Health risk assessment of process-related contaminants in bread.

Bread constitutes a popular and even daily component of human diet world-wide, Iran included. However, there are concerns that various processing methods such as frying and baking could result in the production of potentially a source of known carcinogens such as acrylamide (AA) and benzo(a)pyrene (BaP). The present study tried to perform a risk assessment on seven categories of bread consumers, based on age and gender, calculating the Target Hazard Quotient (THQ), Incremental Lifetime Cancer Risk (ILCR), and Margin of Exposure (MOE) related to the dietary intake of AA and BaP. AA and BaP were analyzed in 87 bread samples using LC-MS/MS and GC-MS/MS, respectively. The results indicated that more than 94% (mean concentration: 25.9 ng/g) and about 20% (mean concentration: 1.98 ng/g) of the samples were contaminated with AA and BaP, respectively. The THQ of AA intake through bread consumption for seven categories was in the following decreasing order: semi-industrial Sangak bread of Shiraz (SIS-Sh)> traditional Sangak bread of Shiraz (TS-Sh)> traditional Sangak bread of Tehran (TS-Th)> commercial bread of Tehran (C-Th). The non-neoplastic and neoplastic MOE for AA (˂10,000) indicates a high risk of exposure for all people in Tehran and Shiraz through the consumption of all tested bread. Due to the consumption of TS, SIS, and C bread, the BaP MOE for all people in Tehran and Shiraz was >10 000, which shows a low health risk for consumers. Our findings showed that ILCR for AA in seven classes of people who had TS-Sh and TS-Th was remarkably higher than ILCR for all categories that consumed the C-Th. BaP ILCR for the people who ingested TS-Th, TS-Sh, SIS-Th, and SIS-Sh was about 2-4 times higher than people who had C-Th. This study indicates that bread is the main source of AA and BaP intake in Iran, which might elevate the cancer risk.

Occurrence of Aflatoxins in Commercial Cereal-based Baby Foods in Iran: A Probabilistic Risk Assessment to Health.

The aim of this study was to assess the occurrence of Aflatoxins (AFs) including B1, B2, G1 and G2 in commercial cereal-based baby foods by HPLC-FLD method in Iran and related risk assessment in three baby age groups (6-12, 12-18, and 18-24 months) using Monte Carlo simulation approach. Results showed an occurrence ranging from 20% to 60% for B1, B2, and G2 aflatoxins, while AFG1 was not detected in any assessed samples. Exposure and risk assessment was estimated to be two groups (aflatoxin B1 and total aflatoxins). The highest estimated dietary exposure to both AFB1 and total AFs was estimated for 6-12 months aged babies, representing 5.81 ng/kg BW/day and 8.55 ng/kg BW/day, respectively. Overall, the margin of exposures to AFB1 and total AFs were lower than 10,000 in all age groups, indicating a health concern about AFB1 and total AFs exposure through cereal-based baby food consumption. High cancer risk for high consumers (P95) of baby food was also estimated in all age groups, calling for immediate intervention due to serious claims that AFB1, is a highly carcinogenic component, causes hepatocellular carcinoma. Risk ranking results indicated the presence of AFB1 is classified as high risk for babies who consume cereal-based foods, which demands the attention of risk managers to reduce or eliminate this risk for the most vulnerable sector of society, whose aged <24 months.

Exposure to multiple mycotoxins in domestic and imported rice commercially traded in Tehran and possible risk to public health.

