupees and 0.17\u0026nbsp;million rupees, respectively \u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e. The reliance on untreated wastewater for irrigation in Pakistan poses a critical threat to vegetable safety, particularly spinach, a nutrient-rich crop, highlighting the urgent need for sustainable water management to mitigate heavy metal contamination and ensure food security.\u003c/p\u003e \u003cp\u003eThe other commonly cultivated vegetables is Coriander, which is a tropical crop, is widely grown in Pakistan and is valued for its fresh leaves and seeds, which are rich in essential oils, vitamins, and nutrients \u003csup\u003e\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u003c/sup\u003e. The plant thrives in low temperatures and frost-free conditions, and its optimal growth range is 20 \u0026ordm;C to 25 \u0026ordm;C \u003csup\u003e\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u003c/sup\u003e. Due to a lack of freshwater, farmers in Pakistan are forced to irrigate their fields with sewage and industrial effluent, which contain a high concentration of dangerous metals and metallic compounds \u003csup\u003e\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e\u003c/sup\u003e. Coriander, a nutrient-rich tropical crop widely grown in Pakistan, faces contamination risks from wastewater irrigation, highlighting the urgent need for sustainable water management practices.\u003c/p\u003e \u003cp\u003ePlants absorb metal from the soil and water, which later affect humans or cattle when they eat the plants. After heavy metals enter the food chain, it is difficult to take them out, leading to long-term adverse effects and health concerns for both humans and animals consuming contaminated vegetables \u003csup\u003e\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u003c/sup\u003e. Heavy metals causing long-term adverse effects and health concerns for both humans and animals consuming contaminated vegetables pose a serious risk to the environment and the health of the human beings \u003csup\u003e\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u003c/sup\u003e. Consuming contaminated vegetables over an extended period may cause serious health issues with the liver, kidneys, circulatory and neurological systems, and bones \u003csup\u003e\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u003c/sup\u003e. Vegetables may accumulate higher concentrations of heavy metals if they are cultivated near sources of pollution or in locations with hazardous metal contamination \u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u003c/sup\u003e. The toxicity level of these metals can be increased when the crops or vegetables grown through contaminated waste water \u003csup\u003e\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/sup\u003e. Lead, a heavy metal, is concentrated in human bones and disrupts the normal maturation of erythroid components in the bone marrow, causing neurological and behavioral consequences in children \u003csup\u003e\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u003c/sup\u003e. Cadmium, the most mobile heavy metal in soil and plants, has the highest potential to spread through the food chain, damaging the kidney, liver, lung, and bones \u003csup\u003e\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u003c/sup\u003e. Cobalt exposure increases damage to fats in the liver and disrupts the liver's antioxidant defense system \u003csup\u003e\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u003c/sup\u003e. Another, important heavy metal Chromium, a necessary dietary vitamin, can be contaminated by consuming contaminated food, inhalation, and skin absorption \u003csup\u003e\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u003c/sup\u003e. Similarly, Copper poisoning is a serious concern, with most cases being unintentional \u003csup\u003e\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e\u003c/sup\u003e. To mitigate the risks associated with untreated wastewater for irrigation in Pakistan, measures such as wastewater treatment protocols, safer irrigation practices, and continuous monitoring of water quality are essential. Therefore, the current study was conducted for the assessment and quantification of heavy metals in wastewater irrigated Spinach (\u003cem\u003eSpinacia oleracea L.\u003c/em\u003e) and Coriander (\u003cem\u003eCoriandrum sativum L.\u003c/em\u003e).\u003c/p\u003e"},{"header":"2. Materials and methods","content":"\u003ch3\u003e2.1 Study area\u003c/h3\u003e\n\u003cp\u003eGujranwala division, the Punjab Province of Pakistan is situated at (31\u0026deg;32\u0026prime;\u0026ndash;32\u0026deg;33\u0026prime; N and 73\u0026deg;11\u0026prime;\u0026ndash;74\u0026deg;28\u0026prime; E) with a total land area of 3,622 km\u003csup\u003e2\u003c/sup\u003e as shown in figure 1. Gujranwala has an economy based primarily on the use of pesticides and fertilizers, which are long-term agricultural practices used to boost crop productivity. The majority of land conversion in the study region is from agricultural land to residential housing societies \u003csup\u003e19\u003c/sup\u003e. The climatic conditions from June to September, the summertime temperature ranges from 27 to 47\u0026deg;C. There are 5-19\u0026deg;C of temperature variation during the winter, which spans from November to March \u003csup\u003e20\u003c/sup\u003e.\u0026nbsp;\u003c/p\u003e\n\u003ch3\u003e2.2 Collection of Samples\u003c/h3\u003e\n\u003cp\u003eThe\u0026nbsp;samples were collected from three different sites from district Gujranwala. The sampling process included the collection of spinach and coriander along with the soil from each of the three sites. 3000g of each vegetable, 100g soil sample and 100ml wastewater was collected from the same field by individual sampling. Materials such as organic trash, boulders, and gravels manually extracted from the soil and then collected in plastic bags. Water sample was cleaned with deionized water to get rid of any impurities, and was labelled 1-liter plastic bottle that has been soaked in 10% HNO\u003csub\u003e3\u003c/sub\u003e for a full day. Before being transported to the lab, it was kept at 4\u0026deg;C for additional tests, the water sample was placed on ice for further Atomic Absorption Spectroscopy (AAS) \u003csup\u003e2\u003c/sup\u003e.\u0026nbsp;\u003c/p\u003e\n\u003ch3\u003e2.3 Processing of Vegetables\u003c/h3\u003e\n\u003cp\u003eThe vegetable samples were washed by removing the dirt or soil with tap water. The leaves were carefully rinsed under running water and distilled water was used to completely rinse the sample in order to get rid of any remaining cleaning solution and outside impurities. The samples were washed, and any surplus water was patted dry with fresh paper towels, and were dried under the shade at room temperature as shown in figure 2 and 3 \u003csup\u003e13\u003c/sup\u003e.\u003c/p\u003e\n\u003ch3\u003e2.4 Homogenization\u003c/h3\u003e\n\u003cp\u003eTo ensure a representative sample for examination, the samples were homogenized once they have dried. To ensure that the heavy metals in the sample are distributed evenly, the leaves were ground into a fine powder using a blender as shown in figure 4. Between samples, the homogenization apparatus was carefully cleaned to prevent contamination \u003csup\u003e13\u003c/sup\u003e.\u003c/p\u003e\n\u003ch3\u003e2.5 Sample digestion\u003c/h3\u003e\n\u003cp\u003eOne gram of each oven-dried and ground vegetable part or soil was digested at 80\u0026deg;C in a 15 ml tri-acid mixture containing HNO\u003csub\u003e3\u003c/sub\u003e, H\u003csub\u003e2\u003c/sub\u003eSO\u003csub\u003e4\u003c/sub\u003e, and HClO\u003csub\u003e4\u003c/sub\u003e in a 4.5:1:1 ratio to obtain a clear solution. The solution was then \u0026nbsp;filtered, and the resulting filtrate was adjusted to a volume of 50 ml by adding distilled water \u003csup\u003e2\u003c/sup\u003e. A 50 ml wastewater sample was digested at 80\u0026deg;C with 10 ml of HNO\u003csub\u003e3\u003c/sub\u003e concentration to produce a transparent solution. The clear solution then underwent additional filtering, and distilled water was added to the filtrate to bring its volume to 50 ml \u003csup\u003e22\u003c/sup\u003e.