Next Article in Journal
Nutritionally Optimized, Culturally Acceptable, Cost-Minimized Diets for Low Income Ghanaian Families Using Linear Programming
Next Article in Special Issue
Protective Effect of Aplysin Supplementation on Intestinal Permeability and Microbiota in Rats Treated with Ethanol and Iron
Previous Article in Journal
Consumption of 100% Pure Fruit Juice and Dietary Quality in French Adults: Analysis of a Nationally Representative Survey in the Context of the WHO Recommended Limitation of Free Sugars
Previous Article in Special Issue
Effects of Consuming a Low Dose of Alcohol with Mixers Containing Carbohydrate or Artificial Sweetener on Simulated Driving Performance
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:

Association between Urinary Aflatoxin (AFM1) and Dietary Intake among Adults in Hulu Langat District, Selangor, Malaysia

Siti Husna Sulaiman
Rosita Jamaluddin
* and
Mohd Redzwan Sabran
Department of Nutrition and Dietetics, Faculty of Medical and Health Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia
Author to whom correspondence should be addressed.
Nutrients 2018, 10(4), 460;
Submission received: 27 February 2018 / Revised: 4 April 2018 / Accepted: 4 April 2018 / Published: 7 April 2018
(This article belongs to the Special Issue Nutrition Solutions for a Changing World)


Aflatoxin is a food contaminant and its exposure through the diet is frequent and ubiquitous. A long-term dietary aflatoxin exposure has been linked to the development of liver cancer in populations with high prevalence of aflatoxin contamination in foods. Therefore, this study was conducted to identify the association between urinary aflatoxin M1 (AFM1), a biomarker of aflatoxin exposure, with the dietary intake among adults in Hulu Langat district, Selangor, Malaysia. Certain food products have higher potential for aflatoxin contamination and these were listed in a Food Frequency Questionnaire, which was given to all study participants. This allowed us to record consumption rates for each food product listed. Concomitantly, urine samples were collected, from adults in selected areas in Hulu Langat district, for the measurement of AFM1 levels using an ELISA kit. Of the 444 urine samples collected and tested, 199 were positive for AFM1, with 37 of them exceeding the limit of detection (LOD) of 0.64 ng/mL. Cereal products showed the highest consumption level among all food groups, with an average intake of 512.54 g per day. Chi-square analysis showed that consumption of eggs (X2 = 4.77, p = 0.03) and dairy products (X2 = 19.36, p < 0.01) had significant associations with urinary AFM1 but both food groups were having a phi and Cramer’s V value that less than 0.3, which indicated that the association between these food groups’ consumption and AFM1 level in urine was weak.

1. Introduction

Foodborne disease is a global concern, as the Centers for Disease Control and Prevention [1] estimates that each year there are 48 million people who are affected, 128,000 of whom are hospitalized and 3000 die. Foodborne diseases are commonly caused by bacteria, viruses, parasites, harmful toxins and chemicals. Mycotoxins, one of the etiologic agents of foodborne diseases, are produced by fungi which can cause mycotoxicosis [2]. There are many mycotoxins that have the potential to contaminate food products and agricultural commodities. Aflatoxin is produced by Aspergillus species of fungi such as Aspergilllus flavus, A. parasiticus and A. nomius [3]. These fungi are found in the warm and humid climates prevailing in the tropical and sub-tropical geographic latitudes [4]. In addition, improper food storage and production procedures promote the growth of these fungi, and subsequently aflatoxin contamination, in many agricultural commodities.
The most common aflatoxin metabolites found in foodstuff are aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1), and aflatoxin G2 (AFG2) [5]. Of these, AFB1 is the most toxic as it has been classified by the International Agency for Research on Cancer (IARC) as Group 1 carcinogen [6]. Besides their occurrence in the foods and agricultural commodities, aflatoxin can be detected in biological samples, resulting from exposure through the diet. In fact, the assessment of human and animal exposure to aflatoxin, through the detection of aflatoxin biomarkers in biological samples such as in serum and urine, is significant to determine the extent and rate of aflatoxin exposure.
The metabolism of aflatoxins, particularly AFB1 in the liver produces several metabolites such as AFB1-lysine adduct [7], AFB1-N7-guanine adduct [8], and urinary AFM1 [9], and these biomarkers have been used in many epidemiological studies reported in the literature [7,8,9,10] assessing the extent of human exposure to aflatoxin. For example, the detection of urinary AFM1 is a good indicator to determine recent aflatoxin exposure of 1–3 days. In addition, Zhu et al. [10] found a good correlation between total dietary AFB1 and the excretion of AFM1 in urine.
In Malaysia, the occurrence of aflatoxin in foodstuffs such as cereals, peanuts, spices and their products has been reported. In a review by Mohd-Redzwan et al. [11], on the occurrence of aflatoxin in Malaysia, the authors highlighted the use of aflatoxin biomarkers as a potential tool to provide a better assessment of aflatoxin exposure as different individuals might be exposed at different rates. Indeed, it is of great significance to find the association between aflatoxin biomarkers and human dietary consumption to reflect aflatoxin exposure from the diet. Hence, this research was conducted to determine the association between a urinary AFM1 biomarker and food consumption among adults in Hulu Langat district, Selangor, Malaysia.

