Trace Elemental and Heavy Metal levels in frequently consumed local Vegetables of three Chronic Kidney Disease prevalent Villages

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Narayana Murthy, P. Balarama Swamy Yadav, P Balarama Swamy Yadav This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-2737719/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 09 Jul, 2023 Read the published version in Biological Trace Element Research → Version 1 posted 7 You are reading this latest preprint version Abstract The present study reports the Trace Elemental and Heavy Metal (24 Elements) levels in six ( Capsicum frutescence L., Carica papaya L., Momordica charantia L. , Moringa oliefera Lam. , Musa Sapientum L., and Solanum melongena L.) Local and frequently consumed vegetables from three locations which are severely affected Villages by Chronic Kidney Disease (CKD) in Kaviti Mandal, in Uddanam area of Srikakulam district. The samples are collected from these three Villages and are subjected to ICP-MS analysis. To test a group of 24 Elements viz., Li, Be, Al, Sc, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Ga, As, Se, Rb, Sr, Ag, Cd, Cs, Ba, TI, Rb, and U for their levels, the obtained levels of each Element is compared with the WHO/FAO permissible values. In these 24 Elements, as per FAO/WHO standards 16 Elements shows severe affect on kidneys and the remaining 8 (Mn, Co, Ni, Cu, Zn, Se, Sr, and TI) cause other health problems when they are in high concentration[1-5]. The results reveal that Ba is in high concentration (2.51 times) in all the tested samples and Pb is in high concentration (1.28 times) in 11 samples, Ag and Fe are in one sample each. Among the three locations highest Ba concentration is observed in S 1 ( Capsicum ) of L 2 followed by S 5 ( Musa ) and S 1 ( Capsicum ) of L 1 . The highest Pb concentration is present in S 1 ( Capsicum ) of L 3 followed by S 1 ( Capsicum ) of L 2 . The results reveal that out of the six vegetables tested, Capsicum showed high concentrations of Ba and Pb. The non-uniformity of levels of Trace Elements and Heavy Metals with regard to location and samples may be due to soil and ground water. Chronic Kidney Disease ICP-MS Trace Elements Vegetables Uddanam area Figures Figure 1 Figure 2 Introduction Uddanam is a region covering few parts of the seven Revenue Mandals or Tahasils (Ichhapuram, Kanchiri, Kaviti, Mandatha, Paratha, Sompeta and Vajrapkotlu) in the Srikakulam district of Andhra Pradesh, India. The area is a lush green environment with rich Coconut and Cashew nut orchards along the Coastline of the Bay of Bengal. However, it is known the region has a high incidence of CKD. Ravi raju et al [6] reported 15.9% CKD in their pilot study and 18.23% in their final study of age group above 18 years in Uddanam area, which is high incidence of CKD. Thousands of people are still suffering from this disease; till now, the reason(s) behind the cases of kidney failure are unknown and many investigations are being conducted to explore the cause(s) of this CKD, Which is specific to this area. Worldwide, 697.5 million people suffering from chronic kidney disease, of which 115.1 million are Indians [7]. According to the World Health Organization (WHO), Uddanam is one of three regions in the world with a high CKD incidence of 18.23% [6], the other two in Sri Lanka and Nicaragua. In 2015, it is estimated that over 4,500 people have died in the Uddanam area over the past decade, with about 34,000 suffering from kidney disease. Nearly every family in the region has at least one person diagnosed with the disease. These cases were first recognized in his early 1990s, and the phenomenon was discussed at the International Nephrology Conference in Hong Kong (2013) and named Uddanam nephropathy [8]. In 2016, the Indian Council of Medical Research partnered with Harvard University, Baba Atomic Research Center (Mumbai), Health University (Andhra Pradesh) and Andhra Medical College (Vishakapatnam) to study the causes of CKD. Few studies have been done on the analysis of Trace Elements in edible plant parts from the Uddanam area, such as rice and finger millet using the EDXRF technique, indicates that Cu and Zn Elements are in high concentration in both the samples and Phosphorus is more in Rice and Se is more in Finger millet [12, 13]. Studies on analysis of Trace Elements of soil and water samples have been carried out [9-11], and showed that the water and soil in the villages of Uddanam contain high concentration of minerals of Ferrus, Manganese, Copper and Zinc [11]. Khandare et al [14] identified excessive concentrations of Li, Si and Sr in the groundwater of Uchapally village of Nellore district, Andhra Pradesh, India; which is also a CKD prevalent village, indicating presence of some Elements and Heavy Metals in high concentration. Studies on Trace Elemental analysis of plant edible parts are few viz. Benerjee et al (2010) noticed excess Lead content in all fruit and vegetable samples collected from the districts of West Bengal [15], Mamatha et al (2014) identified excessive levels of Cadmium and Lead in Leafy vegetables used by local people in Pulivendula, Kadaba district of Andhra Pradesh [16]. Swapnapriya et al (2014) identified high levels of Copper, Chromium, Iron and Lead in leaves of Sorrel ( Rumex sps.) in the Musi River basin of Hyderabad [17]. Swapna Gupta et al (2013) found high levels of Chromium, Zinc, Copper and Lead in leafy Vegetables collected in the city of Raipur [18]. So far, many studies indicate that the water and soil of the Uddanam region contain Trace Elements in high concentration than permissible values. There are no attempts on Trace Elemental analysis in all the local plant edible parts that are frequently consumed by the people of this region. Thus, the present study is taken up. Materials and Methods Study area Geographically, Uddanam is located at 19.0167 degrees north latitude and 84.6833 degrees east longitude, with an average elevation of 41 meters above sea level. It consists of 7 revenue Mandals (Ichapuram, Kanchili, Kaviti, Mandasa, Palasa, Sompeta, and Vajrapukotturu). We have chosen Kaviti revenue Mandal because of high prevalence of CKD. We selected three revenue villages out of 21 revenue Villages in Kaviti Mandal based on the patients number and data obtained from the Community Hospital Sompeta. These three villages are Kusmapuram (L1), Baraka (L2) and Bolivanka (L3) (Figure 1-A, B, C, D). Sample collection and Preparation We identified six locally cultivated vegetables for Trace Elemental and Heavy Metal analysis (Table 1), which are grown in their Coconut orchards as inter crops and are frequently consumed. In which 2 are belong to the family Solanaceae, and one each from Caricaceae, Cucurbitaceae, Moringaceae and Musaceae (Table 1). Table 1. Frequently consumed local Vegetables used in the study Sample No. Botanical Name Family English name Vernacular name S 1 S 2 S 3 S 4 S 5 S 6 Capsicum frutescence L. Carica papaya L. Momordica charantia L. Moringa oliefera Lam. Musa sapientum L. Solanum melongena L. Solanaceae Caricaceae Cucurbitaceae Moringaceae Musaceae Solanaceae Chilli Papaya Bitter gourd Drumsticks Plantain Brinjal Pachchi mirapa kaya Boppayi kaya Kakara kaya Munaga Kada Arati kaya Vanga kaya The fresh samples of the selected vegetables collected from the fields directly, from the three Villages are kept in zip locked bags and are brought to the laboratory. The samples are washed twice with distilled water and then subjected to Acid Digestion process, i.e. fast and accurate Wet Digestion method [19] is followed for the digestion of samples to digest all organic matter in the samples and placed in ICP-MS to obtain the Trace Elemental levels. Procedure We used the ICP-MS instrument (Agilent Technologies- Model 7700) available in Centre for Studies on Bay of Bengal, Andhra University, Visakhapatnam. The Sample preparation for ICP-MS analysis followed here is based on the oxidation and solubility of organic matter in plants when H 2 O 2 (33%) is added directly to the concentrated HNO 3 and heated for a short time. The instrument was switched on and is first run with the Standard Reference Materials of 0 PPB, 0.5 PPB, 1 PPB, 5 PPB, 10 PPB, 25 PPB, 50 PPB, and 100 PPB then ran the samples and obtained the levels of 24 Elements (Li, Be, Al, Sc, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Ga, As, Se, Rb, Sr, Ag, Cd, Cs, Ba, TI, Rb, and U). ICP-MS instrument (Agilent Technologies - Model 7700) available at the Center for studies on Bay of Bengal, Andhra University, Visakhapatnam was used for analysis. The following sample preparation for ICP-MS analysis is based on the oxidation and solubility of organic matter in plants when H 2 O 2 (33%) is added directly to concentrated HNO 3 and heated for short time. First we turn on the instrument and run with standard reference materials of 0 PPB, 0.5 PPB, 1 PPB, 5 PPB, 10 PPB, 25 PPB, 50 PPB, and 100 PPB, then followed by samples to find 24 Elements. (Li, Be, Al, Sc, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Ga, As, Se, Rb, Sr, Ag, Cd, Cs, Ba, TI, Rb and U). Wet Digestion Modified HNO 3 /H 2 O 2 wet digestion method (Pequerul. et al. 1993) [19] was followed. 