\u0026nbsp;\u003c/p\u003e\n\u003ch3\u003e2.6 Atomic Absorption Spectroscopy (AAS) for Heavy metals detection\u003c/h3\u003e\n\u003cp\u003eHeavy metals concentrations in vegetables, soil, and water was detected through Flame Atomic Absorption Spectrophotometer (FAAS) \u003csup\u003e2\u003c/sup\u003e. Flame Atomic absorption is a quantitative technique for analyzing heavy metals. This technique uses light with a certain wavelength that is released by an element radiation source from a sample to determine the heavy metals concentrations. Using standard solutions with known metal concentrations, the instrument creates a calibration curve from which the absorbance of the unknown sample is compared. Parts per million (ppm) is usual reports of the metal\u0026apos;s concentration in the sample, and this comparison makes quantitative reporting possible \u003csup\u003e23\u003c/sup\u003e. The essential parameters required for conducting atomic absorption spectroscopy analysis of several heavy metals. Each element such as lead, chromium, cobalt, copper, and cadmium which was characterized by its specific wavelength for optimal detection. The slit width, which affects spectral resolution, was uniform for most elements at 0.7 nm, except for cobalt with a narrower slit of 0.2 nm. An acetylene flame was employed across all analyses to achieve the necessary high temperatures for sample atomization as shown in table 1. These parameters are crucial for ensuring accurate and consistent measurements of heavy metal concentrations in the samples analyzed using FAAS \u003csup\u003e24\u003c/sup\u003e.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 1. Performance Parameters of Determining the Elements by FAAS.\u003c/strong\u003e\u003c/p\u003e\n\u003cdiv align=\"\"\u003e\n \u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 97px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eHeavy Metals\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 100px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eWavelength (nanometer)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 100px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSlit (nanometer)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 97px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eFlame type\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 97px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eFlow (liters per minute)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 104px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eBurner. Height\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e(millimeters)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 97px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLead\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 100px;\"\u003e\n \u003cp\u003e283\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 100px;\"\u003e\n \u003cp\u003e0.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 97px;\"\u003e\n \u003cp\u003eAcetylene\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 97px;\"\u003e\n \u003cp\u003e2.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 104px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 97px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eChromium\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 100px;\"\u003e\n \u003cp\u003e357\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 100px;\"\u003e\n \u003cp\u003e0.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 97px;\"\u003e\n \u003cp\u003eAcetylene\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 97px;\"\u003e\n \u003cp\u003e2.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 104px;\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 97px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;Cobalt\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 100px;\"\u003e\n \u003cp\u003e240\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 100px;\"\u003e\n \u003cp\u003e0.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 97px;\"\u003e\n \u003cp\u003eAcetylene\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 97px;\"\u003e\n \u003cp\u003e1.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 104px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 97px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCopper\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 100px;\"\u003e\n \u003cp\u003e324\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 100px;\"\u003e\n \u003cp\u003e0.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 97px;\"\u003e\n \u003cp\u003eAcetylene\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 97px;\"\u003e\n \u003cp\u003e1.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 104px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 97px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCadmium\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 100px;\"\u003e\n \u003cp\u003e228\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 100px;\"\u003e\n \u003cp\u003e0.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 97px;\"\u003e\n \u003cp\u003eAcetylene\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 97px;\"\u003e\n \u003cp\u003e1.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 104px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003ch3\u003e2.7 Statistical analysis\u003c/h3\u003e\n\u003cp\u003eA t-test statistical analysis was used in heavy metal detection analysis to assess if the mean concentration of a heavy metal in a sample differs significantly from a hypothesized value, such as regulatory limits, based on calculated t-values and p-values \u003csup\u003e25\u003c/sup\u003e.\u0026nbsp;\u003c/p\u003e\n\u003ch3\u003e2.8 Bio-Concentration Factor\u003c/h3\u003e\n\u003cp\u003eIn order to assess the heavy metal transfer capability and vegetable yield based on soil metal content, the bio-transfer factor Bio-Concentration Factor (BCF) of the samples was determined. The calculation was performed using the following formula, BCF is calculated as follows,\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eBCF = Cv/Cs\u003c/p\u003e\n\u003cp\u003eWhere Cv represents the metal concentration found in the veggies, and Cs represents the metal concentration found in the soil \u003csup\u003e26\u003c/sup\u003e. Values greater than 1, BCF (Bioconcentration Factor) indicate higher BCR (Bioconcentration Ratio) that the plant is a potential heavy metal and vice versa.\u003c/p\u003e"},{"header":"3. Results","content":"\u003ch3\u003e3.1 Detection of heavy metals\u0026nbsp;\u003c/h3\u003e\n\u003cp\u003e\u0026nbsp;The concentrations of Cd, Cr, Cu, Co, and Pb in spinach, coriander, soil, and wastewater samples were measured in mg/kg for solid samples (spinach, coriander, and soil) and mg/L for liquid samples (wastewater). Some vegetables, including coriander and spinach, are eaten in the form of leaves. Thus, the number of heavy metals in each leaf of a particular vegetable along with their soil samples were determined.\u003c/p\u003e\n\u003cp\u003eThe results represented in table 2 shows the concentrations of various heavy metals such as cobalt, copper, cadmium, chromium, and lead in spinach samples and their corresponding soil samples. Cobalt concentrations in the spinach samples varied from 0.2611 to 0.3412 mg/kg. The levels of copper, which range from 10.215 to 11.313 mg/kg, were noticeably higher. Much smaller amounts of cadmium, between 0.0131 and 0.0267 mg/kg, were recorded in the tested samples. Lead values range from 0.4353 to 0.54132 mg/kg, whereas chromium concentrations recorded in the range of 2.4147 to 2.6713 mg/kg. In contrast, the soil samples have greater cobalt contents, ranging from 0.4471 to 0.5132 mg/kg. The soil had substantially lower levels of copper, ranging from 3.0722 to 3.2013 mg/kg, than the spinach. However, the soil had much higher concentrations of cadmium, ranging from 0.361 to 0.4652 mg/kg, suggesting that a large amount of Cd is retained in the soil instead of being absorbed by the spinach. The amounts of chromium in the soil, which ranges from 2.4512 to 2.6306 mg/kg, were comparatively similar to those in spinach. The soil has greater lead (Pb) values than the spinach samples, ranging from 0.6214 to 0.7415 mg/kg. These results showed that the spinach absorbs certain metals more effectively (e.g., Cu) while other metals (e.g., Cd and Pb) tend to retained in higher concentrations by the soil.