2. Materials and Methods

2.1. Study Respondents

The study respondents, including both male and female, ranged in age from 18 to 60 years old, and resided in Hulu Langat district, which is the fifth largest (out of nine total districts) in Selangor, Malaysia. This district consists of six sub-districts (Beranang, Cheras, Hulu Langat, Hulu Semenyih, Semenyih and Kajang). Hulu Langat district was chosen due to desirable density of ethnic population and its proximity to Universiti Putra Malaysia (UPM). A total of 468 respondents underwent a screening examination that included a medical history. All of them gave informed consent and the study was approved by the Ethics Committee for Research Involving Humans at UPM (FPSK (EXP16) P047).
Of the 468 screened respondents, 455 met the following study criteria: (1) in good health; (2) not taking any medications or supplements; (3) not smoking; (4) not following a restricted diet; and (5) not pregnant and not in postpartum period. For the analyses conducted in the current study, 11 respondents were excluded for the following reasons: missing FFQs (n = 5), aged more or less than the required range (n = 2), and non-returnable urine container (n = 4).

2.2. Dietary Assessment

Dietary consumption data were collected using an FFQ [12] and took five months to compile. The elaboration of food groups in the FFQ was obtained from previous studies [4,13]. All respondents completed a self-administered FFQ that consisted of 197 food items that are susceptible to aflatoxin contamination (e.g., cereals products, nuts and legumes, eggs, and dairy products) [4,13]. Typical portion sizes for each food item are considered medium-sized, based on the Malaysian Food Serving Size Album [14] and the list of food item weights in household measures [15]. The amount of food intake (g/day) was calculated based on Norimah and colleagues’ formula [12].
For further statistical analysis such as chi-square analysis, all food groups were divided into two groups, low and high, based on the median total of intakes.

2.3. Urine Analyses

Fifteen-milliliter morning urine samples were collected in containers, and then delivered to UPM on the same day using a specific ice box. These samples were kept frozen at −80 °C in the Nutrition Laboratory 3, Faculty of Medicine and Health Sciences, UPM until their analyses began.
Quantification of urinary AFM1 was undertaken following the protocol of Mohd-Redzwan et al. [16]. The level of AFM1 was analyzed using an ELISA kit specifically designed for the determination of urinary AFM1 (Helica Biosystems, Inc., Santa Ana, CA, USA). The debris and precipitate were removed through centrifugation at 3000× g for five minutes (Kubota Centrifuge Model 2810, Tokyo, Japan), and supernatant was used for the determination of AFM1 according to manufacturer instructions. The protocol’s washing step involved the use of an automated microplate washer (Drop ELISA Washer, RADM, Roma, Italy). A microplate reader (SIRIO S Microplate Reader, RADM, Roma, Italy) was used to measure absorbance at a wavelength of 450 nm. As for method validation, urine samples were spiked with 1.5 ng/mL AFM1 standard and the samples were processed as per protocol.
Before statistical analyses were conducted, the limit of detection (LOD) was calculated based on the lowest concentration of AFM1 obtained from the measurement of urine samples. The concentrations were first extrapolated from the standard curve of AFM1. The r2 of the standard curve was 0.998 and LOD was calculated using the formula by Shrivastava and Gupta [17]:
LOD = 3.3σ/s
Where, σ is the standard deviation of the y-intercept of the regression line and s is the slope of calibration curve.
The calculated LOD was found to be 0.64 ng/mL. The AFM1 readings were then segregated into two groups, positive and negative, based on the presence or absence of detectable AFM1. Then, Chi-square statistical analysis was conducted for each reading. Negative samples were categorized as simple with no detectable level of AFM1, whereas positive samples were categorized as samples with detectable level of AFM1.

2.4. Statistical Analysis

Basic demography characteristics were presented as mean ± standard deviation when the data were continuous, and they were presented in contingency tables if the data were binary or categorical. The intake of potentially aflatoxin-contaminated food was presented as average grams per day (g/day). Our use of Chi-square was to determine the association between urinary AFM1 biomarker level and dietary intake. The level of significance was accepted as p < 0.05. The strength of association between variables was determined through Phi and Cramer’s V value [18].