2 gr. sample is taken, kept in a Teflon beaker, mixed with 8 ml HNO 3 and left overnight. Heat the solution on a hot plate at 120-125 °C for 1 hour and add 4 mL H 2 O 2 (33%) once or twice until the organic matter completely digested. Then we added H 2 O 2 until the solution is clear or colorless (indicating complete digestion of organic matter). Place the residue in a dry place at 80° C., shake well, cool to room temperature and filter through Whattman filter paper. The solution is then diluted to a known volume (50 mL) with Milli-Q water. These digested solutions are stored in high-density polyethylene bottles (100 mL) at room temperature for further use. Results and Discussion Samples from three neighboring villages are analyzed for Trace Elements/ Heavy Metals using ICP-MS instrument. The values obtained are compared with the limits recommended by FAO/WHO (1990, 1995, 1997, 2006, 2008, 2009, 2010, 2011, 2012, 2013, 2014, 2015, 2016, 2017, 2018, and 2019) and the Food Safety and Standards (Contaminants, Toxins and Residues) Regulations (2011) [1-5]. A total of 24 Elemental levels in 18 vegetable samples from 3 different locations (L 1 –L 3 ) were obtained and are shown in Table 2. Out of the 18 samples tested, all tested samples were found high concentrations of Ba and 11 samples had high concentrations of Pb. Fe and Ag are high in each high one sample and the remaining. Table 2. Trace Elemental and Heavy Metal levels in vegetable samples by ICP-MS analysis. Element Li Be Al Sc V Cr Mn Fe Co Ni Cu Zn P.V. 5.6 8.4 60 22 50 2.4 15 50 0.14 5.0 3.5 50 S 1 L 1 L 2 L 3 L 1 L 2 L 3 L 1 L 2 L 3 L 1 L 2 L 3 L 1 L 2 L 3 L 1 L 2 L 3 0.043 0.046 0.019 0.007 0.01 0.016 0.022 0.011 0.014 0.014 0.005 0.024 0.068 0.032 0.026 0.023 0.017 0.02 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 9.96 56.03 44.63 8.76 7.68 10.25 34.86 7.82 43.46 7.31 11.05 16.85 15.26 9.66 9.4 7.57 8.54 24.94 5.547 5.106 6.810 5.108 3.228 6.735 4.926 4.279 7.720 5.192 2.114 3.340 7.183 3.82 6.83 3.708 6.06 6.02 0.035 0.103 0.029 0.018 0.016 0.429 0.027 0.016 0.017 0.019 0.016 0.049 0.029 0.029 0.018 0.014 0.021 0.014 0.535 1.139 0.454 0.613 0.439 0.65 0.509 0.392 0.542 0.504 0.483 0.878 0.482 0.419 0.525 0.498 0.557 0.461 0.376 3.245 1.035 0.523 0.562 0.865 0.426 0.26 2.51 0.874 0.359 0.693 13.59 0.424 0.428 0.484 16.20 0.747 9.082 30.33 27.32 7.31 6.64 216.3* 10.33 6.199 6.22 5.76 5.89 27.41 13.95 7.53 6.24 4.51 8.78 5.61 0.013 0.025 0.025 0.01 0.011 0.02 0.005 0.005 0.01 0.009 0.01 0.005 0.042 0.006 0.014 0.011 0.02 0.003 0.348 0.434 2.013 0.297 0.329 0.314 0.295 0.237 0.419 0.455 0.283 0.458 0.376 0.2 0.23 0.26 0.407 0.568 0.644 1.227 0.895 0.02 0.234 0.099 0.45 0.135 0.031 0.082 0.132 0.214 0.339 0.045 0 0.0008 0.973 0.196 10.19 15.62 18.77 6.83 7.52 6.41 5.96 5.22 8.65 7.09 4.30 10.39 23.59 3.90 8.45 11.58 8.78 8.89 S 2 S 3 S 4 S 5 S 6 contd… Element Ga As Se Rb Sr Ag Cd Cs Ba Tl Pb U P.V. 3.0 0.18 1.5 5.0 10 0.175 1.5 90 12.6 1.5 0.3 7 S 1 L 1 L 2 L 3 L 1 L 2 L 3 L 1 L 2 L 3 L 1 L 2 L 3 L 1 L 2 L 3 L 1 L 2 L 3 0.003 0.013 0.005 0.001 0.002 0.014 0.005 0.002 0.003 0.002 0.003 0.007 0.0060.002 0.003 0.002 0.002 0.003 0.01 0.031 0.008 0.005 0.002 0.011 0.005 0.003 0.004 0.005 0.004 0.013 0.009 0.005 0.005 0.007 0.015 0.005 0.006 0.133 0.039 0.036 0.037 0.039 0.028 0.007 0.037 0.017 0.005 0.043 0.03 0 0.015 0.007 0.021 0.027 0.587 1.107 0.587 0.735 0.823 0.92 0.54 0.489 0.879 0.531 0.949 1.158 1.92 0.463 0.266 0.504 2.356 1.01 1.395 2.832 0.887 0.574 0.63 0.552 0.605 0.72 0.617 1.137 0.842 1.143 1.847 0.882 0.705 0.698 3.258 0.928 0.048 0.124 0.047 0.057 0.122 0.027 0.026 0.033 0.026 0.035 0.388* 0.172 0.048 0.056 0.087 0.104 0.04 0.066 0.006 0.013 0.026 0.009 0.011 0.032 0.01 0.011 0.004 0.008 0.009 0.015 0.018 0.005 0.019 0.007 0.01 0.004 0.0005 0.002 0.0001 0.0001 0.0004 0.0003 0 0 0 0.0001 0.0003 0.0003 0.001 0 0 0 0 0.0001 36.67* 43.92* 27.65* 30.12* 33.73* 21.80* 31.60* 32.29* 26.95* 32.44* 29.23* 23.82* 42.22* 33.34* 33.85* 30.07* 33.64* 26.89* 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.372* 0.798* 0.972* 0.2 0.28 0.327* 0.415* 0.511* 0.252 0.254 0.214 0.416* 0.353* 0.304* 0.460* 0.177 0.316* 0.299 0 0.01 0.0008 0 0 0.0001 0.003 0 0 0 0 0 0 0 0 0 0 0 S 2 S 3 S 4 S 5 S 6 S 1 -S 6 : Samples 1-6; P.V. - Permissible Values (mg/kg or ppm) as per WHO/FAO. L 1 (Location 1) = Kusumapuram , L 2 ( Location 2)= Varaka, L 3 (Location 3) = Borivanka * = Trace Elements and Heavy Metals in more concentrations than the WHO/FAO standards that are harmful to kidney; The results reveal that two Elements i.e. Ba in high concentration in Capsicum frutescence raw fruit (36.67- L 1 , 43.92- L 2 and 27.65- L 3 mg/kg) and Pb (0.372- L 1 , 0.798- L 2 and 0.972- L 3 ). In Carica papaya raw fruit three Elements i.e. Fe in high concentrations (216.3- L 3 mg/kg), Ba (30.12- L 1 , 33.73- L 2 and 21.8- L 3 mg/kg) and pb (0.327- L 3 ). In Momordica charantia raw fruit two Elements i.e. Ba, in high concentrations (31.6- L 1 , 32.29- L 2 and 26.95- L 3 mg/kg) and Pb (0.415- L 1 , 0.511- L 2 ). In Moringa oliefera raw fruit two Elements are in high i.e. Ba (32.44- L 1 , 29.23- L 2 and 23.82- L 3 mg/kg), Pb (0.416- L 3 ) and Ag (L 2 -0.388). In Musa sapientum two Elements i.e. Ba in high concentration (42.22- L 1 , 33.34- L 2 and 33.85- L 3 mg/kg) and Pb (0.353- L 1 , 0.304- L 2 and 0.460- L 3 ). In Brinjal two Elements i.e. Ba (30.07- L 1 , 33.64- L 2 and 26.89- L 3 mg/kg) and Pb (0.316- L 2 ) are in high concentration (Table 2). It was found that the highest concentration of Fe is present among all the 18 tested samples in Carica papaya of Borivanka, highest concentration of Ba observed in Capsicum frutescence of Varaka followed by Musa sapientum L. of Kusumapuram, and then highest concentration of Pb seen in Chillies of Borivanka followed by Capsicum frutescence of Varaka (Table 2). Highest Pb concentration was found in Chilli, and lowest concentration is in Brinjal. Average highest concentration of Elements identified in Borivanka village followed by Varaka and Kusumapuram (Table 2), and it reflects with the CKD patient intensity. Conclusion In this study we analyzed 18 vegetable samples in 3 locations (villages) six vegetables in each for Trace Elemental levels. In all the three villages Ba and Pb are in high concentrations. Fe and Ag are in high concentration in one sample each, and their excessive consumption may influence the functioning of kidneys [1-6]. On an average Lead concentrations are 1.28 times higher and Ba is 2.51 times higher than the permissible values in the tested locations (Table 2). These excess concentrations of Elements in the studied samples may be absorbed from soil and ground water (naturally present in some locations) or may be due to usage of Chemical pesticides and fertilizers. The study reveals the extent of Trace Elements in local vegetables will enhance the risk of disease intensity, by consuming these vegetables frequently and further research is to be needed in this area to find out the cumulative or additive affects of soil, ground water and local plant edibles and or a single exact reason for CKD in Uddanam area. Among the three locations average Ba and Pb concentration are more (Fig 2a, 2b) in Borivanka (L 3 ) followed by Varaka (L 2 ) and Kusumapuram (L 1 ), and it reflects with the CKD patient intensity. The studies on soil, ground water and all local plant edibles will help in understanding the Serological, Urological, nutritional and Genetic studies of the CKD. The present study opens a new dimension to understand the role of Trace Elements behind the CKD by analyzing the Trace Elemental analysis of all local frequently consumed plant edible parts coupled with soil and ground water analysis. The studies [9, 10 and 11] on soil and water in Uddanam area reveal that Fe, Mn, Cu and Zn Metals are in high concentration. Providing safe water to drinking, establishing more dialysis centers and giving pensions to patients affected by CKD are a huge financial burden for governments in many areas where CKD is prevalent. This can be reduced by taking studies on the soil, water and all edible local plants. The present study opens a new dimension to understand the impact of Trace Elements and Heavy Metals on CKD by analyzing the Trace Element and Heavy Metal content of all commonly consumed plant parts of local food crops along with soil and ground water. Declarations Conflict of interest The authors declare that no conflict of interest. The authors alone are responsible for the content and writing of this paper. There is no any funding for this work. 