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2. Concentration of Heavy Metals Detected in Spinach and Soil Samples.\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" align=\"\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" style=\"width: 73px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eHeavy Metals\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"6\" valign=\"top\" style=\"width: 481px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSamples\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSpinach 1\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;(mg/kg)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSpinach 2\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;(mg/kg)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSpinach 3\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;(mg/kg)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSoil 1\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003emg/L\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSoil 2\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003emg/L\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 80px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSoil 3\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003emg/L\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 73px;\"\u003e\n \u003cp\u003eCo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e0.3412\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.2736\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.2611\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.4471\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.4541\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 80px;\"\u003e\n \u003cp\u003e0.5132\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 73px;\"\u003e\n \u003cp\u003eCu\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e11.1740\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e10.215\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e11.313\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e3.0722\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e3.1312\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 80px;\"\u003e\n \u003cp\u003e3.2013\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 73px;\"\u003e\n \u003cp\u003eCd\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e0.0267\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.0213\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.0131\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.4278\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.361\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 80px;\"\u003e\n \u003cp\u003e0.4652\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 73px;\"\u003e\n \u003cp\u003eCr\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e2.4147\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e2.5631\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e2.6713\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e2.6306\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e2.4512\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 80px;\"\u003e\n \u003cp\u003e2.5713\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 73px;\"\u003e\n \u003cp\u003ePb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e0.4353\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.54132\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.4512\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.7415\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.6341\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 80px;\"\u003e\n \u003cp\u003e0.6214\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eThe table 3 presented the concentrations of various heavy metals such as cobalt, copper, cadmium, chromium, and lead in three coriander samples and their corresponding soil samples. Cobalt values in the coriander samples ranged from 0.1649 to 0.2613 mg/kg. The levels of copper, which ranged from 6.1319 to 6.7688 mg/kg, were noticeably high. Significant concentrations of cadmium were recorded with in the coriander samples, ranging from 0.2146 to 0.3313 mg/kg. However, the range of chromium levels was quite small, which was in the range 0.4121 to 0.5131 mg/kg. Moreover, concentrations of lead in the samples was recorded in the range of 0.3712 to 0.5631 mg/kg. The same heavy metals were present in different amounts in the soil samples. The amounts of cobalt in the soil were similar to those in the coriander plants, ranging from 0.2131 to 0.3126 mg/kg. Interestingly, the soil had greater concentrations of copper, ranging from 6.3791 to 7.2134 mg/kg. The soil contained somewhat more cadmium than the coriander samples (0.3219\u0026ndash;0.3513), suggesting that some cadmium was not properly absorbed by the plant and instead stayed in the soil. Similarly, the soil had greater quantities of chromium than the coriander, ranging from 0.5617 to 0.6301 mg/kg. Lead levels in the soil, which ranged from 0.5131 to 0.8721 mg/kg, were significantly higher than in the coriander samples as shown in figure 5.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 3. Concentration of Heavy Metals Detected in Coriander and Soil Samples.\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" align=\"\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" style=\"width: 73px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eHeavy Metals\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"6\" valign=\"top\" style=\"width: 481px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSamples\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCoriander 1\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e(mg/kg)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCoriander 2\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e(mg/kg)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCoriander 3\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;(mg/kg)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSoil 1\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003emg/L\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSoil 2\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003emg/L\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 80px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSoil 3\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003emg/L\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 73px;\"\u003e\n \u003cp\u003eCo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e0.1649\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.2541\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.2613\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.2521\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.3126\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 80px;\"\u003e\n \u003cp\u003e0.2131\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 73px;\"\u003e\n \u003cp\u003eCu\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e6.7688\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e6.2613\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e6.1319\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e7.1634\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e6.3791\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 80px;\"\u003e\n \u003cp\u003e7.2134\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 73px;\"\u003e\n \u003cp\u003eCd\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e0.3173\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.2146\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.3313\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.3513\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.3412\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 80px;\"\u003e\n \u003cp\u003e0.3219\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 73px;\"\u003e\n \u003cp\u003eCr\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e0.4302\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.5131\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.4121\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.5617\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.6301\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 80px;\"\u003e\n \u003cp\u003e0.6213\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 73px;\"\u003e\n \u003cp\u003ePb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 85px;\"\u003e\n \u003cp\u003e0.4374\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.3712\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.5631\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.7642\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 79px;\"\u003e\n \u003cp\u003e0.8721\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 80px;\"\u003e\n \u003cp\u003e0.5131\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003ch3\u003e3.2 Heavy metals detection in Wastewater\u003c/h3\u003e\n\u003cp\u003eThe results regarding heavy metal concentration in waste water samples are shown in figure 3.2. The amount of cobalt (Co) in the wastewater was 0.0721 mg/L, more above the 0.05 mg/L WHO acceptable limit. In small doses, cobalt (Co) is a necessary as trace element, but higher concentrations can be hazardous. The concentration of copper in the wastewater was 0.2183 mg/L, which was well below the WHO permissible limit of 2.0 mg/L. The wastewater sample had 0.0232 mg/L of cadmium (Cd), which was more than the 0.01 mg/L WHO acceptable limit. The amount of chromium (Cr) in the wastewater was 0.1173 mg/L, more than twice the 0.05 mg/L WHO acceptable level. The concentration in the wastewater indicated that there is a serious pollution issue that requires attention. The wastewater has 0.2255 mg/L of lead (Pb), more than twice the WHO recommended limit of 0.1 mg/L as shown in figure 6.\u003c/p\u003e\n\u003ch3\u003e3.3 Descriptive statistical analysis of heavy metals in spinach samples\u003c/h3\u003e\n\u003cp\u003eDescriptive data of spinach samples are shown in figure 7. Including the concentrations of heavy metals and comparing them with World Health Organization (WHO) limits. The average (mean) concentration was 0.29 mg/kg for cobalt. The World Health Organization (WHO) has set a cobalt limit of 0.1 mg/kg in specific dietary and environmental situations in an effort to protect human health. The average content of copper in spinach was determined to be 10.9 mg/kg. These amounts of copper are far below the 73.3 mg/kg WHO guideline. With an average of 0.02 mg/kg, the amounts of cadmium in spinach are below the WHO limit, which is 0.1 mg/kg. Spinach has an average chromium content of 2.54 mg/kg. The average content of chromium is marginally higher than the 2.3 mg/kg WHO limit. The mean concentration of lead was 0.47 mg/kg. These levels are higher than the 0.3 mg/kg lead limit set by the World Health Organization (WHO).\u003c/p\u003e\n\u003ch3\u003e3.4 Descriptive statistical analysis of heavy metals in spinach soil samples\u003c/h3\u003e\n\u003cp\u003eDescriptive data of spinach soil samples are shown in figure 8. Including the concentrations of heavy metals and comparing them with World Health Organization (WHO) limits. The average level of cobalt (Co) in the soil used to grow spinach was 0.47 mg/kg. The cobalt (Co) contents in the soil were substantially lower than the WHO standard of 30 mg/kg for cobalt in soil. The average content of copper (Cu) in the spinach soil was 3.13 mg/kg. The copper (Cu) levels in soil were much below the 36 mg/kg WHO standard for copper in soil. This shows that there is no copper contamination in the soil, and the amounts of copper in spinach are safe for consumption. With an average value of 0.41 mg/kg, cadmium (Cd) levels in the soil range from 0.36 mg/kg to 0.46 mg/kg. These levels were higher above the WHO threshold of 0.31 mg/kg.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eThe average amount of chromium (Cr) in the soil was 2.55 mg/kg. The amounts of chromium in soil were far lower than the WHO limit, which was 68 mg/kg. This indicates that chromium (Cr) levels in spinach are mostly a result of the plant\u0026apos;s natural absorption and that chromium contamination of the soil is unlikely. The average lead (Pb) concentration in the soil was 0.66 mg/kg. The lead levels in soil were well below the 85 mg/kg WHO standard for lead in soil. This implies that there was no lead contamination in the soil, and the slightly higher lead levels in spinach could be the result of other causes such irrigation water.\u003c/p\u003e\n\u003ch3\u003e3.5 Descriptive statistical analysis of heavy metals in coriander samples\u003c/h3\u003e\n\u003cp\u003eDescriptive data of coriander samples are shown in figure 9. Including the concentrations of heavy metals and comparing them with World Health Organization (WHO) limits.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eCoriander had cobalt (Co) values on average 0.22 mg/kg. The average amount of cobalt in coriander surpasses the safety threshold of 0.1 mg/kg set by the WHO for cobalt. The average level of chromium (Cr) in coriander was 0.45 mg/kg. These amounts of copper (Cu) were far below the 73.3 mg/kg WHO guideline. The average level of copper (Cu) in coriander was 6.38 mg/kg, with a range of 0.56 to 0.51 mg/kg. These amounts of copper (Cu) are far below the 2.3 mg/kg WHO guideline. With an average of 0.28 mg/kg, the average level of cadmium (Cd) in coriander exceeds the safety threshold of 0.1 mg/kg as shown in table 4.8, which is the limit set by the WHO. The average lead (Pb) content in coriander was 0.45 mg/kg. The average level of lead in coriander surpasses the 0.3 mg/kg WHO standard for lead.\u003c/p\u003e\n\u003ch3\u003e3.6 Descriptive statistical analysis of heavy metals in coriander soil samples\u003c/h3\u003e\n\u003cp\u003eDescriptive data of coriander soil samples are shown in figure 10. It showed the concentrations of heavy metals and comparing them with World Health Organization (WHO) limits. The average value of cobalt (Co) in the soil used to grow coriander was 0.25 mg/kg. The cobalt contents in the soil were substantially lower than the WHO standard of 30 mg/kg for cobalt in soil.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eThe average content of copper (Cu) in the coriander soil was 6.91 mg/kg. However, the copper levels in soil were much lower to 36 mg/kg of standard for copper in soil. Moreover, the average amount of cadmium (Cd) in the soil was 0.33 mg/kg. The WHO has set a limit of 0.31 mg/kg of cadmium. Similarly, the levels of cadmium in the soil are somewhat higher than this threshold. The average contents of chromium (Cr) in the soil was 0.60 mg/kg. The amounts of chromium in soil were far lower than the WHO limit, which is 68 mg/kg.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eThe average lead (Pb) concentration in the soil was 0.71 mg/kg. The lead levels in soil were well below the 85 mg/kg WHO standard for lead in soil.\u0026nbsp;\u003c/p\u003e\n\u003ch3\u003e3.7 One sample T-test statistical analysis of spinach samples\u003c/h3\u003e\n\u003cp\u003eT-test showed the t value, df and P value of each heavy metal in spinach as shown in table 4. In spinach samples, Co, Cu, Cd and Pb showed significant differences as their P values were less than 0.05, which means that concentration of these heavy metals were different than WHO standards. while Cr showed insignificant results with p-value 0.07, which means that the concentrations of Cr were within limit of WHO standards.\u003cstrong\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 4. T-Test Results for Heavy Metal Concentrations in Spinach samples.