3. Results

3.1. Socio-Demographic Characteristics

Table 1 represents the socio-demographic characteristics of respondents. Out of 444 respondents, 246 were females and 198 were males. Most of the respondents were single (n = 323, 72.1%) with a median age of 24 years old. About half of the respondents were Chinese (n = 211, 47.5%) and most of them had tertiary educational level. Besides, most of the respondents had personal income of less than RM1500 (USD 384.87) while the total household income was RM3500 (USD 898.03) and above.

3.2. Dietary Intake

Table 2 represents the median intake based on food group. Cereal products were consumed at the highest rate, while eggs had the lowest intake rate, among the respondents.
Table A1 (Appendix A) shows the means, standard deviations, medians as well as ranges for the daily intakes of potentially aflatoxin-contaminated foods assessed by the FFQs. The top three highly consumed food items were white rice (270.79 g/day), flavored rice (40.54 g/day) and chocolate-flavored milk (38.11 g/day). The two rice items come from the cereal products food group, and the milk is an item from the dairy product food group.

3.3. Urinary AFM1 Biomarker Level

Negative samples were categorized as samples with no detectable level of AFM1, whereas positive samples were categorized as samples with detectable level of AFM1 through the extrapolation from the standard curve. Of 444 analyzed urine samples, 199 samples were positive for AFM1 while the rest (n = 245) were negative. From those with positive AFM1, 37 of them exceeded the limit of detection (LOD) of 0.64 ng/mL. Those 37 samples had urinary AFM1 ranging from 0.65 to 5.34 ng/mL, with an average of 1.23 ng/mL.

3.4. Association between Urinary AFM1 Biomarker Level and Socio-Demographic Factors and Dietary Intake

Our findings (Appendix Table A2) indicate several significant association between socio-demographic factors, ethnicity (p < 0.01), age (p < 0.05) and household income (p < 0.01), and urinary AFM1 among respondents in Hulu Langat district, Selangor, Malaysia. There were also weak associations between urinary AFM1 and consumption of eggs (X2 = 4.77, p = 0.03) and dairy products (X2 = 19.36, p < 0.01), whereas no other food groups showed any significant associations (Table 3).

