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Incorporating bioaccessibility into human health risk assessment of heavy metals in rice (Oryza sativa L.): A probabilistic-based analysis Tianyuan Li, Yinxian Song, Xuyin Yuan, Jizhou Li, Junfeng Ji, Xiaowen Fu, Qiang Zhang, and Shuhai Guo J. Agric. Food Chem., Just Accepted Manuscript • DOI: 10.1021/acs.jafc.8b01525 • Publication Date (Web): 11 May 2018 Downloaded from http://pubs.acs.org on May 11, 2018

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Journal of Agricultural and Food Chemistry

Incorporating bioaccessibility into human health risk assessment of heavy metals in rice (Oryza sativa L.): A probabilistic-based analysis

Tianyuan Li †, ‡, Yinxian Song

‡, *

, Xuyin Yuan ‡, Jizhou Li ‡, Junfeng Ji ⸹, Xiaowen

Fu †, Qiang Zhang †, *, Shuhai Guo†



Shandong Provincial Key Laboratory of Applied Microbiology, Ecology Institute,

Qilu University of Technology (Shandong Academy of Science), Jinan 250014, Shandong Province, P.R. China. ‡

Key Laboratory of Integrated Regulation and Resource Development on Shallow

Lake of Ministry of Education, College of Environment, Hohai University, Nanjing 210098, Jiangsu Province, P.R. China. ⸹

Key Laboratory of Surficial Geochemistry, Ministry of Education, School of Earth

Sciences and Engineering, Nanjing University, Nanjing 210046, Jiangsu Province, P.R. China.

*Corresponding author. E-mail address: [email protected] (Y. X. Song), during review procedure; [email protected] (Q. Zhang), after acceptance.

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ABSTRACT: A systematic investigation into total and bioaccessible heavy metal

2

concentrations in rice grains harvested from heavy metal-contaminated regions was

3

carried out to assess the potential health risk to local residents. Arsenic, Cr, Cu, Pb

4

and Zn concentrations were within acceptable levels while Cd and Ni concentrations

5

appeared to be much higher than other studies. The bioaccessibity of As, Cd and Ni

6

was high (> 25%) and could be well predicted from their total concentrations. The

7

non-carcinogenic risk posed by As and Cd was significant. The carcinogenic risk

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posed by all bioaccessible heavy metals at the 5th percentile was 10-fold higher than

9

the acceptable level, and Cd and Ni were the major contributors. The contribution of

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each metal to the combined carcinogenic risk indicates that taking pertinent

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precautions for different types of cancer, aimed at individuals with different levels of

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exposure to heavy metals, will greatly reduce morbidity and mortality rates.

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KEYWORDS: rice, heavy metals, bioaccessibility, health risk assessment, Monte

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Carlo simulation

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INTRODUCTION

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Due to rapid industrialization and urbanization, heavy metal contamination in

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soil and crops has received great attention worldwide, especially in developing

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countries such as China and India. Rice, the staple food for half of the world’s

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population, readily accumulates heavy metals such as As and Cd. Therefore,

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consumption of rice can be a significant pathway by which these toxic metals enter

21

the human body, causing related health disorders, including various forms of cancer

22

1-3

23

risks from ingesting heavy metal-contaminated rice, especially in developing

24

countries where intensive farming and industrial activities co-exist.

. Hence, there is a great need to more accurately and realistically assess the health

25

In previous studies, researchers have made a great effort to ensure that health risk

26

assessments (HRA) reflect the actual conditions, for example by using probabilistic

27

risk assessment (PRA) instead of the traditional deterministic assessment 4. PRA

28

characterizes uncertainty by calculating the risks based on the statistical distribution

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of the site-specific exposure parameters. It is also important to include the appropriate

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parameters in risk assessment. For example, when performing risk assessment of

31

foodstuffs, bioaccessible metal concentrations are more important than total metal

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content. Therefore, in vitro studies have been performed to estimate the amount of

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heavy metal absorption into human systemic circulation through ingestion

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previous study demonstrated that in vitro assays could provide a good prediction of in

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vivo

36

Volksgezondheid en Milieu) in vitro digestion model has been created to aid in

arsenic

bioavailability7.

Additionally,

the

RIVM

1

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. A

voor

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determining bioavailability and estimating health risks associated with heavy metal

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consumption from rice 8. Several HRA studies have been conducted using either PRA

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or bioaccessible metal concentrations9-11, and the combination of these two methods

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may produce a more accurate assessment result. However, studies in this area are

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lacking.

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The main objective of this study was to gives an accurate and realistic estimation

43

of the risk of heavy metals in rice grains to human health in southern Jiangsu Province,

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one of the most developed regions in China. To achieve this, total and bioaccessible

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fraction (BAF) of heavy metals in rice grain were determined. Both the carcinogenic

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risks (CR) and non-carcinogenic risks (NCR) to the population in the study region

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(adults and children) from ingestion of heavy metal-contaminated rice were

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determined using an in vitro digestion model combined with Monte Carlo simulations.

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This risk assessment will deepen our understanding of the level of heavy metal

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contamination in rice and the exposure risks to populations living in polluted areas in

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other parts of the world.