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Cite Share Download PDF Status: Published Journal Publication published 09 Jul, 2023 Read the published version in Biological Trace Element Research → Version 1 posted Editorial decision: Major revision 30 May, 2023 Reviews received at journal 28 Mar, 2023 Reviewers agreed at journal 28 Mar, 2023 Reviewers invited by journal 27 Mar, 2023 Editor assigned by journal 27 Mar, 2023 Submission checks completed at journal 27 Mar, 2023 First submitted to journal 26 Mar, 2023 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. 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Narayana Murthy","email":"","orcid":"","institution":"Andhra University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"G.","middleName":"Narayana","lastName":"Murthy","suffix":""},{"id":186698734,"identity":"44ad493a-3eb7-48f1-8a43-758a67c53b4c","order_by":1,"name":"P. Balarama Swamy Yadav","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA6ElEQVRIiWNgGAWjYDCCAwwMzFAG4wMgxcNHihZmA5AWNlK0sEmAaIJa+G4fYHtc8OewPN/xM2aVX3PsZNgYmB8+uoFHi+S5BHbjmW2HDWeeyTG7LbstGegwNmPjHDxaDM4wsEnzNhxm3HAgLe225DZmoBYeNmmCWnj+HLbfcP5ZWrHktnpitbAdTtxwI/kY48dthwlrkTzD2A70S3ryzBuPD0szbjvOw8ZMwC98Z5iPAUPM2rbvfGLjx5/bqu352ZsfPsanhYGBsQ1INIOZzDxgEq9yMADFXR1E9w/CqkfBKBgFo2AEAgCl+0oYCwy5XQAAAABJRU5ErkJggg==","orcid":"","institution":"Andhra University","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"P.","middleName":"Balarama Swamy","lastName":"Yadav","suffix":""},{"id":186698735,"identity":"9f172040-cc94-476f-b652-760d49830060","order_by":2,"name":"P Balarama Swamy Yadav","email":"","orcid":"","institution":"Andhra University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"P","middleName":"Balarama Swamy","lastName":"Yadav","suffix":""}],"badges":[],"createdAt":"2023-03-26 10:14:21","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-2737719/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-2737719/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s12011-023-03761-6","type":"published","date":"2023-07-10T01:08:34+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":34926511,"identity":"2291c752-b1c6-47b4-8c66-0fd6452a4fb5","added_by":"auto","created_at":"2023-03-28 14:28:42","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":877012,"visible":true,"origin":"","legend":"\u003cp\u003eSTUDY AREA (Source-Google)\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-2737719/v1/8a483df63cbf55f22236a279.png"},{"id":34926510,"identity":"67cd2f08-1811-43c4-abae-038af49c906f","added_by":"auto","created_at":"2023-03-28 14:28:42","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":222238,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003ea.\u003c/strong\u003e Comparison of Ba levels with WHO/FAO permissible values in all samples in three locations.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eb.\u003c/strong\u003e Comparison of Pb levels with WHO/FAO permissible values in all samples in three locations.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;P.V. = Permissible Values; L\u003csub\u003e1= \u003c/sub\u003eKusumapuram; L\u003csub\u003e2= \u003c/sub\u003eVaraka; L\u003csub\u003e3= \u003c/sub\u003eBorivanka.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-2737719/v1/3870029543ca7c3d8fb616ea.png"},{"id":41737245,"identity":"ec5c89ac-cbd2-4bf4-b9f5-c22963563b4b","added_by":"auto","created_at":"2023-08-18 04:18:35","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":769234,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-2737719/v1/2b10b96b-8a49-4739-9a08-c8d86550955e.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Trace Elemental and Heavy Metal levels in frequently consumed local Vegetables of three Chronic Kidney Disease prevalent Villages","fulltext":[{"header":"Introduction","content":"\u003cp\u003eUddanam is a region covering few parts of the seven Revenue Mandals or Tahasils (Ichhapuram, Kanchiri, Kaviti, Mandatha, Paratha, Sompeta and Vajrapkotlu) in the Srikakulam district of Andhra Pradesh, India.\u0026nbsp;The area is a lush green environment with rich Coconut and Cashew nut orchards along the Coastline of the Bay of Bengal. However, it is known the region has a high incidence of CKD. Ravi raju et al [6] reported 15.9% CKD in their pilot study and 18.23% in their final study of age group above 18 years in Uddanam area, which is high incidence of CKD. Thousands of people are still suffering from this disease; till now, the reason(s) behind the cases of kidney failure are unknown and many investigations are being conducted to explore the cause(s) of this CKD, Which is specific to this area. Worldwide, 697.5 million people suffering from chronic kidney disease, of which 115.1 million are Indians [7].\u003c/p\u003e\n\u003cp\u003eAccording to the World Health Organization (WHO), Uddanam is one of three regions in the world with a high CKD incidence of 18.23% [6], the other two in Sri Lanka and Nicaragua. In 2015, it is estimated that over 4,500 people have died in the Uddanam area over the past decade, with about 34,000 suffering from kidney disease. Nearly every family in the region has at least one person diagnosed with the disease. These cases were first recognized in his early 1990s, and the phenomenon was discussed at the International Nephrology Conference in Hong Kong (2013) and named Uddanam nephropathy [8]. In 2016, the Indian Council of Medical Research partnered with Harvard University, Baba Atomic Research Center (Mumbai), Health University (Andhra Pradesh) and Andhra Medical College (Vishakapatnam) to study the causes of CKD.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eFew studies have been done on the analysis of Trace Elements in edible plant parts from the Uddanam area, such as rice and finger millet using the EDXRF technique, indicates that Cu and Zn Elements are in high concentration in both the samples and Phosphorus is more in Rice and Se is more in Finger millet [12, 13]. Studies on analysis of Trace Elements of soil and water samples have been carried out [9-11], and showed that the water and soil in the villages of Uddanam contain high concentration of minerals of Ferrus, Manganese, Copper and Zinc [11]. Khandare et al [14] identified excessive concentrations of Li, Si and Sr in the groundwater of Uchapally village of Nellore district, Andhra Pradesh, India; which is also a CKD prevalent village, indicating presence of some Elements and Heavy Metals in high concentration.\u003c/p\u003e\n\u003cp\u003eStudies on Trace Elemental analysis of plant edible parts are few viz. Benerjee et al (2010) noticed excess Lead content in all fruit and vegetable samples collected from the districts of West Bengal [15], Mamatha et al (2014) identified excessive levels of Cadmium and Lead in Leafy vegetables used by local people in Pulivendula, Kadaba district of Andhra Pradesh [16]. Swapnapriya et al (2014) identified high levels of Copper, Chromium, Iron and Lead in leaves of Sorrel (\u003cem\u003eRumex\u003c/em\u003e sps.) in the Musi River basin of Hyderabad [17]. Swapna Gupta et al (2013) found high levels of Chromium, Zinc, Copper and Lead in leafy Vegetables collected in the city of Raipur [18]. So far, many studies indicate that the water and soil of the Uddanam region contain Trace Elements in high concentration than permissible values. There are no attempts on Trace Elemental analysis in all the local plant edible parts that are frequently consumed by the people of this region. Thus, the present study is taken up.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cp\u003e\u003cstrong\u003eStudy area\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eGeographically, Uddanam is located at 19.0167 degrees north latitude and 84.6833 degrees east longitude, with an average elevation of 41 meters above sea level. It consists of 7 revenue Mandals (Ichapuram, Kanchili, Kaviti, Mandasa, Palasa, Sompeta, and Vajrapukotturu). We have chosen Kaviti revenue Mandal because of high prevalence of CKD. We selected three revenue villages out of 21 revenue Villages in Kaviti Mandal based on the patients number and data obtained from the Community Hospital Sompeta. These three villages are Kusmapuram (L1), Baraka (L2) and Bolivanka (L3) (Figure 1-A, B, C, D). \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSample collection and Preparation\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe identified six locally cultivated vegetables for Trace Elemental and Heavy Metal analysis (Table 1), which are grown in their Coconut orchards as inter crops and are frequently consumed. In which 2 are belong to the family Solanaceae, and one each from Caricaceae, Cucurbitaceae, Moringaceae and Musaceae (Table 1). \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eTable 1. Frequently consumed local\u0026nbsp;Vegetables used in the study\u003c/p\u003e\n\u003ctable align=\"\" border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"638\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"14.733542319749215%\"\u003e\n \u003cp\u003eSample No.