4. Discussion

Aflatoxins are present ubiquitously in nuts and nuts products, cereals, and spices and herbs, which are widely used among Malaysians as the main ingredients in cooking [3]. Milk and dairy products as well as eggs and meat products are contaminated as animals consume aflatoxin-contaminated feed. The rate of aflatoxin contamination in foodstuffs varies from one products to another. A study in Terengganu, Malaysia [22], found that 19 out of 53 dairy products samples were positive with AFM1, ranging from 3.5 to 100.5 ng/L, and this contamination levels were still safe according to Malaysian Food Regulation 1985 (>50 ng/L) [23]. Malaysians are one of the Asian citizens that consume rice frequently, as a staple food [24]. Soleimany et al. [19] found detectable levels of aflatoxin, ranging from 0.19 to 3.96 ng/g, in rice sold in the Malaysian General Market in Kuala Lumpur. Besides, there was also a study [20] that found that a maize and two rice samples from the Malaysian markets had aflatoxin levels exceeding the European regulatory limits for aflatoxin, i.e., 4 ng/g. Other than that, another study by Shahzad et al. [25] discovered about 35% of the rice samples was positive with aflatoxin contamination, where brown rice showed the highest aflatoxin contamination (12.4 µg/kg). Another foodstuff that is easily contaminated by aflatoxin is nuts and legumes. A study conducted in Penang, Malaysia [21] found 32 out of 196 nuts and its based products samples were positive with aflatoxin ranging from 16 µg/kg to 711 µg/kg for total aflatoxin and this range exceeded the limit value set of 5 µg/kg for all foodstuffs and 15 µg/kg for processed groundnuts based on the Malaysian Food Regulation 1985. On the other hand, the contamination of aflatoxin in egg samples around Malaysia was less reported but its occurrences do exist in other countries. For example, there was a report that documented the presence of aflatoxin in 22 out of 80 eggs samples collected from central areas of Punjab, Pakistan [26]. The level of aflatoxin B1 reported ranged from 0.5 to 3.19 µg/kg for farm eggs and 0.5 to 1.98 µg/kg for domestic eggs. In fact, the level of aflatoxin in eggs commodity depends on the level of other microorganisms on the eggs such as Salmonella spp., Candida spp. and coccidiosis organisms [27]. As the level of these microorganisms increases, the level of aflatoxin in eggs decreases.
The contamination of food commodities by aflatoxin B1, the most toxic of aflatoxin is common in countries with subtropical and tropical climate. Aflatoxin M1 (AFM1) is a metabolite of AFB1 which can be detected in the urine and milk of exposed animals and humans. According to Zhu et al. [10], 1.23–2.18% of ingested AFB1 is found as AFM1 in the urine. The authors found correlation coefficient (r) of 0.65 between AFB1 intake and AFM1 in the urine among population in China. In fact, a preliminary study conducted in Malaysia showed a linear relationship between dietary AFB1 exposure and urinary AFM1, and the population was exposed to 0.0262 µg/day/kg AFB1 through the diet [16]. Other than that, an intervention study [28] also showed a significant correlation between the concentration of urinary AFM1 with the consumption of nut-based products (r = 0.258) in a probiotic intervention study.
The consumption of aflatoxin-contaminated food was higher in countries from Southeast Asia compared to developed countries from the Western Europe [29] and it shows that the occurrence of aflatoxin is higher in developing countries, where the prevalence of aflatoxin contamination in foods and agricultural product is high [30]. A study conducted in Bangladesh showed a significant difference of AFM1 level in two different areas, rural and urban [31]. The authors found that the mean urinary AFM1 level was higher among residents in rural areas (99 ± 71 pg/mL) compared to urban areas (54 ± 15 pg/mL). Besides, the highest AFM1 found in the study was among residents in the 50–60 years age group. In contrast, the present study found that the highest AFM1 levels were among the younger age group residents (≤24 years of age). In Malaysia, a study found that AFB1 exposure ranged from 24.3 to 34 ng/kg body weight/day [32]. Another study, by Mohd-Redzwan et al. [33], found that urinary AFM1 level in a population in Serdang, Selangor was 18.8 ± 28.6 pg/mL, ranging from 2.4 to 100.4 pg/mL, which also differs from our findings (average = 1.23 ng/mL). Urinary AFM1 level among Chinese respondents were 3.20 times higher compared to non-Chinese respondents [21]. Their findings were comparable to a previous study in Penang conducted by Leong et al. [21], which reported high occurrence of aflatoxin among Chinese respondents was related to the higher intake of food commodities that have high risk of being contaminated by aflatoxin-producing fungi such as cereal, eggs, nuts and legumes and dairy products. Leong et al. [21] further added that food preferences based on cultural differences and food choices may be the cause of the significantly high level of aflatoxin biomarkers.