\u003c/strong\u003e\u003c/p\u003e\n\u003cdiv align=\"\"\u003e\n \u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 110px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eHeavy Metals\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 110px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eT-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 110px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eDf\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 110px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eP-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 110px;\"\u003e\n \u003cp\u003eCo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 110px;\"\u003e\n \u003cp\u003e7.7158\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 110px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 110px;\"\u003e\n \u003cp\u003e0.01639*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 110px;\"\u003e\n \u003cp\u003eCu\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 110px;\"\u003e\n \u003cp\u003e-180.78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 110px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 110px;\"\u003e\n \u003cp\u003e0.0000306*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 110px;\"\u003e\n \u003cp\u003eCd\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 110px;\"\u003e\n \u003cp\u003e-20.142\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 110px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 110px;\"\u003e\n \u003cp\u003e0.02456*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 110px;\"\u003e\n \u003cp\u003eCr\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 110px;\"\u003e\n \u003cp\u003e3.3575\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 110px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 110px;\"\u003e\n \u003cp\u003e0.07843\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 110px;\"\u003e\n \u003cp\u003ePb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 110px;\"\u003e\n \u003cp\u003e5.3307\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 110px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 110px;\"\u003e\n \u003cp\u003e0.03344*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cstrong\u003e(*) indicates significant differences (p-value \u0026lt; 0.05), while others are non-significant.\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003ch3\u003e3.8 One sample T-test statistical analysis of spinach soil samples\u003c/h3\u003e\n\u003cp\u003eT-test showed the t value, df and p value of each heavy metal in spinach soil samples as shown in table 5. In spinach soil samples, all heavy metals like Co, Cu, Cd, Cr and Pb showed significant differences as their p values were less than 0.05, which means that concentration of these heavy metals were different than WHO standards.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 5. T-Test Results for Heavy Metal Concentrations in Spinach soil samples.\u003c/strong\u003e\u003c/p\u003e\n\u003cdiv align=\"\"\u003e\n \u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eHeavy Metals\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eT-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eDf\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 114px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eP-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003eCo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003e-1194.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 114px;\"\u003e\n \u003cp\u003e0.00000070148*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003eCu\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003e-72.715\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 114px;\"\u003e\n \u003cp\u003e0.0001891*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003eCd\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003e-73.258\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 114px;\"\u003e\n \u003cp\u003e0.0001863*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003eCr\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003e-879.99\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 114px;\"\u003e\n \u003cp\u003e0.000001291*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003ePb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003e-2213.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 114px;\"\u003e\n \u003cp\u003e0.0000000204*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cstrong\u003e(*) indicates significant differences (p-value \u0026lt; 0.05), while others are non-significant.\u003c/strong\u003e\u003c/p\u003e\n\u003ch3\u003e3.9 One sample T-test statistical analysis of coriander samples\u003c/h3\u003e\n\u003cp\u003eIn coriander samples, Cu and Cr showed significant differences as their p values were less than 0.05, which means that concentration of these heavy metals were different than WHO standards. While Co, Cd and Pb showed insignificant results with p values above than 0.05 as shown in table 6, which indicated the concentrations of these heavy metals were within limit of WHO standards as shown in .\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 6. T-Test Results for Heavy Metal Concentrations in Coriander samples.\u003c/strong\u003e\u003c/p\u003e\n\u003cdiv align=\"\"\u003e\n \u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 103px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eHeavy Metals\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 103px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eT-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 103px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eDf\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 150px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eP-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 103px;\"\u003e\n \u003cp\u003eCo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 103px;\"\u003e\n \u003cp\u003e4.0888\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 103px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 150px;\"\u003e\n \u003cp\u003e0.05493\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 103px;\"\u003e\n \u003cp\u003eCu\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 103px;\"\u003e\n \u003cp\u003e-344.28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 103px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 150px;\"\u003e\n \u003cp\u003e0.000008437*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 103px;\"\u003e\n \u003cp\u003eCd\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 103px;\"\u003e\n \u003cp\u003e-0.60525\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 103px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 150px;\"\u003e\n \u003cp\u003e0.6065\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 103px;\"\u003e\n \u003cp\u003eCr\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 103px;\"\u003e\n \u003cp\u003e-59.443\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 103px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 150px;\"\u003e\n \u003cp\u003e0.0002829*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 103px;\"\u003e\n \u003cp\u003ePb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 103px;\"\u003e\n \u003cp\u003e2.7939\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 103px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 150px;\"\u003e\n \u003cp\u003e0.1078\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cstrong\u003e(*) indicates significant differences (p-value \u0026lt; 0.05), while others are non-significant.\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003ch3\u003e3.10 One sample T-test statistical analysis of coriander soil samples\u003c/h3\u003e\n\u003cp\u003eIn coriander\u0026rsquo;s soil samples, Co, Cu, Cr and Pb showed significant differences as their p values were less than 0.05, which means that concentration of these heavy metals were different than WHO standards. while Cd showed insignificant results with p value 0.08 as shown in table 7, which indicated the concentrations of Cd were within limit of WHO standards.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 7. T-Test Results for Heavy Metal Concentrations in Coriander soil samples.\u003c/strong\u003e\u003c/p\u003e\n\u003cdiv align=\"\"\u003e\n \u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 99px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eHeavy Metals\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 99px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eT-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 100px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eDf\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 133px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eP-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 99px;\"\u003e\n \u003cp\u003eCo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 99px;\"\u003e\n \u003cp\u003e-1027.