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MATERIALS AND METHODS

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Chemicals and reagents. The inorganic reagents were purchased from

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Sinopharm Chemical Reagent Co Ltd. (Shanghai, China). Urea, α-amylase, mucin,

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glucose, hydrochloride, pepsin, pancreatin, bile salts, albumin from bovine serum and

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lipase used for in vitro analysis were purchased from Sigma-Aldrich (St. Louis, MO,

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USA). All reagents were prepared with deionized water obtained from a Milli-Q

58

system (Millipore, Billerica, MA, USA). 2

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Study areas and sampling. Rice samples were collected from Yifeng, Dingshu,

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Ehu, Wangting and Taicang villages, which are located in southern Jiangsu Province,

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China (Figure S1). The longitude and latitude of the study regions ranged from

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119.81°E to 121.13°E and from 31.27°N to 31.54°N. The study areas have a

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subtropical monsoon humid climate. There are numerous heavy metal-associated

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industries such as chemical, battery, ceramic making and foundry industries in these

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areas

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samples (14 samples from each area) were randomly collected. At least five

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sub-samples per sampling site were mixed together. All rice samples collected were of

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the Indica variety. More detailed information about the study areas and sampling

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methods was reported in our previous study 12.

12

. In late July 2015, when rice plants reached maturity, a set of 70 rice grain

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Sample pretreatment. Raw rice samples were used to study metal

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concentrations and bioaccessibility. Rice grains were oven dried at 105 ℃ for 1 h,

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and then samples were dried at 70 ⸹ to constant weight. In order to avoid the

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influence of particle size on metal bioaccessibility and to avoid cross-contamination,

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raw rice samples were ground manually with a pestle and mortar. Then, the rice

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samples were passed through a 0.25 mm mesh sieve and kept in sealed plastic bags at

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4 ⸹ prior to further analysis.

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Determination of total heavy metal concentrations in rice. Total

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concentrations of heavy metals were determined in a 1.000 g rice sample that was

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digested with a mixture of HClO4 and HNO3 in beaker placed on the electric heating

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plate for 6 hours. The heavy metal concentrations in these digests were determined by 3

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ICP-MS (Inductively Coupled Plasma Mass Spectrometry, XSERIES, Thermo

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Electron, USA). Quality assurance and quality control (QA/QC) for metals in rice

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samples were estimated by determining the metal contents of blank and duplicate

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samples as well as of certified reference materials (GSB-1). The recovery of GSB-1

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was 93-105%. The relative difference values for all replicates were less than 5%.

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Bioaccessibility of heavy metals in rice grain. The BAF of heavy metals in rice

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grain was determined using the RIVM in-vitro digestion model, which includes

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models for three compartments: mouth, stomach and small intestine. Digestive juices

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were prepared artificially as described in a previous in vitro study

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constituents of the digestive juices are presented in Table S1. Briefly, the simulated

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digestive process was initiated by adding 6 mL of artificial saliva to 4.5 g of rice

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sample to represent digestion in the mouth. Next, 12 mL of artificial gastric juice was

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added, and the mixture was shaken to represent digestion in the stomach. Finally, 12

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mL of duodenal juice, 6 mL of bile juice and 2 mL of NaHCO3 solution was added to

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the mixture to represent digestion in the small intestine. The mixture was incubated at

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37±2 ⸹ in a shaker (55 rpm) during in vitro digestion. The incubation time was 5 min

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for mouth and 2 hours each for stomach and small intestine. The pH of the mixture

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was adjusted to 6.8, 2.0 and 7.0 using HCl (37%) or NaOH (0.1 M) when simulating

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digestion in the mouth, stomach and small intestine, respectively. When in vitro

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digestion was complete, the mixture was centrifuged at 2750 g for 5 min. The

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supernatant was filtered through 0.45 µm filter paper. Heavy metal concentrations

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were measured using ICP-MS. The kinetic energy discrimination (KED) mode (He as 4

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. The

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the reaction gas) was used to eliminate the mass spectrometric determination of

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multi-atom spectra interference. The results from analysis of standard reference

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material were satisfactory. QC for the in vitro experiment was assessed by using

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duplicate and blank samples. In addition, heavy metal contents in both the soluble

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fraction and pellet were calculated from liquid volumes, pellet weights and measured

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metal concentrations. The sum of the heavy metal contents in the supernatant and in

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the pellet was compared with total amount of metal introduced into the in vitro system.

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The recovery of the As, Cd, Pb, Cr, Cu, Zn and Ni was 88.3%, 96.2%, 95.9%, 99.1%,

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101.2%, 92.5% and 102.6%, respectively. The bioaccessibility of each metal was

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calculated as the percentage of the metal contents in the soluble fraction relative to the

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total concentration in rice grains.

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Risk assessment

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Exposure analysis. The exposure to heavy metals in rice can be characterized by

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the average daily dose (ADD, mg/kg/day), which is calculated by multiplying the rate

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of consumption (ingestion rate, IR) and the heavy metal concentration (Cm) and then

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dividing by body weight (BW). BW used in this study was based on questionnaire

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answers from a survey performed in the study area. The BW values for adults (18

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years old or over) and children (7-18 years old) were 59.80±7.19 and 38.9±4.81 kg,

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respectively. BW follows a normal distribution. IR for survey participants (adults and

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children) was calculated based on the amount of rice consumed per meal (g/meal) and

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the frequency of rice consumption (meals/day). The daily IR averaged over one year

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was reported as the daily intake. Survey answers were validated by asking some 5

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logical questions with different wording and cross-checking for inconsistencies

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among the answers

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362.80±30.81 and 236.23±12.53 g/day per person, respectively. IR for both adults and

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children followed the normal distribution. The 5th percentile, median (50th percentile)

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and 95th percentile of the daily exposure levels were used to represent ADD for low,

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average and high consumers, respectively 16.