In our study, 199 respondents had positive occurrence of urinary AFM1, who primarily consumed cereal-based products such as white rice (270.77 ± 197.96 g/day). This finding in agreement with the average intake of white rice among Malaysians of 275.03 g/day as reported by the Food Consumption Statistics of Malaysia [34]. Hence, it is postulated that high consumption of cereals, especially white rice, could be associated with high percentage of AFM1 occurrence in urine among the respondents. For example, previous studies reported a significant correlation between the occurrence of aflatoxin biomarkers in humans and consumption of cereal-based products [35,36]. In the present study, no significant association was detected between urinary AFM1 with the consumption of cereal products. However, respondents who consumed more than 22.71 g/day of egg were more likely to have positive occurrence of AFM1 in the urine, as shown in Table 3.
To the best of our knowledge, no associations have been reported in the literature on the intake of eggs and the occurrence of aflatoxin biomarkers in biological samples such as in urine and serum. The presence of aflatoxin metabolite in eggs can be explained when the poultry animals and birds are feeding on aflatoxin-contaminated feed. Aflatoxins are then metabolized by the liver, where the end-products of the metabolism such as AFB1-lysine adduct [7], AFB1-N7-guanine adduct [8], and urinary AFM1 [9] are produced. Aflatoxin contamination of eggs has been reported [37]. In fact, the first aflatoxicosis case related to aflatoxin contamination was reported in turkeys and ducklings in the 1960s, where millions died as a result of consuming AFB1-contaminated feeds [38].
The present study also found significant association between dairy product consumption and occurrence of urinary AFM1 among respondents. Respondents whose daily intake of dairy products exceeded 77.68 g/day were determined to be more likely to have detectable levels of urinary AFM1. This finding in agreement with a study conducted by Mohd-Redzwan et al. where the level of urinary AFM1 was significantly higher (2.67 ± 2.27 ng/mL) among respondents with high intake of dairy products than those with low intake [16]. In Malaysia, the permissible level of aflatoxin in food and agricultural commodities is regulated in Malaysian Food Regulations 1985 [23] which permits a maximum level of aflatoxin in dairy products of 0.5 µg/kg, similar to the maximum level suggested by Codex Alimentarius Commission (CAC) for the international reference [39].
Although the other food groups examined in this study did not show significant association with the occurrence of urinary AFM1, it should be noted that these food groups (e.g., cereal-based products, and nuts and legumes) are susceptible to fungal infection and aflatoxin contamination. Most of the occurrences were related to the consumption of aflatoxin-contaminated staple food. For example, the largest outbreak of human aflatoxicosis reported so far occurred in Kenya during 2004 [40]. This case reported as high as 8000 µg/kg of aflatoxin in maize and this contamination led to 125 deaths. Other than maize, rice is another staple food cultivated in subtropical areas that is highly exposed to aflatoxin. The Food and Agriculture Organization (FAO) reported that 15% of the harvested rice is damaged annually due to poor storage conditions, which encourage the growth of mycotoxin-producing fungi [41]. Ephrem [42] found that contamination of aflatoxin in rice was as high as 2830 µg/kg, which is higher than the levels of contamination in wheat and maize, as reported in several cases. Ephrem [42] also emphasized that some of the foods may lack of pre-harvest storage practice. Thus, the growth of aflatoxin-producing fungi occurs and allows aflatoxin contamination. In addition, food groups such as nuts and legumes are suitable for the growth of aflatoxin producing fungi due to high fat contents of this food group [42]. A study in Malaysia found that 16.33% of the collected nuts and nut-based products samples were contaminated by aflatoxins, ranging from 16.6 µg/kg to as high as 711 µg/kg [43]. This level of contamination is higher than several studies conducted in Pakistan (less than 15 µg/kg) [44] and Iran (less than 16 µg/kg) [45].
Most mycotoxins are stable during food processing [46] so it can even show up in food products such as peanut butter and many processed products. However, certain food processing techniques can reduce the toxicity of aflatoxin in food products through physical removal and chemical or enzymatic decontamination [46]. That must be one of the reasons for the significant association between AFM1 levels and the consumption of eggs and dairy products discovered from this study. The highest consumption of food from food group of eggs and dairy products were hen eggs (mean ± standard deviation = 33.51 ± 44.64) and chocolate-flavored milk (mean ± standard deviation = 38.07 ± 92.71). Both food items are subjected to one or more processing steps before consumption. For example, the processes involved in producing flavored milk are pasteurization, sterilization and boiling, which reportedly can reduce the concentration of aflatoxin in milk [47]. Previous studies also mentioned that boiling, sterilization and pasteurization reduce aflatoxin in dairy products by 14.5%, 12.2% and 7.6%, respectively [48,49]. Eggs also undergo pasteurization and cooking processes such as boiling, frying, poaching, and steaming before they are consumed. Thus, the aflatoxin concentration in eggs can be reduced too. Food processing that applies heat such as boiling, roasting and steaming can reduce aflatoxin in food by 50–70% [50]. These high temperature processes are not the best solution to reduce aflatoxin levels in certain foods, such as legumes as high heat can deteriorate their nutrients and vitamins.