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"28.369905956112852%\"\u003e\n \u003cp\u003eBotanical Name\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.4576802507837%\"\u003e\n \u003cp\u003eFamily\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"17.55485893416928%\"\u003e\n \u003cp\u003eEnglish name\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"22.884012539184955%\"\u003e\n \u003cp\u003eVernacular name\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"14.733542319749215%\"\u003e\n \u003cp\u003eS\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eS\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eS\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eS\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eS\u003csub\u003e5\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eS\u003csub\u003e6\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"28.369905956112852%\"\u003e\n \u003cp\u003e\u003cem\u003eCapsicum frutescence\u0026nbsp;\u003c/em\u003eL.\u003c/p\u003e\n \u003cp\u003e\u003cem\u003eCarica papaya\u0026nbsp;\u003c/em\u003eL.\u003c/p\u003e\n \u003cp\u003e\u003cem\u003eMomordica charantia\u0026nbsp;\u003c/em\u003eL.\u003c/p\u003e\n \u003cp\u003e\u003cem\u003eMoringa\u0026nbsp;\u003c/em\u003e\u003cem\u003eoliefera\u0026nbsp;\u003c/em\u003eLam.\u003c/p\u003e\n \u003cp\u003e\u003cem\u003eMusa sapientum\u0026nbsp;\u003c/em\u003eL.\u003c/p\u003e\n \u003cp\u003e\u003cem\u003eSolanum melongena\u0026nbsp;\u003c/em\u003eL.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.4576802507837%\"\u003e\n \u003cp\u003eSolanaceae\u003c/p\u003e\n \u003cp\u003eCaricaceae\u003c/p\u003e\n \u003cp\u003eCucurbitaceae\u003c/p\u003e\n \u003cp\u003eMoringaceae\u003c/p\u003e\n \u003cp\u003eMusaceae\u003c/p\u003e\n \u003cp\u003eSolanaceae\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"17.55485893416928%\"\u003e\n \u003cp\u003eChilli\u003c/p\u003e\n \u003cp\u003ePapaya\u003c/p\u003e\n \u003cp\u003eBitter gourd\u003c/p\u003e\n \u003cp\u003eDrumsticks\u003c/p\u003e\n \u003cp\u003ePlantain\u003c/p\u003e\n \u003cp\u003eBrinjal\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"22.884012539184955%\"\u003e\n \u003cp\u003ePachchi mirapa kaya\u003c/p\u003e\n \u003cp\u003eBoppayi kaya\u003c/p\u003e\n \u003cp\u003eKakara kaya\u003c/p\u003e\n \u003cp\u003eMunaga Kada\u003c/p\u003e\n \u003cp\u003eArati kaya\u003c/p\u003e\n \u003cp\u003eVanga kaya\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe fresh\u0026nbsp;samples of the selected vegetables collected from the fields directly,\u0026nbsp;from the three Villages are kept in zip locked bags and are brought to the laboratory. The samples are washed twice with distilled water and then subjected to Acid Digestion process, i.e. fast and accurate Wet Digestion method [19] is followed for the digestion of samples to digest all organic matter in the samples and placed in\u0026nbsp;ICP-MS to obtain the Trace Elemental levels.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eProcedure\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe used the ICP-MS instrument (Agilent Technologies- Model 7700) available in Centre for Studies on Bay of Bengal, Andhra University, Visakhapatnam.\u0026nbsp;The Sample preparation for ICP-MS analysis followed here is based on the oxidation and solubility of organic matter in plants when H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e (33%) is added directly to the concentrated HNO\u003csub\u003e3\u003c/sub\u003e and heated for a short time.\u0026nbsp;The instrument was switched on and is first run with the Standard Reference Materials of 0 PPB, 0.5 PPB, 1 PPB, 5 PPB, 10 PPB, 25 PPB, 50 PPB, and 100 PPB then ran the samples and\u0026nbsp;obtained the levels of 24 Elements (Li, Be, Al, Sc, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Ga, As, Se, Rb, Sr, Ag, Cd, Cs, Ba, TI, Rb, and U).\u003c/p\u003e\n\u003cp\u003eICP-MS instrument (Agilent Technologies - Model 7700) available at the Center for studies on Bay of Bengal, Andhra University, Visakhapatnam was used for analysis. The following sample preparation for ICP-MS analysis is based on the oxidation and solubility of organic matter in plants when H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e (33%) is added directly to concentrated HNO\u003csub\u003e3\u0026nbsp;\u003c/sub\u003eand heated for short time. First we turn on the instrument and run with standard reference materials of 0 PPB, 0.5 PPB, 1 PPB, 5 PPB, 10 PPB, 25 PPB, 50 PPB, and 100 PPB, then followed by samples to find 24 Elements. (Li, Be, Al, Sc, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Ga, As, Se, Rb, Sr, Ag, Cd, Cs, Ba, TI, Rb and U).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eWet Digestion\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eModified HNO\u003csub\u003e3\u003c/sub\u003e/H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e wet digestion method (Pequerul. et al. 1993) [19] was followed. 2 gr. sample is taken, kept in a Teflon beaker, mixed with 8 ml HNO\u003csub\u003e3\u003c/sub\u003e and left overnight. Heat the solution on a hot plate at 120-125 \u0026deg;C for 1 hour and add 4 mL H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e (33%) once or twice until the organic matter completely digested. Then we added H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u0026nbsp;\u003c/sub\u003euntil the solution is clear or colorless (indicating complete digestion of organic matter). Place the residue in a dry place at 80\u0026deg; C., shake well, cool to room temperature and filter through Whattman filter paper. The solution is then diluted to a known volume (50 mL) with Milli-Q water. These digested solutions are stored in high-density polyethylene bottles (100 mL) at room temperature for further use.\u0026nbsp;\u003c/p\u003e"},{"header":"Results and Discussion","content":"\u003cp\u003eSamples from three neighboring villages are analyzed for Trace Elements/ Heavy Metals using ICP-MS instrument. The values obtained are compared with the limits recommended by FAO/WHO (1990, 1995, 1997, 2006, 2008, 2009, 2010, 2011, 2012, 2013, 2014, 2015, 2016, 2017, 2018, and 2019) and the Food Safety and Standards (Contaminants, Toxins and Residues) Regulations (2011) [1-5]. A total of 24 Elemental levels in 18 vegetable samples from 3 different locations (L\u003csub\u003e1\u003c/sub\u003e\u0026ndash;L\u003csub\u003e3\u003c/sub\u003e) were obtained and are shown in Table 2. Out of the 18 samples tested, all tested samples were found high concentrations of Ba and 11 samples had high concentrations of Pb. Fe and Ag are high in each high one sample and the remaining.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2.\u003c/strong\u003e Trace Elemental and Heavy Metal levels in vegetable samples by ICP-MS analysis.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width: 19.4161%;\" valign=\"top\" width=\"9.972677595628415%\"\u003e\n \u003cp\u003eElement\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 4.299%;\" valign=\"top\" width=\"7.6502732240437155%\"\u003e\n \u003cp\u003eLi\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 2.7817%;\" valign=\"top\" width=\"5.8743169398907105%\"\u003e\n \u003cp\u003eBe\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 4.8047%;\" valign=\"top\" width=\"7.6502732240437155%\"\u003e\n \u003cp\u003eAl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 4.8047%;\" valign=\"top\" width=\"7.6502732240437155%\"\u003e\n \u003cp\u003eSc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 4.8047%;\" valign=\"top\" width=\"7.6502732240437155%\"\u003e\n \u003cp\u003eV\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 4.8047%;\" valign=\"top\" width=\"7.6502732240437155%\"\u003e\n \u003cp\u003eCr\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 4.8047%;\" valign=\"top\" width=\"7.6502732240437155%\"\u003e\n \u003cp\u003eMn\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5.8162%;\" valign=\"top\" width=\"7.6502732240437155%\"\u003e\n \u003cp\u003eFe\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 4.8047%;\" valign=\"top\" width=\"7.6502732240437155%\"\u003e\n \u003cp\u003eCo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 4.8047%;\" valign=\"top\" width=\"7.6502732240437155%\"\u003e\n \u003cp\u003eNi\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5.8162%;\" valign=\"top\" width=\"7.6502732240437155%\"\u003e\n \u003cp\u003eCu\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 4.8047%;\" valign=\"top\" width=\"7.6502732240437155%\"\u003e\n \u003cp\u003eZn\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width: 34.6446%;\" valign=\"top\" width=\"8.891928864569083%\"\u003e\n \u003cp\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; P.V.