. The estimated IR values for adults and children were

15

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Non-carcinogenic risks. The potential human health risk from consumption of

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rice was assessed based on the hazard quotient (HQ), which was provided by the US

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EPA 17. An HQ value Cu > Ni > Cr > Cd > As > Pb.

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Total concentrations of Cr, Cu, Pb and Zn (average values) were lower than the

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recommended maximum permissible level (MPL) suggested by China, while the Cd

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concentration was about 1.5 times greater than the MPL

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concentrations of 1.43% and 4.29% of the samples, respectively, exceeded the MPL

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while the Cu and Zn concentrations of all samples were lower than the MPL. The As

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concentration in rice samples ranged from 0.01 to 0.23 mg/kg, with an average of 0.13

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mg/kg. Approximately 92% of the samples had As concentrations < 0.2 mg/kg, the

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Chinese limit for inorganic As in rice 21. Our finding is comparable to that of Li et al.

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who reported a mean As concentration of 0.13 mg/kg in Chinese rice

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concentration in 31.43% of the rice samples surpassed the MPL. The highest Cd

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concentration was 2.77 mg/kg, nearly 14-fold higher than the MPL, and the 75%

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percentile concentration (0.34 mg/kg) was 1.7 times higher than the MPL. Although

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the MPL for Ni in rice has not been established by the Chinese government, the mean

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Ni concentration in this study was higher than that previously observed for rice grown

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in the Yangtze River Basin (0.54 mg/kg, n=681) and Jiangsu Province (0.31 mg/kg, 8

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. The Pb and Cr

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. The Cd

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n=50)

. The rice Ni concentration in this study was even higher than that of rice

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grown in Hunan Province (0.70 mg/kg, n=309), which is well known as “the

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hometown of nonferrous metals” because mining activities have caused serious heavy

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metal contamination of soil 23. Based on our analysis, we conclude that rice grains in

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the study areas are contaminated the most by Cd and Ni and the least by Cu and Zn.

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Soil heavy metal contamination has become increasingly severe in some areas of

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southern Jiangsu Province, where the sampling sites in this study are located25.

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However, the background value of some metals in these areas are low. For example,

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the background values of As, Cr and Cu in soil were previously found to be 9.4, 75.6

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and 23.4 mg/kg

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activities, the concentrations of As, Cr and Cu are still below the safety threshold 27.

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Thus these concentrations in rice grains were within acceptable levels.

26

. In other words, despite the pollution of soil by anthropogenic

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The situation is different for Cd. The factors responsible for excessive Cd

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accumulation in rice grains are complicated and include water management, farming

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method and the degree of Cd transfer in the soil-rice system

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and Cd accumulation in the soil also contribute to Cd uptake and are likely the main

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factors contributing to the high Cd concentration in rice grains in this study 27. Human

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activities in this region have resulted in severe soil Cd pollution. For example, in

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Dingshu the ceramic industry has flourished over the past several decades, and Cd, 8%

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of which is used for pigments, is an important material in ceramic processing 30.

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According to our previous investigation, the average soil Cd concentration in this

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region was 1.56 mg/kg, which is about 3-fold higher than the safety value 9

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. Soil acidification

12

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Furthermore, the pH value of topsoil in this region has been declining over time,

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indicating increased soil acidification 26. Because Cd transfer from the soil to rice is

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highly influenced by soil pH, with the highest Cd transfer occurring at approximately

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pH 5.5,

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rice.

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increased soil acidification has likely led to increased Cd accumulation in

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Bioaccessibility of heavy metals in rice. The BAF of heavy metals in rice

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grains ranked as follows: Zn (4.02 mg/kg) > Cu (0.86 mg/kg) > Ni (0.24 mg/kg) > Cd

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(0.06 mg/kg) > As (0.05 mg/kg) > Cr (0.03 mg/kg) > Pb (0.02 mg/kg). Thus, the

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concentrations of Zn and Pb were the highest and the lowest, respectively, based on

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analysis of both total and bioaccessible concentrations. Bioaccessibility (the

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percentage of the metal that is bioaccessible) of heavy metals in rice grains ranked as

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follows: As > Cd > Ni > Pb > Zn > Cu > Cr (Table 1). Previous research showed that 23.16-32.34% of As in rice grains was accessible

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32

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chemical form of As largely determines its bioaccessibility

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of inorganic As was reported to be almost 100%, which is much higher than that of

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dimethyl arsenic acid (DMAⅤ, 33%)

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inorganic As was 93.60% of the total As in raw rice, also indicating that inorganic As

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is more bioaccessible than organic As

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rice observed in this study is likely due to the presence of inorganic As 32.

, which is lower than that observed in the present study (37.90±7.89%). The 32-34

. The bioaccessibility

35

. In intestinal extracts, the percentage of

34

. Therefore, the high As bioaccessibility in

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The bioaccessibility of Cd (35.25%) in rice grains in this study was higher than

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previously observed in rice grains collected from mining areas (16.94%) 36 and lower 10

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than in rice grains from the market

. The relatively high Cd bioaccessibility

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observed in this study may due to the high concentration of total Cd in rice. For

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example, a significant relationship was found between Cd bioaccessibility and total

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concentration in oysters

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controlling Cd bioaccessibility in rice remains unknown. Further research should

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focus on the factors (such as subcellular distribution and the chemical form of Cd) and

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the mechanisms affecting Cd bioaccessibility in crops.

38

. But whether total Cd concentration is the main factor

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Mean Ni bioaccessibility was 27.77%, which is much higher than that observed

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in convenience and fast food (4.50-7.75%) 39. Little information is available about Ni

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bioaccessibility in rice grain, and our finding suggests that further studies on Ni

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bioaccessibility and the factors influencing it are needed.