5. Conclusions

Results from this study indicate a low exposure of AFB1 in Hulu Langat district, Selangor, Malaysia through the detection of urinary AFM1. Future study is encouraged to focus on broader areas which cover the whole of Malaysia as well as to examine the extent of aflatoxin exposure among the general population of Malaysia. The measurement of the AFM1 in urine samples indicated recent exposure of aflatoxin AFM1 within the preceding 24 h. Since this biomarker is short-lived, and its measurement in urine samples may vary from day to day as well as the dietary intake of the individual, it may not be suitable for assessing long-term aflatoxin exposure. Thus, it is suggested to use other biological samples such as serum AFB1-lysine adduct and hair, which can represent the long-term effect of aflatoxin exposure [51].


This research was financially supported by UPM Putra Research Grant-Inisiatif Putra Siswazah (GP-IPS) (No: 9500200). Siti Husna S. would like to thank School of Graduate Studies, Universiti Putra Malaysia for the Graduate Research Fellow (GRF) scholarship and the Ministry of Higher Education.

Author Contributions

S.H.S, M.R.S. and R.J. conceived and designed the experiments; S.H.S. performed the experiments; S.H.S. analyzed the data; M.R.S. and R.J. contributed reagents/ materials/ analysis tools; S.H.S. wrote the paper.

Conflicts of Interest

The authors declare no conflict of interest.