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.618331053351573%\"\u003e\n \u003cp\u003e5.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 2.7817%;\" valign=\"top\" width=\"5.882352941176471%\"\u003e\n \u003cp\u003e8.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 4.8047%;\" valign=\"top\" width=\"7.660738714090288%\"\u003e\n \u003cp\u003e60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 4.8047%;\" valign=\"top\" width=\"7.660738714090288%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 4.8047%;\" valign=\"top\" width=\"7.660738714090288%\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 4.8047%;\" valign=\"top\" width=\"7.660738714090288%\"\u003e\n \u003cp\u003e2.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 4.8047%;\" valign=\"top\" width=\"7.660738714090288%\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5.8162%;\" valign=\"top\" width=\"7.660738714090288%\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 4.8047%;\" valign=\"top\" width=\"7.660738714090288%\"\u003e\n \u003cp\u003e0.14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 4.8047%;\" valign=\"top\" width=\"7.660738714090288%\"\u003e\n \u003cp\u003e5.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5.8162%;\" valign=\"top\" width=\"7.660738714090288%\"\u003e\n \u003cp\u003e3.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 4.8047%;\" valign=\"top\" width=\"7.660738714090288%\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 32.3686%;\" valign=\"top\" width=\"4.098360655737705%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003eS\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"6\" style=\"width: 2.2759%;\" valign=\"top\" width=\"4.918032786885246%\"\u003e\n \u003cp\u003eL\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eL\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eL\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eL\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eL\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eL\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eL\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eL\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n 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\u003cp\u003e0.016\u003c/p\u003e\n \u003cp\u003e0.049\u003c/p\u003e\n \u003cp\u003e0.029\u003c/p\u003e\n \u003cp\u003e0.029\u003c/p\u003e\n \u003cp\u003e0.018\u003c/p\u003e\n \u003cp\u003e0.014\u003c/p\u003e\n \u003cp\u003e0.021\u003c/p\u003e\n \u003cp\u003e0.014\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"6\" style=\"width: 4.8047%;\" valign=\"top\" width=\"7.6502732240437155%\"\u003e\n \u003cp\u003e0.535\u003c/p\u003e\n \u003cp\u003e1.139\u003c/p\u003e\n \u003cp\u003e0.454\u003c/p\u003e\n \u003cp\u003e0.613\u003c/p\u003e\n \u003cp\u003e0.439\u003c/p\u003e\n \u003cp\u003e0.65\u003c/p\u003e\n \u003cp\u003e0.509\u003c/p\u003e\n \u003cp\u003e0.392\u003c/p\u003e\n \u003cp\u003e0.542\u003c/p\u003e\n \u003cp\u003e0.504\u003c/p\u003e\n \u003cp\u003e0.483\u003c/p\u003e\n \u003cp\u003e0.878\u003c/p\u003e\n \u003cp\u003e0.482\u003c/p\u003e\n \u003cp\u003e0.419\u003c/p\u003e\n \u003cp\u003e0.525\u003c/p\u003e\n \u003cp\u003e0.498\u003c/p\u003e\n \u003cp\u003e0.557\u003c/p\u003e\n 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\u003cp\u003e0.407\u003c/p\u003e\n \u003cp\u003e0.568\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"6\" style=\"width: 5.8162%;\" valign=\"top\" width=\"7.6502732240437155%\"\u003e\n \u003cp\u003e0.644\u003c/p\u003e\n \u003cp\u003e1.227\u003c/p\u003e\n \u003cp\u003e0.895\u003c/p\u003e\n \u003cp\u003e0.02\u003c/p\u003e\n \u003cp\u003e0.234\u003c/p\u003e\n \u003cp\u003e0.099\u003c/p\u003e\n \u003cp\u003e0.45\u003c/p\u003e\n \u003cp\u003e0.135\u003c/p\u003e\n \u003cp\u003e0.031\u003c/p\u003e\n \u003cp\u003e0.082\u003c/p\u003e\n \u003cp\u003e0.132\u003c/p\u003e\n \u003cp\u003e0.214\u003c/p\u003e\n \u003cp\u003e0.339\u003c/p\u003e\n \u003cp\u003e0.045\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0.0008\u003c/p\u003e\n \u003cp\u003e0.973\u003c/p\u003e\n \u003cp\u003e0.196\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"6\" style=\"width: 4.8047%;\" valign=\"top\" width=\"7.6502732240437155%\"\u003e\n \u003cp\u003e10.19\u003c/p\u003e\n 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\u003cp\u003eS\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 32.3686%;\" valign=\"top\" width=\"100%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003eS\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 32.3686%;\" valign=\"top\" width=\"100%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003eS\u003csub\u003e5\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 32.3686%;\" valign=\"top\" width=\"100%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003eS\u003csub\u003e6\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 32.3686%;\" width=\"4.098360655737705%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 2.2759%;\" width=\"4.918032786885246%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 4.299%;\" width=\"7.6502732240437155%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 2.7817%;\" width=\"5.8743169398907105%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 4.8047%;\" width=\"7.6502732240437155%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 4.8047%;\" width=\"7.6502732240437155%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 4.8047%;\" width=\"7.6502732240437155%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 4.8047%;\" width=\"7.6502732240437155%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 4.8047%;\" width=\"7.6502732240437155%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 5.8162%;\" width=\"7.6502732240437155%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 4.8047%;\" width=\"7.6502732240437155%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 4.8047%;\" width=\"7.6502732240437155%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 5.8162%;\" width=\"7.6502732240437155%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 4.8047%;\" width=\"7.6502732240437155%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003econtd\u0026hellip;\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.556786703601109%\"\u003e\n \u003cp\u003eElement\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.756232686980609%\"\u003e\n \u003cp\u003eGa\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.756232686980609%\"\u003e\n \u003cp\u003eAs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.756232686980609%\"\u003e\n \u003cp\u003eSe\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.756232686980609%\"\u003e\n \u003cp\u003eRb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.756232686980609%\"\u003e\n \u003cp\u003eSr\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.756232686980609%\"\u003e\n \u003cp\u003eAg\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.756232686980609%\"\u003e\n \u003cp\u003eCd\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.756232686980609%\"\u003e\n \u003cp\u003eCs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.756232686980609%\"\u003e\n \u003cp\u003eBa\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.401662049861496%\"\u003e\n \u003cp\u003eTl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.617728531855955%\"\u003e\n \u003cp\u003ePb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.617728531855955%\"\u003e\n \u003cp\u003eU\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.556786703601109%\"\u003e\n \u003cp\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; P.V.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.756232686980609%\"\u003e\n \u003cp\u003e3.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.756232686980609%\"\u003e\n \u003cp\u003e0.18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.756232686980609%\"\u003e\n \u003cp\u003e1.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.756232686980609%\"\u003e\n \u003cp\u003e5.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.756232686980609%\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.756232686980609%\"\u003e\n \u003cp\u003e0.175\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.756232686980609%\"\u003e\n \u003cp\u003e1.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.756232686980609%\"\u003e\n \u003cp\u003e90\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.756232686980609%\"\u003e\n \u003cp\u003e12.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.401662049861496%\"\u003e\n \u003cp\u003e1.