Mycotoxins are secondary fungi metabolites that induce acute and chronic toxic effects in humans and animals. In the present study, nine mycotoxins including aflatoxins (AFB1, AFB2, AFG1 and AFG2), fumonisins (FB1 and FB2), Ochratoxin A (OTA), deoxynivalenol (DON), and zearalenone (ZEN) were determined in one hundred rice samples collected from Tehran using high performance liquid chromatography (HPLC) with fluorescence or photodiode array detector. In addition, possible risk to public health was investigated by assessing dietary exposure through rice consumption, the margin of exposure (MOE), respective risk of cancer and hazard index (HI) of the monitored mycotoxins in children and adults. The higher mean levels were determined for DON (102.22 µg.Kg-1), followed by FB1 (85.00 µg.Kg-1). For the rests of mycotoxins the levels did not exceed 20 µg.Kg-1. The estimated AFB1 intake for the adults and children through rice consumption exceeds the safe levels established for both carriers and non-carriers of hepatitis B virus. The mean and median determined exposure levels of OTA, DON ZEN and FB1, were found lower than the Provisional Maximum Tolerable Daily Intake (PMTDI) value for both adults and children of Tehran that consuming domestic and imported rice. The mean HI for adults and median HI for adults and children were below one, and mean HI for children was close to one. All the mean, median and maximum MoE values were <10,000 in adults and children, indicating a risk due to AFB1 exposure through rice consumption in Tehran. In addition, the calculated mean cancer risk in adult and child populations of Tehran were 0.27 and 0.64 cases per year per 105 individuals, respectively, that shows population in Tehran could be at risk of cancer due to AFB1 exposure through rice consumption as calculated. So further studies are necessary for the monitoring mycotoxins in rice and different food products as well as estimating average dietary exposure and cumulative exposure assessment of mycotoxins for main foods in IR Iran.

Development of a Sensitive and Rapid Method for Determination of Acrylamide in Bread by LC-MS/MS and Analysis of Real Samples in Iran IR.

A new sample preparation procedure and a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method were developed for the quantitative analysis of acrylamide in bread. The method is based on sample extraction in methanol, purification with Carrez solutions and clean- up with Primary Secondary Amine (PSA).The developed method offers an efficient, inexpensive, easy sample preparation and very sensitive procedure for determination of acrylamide in bread. The use of spiked calibration curves for constructing the calibration curve substantially reduced adverse matrix-related effects. Recoveries were between 96 and 105.3%. Good results were obtained with respect to repeatability (RSDs <11%). The limit of detection and quantification of the method was 0.3 and 1 ng/g, respectively, which shows the method is very sensitive. The developed method was used for the determination of acrylamide in 26 traditional bread samples (Sangak) collected from Shiraz. The results showed that about 96% of Sangak bread samples were contaminated with acrylamide that 64.3 and 33.3 of semi-industrial and traditional Sangak bread were higher than benchmark levels (50 µg/kg), respectively. There are a few reports concerning contamination of Sangak bread samples with acrylamide in Iran. Therefore, this method could be used for a comprehensive survey of acrylamide in Sangak bread samples in the country.

Health risk assessment of aflatoxin M1 in infant formula milk in IR Iran.

In this study, two accurate, precise, selective and sensitive methods were developed for determining aflatoxin M1 (AFM1) in infant formula milk using immunoaffinity column clean-up followed by high performance liquid chromatography (HPLC) with fluorescence detection. The validated methods were used for determination of AFM1 in 29 samples of 6 different infant formula milk brands and the risk of AFM1 in infants aged zero to 6 months old was assessed using cancer risk, Margin of Exposure (MOE) and Hazard Index (HI). Only one sample (3.4%) was contaminated with AFM1. Although the results showed that MOE values for the mean and median exposure to AFM1 was <10,000 in infants, the additional cancer risk due to mean and median exposure to AFM1 in infant <6 months were 0.00010 and 0.00012 additional cases per year per 105 individuals, respectively, which indicates no health concern. In addition, HI values for the mean and median exposure to AFM1 for infants were quite below one which indicates no health concern. To the best of our knowledge, this is the first report on risk assessment of AFM1 in infant formula milk consumed by Iranian infants <6 months old, presenting a low risk for the evaluated groups.

Pesticide Residues Analysis in Iranian Fruits and Vegetables by Gas Chromatography-Mass Spectrometry.

Pesticide residues in fruits and vegetables are one of the highest concerns of consumers who need food safety. In this study, forty-eight pesticide residues from different chemical structures including organochlorine, organophosphorus, organonitrogen, dicarboximides, strobilurin, triazine, pyrethroids, and other chemical groups. In 85 fruits and vegetables were determined and confirmed by GC-MS. The pesticide was extracted with ethyl-acetate, then, the extracts cleaned using high performance gel permeation column chromatography (GPC) and solid phase column (SPE). The mean recoveries of the pesticides were between 81 and 136%. The reproducibility of the relative standard deviation values was 2.1% and 14.8%. Pesticide residues were more frequently found in vegetables (65.5%) than in fruits (26.7%). The limits of detection and quantification of pesticide residues for the method were ranged from 0.003 to 0.06 µg/g and between 0.01 to 0.1 µg/g respectively. The analyzed samples did not contain residues from the monitored pesticides that were higher than the accepted maximum residue limits (MRLs) as adapted by the FAO/WHO Codex Alimentarius Commission.