Appendix A

Table A1. Intake of aflatoxin-potentially-contaminated food (n = 444).
Table A1. Intake of aflatoxin-potentially-contaminated food (n = 444).
Food GroupFood ItemsMean ± Standard DeviationMedianRange
Cereal productsWhite rice 1270.77 ± 197.96255.090–1431.00
Brown rice14.27 ± 72.630.140–1170.00
Flavored rice (“Nasi briyani”, fried rice) 240.55 ± 103.320.830–1150.00
Rice porridge5.92 ± 16.510.250–166.00
Glutinous rice3.24 ± 14.460.160–190.00
Noodles 329.72 ± 71.2415.570–981.00
Bihun/kway teow/laksa/laksam/loh shi fun a24.70 ± 51.0411.360–590.00
Pasta5.33 ± 15.370.230–163.43
Sagu/ambuyat/linut 4,b1.80 ± 0.950.030–172.00
Bread25.16 ± 31.6813.680–294.00
Whole meal bread8.93 ± 34.270.490–600.00
Bread bun7.83 ± 21.401.000–270.00
Roti canai c27.96 ± 58.9411.070–665.00
Capati d7.77 ± 23.600.660–200.00
Tosai e7.27 ± 16.700.770–160.00
Breakfast cereal4.51 ± 12.780.090–144.00
Cereal grains prepared by water6.87 ± 36.620.020–520.00
Corn3.44 ± 11.060.090–126.86
Wheat7.22 ± 31.090.230–396.00
Barley9.29 ± 46.440.310–624.00
EggsHen eggs 133.51 ± 44.6422.710–484.57
Duck eggs 30.81 ± 7.400.020–146.00
Quail eggs 40.16 ± 1.000.010–13.71
Salted egg 22.02 ± 11.280.040–194.00
Nuts and legumesLegumes (Green bean, kacang kuda, red bean)5.68 ± 18.060.090–230.00
Groundnut 37.39 ± 123.050.020–2600.00
Taufufa f3.00 ± 7.730.240–70.86
Taufu g,137.89 ± 86.800.780–868.57
Fermented soy beans (Tempe)5.68 ± 20.750.120–213.00
Steamed redbean bun2.30 ± 7.920.070–80.00
Peanut butter biscuit/wafer1.44 ± 4.770.020–56.00
Kuih kacang h0.60 ± 2.890.010–38.00
Peanut sauce2.50 ± 11.450.020–172.00
Rempeyek i0.64 ± 4.850.010–90.00
Almond1.02 ± 4.020.020–45.00
Rojak sauce j1.37 ± 6.450.010–75.00
Cashew nut0.83 ± 3.630.040–39.60
Canned braised peanut0.49 ± 3.82 0.010–51.00
Brazil nut 40.16 ±1.280.0010–17.50
Almond biscuit2.46 ± 8.700.050–87.00
Peanut biscuit0.91 ± 3.180.020–38.40
Pistachio0.32 ± 1.600.0040–22.86
Chestnut0.92 ± 9.510.020–184.00
Hazelnut biscuit0.58 ± 3.840.0040–60.00
Peanut soup6.44 ± 25.750.160–215.00
Ice cream with nuts4.07 ± 11.540.110–138.00
Walnut0.90 ± 5.190.020–84.00
Cashew nut biscuit0.23 ± 1.620.0030–30.00
Almond powder0.24 ± 1.400.0040–15.01
Roasted hazelnut0.26 ± 1.540.0020–21.43
Peanut slice0.19 ± 1.710.0020–34.00
Ais kacang/ABC k,220.93 ± 68.691.120–1150.00
Cake or bread with nuts3.40 ± 8.590.150–72.20
Dairy productsCow’s fresh milk 232.85 ± 67.360.580–450.00
Goat’s fresh milk2.43 ± 18.100.040–300.00
Chocolate flavoured milk 138.07 ± 92.710.520–855.04
Strawberry flavoured milk12.59 ± 51.190.130–500.00
Coffee flavoured milk 317.70 ± 62.580.140–500.00
Full cream yogurt6.48 ± 35.950.660–300.00
Low fat yogurt8.35 ± 25.270.180–300.00
No fat yogurt1.89 ± 16.210.100–300.00
Full cream yogurt with fruits3.47 ± 23.260.200–300.00
Low fat yogurt with fruits2.51 ± 17.270.350–342.86
No fat yogurt with fruits2.57 ± 21.820.170–300.00
Full cream yogurt drink0.92 ± 4.910.060–42.86
Low fat yogurt drink2.76 ± 16.120.310–300.00
Regular powdered milk2.12 ± 6.080.010–42.00
No fat powdered milk0.12 ± 1.390.010–28.00
Evaporated milk unsweetened13.04 ± 84.810.070–1342.29
Cheddar cheese0.86 ± 3.460.010–36.00
Mozarella0.24 ± 1.300.020–15.00
Swiss cheese0.13 ± 1.600.010–30.00
Parmesan0.18 ± 1.660.010–30.00
Ricotta0.04 ± 0.250.020–2.14
Cottage cheese0.04 ± 0.250.010–4.29
American cheese 40.02 ± 0.140.010–2.14
Full cream pasteurized milk5.13 ± 29.510.280–450.00
Low fat pasteurized milk1.82 ± 12.850.140–150.00
High calcium pasteurized milk1.91 ± 11.510.150–150.00
No fat pasteurized milk0.61 ± 4.420.020–42.86
Full cream sterilized milk1.27 ± 8.900.070–150.00
Low fat sterilized milk2.16 ± 16.340.120–150.00
No fat sterilized milk0.32 ± 2.250.010–21.43
Cultured milk (Yakult, Vitagen, Sustgen)9.69 ± 38.880.710–535.71
3 in 1 (Milo, Nescafe, Nestum)12.12 ± 41.010.320–560.00
Sweetened condensed milk2.35 ± 9.200.100–76.00
a Bihun/kway teow/laksa/laksam/loh shi fun = variety type of noodles consumed by Malaysian; b Sagu/ambuyat/linut = starchy food extracted from a tropical palm stems (especially Metroxylon sagu spp.); c Roti canai = oiled flatbread; d Capati = wholemeal, flat pancake-like bread; e Tosai = Indian pancake; f Taufufa = Soy bean pudding; g Taufu = soy bean curd; h Kuih kacang = mung bean fritters; i Rempeyek = deep-fried savory Javanese cracker; j Rojak sauce = Spicy, thick, brown sauce with crushed peanut eaten with Malaysian fruits salad; k Ais kacang/ABC = Sweet shaved ice with beans. 1 The highest consumption of food; 2 the second highest consumption of food; 3 the third highest consumption of food; 4 the lowest consumption of food.
Table A2. AFM1 level based on socio-demographic factors (n = 37).
Table A2. AFM1 level based on socio-demographic factors (n = 37).
Socio-Demographic FactorsnAFM1 Level (ng/mL) Mean± SEM ap-Value bRange
Gender 0.810.65–5.34 ng/mL
 Male141.04 ± 0.10
 Female231.35 ± 0.23
Marital status 0.19
 Married151.31 ± 0.21
 Single/Divorced/Others221.18 ± 0.21
Ethnic 0.01 **
 Chinese181.16 ± 0.14
 Non-Chinese191.30 ± 0.26
Educational level 0.62
 Low130.95 ± 0.12
 High241.39 ± 0.22
Personal income 0.75
 ≤RM1500151.00 ± 0.10
 ≥RM1501221.39 ± 0.24
Household income 0.01 **
 ≤RM150030.84 ± 0.16
 ≥RM1501341.27 ± 0.16
Age 0.05 *
 24 and below271.36 ± 0.20
 25 and above100.90 ± 0.05
a Standard error mean, b Obtained from Chi-square analysis, * p < 0.05, ** p < 0.01.