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 100px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 133px;\"\u003e\n \u003cp\u003e0.00000009473*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 99px;\"\u003e\n \u003cp\u003eCu\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 99px;\"\u003e\n \u003cp\u003e-101.65\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 100px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 133px;\"\u003e\n \u003cp\u003e0.00008628*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 99px;\"\u003e\n \u003cp\u003eCd\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 99px;\"\u003e\n \u003cp\u003e3.265\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 100px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 133px;\"\u003e\n \u003cp\u003e0.08251\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 99px;\"\u003e\n \u003cp\u003eCr\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 99px;\"\u003e\n \u003cp\u003e-3137\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 100px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 133px;\"\u003e\n \u003cp\u003e0.000000001016*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 99px;\"\u003e\n \u003cp\u003ePb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 99px;\"\u003e\n \u003cp\u003e-792.53\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 100px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 133px;\"\u003e\n \u003cp\u003e0.0000001592*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cstrong\u003e(*) indicates significant differences (p-value \u0026lt; 0.05), while others are non-significant.\u003c/strong\u003e\u003c/p\u003e\n\u003ch3\u003e3.11 Bio-concentration Factor (BCF) of heavy metals from soil to vegetables\u003c/h3\u003e\n\u003cp\u003eThe average bio-concentration factor values for each heavy metal (Co, Cd, Cr, Cu, Pb) in spinach and coriander samples are shown in table 8. Spinach has an average cobalt concentration of 0.291967 mg/kg. The average cobalt concentration in the soil used to grow spinach was 0.471467 mg/kg. For cobalt (Co) in spinach, the bio-concentration ratio (BCR) is 0.619273. This value suggests that spinach had a moderate soil-based cobalt accumulation. Coriander had an average cobalt content of 0.226767 mg/kg. The average content of cobalt (Co) in the soil used to grow coriander was 0.259267 mg/kg. However, in coriander, the BCR for cobalt (Co) has been determined as 0.874646. This indicates that coriander is somewhat more capable than spinach of absorbing cobalt from the soil. Spinach has an average copper concentration of 10.900667 mg/kg. The average concentration of copper (Cu) in the soil used to grow spinach was 3.1349 mg/kg. It is determined that spinach had a higher BCR of 3.477198 for copper. This suggests that spinach is an extremely effective way for the plant to absorb copper (Cu) from the soil. Coriander has an average copper (Cu) content of 6.387333 mg/kg. The average content of copper (Cu) in the soil used to cultivate coriander was 6.918633 mg/kg. In coriander, the BCR for copper (Cu) was determined to be 0.923207. This indicates that spinach is somewhat more capable than coriander of absorbing cobalt (Co) from the soil. Cadmium (Cd) concentration in spinach was 0.020367 mg/kg on average. The average level of cadmium (Cd) in the soil used for the growth of spinach was 0.418 mg/kg. The BCR for spinach\u0026apos;s cadmium content is 0.048724. This suggests that spinach absorbs less cadmium (Cd) from the soil. Cadmium (Cd) content in coriander is 0.287733 mg/kg on average.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe average quantity of cadmium (Cd) exhibited by the soil for growing coriander is 0.338133 mg/kg. It is determined that the BCR for cadmium in coriander is 0.850946 which indicates that coriander is somewhat more capable than spinach of absorbing cadmium from the soil. Spinach has an average chromium (Cr) concentration of 2.5497 mg/kg. Likewise, the mean concentration values of chromium (Cr) in the soil used to grow spinach was 2.551033 mg/kg. For spinach, the BCR for chromium is 0.999477. This suggests that spinach takes up a moderate amount of chromium (Cr) from the soil. Coriander had an average chromium concentration of 0.4518 mg/kg. The average amount of chromium (Cr) in the soil used to cultivate coriander was 0.604367 mg/kg. It is calculated that the BCR for chromium (Cr) in coriander was 0.747559 which implies that coriander absorbs chromium (Cr) from the soil in a moderate amount which indicated that spinach is slightly more capable than coriander of absorbing chromium from the soil. Spinach has an average lead concentration of 0.47594 mg/kg. The average lead (Pb) concentration in the soil used to grow spinach was 0.665667 mg/kg. The calculated BCR for lead (Pb) in spinach was 0.714982. This suggests that spinach absorbs lead (Pb) from the soil to a moderate extent. Coriander has an average lead (Pb) content of 0.457233 mg/kg. Lead (Pb) concentrations in the coriander-growing soil average 0.716467 mg/kg. Coriander\u0026apos;s BCR for lead (Pb) was determined to be 0.638178. This implies that coriander, like spinach, accumulates lead (Pb) from the soil in a moderate amount indicated its more capability than coriander in absorbing lead from the soil.\u003cstrong\u003e\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 8.\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;Bio-concentration Ratio for each heavy metal from soil to vegetables\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"630\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eHeavy metals\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAverage conc. in Spinach (mg/kg)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAverage conc. in soil (mg/kg)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eBCR Soil to Spinach\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 102px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAverage conc. in Coriander (mg/kg)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 90px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAverage conc. in soil (mg/kg)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 90px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eBCR Soil to\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eCoriander\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003eCo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e0.2919\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e0.4714\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e0.6192\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 102px;\"\u003e\n \u003cp\u003e0.2267\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 90px;\"\u003e\n \u003cp\u003e0.2592\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 90px;\"\u003e\n \u003cp\u003e0.8746\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003eCu\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e10.900\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e3.1349\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e3.4771\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 102px;\"\u003e\n \u003cp\u003e6.3873\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 90px;\"\u003e\n \u003cp\u003e6.918\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 90px;\"\u003e\n \u003cp\u003e0.9232\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003eCd\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e0.0203\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e0.4180\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e0.0487\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 102px;\"\u003e\n \u003cp\u003e0.2877\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 90px;\"\u003e\n \u003cp\u003e0.3381\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 90px;\"\u003e\n \u003cp\u003e0.8509\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003eCr\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e2.5497\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e2.5510\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e0.99947\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 