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We found a positive linear correlation between the total concentrations and BAF 36, 39

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of As, Cd and Ni; thus, consistent with previous studies

, BAF for these metals

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could be well predicted by total concentrations in rice (Figure S2).

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Health risk assessment of heavy metals in rice

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Non-carcinogenic risk. It is crucial to conduct HRA to assess human health

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risks due to heavy metal contamination of food 40. BAF concentrations were used to

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conduct ADD, and the results were compared to those based on total heavy metal

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concentrations (Figure S3). In order to assess the risks more precisely, PRA, which

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provides a detailed risk distribution, was used (Figure 1 and Table 2). The highest HQ

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for the local residents was found for As at P95 (HQ=2.20 and 1.83 for children and

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adults, respectively), followed by Cd (HQ=1.97 and 1.53 for children and adults, 11

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respectively). The HQ value of As for children (HQ =1.18) at P50 was still greater

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than 1, indicating that exposure to As and Cd in particular via rice ingestion poses

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great NCR to local residents. The HQ values for the remaining heavy metals were less

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than 1, indicating they pose no potential NCR to the population. P50 for combined

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HQ (HI) was 1.74 and 1.36 for children and adults, respectively. HI for children at P5

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was 0.71, which is very close to the maximum admissible, indicating that most

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residents may be exposed to metals at hazard levels from consumption of locally

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produced rice. The situation was even worse for the high rice consumption population

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(HI=5.06 and 4.08 for children and adults, respectively, at P95). In addition, result

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showed that HI value based on total metal contents in rice were nearly 4 folds higher

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than that based on bioaccessible metal contents at different percentiles. Our results

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suggest that NCR based on PRA combined with bioaccessible heavy metal

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concentrations in rice could give a more accurate and precise risk assessment. The

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health risk posed by Pb, Cr, Cu, Zn and Ni is negligible, while exposure to As and Cd

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via rice ingestion poses a great threat to the heath of local residents.

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Carcinogenic risk. For CR assessment based on total metal concentrations in

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rice, all Riski values for individual metals, were greater than the acceptable range

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(1E-06 to 1E-04) specified by the US EPA except for Pb (Table S2). When assessing

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CR with BAF, the CR posed by Cr was also considered acceptable at P5 for both

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adults and children, and at P50 for adults only. The highest Riski for both adults and

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children was found for Cd (P95=1.71E-01 for children and 1.36E-01 for adults). Even

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though the Riski was assessed based on bioaccessible Cd concentration in rice, the 12

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Riski value (P95) was still much higher than the acceptable value (Riski for Cd > 0.02

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for both adults and children). The results for Ni were also striking: when CR was

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assessed based on total and bioaccessible concentrations, CR for Ni was about 100

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and 27 times higher than the acceptable level (Median) for children and adults,

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respectively. The RISK value for all heavy metals was much higher than the

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acceptable range (> 1.00E-03), even at P5, irrespective of whether CR was assessed

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based on total or bioaccessible metal concentrations. This result indicates that heavy

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metal exposure from rice ingestion poses a great CR to the local residents (Table S2

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and Figure S4).

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The contribution of each metal to the RISK showed great variation between

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different Monte-Carlo-derived percentiles. The contribution of Ni, As and Cr to RISK

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at P5, Median and P95 decreased, while Cd showed the opposite trend. In addition,

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the contribution of each metal to RISK is different when CR was assessed based on

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total concentrations and BAF. For example, the contribution of As to RISK was

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higher when CR was assessed based on BAF than when assessed based on total As

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(Figure 2). This observation illustrates that different types of cancer may occur in

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populations with different levels of rice consumption. For example, chronic exposure

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to cadmium is related to lung cancer and breast cancer 41 while As can cause ovarian

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and kidney cancer

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more likely to suffer from lung and breast cancer while those with low rice

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consumption have a high risk of getting ovarian and kidney cancer. Hence, taking

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pertinent precautions for different types of cancer, aimed at individuals with different

42, 43

, indicating that individuals with high rice consumption are

13

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heavy metal exposure, will greatly reduce the morbidity and mortality rate.

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Although CR assessed based on the total and bioaccessible heavy metal

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concentrations in rice grains showed differences in the contributions of different

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metals to RISK at different percentiles (Figure 2), both analyses suggest that local

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residents have likely been exposed to high levels of heavy metals from ingesting rice

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produced in the study areas, which poses a high CR. This is consistent with previous

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research demonstrating that southern Jiangsu Province has one of the highest cancer

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incidence rates in China

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been reduced by the following factors: 1. The Grain Purchase Policy issued by the

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State Administration of Grain of the People’s Republic of China may greatly reduce

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the cancer incidence

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(the floor price policy for grain) by the government. In addition, southern Jiangsu

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Province is one of the most developed regions in China, and the convenient transport

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and logistical conditions allows more frequent movement of the population and

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cereals (such as rice). Together these factors could effectively reduce the consumption

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of locally produced rice. 2. Proper medical care and the high level of income and

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education of the local residents can reduce the cancer morbidity and mortality rate.

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Nevertheless, the situation will get worse if heavy metal pollution occurs in less

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developed areas or if the pollution range gets wider. Hence, proper measures such as

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soil remediation to prevent metal transfer from soil to the human body are urgently

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needed. Soil threshold values for heavy metal concentrations should also be improved

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based on accurate health risk assessments to ensure food security. In addition, rice

44

. To some extent, cancer incidence in the study areas has

45

. Unqualified cereals must be purchased at the original price

14

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consumption is only one of the exposure routes to humans, and exposure to heavy

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metals through vegetables and drinking water is also an important health threat to

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populations in metal-contaminated regions. Thus further research considering

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multiple types of exposure to bioaccessible metals should be conducted.