Aflatoxin M1 in raw, pasteurized and UHT milk marketed in Iran.

Aflatoxin M1 (AFM1) is a 2B Group human carcinogens agent and introduces into the human diet by milk consumption. In the present study 257 raw, 143 pasteurized and 61 UHT milk samples were collected from markets in 117 cities throughout Iran, during winter 2017 and AFM1 contents were determined using a HPLC-FLD method after immunoaffinity clean-up. Mean recovery rates and relative standard deviations of the method were 90.6 ± 5.7%. AFM1 was detected in 252 of 461 (54.7%) samples (mean 31.3 ± 8.8 ng/L), ranging from <10 to 150 ng/L. Three samples (0.6%) exceeded the Iranian maximum limit of 100 ng/L. The mean AFM1 content was significantly higher in raw milk samples obtained from humid climate areas. The extrapolated concentration of AFB1 in dairy cattle feed based on the AFM1 value of milk was 1.96 µg/kg. The AFM1 contamination of milk seems not to be a serious public health concern in Iran.

Health risk assessment of acrylamide in bread in Iran using LC-MS/MS.

Acrylamide is a chemical, often present in bread, legally classified as carcinogen, mutagen and reproductive toxicant. Since bread is consumed both world-wide and in Iran, determination of acrylamide in different types of breads is of high interest. In the present study, acrylamide was monitored in 56 Sangak and 30 industrial bread samples collected from Tehran and Shiraz, using LC-MS/MS (LOQ = 1 ng/g). In addition, the noncarcinogenic risk (target hazard quotient-THQ) and carcinogenic risk (incremental lifetime cancer risk-ILCR) due to ingestion of acrylamide through bread consumption in children and adults were assessed. Acrylamide was detected in more than 90% of the samples tested. The average daily intake of acrylamide in Iran based on exclusive consumption of Sangak bread, was estimated at 145 ng/kg bw/day. Based on the THQ for bread acrylamide in adults and children, the decreasing risk order was: Shiraz semi-industrial Sangak, Shiraz traditional Sangak, Tehran traditional Sangak, Tehran industrial bread. The ILCR of bread acrylamide calculated for adults and children was higher than the permissible lifetime carcinogenic risk value established by USEPA (1.00E-5). Results show that bread is a major source of acrylamide intake by people in Iran and all consumers regardless of age could be at elevated carcinogenic risk.

Exposure to Aflatoxin M1 through Milk Consumption in Tehran Population, Iran.

Milk would be contaminated with Aflatoxin M1 (AFM1), if it was obtained from lactating animal which fed with feedstuffs containing Aflatoxin B1 (AFB1). AFM1 is classified as group 2B, possibly carcinogenic to humans and its exposure to AFM1 through milk consumption is a public concern. The purpose of this study was to determine the AFM1 exposure through liquid milk consumption for adult consumers in Tehran. Forty-five samples including raw, pasteurized, and UHT milk samples were collected from markets in different cities of Tehran province in January and February 2017. The AFM1 was determined by HPLC method after immunoaffinity column clean up. Also, the milk intake was calculated using household budget survey. Finally, the daily intake of AFM1 through milk consumption was estimated using a deterministic approach. From total 45 samples, AFM1 was detected in 36 (80%) samples, although none of the analyzed samples were exceeded Iran legal limit of 0.1 µg/kg. On the basis of the average milk intake, the mean daily exposure to AFM1 was estimated between 0.03 ng/ Kg BW per day (lower bound estimate) and 0.06 ng/ Kg BW per day (upper bound estimate) and the 95th percentile daily exposure was calculated at 0.14 ng/ Kg BW per day. According to these values, it should be expected that the adults of Tehran population are not exposed to a significant risk of Hepatocarcinoma associated with AFM1 intake through milk consumption.