102px;\"\u003e\n \u003cp\u003e0.45180\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 90px;\"\u003e\n \u003cp\u003e0.6043\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 90px;\"\u003e\n \u003cp\u003e0.7475\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003ePb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e0.4759\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e0.6656\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e0.71498\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 102px;\"\u003e\n \u003cp\u003e0.4572\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 90px;\"\u003e\n \u003cp\u003e0.7164\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 90px;\"\u003e\n \u003cp\u003e0.6381\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e"},{"header":"4. Discussion","content":"\u003cp\u003ePakistan is considered as a developing country with numerous serious problems that have a negative impact on people's health. One of the main issues facing by the human beings in the current situation is the lack of fresh water availability for humans and cultivation of crops and vegetables, which eventually affects the nation's agriculture sector by making it difficult to meet the fundamental needs of agriculturally produced food to feed the expanding population. In order to get rid of this problem, farmers are growing vegetables in peri-urban regions by mindlessly using untreated sewage or industrial water (Jabeen et al., 2020). In this study, a survey was conducted in 2023 and sites were selected where vegetables have been irrigated by wastewater of industry and sewage. Later on wastewater has been collected from the field at Gujranwala for the analysis of heavy metal concentration in waste water and compared with WHO limits. Our findings on assessment and quantification showed serious concerns about cobalt (Co), cadmium (Cd), chromium (Cr), and lead (Pb) levels over the WHO permitted limits by the examination of heavy metal concentrations in the wastewater. Both lead and chromium can cause serious health problems, when their levels are more than double the recommended limits and decisive action is essential in order to reduce these contaminants to assure the environmental and public health's safety. Other researchers in Pakistan additionally investigated higher levels of heavy metals in wastewater and discovered abnormal quantities. Jabeen et al., (2020), assessed higher contents of Manganese and Nickel in wastewater in Faisalabad, Pakistan. Similarly, Rehman et al., (2019), reported higher quantities of heavy metals such as Cd and Manganese in wastewater in district Sahiwal, Pakistan, where wastewater has been applied for irrigation. The analysis of heavy metal concentrations in spinach, coriander, and their respective soils reveals several significant findings. The quantification of heavy metals in the spinach samples varied, with lead, chromium, and cobalt concentrations above WHO recommendations. The cobalt levels were higher above the WHO limit of 0.1 mg/kg, ranging from 0.26 to 0.34 mg/kg on average. The average chromium concentration was 2.54 mg/kg, which is marginally more than the 2.3 mg/kg of WHO standard. With a mean of 0.47 mg/kg, lead levels in spinach ranged from 0.43 to 0.54 mg/kg, exceeding the WHO recommendation of 0.3 mg/kg. Our findings were supported by Kalhoro et al., (2023), who reported higher level of six heavy metals such as Cr, Ni, Cd, Pb, As and Hg in cabbage, cauliflower, turnip and radish. Similarly, Iqbal et al., (2020), found higher levels of heavy metals including Fe, Pb and Cr in sponge gourd in Pakistan. In contrast, levels of cadmium and copper in spinach's were within acceptable limits. The values of cadmium were between 0.01 and 0.02 mg/kg, which were below the WHO limit of 0.1 mg/kg, and copper varied from 10.2 to 11.3 mg/kg, both significantly below the WHO recommendation of 73.3 mg/kg. Lead, copper, chromium, and cobalt levels in the spinach soil were all much below WHO recommendations, suggesting that plant uptake of the metals rather than soil contamination is most likely to be responsible for the increased levels in spinach. The WHO standard of 0.31 mg/kg for cadmium is exceeded by the average of 0.41 mg/kg in the soil, indicating possible pollution and the need for soil remediation to reduce health concerns. Mahmood et al., (2020), investigated higher level of heavy metals such as Fe, Mg, Na, Ca and K in soil due to waste water irrigation.. Leblebici et al., (2020), also assessed higher level of heavy metals such as Zn and Mn in soil where waste water was applied. Aftab et al., (2023), found higher levels of heavy metals like Cr and Co in crops irrigated with waste water. Lead, cadmium, and cobalt levels in coriander samples exceeded the WHO recommendations. The mean cobalt content was 0.22 mg/kg, exceeding the WHO threshold of 0.1 mg/kg. Our investigations indicated Lead concentrations which varied from 0.37 to 0.56 mg/kg, with an average of 0.45 mg/kg, exceeding the WHO limit of 0.3 mg/kg. Moreover, cadmium levels averaged 0.28 mg/kg, exceeding the WHO threshold of 0.1 mg/kg. Our findings are in agreement with Ugulu et al., (2023), who investigated higher level of heavy metals of cadmium, cobalt, chromium, copper, lead in coriander when irrigated with waste water. Similarly, Hussain et al., (2022), showed higher levels of Cd in coriander while lower level of Mn accumulation as compared to WHO limits. On the other hand, coriander has safe copper levels of 6.38 mg/kg, below the 73.3 mg/kg WHO recommendation. However, Lead, chromium, copper, and cobalt concentrations in the coriander soil were all found to be substantially below the corresponding WHO standards, indicating no evidence of pollution. Whereas, the average level of cadmium in the coriander soil was 0.33 mg/kg, which is somewhat higher than the WHO guideline of 0.31 mg/kg. Aftab et al., (2023), found lower levels of heavy metals like Ni, Cu, Pb, and Mn concentrations which were in line with WHO standards in coriander. To determine the statistical significance differences, the t-value, degrees of freedom (df), and associated p-value for each heavy metal were computed. A p-value less than the conventional significance level (typically 0.05) suggests strong evidence against the null hypothesis, leading to its rejection. The results for chromium (Cr) were non-significant (p\u0026thinsp;=\u0026thinsp;0.07843), meaning that the amount of the heavy metal in spinach was within WHO-acceptable limits. The quantities of heavy metals in relation to WHO guidelines were revealed by the one-sample t-test studies performed on coriander, spinach, and soil samples. Significant variations were found in the amounts of copper and chromium in coriander samples, suggesting levels exceeding WHO guidelines, but acceptable ranges were found for cobalt, cadmium, and lead. Ali et al., (2021) demonstrated a significant difference in soil samples for Pb, Cr, Cd, Ni, Zn and Cu at p\u0026thinsp;\u0026le;\u0026thinsp;0.001, for Mn at p\u0026thinsp;\u0026le;\u0026thinsp;0.05 while no significant difference was observed for Fe respectively. Also exhibited the highest mean value for Pb, Cr, Cd and Zn in vegetables. Hussain \u0026amp; Qureshi, (2020), showed results indicate significant differences that iron (Fe) was highest in lettuce followed by spinach, and Zn and Cr were second and third element in the vegetables. Eggplant and radish showed non-significant differences the lowest concentrations of heavy metals. Ahmed et al., (2022), showed similar results by assessing higher accumulation of heavy metals like chromium and cadmium as compared to WHO limits, in green leafy vegetables along with their soils. Similarly, Shahid et al., (2019), showed similar results which indicated significant differences in Cu and Zn in cucumber and chili. The heavy metal Bioconcentration Ratios (BCRs) for soil, spinach, and coriander show distinct patterns of accumulation that emphasize the bioindicators' importance in contaminated agricultural settings. Our research findings showed that Spinach