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ABBREVIATIONS USED

324

Bioaccessible fraction of heavy metal concentration

325

Health Risk Assessment

326

Probabilistic Risk Assessment

327

Hazard Quotient

328

Hazard Index

329

Maximum Permissible Levels

330

Rijksinstituut voor Volksgezondheid en Milieu

331

Carcinogenic Risk

332

Non-Carcinogenic Risk

333

Carcinogenic risk value for single element

334

Combined carcinogenic risk value of each element

335

ACKNOWLEDGEMENT

BAF

HRA PRA

HQ HI MPL RIVM

CR NCR Risk RISK

336

We sincerely acknowledge Hongyan Chen, Xueqiang Zhao, Hailong Chen, Qing

337

Liu and Sun Hu, for their contribution to field sampling. We also acknowledge Qing

338

Chang from Geological Survey of Jiangsu Province, for her precious contribution to

339

analytical work.

340

SUPPORTING INFORMATION 15

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Brief statement in non-sentence format listing the contents of the material

342

supplied as Supporting Information. Supporting information description are as

343

follows:

344 345

Table S1. Constituents of the Various Synthetic Digestive Juices Used for RIVM In Vitro Digestion.

346

Table S2. Carcinogenic Risk (Risk) and ∑Risk (RISK) From Rice Ingestion

347

Calculated Based on Total and Bioaccessible Heavy Metal Concentrations Using

348

Monte-Carlo Simulation.

349

Figure S1. The locations of the sites where rice grains were sampled. Five

350

regions with severe soil heavy metal pollution in southern Jiangsu Province, China

351

were selected.

352

Figure S2. Correlation between the bioaccessible As, Cd and Ni fractions and

353

total concentrations in rice grains. The significant positive correlations indicate that

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bioaccessible As, Cd and Ni concentrations can be predicted from their total

355

concentrations in rice grains. The dashed lines represent the 95% confidence intervals.

356

Figure S3. Probability density functions and best fitted distributions of the

357

average daily dose (ADD) of heavy metals from rice ingestion in the study region.

358

ADD values calculated based on bioaccessible and total heavy metal concentrations

359

are compared. ADD was evaluated using a Monte Carlo in crystal ball with 10000

360

iterations.

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Figure S4. Probability density functions (A) and cumulative probability (B) of

362

the carcinogenic risk of all heavy metals (RISK) from rice ingestion in the study

363

region. RISK values calculated based on bioaccessible and total heavy metal

364

concentrations are compared. RISK was evaluated using a Monte Carlo simulation in

365

Crystal Ball with 10000 iterations. 16

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REFERENCES

368

1.

Lin, Y.; Ma, J.; Zhang, Z.; Zhu, Y.; Hou, H.; Zhao, L.; Sun, Z.; Xue, W.; Shi, H.,

369

Linkage between human population and trace elements in soils of the Pearl River

370

Delta: Implications for source identification and risk assessment. Science of the

371

Total Environment 2017, 610-611, 944-959.

372

2.

Chervona, Y.; Arita, A.; Costa, M., Carcinogenic metals and the epigenome:

373

understanding the effect of nickel, arsenic, and chromium. Metallomics 2012, 4,

374

(7), 619-627.

375

3.

Liu, C.P.; Luo, C. L.; Gao, Y.; Li, F. B.; Lin, L. W.; Wu, C. A.; Li, X. D., Arsenic

376

contamination and potential health risk implications at an abandoned tungsten

377

mine, southern China. Environmental Pollution 2010, 158, (3), 820-826.

378

4.

Kim, E.; Moon, J. K.; Choi, H.; Kim, J. H., Probabilistic exposure assessment for

379

applicators during treatment of the fungicide kresoxim-methyl on apple orchard

380

by speed sprayer. Journal of Agricultural & Food Chemistry 2015, 63, (48),

381

53-66.

382

5.

Juhasz, A. L.; Smith, E.; Weber, J.; Rees, M.; Rofe, A.; Kuchel, T.; Sansom, L.;

383

Naidu, R., Comparison of in vivo and in vitro methodologies for the assessment

384

of arsenic bioavailability in contaminated soils. Chemosphere 2007, 69, (6),

385

961-966.

386

6.

Kong, F.; Oztop, M. H.; Singh, R. P.; McCarthy, M. J., Physical changes in white

387

and brown rice during simulated gastric digestion. Journal of Food Science 2011,

388

76, (6), E450-E457. 18

ACS Paragon Plus Environment

Page 20 of 33

Page 21 of 33

Journal of Agricultural and Food Chemistry

389

7.

Juhasz, A. L.; Weber, J.; Smith, E.; Naidu, R.; Rees, M.; Rofe, A.; Kuchel, T.;

390

Sansom, L., Assessment of four commonly employed in vitro arsenic

391

bioaccessibility assays for predicting in vivo relative arsenic bioavailability in

392

contaminated soils. Environmental Science & Technology 2009, 43, (24),

393

9487-9494.

394

8.

Chm, V.; De, K. E. V.; Cjm, R., Development and applicability of an in vitro

395

digestion model in assessing the bioaccessibility of contaminants from food.

396

Rijksinstituut Voor Volksgezondheid En Milieu Rivm 2007.