The Effects of House Cooking Process on Residue Concentrations of 41 Multi-Class Pesticides in Rice.

In current study, the effects of Iranian rice cooking method (Kateh) on residue levels of 41 pesticides were investigated. The pesticides were selected according to Iranian National Standards Organization (INSO) regulations and covered 18 permitted and 23 banned pesticides belonging to different chemical classes such as organophosphate, triazole, and carbamate. A 250 g portion of rice sample was soaked in 2.5 L spiked tap water containing studied pesticides at final concentration 2 µg/mL and then, the effects of washing and cooking were investigated. The pesticides were analyzed simultaneously in a single run using positive electrospray ionisation with multiple reaction monitoring (MRM) after extraction with QuEChERS method. The results showed that washing removed different portions of pesticide residues in the range between 12.0-88.1%. Washing effect was not associated with the water solubility of the pesticides but amount of residue binding to rice matrix was a major factor for residue reduction. In Iranian method of rice preparation, cooking process includes boiling and steam cooking. In this study, the amount of the pesticide residues was decreased in the range of 20.7-100%. Under these conditions, volatilization, hydrolysis, and thermal degradation caused the reduction of the pesticide residues.

Exposure Assessment for Some Pesticides through Rice Consumption in Iran Using a Multiresidue Analysis by GC-MS.

In communities which consume rice as main food, importance of risk assessment for contaminants is always taken into consideration by health authorities. The present study is an attempt for monitoring of 56 pesticides from different chemical groups in rice samples collected from local markets in Tehran and estimation of daily intake of interested pesticides through this monitoring. A valid method based on spiked calibration curves and QuEChERS sample preparation was developed for determination of pesticides residue in rice by GC/MS. The analytical results of the proposed method were in good agreement with the proficiency test (FAPAS 0969). One-hundred-thirty-five rice samples were analyzed and 11 pesticide residues were found in 10.4% of the samples. Of which 5.2% were contaminated with unregulated pesticides. None of the samples, which were contaminated with regulated pesticides, had contamination higher than maximum residue limit. The mean estimated dose (ED) was calculated with respect of mean of contamination and mean daily consumption of rice. ED of the found pesticides is much lower than the related ADIs.

Simultaneous Determination of 13 Priority Polycyclic Aromatic Hydrocarbons in Tehran's Tap Water and Water for Injection Samples Using Dispersive Liquid-Liquid Micro Extraction Method and Gas Chromatography-Mass Spectrometry.

Polycyclic aromatic hydrocarbons (PAHs) are classified as persistent and carcinogenic organic pollutants. PAHs contamination has been reported in water. Many of relevant regulatory bodies such as EU and EPA have regulated the limit levels for PAHs in drinking water. In this study, 13 priority polycyclic aromatic hydrocarbons (PAHs) were determined in tap water samples of Tehran and water for injection. Dispersive liquid-liquid microextraction procedure combined with gas chromatography-mass spectrometry was used for the extraction and determination of PAHs in the samples. Under the optimized conditions, the range of extraction recoveries and relative standard deviations (RSDs) of PAHs in water using internal standard (anthracene-d10) were in the range of 71-90% and 4-16%, respectively. Limit of detection for different PAHs were between 0.03 and 0.1 ngmL(-1). The concentration of PAHs in all tap water as well as water for injection samples were lower than the limit of quantification of PAHs. This is the first study addressing the occurrence of PAHs in water for injection samples in Iran using dispersive liquid-liquid micro extraction procedure combined with gas chromatography-mass spectrometry.

Method Development for Simultaneous Determination of 41 Pesticides in Rice Using LC-MS/MS Technique and Its Application for the Analysis of 60 Rice Samples Collected from Tehran Market.