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.617728531855955%\"\u003e\n \u003cp\u003e0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.617728531855955%\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"4.149377593360996%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003eS\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"6\" valign=\"top\" width=\"5.532503457814661%\"\u003e\n \u003cp\u003eL\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eL\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eL\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eL\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eL\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eL\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eL\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eL\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eL\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eL\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eL\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eL\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eL\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eL\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eL\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eL\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eL\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003eL\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"6\" valign=\"top\" width=\"7.745504840940526%\"\u003e\n \u003cp\u003e0.003\u003c/p\u003e\n \u003cp\u003e0.013\u003c/p\u003e\n \u003cp\u003e0.005\u003c/p\u003e\n \u003cp\u003e0.001\u003c/p\u003e\n \u003cp\u003e0.002\u003c/p\u003e\n \u003cp\u003e0.014\u003c/p\u003e\n \u003cp\u003e0.005\u003c/p\u003e\n \u003cp\u003e0.002\u003c/p\u003e\n \u003cp\u003e0.003\u003c/p\u003e\n \u003cp\u003e0.002\u003c/p\u003e\n \u003cp\u003e0.003\u003c/p\u003e\n \u003cp\u003e0.007\u003c/p\u003e\n \u003cp\u003e0.0060.002\u003c/p\u003e\n \u003cp\u003e0.003\u003c/p\u003e\n \u003cp\u003e0.002\u003c/p\u003e\n \u003cp\u003e0.002\u003c/p\u003e\n \u003cp\u003e0.003\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"6\" valign=\"top\" width=\"7.745504840940526%\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003cp\u003e0.031\u003c/p\u003e\n \u003cp\u003e0.008\u003c/p\u003e\n \u003cp\u003e0.005\u003c/p\u003e\n \u003cp\u003e0.002\u003c/p\u003e\n \u003cp\u003e0.011\u003c/p\u003e\n \u003cp\u003e0.005\u003c/p\u003e\n \u003cp\u003e0.003\u003c/p\u003e\n \u003cp\u003e0.004\u003c/p\u003e\n \u003cp\u003e0.005\u003c/p\u003e\n \u003cp\u003e0.004\u003c/p\u003e\n \u003cp\u003e0.013\u003c/p\u003e\n \u003cp\u003e0.009\u003c/p\u003e\n \u003cp\u003e0.005\u003c/p\u003e\n \u003cp\u003e0.005\u003c/p\u003e\n \u003cp\u003e0.007\u003c/p\u003e\n \u003cp\u003e0.015\u003c/p\u003e\n \u003cp\u003e0.005\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"6\" valign=\"top\" width=\"7.745504840940526%\"\u003e\n \u003cp\u003e0.006\u003c/p\u003e\n \u003cp\u003e0.133\u003c/p\u003e\n \u003cp\u003e0.039\u003c/p\u003e\n \u003cp\u003e0.036\u003c/p\u003e\n \u003cp\u003e0.037\u003c/p\u003e\n \u003cp\u003e0.039\u003c/p\u003e\n \u003cp\u003e0.028\u003c/p\u003e\n \u003cp\u003e0.007\u003c/p\u003e\n \u003cp\u003e0.037\u003c/p\u003e\n \u003cp\u003e0.017\u003c/p\u003e\n \u003cp\u003e0.005\u003c/p\u003e\n \u003cp\u003e0.043\u003c/p\u003e\n \u003cp\u003e0.03\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0.015\u003c/p\u003e\n \u003cp\u003e0.007\u003c/p\u003e\n \u003cp\u003e0.021\u003c/p\u003e\n \u003cp\u003e0.027\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"6\" valign=\"top\" width=\"7.745504840940526%\"\u003e\n \u003cp\u003e0.587\u003c/p\u003e\n \u003cp\u003e1.107\u003c/p\u003e\n \u003cp\u003e0.587\u003c/p\u003e\n \u003cp\u003e0.735\u003c/p\u003e\n \u003cp\u003e0.823\u003c/p\u003e\n \u003cp\u003e0.92\u003c/p\u003e\n \u003cp\u003e0.54\u003c/p\u003e\n \u003cp\u003e0.489\u003c/p\u003e\n \u003cp\u003e0.879\u003c/p\u003e\n \u003cp\u003e0.531\u003c/p\u003e\n \u003cp\u003e0.949\u003c/p\u003e\n \u003cp\u003e1.158\u003c/p\u003e\n \u003cp\u003e1.92\u003c/p\u003e\n \u003cp\u003e0.463\u003c/p\u003e\n \u003cp\u003e0.266\u003c/p\u003e\n \u003cp\u003e0.504\u003c/p\u003e\n \u003cp\u003e2.356\u003c/p\u003e\n \u003cp\u003e1.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"6\" valign=\"top\" width=\"7.745504840940526%\"\u003e\n \u003cp\u003e1.395\u003c/p\u003e\n \u003cp\u003e2.832\u003c/p\u003e\n \u003cp\u003e0.887\u003c/p\u003e\n \u003cp\u003e0.574\u003c/p\u003e\n \u003cp\u003e0.63\u003c/p\u003e\n \u003cp\u003e0.552\u003c/p\u003e\n \u003cp\u003e0.605\u003c/p\u003e\n \u003cp\u003e0.72\u003c/p\u003e\n \u003cp\u003e0.617\u003c/p\u003e\n \u003cp\u003e1.137\u003c/p\u003e\n \u003cp\u003e0.842\u003c/p\u003e\n \u003cp\u003e1.143\u003c/p\u003e\n \u003cp\u003e1.847\u003c/p\u003e\n \u003cp\u003e0.882\u003c/p\u003e\n \u003cp\u003e0.705\u003c/p\u003e\n \u003cp\u003e0.698\u003c/p\u003e\n \u003cp\u003e3.258\u003c/p\u003e\n \u003cp\u003e0.928\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"6\" valign=\"top\" width=\"7.745504840940526%\"\u003e\n \u003cp\u003e0.048\u003c/p\u003e\n \u003cp\u003e0.124\u003c/p\u003e\n \u003cp\u003e0.047\u003c/p\u003e\n \u003cp\u003e0.057\u003c/p\u003e\n \u003cp\u003e0.122\u003c/p\u003e\n \u003cp\u003e0.027\u003c/p\u003e\n \u003cp\u003e0.026\u003c/p\u003e\n \u003cp\u003e0.033\u003c/p\u003e\n \u003cp\u003e0.026\u003c/p\u003e\n \u003cp\u003e0.035\u003c/p\u003e\n \u003cp\u003e0.388*\u003c/p\u003e\n \u003cp\u003e0.172\u003c/p\u003e\n \u003cp\u003e0.048\u003c/p\u003e\n \u003cp\u003e0.056\u003c/p\u003e\n \u003cp\u003e0.087\u003c/p\u003e\n \u003cp\u003e0.104\u003c/p\u003e\n \u003cp\u003e0.04\u003c/p\u003e\n \u003cp\u003e0.066\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"6\" valign=\"top\" width=\"7.745504840940526%\"\u003e\n \u003cp\u003e0.006\u003c/p\u003e\n \u003cp\u003e0.013\u003c/p\u003e\n \u003cp\u003e0.026\u003c/p\u003e\n \u003cp\u003e0.009\u003c/p\u003e\n \u003cp\u003e0.011\u003c/p\u003e\n \u003cp\u003e0.032\u003c/p\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003cp\u003e0.011\u003c/p\u003e\n \u003cp\u003e0.004\u003c/p\u003e\n \u003cp\u003e0.008\u003c/p\u003e\n \u003cp\u003e0.009\u003c/p\u003e\n \u003cp\u003e0.015\u003c/p\u003e\n \u003cp\u003e0.018\u003c/p\u003e\n \u003cp\u003e0.005\u003c/p\u003e\n \u003cp\u003e0.019\u003c/p\u003e\n \u003cp\u003e0.007\u003c/p\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003cp\u003e0.004\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"6\" valign=\"top\" width=\"7.745504840940526%\"\u003e\n \u003cp\u003e0.0005\u003c/p\u003e\n \u003cp\u003e0.002\u003c/p\u003e\n \u003cp\u003e0.0001\u003c/p\u003e\n \u003cp\u003e0.0001\u003c/p\u003e\n \u003cp\u003e0.0004\u003c/p\u003e\n \u003cp\u003e0.0003\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0.0001\u003c/p\u003e\n \u003cp\u003e0.0003\u003c/p\u003e\n \u003cp\u003e0.0003\u003c/p\u003e\n \u003cp\u003e0.001\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"6\" valign=\"top\" width=\"7.745504840940526%\"\u003e\n \u003cp\u003e36.67*\u003c/p\u003e\n \u003cp\u003e43.92*\u003c/p\u003e\n \u003cp\u003e27.65*\u003c/p\u003e\n \u003cp\u003e30.12*\u003c/p\u003e\n \u003cp\u003e33.73*\u003c/p\u003e\n \u003cp\u003e21.80*\u003c/p\u003e\n \u003cp\u003e31.60*\u003c/p\u003e\n \u003cp\u003e32.29*\u003c/p\u003e\n \u003cp\u003e26.95*\u003c/p\u003e\n \u003cp\u003e32.44*\u003c/p\u003e\n \u003cp\u003e29.23*\u003c/p\u003e\n \u003cp\u003e23.82*\u003c/p\u003e\n \u003cp\u003e42.22*\u003c/p\u003e\n \u003cp\u003e33.34*\u003c/p\u003e\n \u003cp\u003e33.85*\u003c/p\u003e\n \u003cp\u003e30.07*\u003c/p\u003e\n \u003cp\u003e33.64*\u003c/p\u003e\n \u003cp\u003e26.89*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"6\" valign=\"top\" width=\"5.394190871369295%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"6\" valign=\"top\" width=\"7.607192254495159%\"\u003e\n \u003cp\u003e0.372*\u003c/p\u003e\n \u003cp\u003e0.798*\u003c/p\u003e\n \u003cp\u003e0.972*\u003c/p\u003e\n \u003cp\u003e0.2\u003c/p\u003e\n \u003cp\u003e0.28\u003c/p\u003e\n \u003cp\u003e0.327*\u003c/p\u003e\n \u003cp\u003e0.415*\u003c/p\u003e\n \u003cp\u003e0.511*\u003c/p\u003e\n \u003cp\u003e0.252\u003c/p\u003e\n \u003cp\u003e0.254\u003c/p\u003e\n \u003cp\u003e0.214\u003c/p\u003e\n \u003cp\u003e0.416*\u003c/p\u003e\n \u003cp\u003e0.353*\u003c/p\u003e\n \u003cp\u003e0.304*\u003c/p\u003e\n \u003cp\u003e0.460*\u003c/p\u003e\n \u003cp\u003e0.177\u003c/p\u003e\n \u003cp\u003e0.316*\u003c/p\u003e\n \u003cp\u003e0.299\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"6\" valign=\"top\" width=\"7.607192254495159%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003cp\u003e0.0008\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0.0001\u003c/p\u003e\n \u003cp\u003e0.003\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"100%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003eS\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"100%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003eS\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"100%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003eS\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"100%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003eS\u003csub\u003e5\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"100%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003eS\u003csub\u003e6\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eS\u003csub\u003e1\u003c/sub\u003e-S\u003csub\u003e6\u003c/sub\u003e: Samples 1-6; P.V. - Permissible Values (mg/kg or ppm) as per WHO/FAO.