397

9.

Yang, Y.; Wang, M.; Chen, W.; Li, Y.; Chi, P., Cadmium accumulation risk in

398

vegetables and rice in southern china: insights from solid-solution partitioning

399

and plant uptake factor. Journal of Agricultural & Food Chemistry 2017, 65,

400

(27), 5463-5469.

401

10. Yu, Y. X.; Li, J. L.; Zhang, X. Y.; Yu, Z. Q.; Van, d. W. T.; Han, S. Y.; Wu, M. H.;

402

Sheng, G. Y.; Fu, J. M., Assessment of the bioaccessibility of polybrominated

403

diphenyl ethers in foods and the correlations of the bioaccessibility with nutrient

404

contents. Journal of Agricultural & Food Chemistry 2010, 58, (1), 301-308.

405

11. Jadán-Piedra, C.; Clemente, M. J.; Devesa, V.; Vélez, D., Influence of

406

physiological gastrointestinal parameters on the bioaccessibility of mercury and

407

selenium from swordfish. Journal of Agricultural & Food Chemistry 2016, 64,

408

(3), 690-698.

409

12. Li, T.; Chang, Q.; Yuan, X.; Li, J.; Ayoko, G. A.; Frost, R. L.; Chen, H.; Zhang,

410

X.; Song, Y.; Song, W., Cadmium transfer from contaminated soils to the human 19

ACS Paragon Plus Environment

Journal of Agricultural and Food Chemistry

411

body through rice consumption in southern Jiangsu Province, China.

412

Environmental Science Processes & Impacts 2017, 19, (6), 843-850.

413

13. Omar, N. A.; Praveena, S. M.; Aris, A. Z.; Hashim, Z., Health Risk Assessment

414

using in vitro digestion model in assessing bioavailability of heavy metal in rice:

415

A preliminary study. Food Chemistry 2015, 188, 46-50.

416

14. Versantvoort, C. H.; Oomen, A. G.; Van de Kamp, E.; Rompelberg, C. J.; Sips, A.

417

J., Applicability of an in vitro digestion model in assessing the bioaccessibility of

418

mycotoxins from food. Food and Chemical Toxicology 2005, 43, (1), 31-40.

419

15. Iarossi, B. G., The power of survey design: a user's guide for managing surveys,

420

interpreting results, and influencing respondents. World Bank Publications 2010.

421

16. Huang, Z.; Pan, X. D.; Wu, P. G.; Han, J. L.; Chen, Q., Health risk assessment of

422

heavy metals in rice to the population in Zhejiang, China. Plos One 2013, 8, (9),

423

75-87.

424

17. Luna, C. M.; Pastori, G. M.; Driscoll, S.; Groten, K.; Bernard, S.; Foyer, C. H.,

425

Drought controls on H2O2 accumulation, catalase (CAT) activity and CAT gene

426

expression in wheat. Journal of Experimental Botany 2005, 56, (411), 417-423.

427

18. US EPA, Office of Solid Waste Emergency Response, waste and cleanup risk

428

assessment, Risk Assessment Guidance for Superfund (RAGS) Part A, 1989, 804,

429

(7), 636-640.

430

19. Li, Z.; Ma, Z.; van der Kuijp, T. J.; Yuan, Z.; Huang, L., A review of soil heavy

431

metal pollution from mines in China: Pollution and health risk assessment.

432

Science of The Total Environment 2014, 468–469, 843-853. 20

ACS Paragon Plus Environment

Page 22 of 33

Page 23 of 33

Journal of Agricultural and Food Chemistry

433

20. Nakazawa, K.; Nagafuchi, O.; Kawakami, T.; Inoue, T.; Yokota, K.; Serikawa, Y.;

434

Cyio, B.; Elvince, R., Human health risk assessment of mercury vapor around

435

artisanal small-scale gold mining area, Palu city, Central Sulawesi, Indonesia.

436

Ecotoxicology and Environmental Safety 2016, 124, 155-162.

437

21. Ministry of Health of the People’s Republic of China (MHPC). National food

438

safety standard, Maximum level of contaminants in foods (GB 2762-2012).

439

Beijing: China Standard Press, 2012 (in Chinese).

440

22. Li, H. B.; Li, J.; Zhao, D.; Li, C.; Wang, X.; Sun, H. J.; Juhasz, A. L.; Ma, L. Q.,

441

Arsenic relative bioavailability in rice using a mouse arsenic urinary excretion

442

bioassay and its application to assess human health risk. Environmental Science

443

& Technology 2017, 51, (8), 4689-4696.

444

23. Cao, Z.; Mou, R.; Cao, Z.; Lin, X.; Xu, P.; Chen, Z.; Zhu, Z.; Chen, M., Nickel

445

in milled rice (Oryza sativa L.) from the three main rice-producing regions in

446

China. Food Additives & Contaminants Part B Surveillance 2016, 10, (1), 69-82.

447

24. Wang, W.; Zhang, Z.; Yang, G.; Wang, Q., Health risk assessment of Chinese

448

consumers to nickel via dietary intake of foodstuffs. Food Additives &

449

Contaminants Part A Chemistry Analysis Control Exposure & Risk Assessment

450

2014, 31, (11), 1861-1871.

451

25. Xing, P. G.; Min, C. J.; Qin, G. J.; Liang, L. S.; Wei, Z. J., On the change of

452

status of some heavy metal elements in soil environment under intensive

453

economical development. Resources & Enuironment in the Yangtza Basin 2000.