A multi-residue method for simultaneous determination of 41 LC-amenable pesticides in rice, belonging to different chemical classes has been developed in Iran by LC-MS/MS. For the first time the pesticides were analyzed simultaneously in a single run using positive electrospray ionization with multiple reaction monitoring (MRM) after extraction with slightly modified QuEChERS method. The calibration curve for each analyte was linear over the concentration range of 0.02-1.0 µg/g with a correlation coefficient range between 0.993 and 0.999. The LOQ and LOD were .025 µg/g and 0.008 µg/g respectively, for all 41 pesticides and the mean recoveries obtained for three fortification levels (0.025, 0.08 and 0.250 µg/g) were 71-119% with satisfactory precision (RSD<20%). The developed method was used to investigate the occurrence of pesticides in 30 domestic and 30 imported rice samples collected from Tehran market. Five compounds were detected in 11 domestic and 9 imported positive samples in concentration range from 0.032 µg/g to 0.081 µg/g and 0.028 µg/g to 0.074 µg/g, respectively. With the exception of prohibited pesticides, phosphamidon and TCMTB, three permitted pesticides, cinosulfuron, triadimenol and tricyclazole, found in positive rice samples were below MRLs established by Institute of Standards and Industrial Research of Iran (ISIRI).

Effect of cooking process on the residues of three carbamate pesticides in rice.

A gas chromatography mass spectrometry with spike calibration curve method was used to quantify three carbamate pesticides residue in cooked white rice and to estimate the reduction percentage of the cooking process duration. The selected pesticides are three carbamate pesticides including carbaryl and pirimicarb that their MRL is issued by "The Institute of Standards of Iran" and propoxur which is used as a widely consumed pesticide in rice. THE ANALYTICAL METHOD ENTAILED THE FOLLOWING STEPS: 1- Blending 15 g cooked sample with 120 mL acetonitrile for 1 min in solvent proof blender, 2- Adding 6 g NaCl and blending for 1 min, 3- Filtering upper layer through 25 g anhydrous Na2SO4, 4- Cleaning up with PSA and MgSO4, 5- Centrifuging for 7 min, 6- Evaporating about 0.3 mL and reconstituting in toluene till 1 mL, 7- Injecting 2 µL extract into GC/MS and analyzing by single quadruple selected ion monitoring GC/MS-SQ-SIM. The concentration of pesticides and the percentage of pesticides amounts after the cooking were determined by gas chromatography mass-spectrometry (GC/MS) using with interpolation of the relative peak areas for each pesticide to internal standard peak area in the sample on the spiked calibration curve. Calibration curve was linear over the range of 25 to 1000 ng/g, and LOQ was 25 ng/g for all three pesticides. The percent of loss for the three pesticides were 78%, 55% and 35% for carbaryl, propoxur and pirimicarb respectively. Different parameters such as vapor pressure, boiling point, and suspect ability of the compound to hydrolysis, could be responsible for the losing of pesticides during the cooking process.

Fumonisin production by Fusarium species isolated from freshly harvested corn in Iran.

Fifty-one strains of Fusarium verticillioides and F. proliferatum isolated from corn collected from four different geographic areas in Iran, namely Fars, Khuzestan, Kermanshah and Mazandaran (an endemic oesophageal cancer (OC) area) were evaluated for their ability to produce fumonisins B1 (FB1), B2 (FB2) and B3 (FB3) in corn culture. Fumonisin levels were determined by high-performance liquid chromatography. All tested strains of F. verticillioides and F. proliferatumproduced fumonisins within a wide range of concentrations, 197-9661 microg/g, 18-1974 microg/g, and 21-1725 microg/g for FB1, FB2, and FB3, respectively. The highest mean concentrations of FB1, FB2, and FB3 were 3897, 806 and 827 microg/g, respectively. Overall, 61% of the F. verticillioides and F. proliferatum strains produced higher levels of FB3 than FB2. The mean ratios of FB1:FB2, FB1:FB3 and FB1:total fumonisins were 8, 7 and 0.7 for F. verticillioides and 5.7, 10.7 and 0.7 for F. proliferatum, respectively. Significant differences in some of the meteorological data (rainfall, relative humidity and minimum temperature) from the four provinces were observed. Fumonisin levels produced by F. verticillioides strains isolated from Khuzestan province (tropical zone) were significantly (P < 0.01) higher than the other three provinces. This is the first report of the fumonisin-producing ability of F.verticillioides and F. proliferatum strains isolated from corn harvested from different geographic areas in Iran.