\u003c/p\u003e\n\u003cp\u003eL\u003csub\u003e1\u003c/sub\u003e (Location 1) = Kusumapuram , \u0026nbsp; L\u003csub\u003e2\u003c/sub\u003e ( Location 2)= Varaka, \u0026nbsp; L\u003csub\u003e3\u003c/sub\u003e (Location 3) = Borivanka\u003c/p\u003e\n\u003cp\u003e* = Trace Elements and Heavy Metals in more concentrations than the WHO/FAO standards that are harmful to kidney;\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe results reveal that two Elements i.e. Ba \u0026nbsp;in high concentration in \u003cem\u003eCapsicum frutescence\u0026nbsp;\u003c/em\u003eraw fruit \u0026nbsp;(36.67- L\u003csub\u003e1\u003c/sub\u003e, 43.92- L\u003csub\u003e2\u003c/sub\u003e and 27.65- L\u003csub\u003e3\u003c/sub\u003e mg/kg) and Pb (0.372- L\u003csub\u003e1\u003c/sub\u003e, 0.798- L\u003csub\u003e2\u003c/sub\u003e and 0.972- L\u003csub\u003e3\u003c/sub\u003e). In \u003cem\u003eCarica papaya\u003c/em\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003eraw fruit three Elements i.e. Fe in high concentrations (216.3- L\u003csub\u003e3\u003c/sub\u003e mg/kg), Ba (30.12- L\u003csub\u003e1\u003c/sub\u003e, 33.73- L\u003csub\u003e2\u003c/sub\u003e and 21.8- L\u003csub\u003e3\u003c/sub\u003e mg/kg) and pb (0.327- L\u003csub\u003e3\u003c/sub\u003e). In \u003cem\u003eMomordica charantia\u0026nbsp;\u003c/em\u003eraw fruit two Elements i.e. Ba, in high concentrations (31.6- L\u003csub\u003e1\u003c/sub\u003e, 32.29- L\u003csub\u003e2\u003c/sub\u003e and 26.95- L\u003csub\u003e3\u003c/sub\u003e mg/kg) and Pb (0.415- L\u003csub\u003e1\u003c/sub\u003e, 0.511- L\u003csub\u003e2\u003c/sub\u003e). In \u003cem\u003eMoringa\u0026nbsp;\u003c/em\u003e\u003cem\u003eoliefera\u0026nbsp;\u003c/em\u003eraw fruit two Elements are in high i.e. Ba (32.44- L\u003csub\u003e1\u003c/sub\u003e, 29.23- L\u003csub\u003e2\u003c/sub\u003e and 23.82- L\u003csub\u003e3\u003c/sub\u003e mg/kg), Pb (0.416- L\u003csub\u003e3\u003c/sub\u003e) and Ag (L\u003csub\u003e2\u003c/sub\u003e-0.388). In \u003cem\u003eMusa sapientum\u0026nbsp;\u003c/em\u003e two Elements i.e. Ba in high concentration (42.22- L\u003csub\u003e1\u003c/sub\u003e, 33.34- L\u003csub\u003e2\u003c/sub\u003e and 33.85- L\u003csub\u003e3\u003c/sub\u003e mg/kg) and Pb (0.353- L\u003csub\u003e1\u003c/sub\u003e, 0.304- L\u003csub\u003e2\u003c/sub\u003e and 0.460- L\u003csub\u003e3\u003c/sub\u003e). In Brinjal two Elements i.e. Ba (30.07- L\u003csub\u003e1\u003c/sub\u003e, 33.64- L\u003csub\u003e2\u003c/sub\u003e and 26.89- L\u003csub\u003e3\u003c/sub\u003e mg/kg) and Pb (0.316- L\u003csub\u003e2\u003c/sub\u003e) are \u0026nbsp;in high concentration (Table 2).\u003c/p\u003e\n\u003cp\u003eIt was found that the highest concentration of Fe is present among all the 18 tested samples in \u0026nbsp;\u003cem\u003eCarica papaya\u003c/em\u003e of Borivanka, highest concentration of Ba observed in \u003cem\u003eCapsicum frutescence\u003c/em\u003e of Varaka followed by \u003cem\u003eMusa sapientum\u0026nbsp;\u003c/em\u003eL. of Kusumapuram, and then highest concentration of Pb seen \u0026nbsp;in \u0026nbsp;Chillies of Borivanka followed by \u003cem\u003eCapsicum frutescence\u003c/em\u003e of Varaka (Table 2). Highest Pb concentration was found in Chilli, and lowest concentration is in Brinjal. Average highest concentration of Elements identified in Borivanka village followed by Varaka and Kusumapuram (Table 2), and it reflects with the CKD patient intensity.\u0026nbsp;\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eIn this study we analyzed 18 vegetable samples in 3 locations (villages) six vegetables in each for Trace Elemental levels. In all the three villages Ba and Pb are in high concentrations. Fe and Ag are in high concentration in one sample each, and their excessive consumption may influence the functioning of kidneys [1-6]. On an average Lead concentrations are 1.28 times higher and Ba is 2.51 times higher than the permissible values in the tested locations (Table 2). \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThese excess concentrations of Elements in the studied samples may be absorbed from soil and ground water (naturally present in some locations) or may be due to usage of Chemical pesticides and fertilizers. The study reveals the extent of Trace Elements in local vegetables will enhance the risk of disease intensity, by consuming these vegetables frequently and further research is to be needed in this area to find out the cumulative or additive affects of soil, ground water and local plant edibles and or a single exact reason for CKD in Uddanam area. Among the three locations average Ba and Pb concentration are more (Fig 2a, 2b) in Borivanka (L\u003csub\u003e3\u003c/sub\u003e) followed by Varaka (L\u003csub\u003e2\u003c/sub\u003e) and Kusumapuram (L\u003csub\u003e1\u003c/sub\u003e), and it reflects with the CKD patient intensity.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe studies on soil, ground water and all local plant edibles will help in understanding the Serological, Urological, nutritional and Genetic studies of the CKD. The present study opens a new dimension to understand the role of Trace Elements behind the CKD by analyzing the Trace Elemental analysis of all local frequently consumed plant edible parts coupled with soil and ground water analysis. The studies [9, 10 and 11] on soil and water in Uddanam area reveal that Fe, Mn, Cu and Zn Metals are in high concentration. Providing safe water to drinking, establishing more dialysis centers and giving pensions to patients affected by CKD are a huge financial burden for governments in many areas where CKD is prevalent. This can be reduced by taking studies on the soil, water and all edible local plants. The present study opens a new dimension to understand the impact of Trace Elements and Heavy Metals on CKD by analyzing the Trace Element and Heavy Metal content of all commonly consumed plant parts of local food crops along with soil and ground water.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eConflict of interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that \u0026nbsp;no conflict of interest. The authors alone are responsible for the content and writing of this paper. There is no any funding for this work.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors are thankful to The Director, Center for Studies on Bay of Bengal (CSBoB), Andhra University, Visakhapatnam, for giving permission to use ICP-MS facility to carry out this work.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eCodex Alimentarius FAO \u0026amp; WHO. General Standard for Contaminants and Toxins in Food and Feed (CXS 193-1995). Adopted in 1995 Revised in 1997, 2006, 2008, 2009 Amended in 2010, 2012, 2013, 2014, 2015, 2016, 2017, 2018 and 2019: 39-50\u003cstrong\u003e\u003cem\u003e \u003c/em\u003e\u003c/strong\u003e\u003cem\u003ehttps//doi.CXS 193-1995\u003c/em\u003e\u003c/li\u003e\n\u003cli\u003eToxicological Profile for Silver. Agency for Toxic Substances and Disease Registry, U.S. Public Health Service. 1990: 8-18\u003cem\u003ehttps://www.atsdr.cdc.gov/toxprofiles/tp146.pdf\u003c/em\u003e\u003c/li\u003e\n\u003cli\u003eProvisional peer reviewed toxicity values for Lithium (CASRN 7439-93-2). Superfund Health Risk Technical Support Center, National Center for Environmental Assessment: Office of Research and Development. U.S. Environmental Protection Agency, Cincinnati, OH, 45268; 2008: 4-7\u003cem\u003ehttps://cfpub.epa.gov/ncea/pprtv/documents/Lithium.pdf\u003c/em\u003e\u003c/li\u003e\n\u003cli\u003eCodex Alimentarius Commission, Joint FAO / WHO Food Standards Programme. Codex Committee on contaminants in Foods. 10\u003csup\u003eth\u003c/sup\u003e Session Rotterdam, The Netherlands 4-8; (Prepared by Japan and The Netherlands). 2016: 6, 8, 13, 17, 88, 142, 145 and 148\u003cem\u003e \u003c/em\u003e\u003cem\u003ehttps//doi.CF/10 INF/1\u003c/em\u003e\u003c/li\u003e\n\u003cli\u003eFood Safety and Standards (contaminants, toxins and residues) regulations, 2011 chapter-I General. 2017 and 2019: 3, 14, 20, 32, 38, 43, 54, 60 and 72\u003cem\u003e \u003c/em\u003e \u003cem\u003ehttps//doi.Version \u0026ndash;II (18.08.2017); \u003c/em\u003eand\u003cem\u003e https//doi.Version \u0026ndash;IV (26.04.2019).\u003c/em\u003e\u003c/li\u003e\n\u003cli\u003eTatapudi RR, Rentala S, Gullipalli P, Komarraju AL, Singh AK, Tatapudi VS, Goru KB, Bhimarasetty DM, Narni H (2018) High Prevalence of CKD of Unknown Etiology in Uddanam, India. Kidney Int. Repo. 4(3): 380-389 doi: 10.1016/j.ekir.2018.10.006. PMID: 30899865; PMCID: PMC6409405.