454

26. Liao, Q. L.; Evans, L. J.; Gu, X.; Fan, D. F.; Jin, Y.; Wang, H., A regional 21

ACS Paragon Plus Environment

Journal of Agricultural and Food Chemistry

455

geochemical survey of soils in Jiangsu Province, China: Preliminary assessment

456

of soil fertility and soil contamination. Geoderma 2007, 142, (1–2), 18-28.

457

27. Chen, L.; Zhou, S.; Shi, Y.; Wang, C.; Li, B.; Li, Y.; Wu, S., Heavy metals in

458

food crops, soil, and water in the Lihe River Watershed of the Taihu Region and

459

their potential health risks when ingested. Science of the Total Environment 2017,

460

615, 141-149.

461

28. Wang, J.; Feng, X.; Anderson, C. W.; Qiu, G.; Bao, Z.; Shang, L., Effect of

462

cropping systems on heavy metal distribution and mercury fractionation in the

463

Wanshan mining district, China: implications for environmental management.

464

Environmental Toxicology & Chemistry 2015, 33, (9), 2147-2155.

465

29. Chen, H.; Yuan, X.; Li, T.; Hu, S.; Ji, J.; Wang, C., Characteristics of heavy metal

466

transfer and their influencing factors in different soil–crop systems of the

467

industrialization region, China. Ecotoxicology and Environmental Safety 2016,

468

126, 193-201.

469

30. Obaid F.; Annette A.; Scott W.; Pam T.; Kim J. Agency for Toxic Substances and

470

Diseases Registry (ATSDR), 2012. Toxicological profile for cadmium. URL

471

(http://www.atsdr.cdc.gov/toxprofiles/tp.asp?id=48&tid=15).

472

31. Rafiq, M. T.; Aziz, R.; Yang, X.; Xiao, W.; Rafiq, M. K.; Ali, B.; Li, T.,

473

Cadmium phytoavailability to rice (Oryza sativa L.) grown in representative

474

Chinese soils. A model to improve soil environmental quality guidelines for food

475

safety. Ecotoxicology & Environmental Safety 2014, 103, (1), 101-107.

476

32. Bolan, S.; Kunhikrishnan, A.; Chowdhury, S.; Seshadri, B.; Naidu, R.; Ok, Y. S., 22

ACS Paragon Plus Environment

Page 24 of 33

Page 25 of 33

Journal of Agricultural and Food Chemistry

477

Comparative analysis of speciation and bioaccessibility of arsenic in rice grains

478

and complementary medicines. Chemosphere 2017, 182, 433-440.

479

33. Meliker, J. R.; Franzblau, A., Major contributors to inorganic arsenic intake in

480

southeastern Michigan. International Journal of Hygiene & Environmental

481

Health 2006, 209, (5), 399-411.

482

34. Sun, G. X.; Van de Wiele, T.; Alava, P.; Tack, F.; Du Laing, G., Arsenic in cooked

483

rice: Effect of chemical, enzymatic and microbial processes on bioaccessibility

484

and speciation in the human gastrointestinal tract. Environmental Pollution 2012,

485

162, 241-246.

486

35. Juhasz, A. L.; Smith, E.; Weber, J.; Rees, M.; Rofe, A.; Kuchel, T.; Sansom, L.;

487

Naidu, R., In vivo assessment of arsenic bioavailability in rice and its

488

significance for human health risk assessment. Environmental Health

489

Perspectives 2006, 114, (2), 1826-1831.

490

36. Yang, L. S.; Zhang, X. W.; Li, Y. H.; Li, H. R.; Wang, Y.; Wang, W. Y.,

491

Bioaccessibility and risk assessment of cadmium from uncooked rice using an in

492

vitro digestion model. Biological Trace Element Research 2012, 145, (1), 81-86.

493

37. Zhuang, P.; Zhang, C.; Li, Y.; Zou, B.; Mo, H.; Wu, K.; Wu, J.; Li, Z.,

494

Assessment of influences of cooking on cadmium and arsenic bioaccessibility in

495

rice, using an in vitro physiologically-based extraction test. Food Chemistry

496

2016, 213, 206-214.

497

38. He, M.; Ke, C. H.; Tian, L.; Li, H. B., Bioaccessibility and health risk assessment

498

of Cu, Cd, and Zn in “colored” oysters. Archives of Environmental 23

ACS Paragon Plus Environment

Journal of Agricultural and Food Chemistry

499

Contamination and Toxicology 2016, 70, (3), 595-606.

500

39. Cabrera-Vique, C.; Mesías, M.; Bouzas, P. R., Nickel levels in convenience and

501

fast foods: In vitro study of the dialyzable fraction. Science of The Total

502

Environment 2011, 409, (8), 1584-1588.

503

40. Lee, S. W.; Lee, B. T.; Kim, J. Y.; Kim, K. W.; Lee, J. S., Human risk assessment

504

for heavy metals and as contamination in the abandoned metal mine areas, Korea.

505

Environmental Monitoring and Assessment 2006, 119, (1), 233-244.

506

41. Nawrot, T. S.; Martens, D. S.; Hara, A.; Plusquin, M.; Vangronsveld, J.; Roels, H.

507

A.; Staessen, J. A., Association of total cancer and lung cancer with

508

environmental exposure to cadmium: the meta-analytical evidence. Cancer

509

Causes & Control 2015, 26, (9), 1281-1288.

510

42. Arsenic, R.; Braicu, E. I.; Letsch, A.; Dietel, M.; Sehouli, J.; Keilholz, U.;

511

Ochsenreither, S., Cancer-testis antigen cyclin A1 is broadly expressed in ovarian

512

cancer and is associated with prolonged time to tumor progression after

513

platinum-based therapy. Bmc Cancer 2015, 15, (1), 1-11.