exhibits a high BCR and effective copper accumulation, suggesting that it could be a sensitive bioindicator of soil copper contamination. A study conducted by Gupta et al., (2022), assessed significant accumulation of heavy metals like Zn and Mn in vegetables, whereas, spinach has less ability to carry lead, cadmium, and cobalt, but its uptake of chromium is balanced. Likewise, coriander shows moderate cobalt, copper, and cadmium accumulation efficiency which suggested that in contaminated soils, coriander can act as a reliable bioindicator for these heavy metals. in comparison, spinach, accumulates chromium and lead less efficiently, suggesting that it is not as effective as spinach at monitoring these specific elements. These research findings were supported by Oguh \u0026amp; Obiwulu, (2020), who reported low to moderate BCR values for heavy metals like As, Pb, Cd and Hg from soil to vegetables. Ali et al., (2021), also demonstrated similar higher BCR results, the transfer factor for Cr, Pb, Zn, Mn, Ni, Cd and Cu was greater than 0.5 due to contamination caused by discharges and industrial effluents. Understanding these accumulation patterns is crucial for developing effective strategies for monitoring and managing heavy metal contamination to ensure food safety and protect environmental health Additional research is aimed on more vegetables to detect heavy metals concerning with the health hazardous issue on human beings and healthier generation.\u003c/p\u003e"},{"header":"5. Conclusion","content":"\u003cp\u003eThis study concludes that there is a potential risk of heavy metal contamination in vegetables as a result of wastewater irrigation. This study evaluated the presence of heavy metals (Co, Cu, Cd, Cr, Pb) in wastewater, soil, and vegetable samples spinach and coriander. The atomic absorption spectrophotometry, revealed the levels of Pb, Co, Cr, and Cd in wastewater were alarmingly high. Spinach and coriander samples also exhibited heavy metal concentrations exceeding WHO permissible limits, with significant levels of Co, Cr, and Pb detected. Similarly, the one-sample t-test analysis on spinach and coriander samples revealed significant deviations in cobalt, copper, cadmium, and lead concentrations from WHO standards. However, the bioaccumulation factor (BCR) indicated higher Cu accumulation compared to Cr, Cd, Pb, and Co in the vegetables. These findings underscore the critical issue of heavy metal toxicity in urban areas due to wastewater irrigation. Therefore, the study suggested the urgent need for sustainable land and water resource management practices to mitigate heavy metal contamination for human health and food safety.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eCredit authorship contribution statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eNM, and SR:\u003c/strong\u003e Experimentation, Data analysis and manuscript writing; \u003cstrong\u003eMA, and LW:\u003c/strong\u003e Data analysis and Reviewing and editing; \u003cstrong\u003eAM: S\u003c/strong\u003eupervision; \u003cstrong\u003eMA, and AA:\u003c/strong\u003e Visualization; \u003cstrong\u003eAM, KFA, and MA:\u003c/strong\u003e Conceptualization and resources, \u003cstrong\u003eAM, AES, and MMS:\u003c/strong\u003e Analysis, reviewing, revising and editing; All authors have read and approved the final manuscript.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgment\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAuthors are thankful to all the contributors who work and helped during the whole experiment. Cooperation of Assistant Professor Abdul Mateen and Director Dr. Maqsood Ahmed is greatly acknowledged for the supervision, conceiving of the manuscript and financial assistance. The authors extend their appreciation to Researchers Supporting Project number (RSPD2025R561), King Saud University, Riyadh, Saudi Arabia.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll authors have been agreed to the publication of this manuscript.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable. The funding information will be provided if required by the journal. The authors extend their appreciation to Researchers Supporting Project number (RSPD2025R561), King Saud University, Riyadh, Saudi Arabia.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and material:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll the data and material is available in the manuscript file\u0026nbsp;\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eKalhoro A, Mirani AA, Siyal FK, Jatt T, Mahar AR, Iram S, et al. Accumulation of Heavy Metals in Vegetable Food under Wastewater Irrigation. Pak J Zool. 2023;55(1). \u003c/li\u003e\n\u003cli\u003eAndleeb S, Rehman KU, Mahmood A, Elsadek MF, Safa NU, Hussein DS, et al. Human health risk hazards by heavy metals through consumption of vegetables cultivated by wastewater. J King Saud Univ. 2023;35(2):102467. \u003c/li\u003e\n\u003cli\u003eSardar A, Shahid M, Natasha, Khalid S, Anwar H, Tahir M, et al. 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Health risks of heavy metal exposure and microbial contamination through consumption of vegetables irrigated with treated wastewater at Dubai, UAE. Environ Sci Pollut Res. 2020;27:11213\u0026ndash;26. \u003c/li\u003e\n\u003cli\u003eAhmed S, Mahdi MM, Nurnabi M, Alam MZ, Choudhury TR. Health risk assessment for heavy metal accumulation in leafy vegetables grown on tannery effluent contaminated soil. Toxicol Reports. 2022;9:346\u0026ndash;55. \u003c/li\u003e\n\u003cli\u003eGupta N, Yadav KK, Kumar V, Prasad S, Cabral-Pinto MMS, Jeon BH, et al. Investigation of heavy metal accumulation in vegetables and health risk to humans from their consumption. Front Environ Sci. 2022;10:791052. \u003c/li\u003e\n\u003cli\u003eOguh CE, Obiwulu ENO. Human risk on heavy metal pollution and bioaccumulation factor in soil and some edible vegetables around active auto-mechanic workshop in Chanchaga Minna Niger state, Nigeria. Ann Ecol Environ Sci. 2020;4(1):12\u0026ndash;22. \u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"University of Sailkot","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Heavy metals, wastewater, vegetable, spectrometry, contamination","lastPublishedDoi":"10.21203/rs.3.rs-6150572/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6150572/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eVegetables are vital for human nutrition but can accumulate heavy metals, posing risks to public health and the environment, particularly in regions using wastewater for irrigation. This study, conducted in Gujranwala, Pakistan, evaluated heavy metal contamination in wastewater, soil, \u003cem\u003eSpinacia oleracea\u003c/em\u003e L., and \u003cem\u003eCoriandrum sativum\u003c/em\u003e L. using Atomic Absorption Spectrometry (AAS). Results revealed significant levels of lead (0.2255 mg/L), cobalt (0.0721 mg/L), chromium (0.1173 mg/L), and cadmium (0.0232 mg/L) in wastewater. Spinach and coriander samples exhibited heavy metal concentrations exceeding World Health Organization (WHO) limits, including chromium (11.313 mg/kg), lead (0.541 mg/kg), and cadmium (0.331 mg/kg). Soil samples also showed high cadmium levels. The findings underscore the urgent need for sustainable irrigation practices and land management to mitigate heavy metals bioaccumulation and safeguard food safety.\u003c/p\u003e","manuscriptTitle":"Assessment of Heavy Metals Contamination in Spinacia oleracea L. and Coriandrum sativum L. Irrigated with Wastewater: Implications for Food Safety and Environmental Health","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-03-05 06:48:52","doi":"10.21203/rs.3.rs-6150572/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"58ef1f91-09c4-4878-9d23-293bcee2fa04","owner":[],"postedDate":"March 5th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":45152039,"name":"Plant Physiology and Morphology"}],"tags":[],"updatedAt":"2025-03-05T06:48:52+00:00","versionOfRecord":[],"versionCreatedAt":"2025-03-05 06:48:52","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6150572","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6150572","identity":"rs-6150572","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}