\u003c/li\u003e\n\u003cli\u003ePaul Cockwell, Lori-Ann Fisher (2020) The global burden of Chronic Kidney Disease. The Lancet 395 (10225): 662-664 \u003cem\u003ehttps://doi.org/10.1016/S0140-6736(19)32977-0\u003c/em\u003e.\u003c/li\u003e\n\u003cli\u003ePraveen Gadde, Suresh Sanikommu, Ramesh Manumanthu, Anitha Akkaloori (2020) Uddanam Nephropathy in India: A challenge for Epidemiologists. Bulletin of the World Health Organization 95: 848-849\u003cem\u003ehttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5710086/\u003c/em\u003e\u003c/li\u003e\n\u003cli\u003eSatyanarayana G, Ramadasu P, Padmavathi Devi P, Prasad N.V.B.S.S, Nageswara Rao G (2017) Ground water quality assessment in Uddanam region, Costal Srikakulam, Andhra Pradesh, India. International Journal of Pharmaceuticals and Drug Analysis 4: 116-128 \u003cem\u003ehttps://www.ijpda.com/index.php/journal/article/view/277 \u003c/em\u003e\u003c/li\u003e\n\u003cli\u003eSatyanarayana G, Ramadasu P, Padmavathi Devi P, Prasad N.V.B.S.S, Nageswara Rao G (2017) Ground water quality interpretation in coastal Srikakulam, Andhra Pradesh by principal component analysis. 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International Journal of Science and Research 4: 1062-1066 \u003cem\u003ehttps://www.ijsr.net/get_abstract.php?paper_id=SUB151409\u003c/em\u003e\u003c/li\u003e\n\u003cli\u003eRaju TP, Giridhar Babu N, Srinivasu Ch Ch, Ramanamam V, Ravi Shankar MS, Ram SS, Sudarshan M, Lakshmi Narayana PV, Lakshmana Das N (2016) Trace Elemental analysis of Finger millet samples from Kidney Effected area using EDXRF Technique. International Journal of Scientific Research in Science, Engineering and Technology 2: 174-178 \u003cem\u003ehttps://ijsrset.com/IJSRSET162132\u003c/em\u003e \u003c/li\u003e\n\u003cli\u003eKhandare AL, Reddy YS, Balakrishna N, Rao GS, Gangadhar T, Arlappa N (2015) Role of drinking water with high Silica and Strontium in Chronic Kidney Disease: An Exploratory Community-based Study in an Indian Village. Indian Journal of Community Health 27: 95-102 \u003cem\u003ehttps://www.iapsmupuk.org/journal/index.php/IJCH/article/view/539\u003c/em\u003e\u003c/li\u003e\n\u003cli\u003eBanerjee D, Bairagi, Himadri, Mukhopadhyay S, Pal A, Bera, Debabrata, Ray Lalitagauri (2010) Heavy metal contamination in fruits and vegetables in two Districts of West Bengal, India. Electronic Journal of Environmental, Agricultural and Food Chemistry 9: 1423 -1432 \u003cem\u003ehttps://www.researchgate.net/publication/215565989_Heavy_metal_contamination_in_\u003c/em\u003e\u003cem\u003efruits_and_vegetables_in_two_districts_of_West_Bengal_India\u003c/em\u003e\u003c/li\u003e\n\u003cli\u003eMamatha P, Salamma S, Swamy AVN, Ravi Prasad Rao B (2014) Quantitative and risk analysis of heavy metals in selected Leafy vegetables. Der Pharma Chemica 6: 179-185 \u003cem\u003eh\u003c/em\u003e\u003cem\u003ettps://www.derpharmachemica.com/pharma-chemica/quantitative-and-risk-analysis-o\u003c/em\u003e\u003cem\u003ef-heavy-metals-in-selected-leafy-vegetables.pdf\u003c/em\u003e\u003c/li\u003e\n\u003cli\u003ePriya E, Sunil G, Shivaiah K, Anil Gaddameedi, Ashish Kumar (2014) Extent of Heavy metal contamination in leafy vegetables, soil and water from surrounding of Musi River, Hyderabad, India. Journal of Industrial Pollution Control 30(2): 289-293\u003cem\u003ehttps://www.researchgate.net/publication/271823133_Extent_of_heavy_metal_contamination_in_leafy_vegetables_soil_and_water_from_surrounding_of_Musi_River_Hyderabad_India\u003c/em\u003e\u003c/li\u003e\n\u003cli\u003eSapana Gupta, Jena V, Jena S, Neda Davic, Natalija Matic, Radojevic D, Solanki JS (2013) Assessment of heavy metal contents of green Leafy vegetables. 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Springer, Dordrecht \u003cem\u003ehttps://doi.org/10.1007/978-94-017-2496-8_1\u003c/em\u003e\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"biological-trace-element-research","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"bter","sideBox":"Learn more about [Biological Trace Element Research](https://www.springer.com/journal/12011)","snPcode":"12011","submissionUrl":"https://submission.nature.com/new-submission/12011/3","title":"Biological Trace Element Research","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Chronic Kidney Disease, ICP-MS, Trace Elements, Vegetables, Uddanam area","lastPublishedDoi":"10.21203/rs.3.rs-2737719/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-2737719/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe present study reports the Trace Elemental and Heavy Metal (24 Elements) levels in six (\u003cem\u003eCapsicum frutescence \u003c/em\u003eL., \u003cem\u003eCarica papaya \u003c/em\u003eL.,\u003cem\u003e Momordica charantia \u003c/em\u003eL.\u003cem\u003e, Moringa oliefera \u003c/em\u003eLam.\u003cem\u003e, Musa Sapientum \u003c/em\u003eL., and \u003cem\u003eSolanum melongena \u003c/em\u003eL.) \u0026nbsp;Local and frequently consumed vegetables from three locations which are severely affected Villages by Chronic Kidney Disease (CKD) in Kaviti Mandal, in Uddanam area of Srikakulam district. The samples are collected from these three Villages and are subjected to ICP-MS analysis. To test a group of 24 Elements viz., Li, Be, Al, Sc, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Ga, As, Se, Rb, Sr, Ag, Cd, Cs, Ba, TI, Rb, and U for their levels, the obtained levels of each Element is compared with the WHO/FAO permissible values. In these 24 Elements, as per FAO/WHO standards 16 Elements shows severe affect on kidneys and the remaining\u0026nbsp; 8 (Mn, Co, Ni, Cu, Zn, Se, Sr, and TI) cause other health problems when they are in high concentration[1-5]. The results reveal that Ba is in high concentration (2.51 times) in all the tested samples and Pb is in high concentration (1.28 times) in 11 samples, Ag and Fe are in one sample each. Among the three locations highest Ba concentration is observed in S\u003csub\u003e1\u003c/sub\u003e (\u003cem\u003eCapsicum\u003c/em\u003e) of L\u003csub\u003e2\u003c/sub\u003e followed by S\u003csub\u003e5\u003c/sub\u003e (\u003cem\u003eMusa\u003c/em\u003e) and S\u003csub\u003e1 \u003c/sub\u003e(\u003cem\u003eCapsicum\u003c/em\u003e) of L\u003csub\u003e1\u003c/sub\u003e. The highest Pb concentration is present in S\u003csub\u003e1\u003c/sub\u003e (\u003cem\u003eCapsicum\u003c/em\u003e) of L\u003csub\u003e3\u003c/sub\u003e followed by S\u003csub\u003e1\u003c/sub\u003e (\u003cem\u003eCapsicum\u003c/em\u003e) of L\u003csub\u003e2\u003c/sub\u003e. The results reveal that out of the six vegetables tested, \u003cem\u003eCapsicum\u003c/em\u003e showed high concentrations of Ba and Pb. The non-uniformity of levels of Trace Elements and Heavy Metals with regard to location and samples may be due to soil and ground water.\u003c/p\u003e","manuscriptTitle":"Trace Elemental and Heavy Metal levels in frequently consumed local Vegetables of three Chronic Kidney Disease prevalent Villages","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2023-03-28 14:28:37","doi":"10.21203/rs.3.rs-2737719/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Major revision","date":"2023-05-30T12:41:05+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2023-03-28T16:23:53+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"0182fcfe-c998-4ac5-81d0-92ad4d5b7f5d","date":"2023-03-28T15:43:38+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2023-03-27T12:27:04+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2023-03-27T12:10:16+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2023-03-27T06:04:46+00:00","index":"","fulltext":""},{"type":"submitted","content":"Biological Trace Element Research","date":"2023-03-26T10:01:41+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"biological-trace-element-research","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"bter","sideBox":"Learn more about [Biological Trace Element Research](https://www.springer.com/journal/12011)","snPcode":"12011","submissionUrl":"https://submission.nature.com/new-submission/12011/3","title":"Biological Trace Element Research","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"9b7020fa-fe30-4079-a7d8-8591bca13be5","owner":[],"postedDate":"March 28th, 2023","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2023-08-18T04:10:31+00:00","versionOfRecord":{"articleIdentity":"rs-2737719","link":"https://doi.org/10.1007/s12011-023-03761-6","journal":{"identity":"biological-trace-element-research","isVorOnly":false,"title":"Biological Trace Element Research"},"publishedOn":"2023-07-10 01:08:34","publishedOnDateReadable":"July 10th, 2023"},"versionCreatedAt":"2023-03-28 14:28:37","video":"","vorDoi":"10.1007/s12011-023-03761-6","vorDoiUrl":"https://doi.org/10.1007/s12011-023-03761-6","workflowStages":[]},"version":"v1","identity":"rs-2737719","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-2737719","identity":"rs-2737719","version":["v1"]},"buildId":"-HB7Z8yhvgn0wM9Nzuekk","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}