514

43. Saint-Jacques, N.; Brown, P.; Nauta, L.; Boxall, J.; Parker, L.; Dummer, T. J. B.,

515

Estimating the risk of bladder and kidney cancer from exposure to low-levels of

516

arsenic in drinking water, Nova Scotia, Canada. Environment International 2017,

517

110, (32), 95-104.

518

44. Lu, Y.; Song, S.; Wang, R.; Liu, Z.; Meng, J.; Sweetman, A. J.; Jenkins, A.;

519

Ferrier, R. C.; Li, H.; Luo, W., Impacts of soil and water pollution on food safety

520

and health risks in China. Environment International 2015, 77, (1), 5-15. 24

ACS Paragon Plus Environment

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Page 27 of 33

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45. URL (http://www.chinagrain.gov.cn/n787423/c938910/content.html). Important

522

works relevant to grain quality in 2016 issued by the State Administration of

523

Grain of People’s Republic of China.

524

FUNDING SOURCES

525

This study was supported by the the Natural Science Foundation of Shandong

526

Province (ZR2016YL002), Research Project (Youth Fund) of Shandong Academy of

527

Science (Grant No. 2018QN0023), the Key Research and Development Project of

528

Jiangsu Province, China (Grant No. BE2015708), the National Natural Science

529

Foundation of China (Grant No. 41601540), the Foundation Research Project of

530

Jiangsu Province (Grant No. BK20160859) and the Fundamental Research Funds for

531

the Central Universities (Grant No. 2014B18514).

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FIGURE CAPTIONS

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Figure 1. Probability density functions (A) and cumulative probability (B) of the hazard

534

index (HI) of heavy metals from rice ingestion in the study region. HI values calculated based

535

on bioaccessible and total heavy metal concentrations are compared. HI was evaluated using a

536

Monte Carlo simulation in Crystal Ball with 10000 iterations.

537

Figure 2. The relative contribution of each metal to the RISK (sum of carcinogenic

538

risk due to each metal) at Monte Carlo-derived P5 (5th percentile), Median (50th

539

percentile) and P95 (95th percentile). (A) Carcinogenic risk for children based on

540

total metal concentrations; (B) Carcinogenic risk for adults based on total metal

541

concentrations; (C) Carcinogenic risk for children based on bioaccessible metal

542

concentrations; (D) Carcinogenic risk for adults based on bioaccessible metal

543

concentrations.

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Table 1 Descriptive Statistics for Total Concentrations (mg kg -1) and Bioaccessibility (%) of Heavy Metals in Rice Grains (n=70). As

Cd

Pb

Cr

Cu

Zn

Ni

Total Heavy Metal

0.13±0.05

0.30±0.52

0.09±0.04

0.44±0.23

3.94±0.88

18.87±2.94

0.84±0.40

Concentrations (mg kg-1)

[0.01,0.23]

[0.01,2.77]

[0.05,0.29]

[0.13,1.22]

[1.54,6.17]

[14.00,30.10]

[0.38,2.62]

Probabilistic Distributionsa

Logistic

Lognormal

Lognormal

Lognormal

Normal

Lognormal

Lognormal

0.2d

0.2

0.2

1.0

10

50

-

Regulated -1 b

Concentrations (mg kg )

37.90±7.89

35.25±20.06 23.94±11.00

9.87±5.66

21.19±11.18

21.67±8.22

27.77±8.05

[18.04,58.64]

[6.67,87.50]

[7.58,48.34]

[3.26,32.04]

[4.02,49.25]

[9.01,40.89]

[18.22,60.08]

Lognormal

Lognormal

Lognormal

Beta

Gamma

Gamma

Lognormal

Bioaccessibility (%) Probabilistic Distributionc

a: The Probabilistic Distributions of Total Heavy Metal Concentrations b: Staple Rice Safety Criteria Issued by the Ministry of Agriculture, P.R. China c: The Probabilistic Distributions of Bioaccessible Heavy Metal Concentrations d: The Maximum Level of Inorganic As in Rice Specified by the Ministry of Agriculture P.R. China.

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Table 2. Hazard Quotient (HQ) and Hazard Index (HI) for Non-Carcinogenic Risk from Rice Ingestion Calculated Based on Bioaccessible Heavy Metals Concentrations Using Monte-Carlo Simulation. P5

Metal

Median

P95

HQ

Children

Adults

Children

Adults

Children

Adults

As

5.10E-01

3.43E-01

1.18E+00

8.97E-01

2.20E+00

1.83E+00

Cd

4.82E-02

3.45E-02

2.06E-01

1.90E-01

1.97E+00

1.53E+00

Pb

1.54E-02

1.07E-02

3.73E-02

2.83E-02

9.00E-02

7.40E-02

Cr

1.10E-04

7.07E-05

1.71E-04

1.31E-04

2.83E-04

2.43E-04

Cu

4.45E-02

3.20E-02

1.36E-01

1.04E-01

4.18E-01

3.35E-01

Zn

4.67E-02

3.24E-02

9.83E-02

7.47E-02

1.96E-01

1.63E-01

Ni

4.48E-02

2.96E-02

8.05E-02

6.20E-02

1.85E-01

1.52E-01

7.10E-01

4.83E-01

1.74E+00

1.36E+00

5.06E+00

4.08E+00

HI (∑ HQ)

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Figure 1

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Figure 2

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