  1. Foodborne Germs and Illnesses. Centers for Disease Control and Prevention. Available online: (accessed on 9 December 2017).
  2. Robert, K.; Matthias, K.; David, S.; Stefan, M.; Ronald, M.; Irene, N. Determination of Mycotoxins in Foods: Current State of Analytical Methods and Limitations. Appl. Microbiol. Biotechnol. 2010, 86, 1595–1612. [Google Scholar] [CrossRef]
  3. Reddy, K.R.N.; Farhana, N.I.; Salleh, B. Occurance of Aspergillus spp. and aflatoxin B1 in Malaysian food used for human consumption. J. Food Sci. 2011, 76, T99–T104. [Google Scholar] [CrossRef] [PubMed]
  4. Leong, Y.H.; Rosma, A.; Latiff, A.A.; Ahmad, N.I. Exposure assessment and risk characterization of aflatoxin B1 in Malaysia. Mycotoxin Res. 2011, 27, 207–214. [Google Scholar] [CrossRef] [PubMed]
  5. Liu, Y.; Wu, F. Global burden of aflatoxin-induced hepatocellular carcinoma: A risk assessment. Environ. Health Perspect. 2010, 118, 818–824. [Google Scholar] [CrossRef] [PubMed]
  6. International Agency for Research on Cancer (IARC). Aflatoxin in some traditional herbal medicines, some mycotoxins, naphthalene and styrene. In IARC Monograph on the Evaluation of Carcinogenic Risk to Humans; IARC: Lyon, France, 2002; Volume 82, pp. 1–599. [Google Scholar]
  7. Mohd Redzwan, S.; Rosita, J.; Mohd Sokhini, A.M.; Nurul Aqilah, A.R.; Wang, J.S.; Zuraini, A. Detection of serum AFM1-lysine adduct in Malaysia and its association with liver and kidney functions. Int. J. Hyg. Environ. Health 2014, 217, 443–451. [Google Scholar] [CrossRef] [PubMed]
  8. Essigmann, J.M.; Croy, R.G.; Nadzan, A.M.; Busby, W.F., Jr.; Reinhold, V.N.; Buchi, G.; Wogan, G.N. Structural identification of the major DNA adduct formed by aflatoxin B1 in vitro. Proc. Natl. Acad. Sci. USA 1977, 74, 1870–1874. [Google Scholar] [CrossRef] [PubMed]
  9. Gorelick, N.J. Risk assessment for aflatoxin: I. Metabolism of aflatoxin B1 by different species. Risk Anal. 1990, 10, 539–559. [Google Scholar] [CrossRef] [PubMed]
  10. Zhu, J.Q.; Zhang, L.S.; Hu, X.; Xiao, Y.; Chen, J.S.; Xu, Y.C.; Fremy, J.; Chu, F.S. Correlation of dietary aflatoxin B1 levels with excretion of aflatoxin M1 in human urine. Cancer Res. 1987, 47, 1848–1852. [Google Scholar] [PubMed]
  11. Mohd Redzwan, S.; Jamaluddin, R.; Abd Mutalib, M.S.; Ahmad, Z. A mini review on aflatoxin exposure in Malaysia: Past, present and future. Front. Microbiol. 2013, 4, 334. [Google Scholar] [CrossRef] [PubMed]
  12. Norimah, A.K.; Safiah, A.K.; Jamal, K.; Harun, S.H.; Zuhaida, H.; Rohida, S.; Fatimah, S.; Norazlin, S.; Poh, B.K.; Kandiah, M.; et al. Food consumption patterns: Findings from the Malaysian Adult Nutrition Survey (MANS). Malays. J. Nutr. 2008, 14, 25–39. [Google Scholar] [PubMed]
  13. Reddy, K.R.N.; Salleh, B. A preliminary study on the occurrence of Aspergillus spp. and aflatoxin B1 in imported wheat and barley in Penang, Malaysia. Mycotoxin Res. 2010, 26, 267–271. [Google Scholar] [CrossRef] [PubMed]
  14. Album Saiz Sajian Makanan Malaysia. Kajian Pengambilan Makanan Malaysia by Ministry of Health (MOH, Putrajaya). Available online: (accessed on 1 December 2017).
  15. Suzana, S.; Aini, M.Y.; Shanita, S.; Rafidah, G.; Roslina, A. Atlas Makanan: Saiz Pertukaran & Porsi: Atlas of Food Exchanges & Portion Sizes, 2nd ed.; MDC Publishers: Kuala Lumpur, Malaysia, 2009; 127p, ISBN 9677011928. [Google Scholar]
  16. Mohd Redzwan, S.; Rosita, J.; Mohd Sokhini, A.B. Screening of aflatoxin M1, a metabolite of aflatoxin B1 in human urine samples in Malaysia: A preliminary study. Food Control 2012, 28, 55–58. [Google Scholar] [CrossRef]
  17. Shrivastava, A.; Gupta, V.B. Methods for the determination of limit of detection and limit of quantitation of the analytical methods. Chron. Young Sci. 2011, 2, 21–25. [Google Scholar] [CrossRef]
  18. Hinkle, D.E.; Wiersma, W.; Jurs, G.J. Applied Statistics for the Behavioral Sciences, 5th ed.; Houghton Mifflin: Boston, MA, USA, 2002; p. 756. ISBN 978-0618124053. [Google Scholar]
  19. Soleimany, F.; Jinap, S.; Abas, F. Determination of mycotoxins in cereals by liquid chromatography tandem mass spectrometry. Food Chem. 2012, 130, 1055–1060. [Google Scholar] [CrossRef]
  20. Soleimany, F.; Jinap, S.; Faridah, A.; Khatib, A. A UPLC-MS/MS for simultaneous determination of aflatoxins, ochratoxin A, zearalenone, DON, fumonisins, T-2 toxin and HT-2 toxin, in cereals. Food Control 2012, 25, 647–653. [Google Scholar] [CrossRef]
  21. Leong, Y.-H.; Rosma, A.; Latiff, A.A.; Nurul Izzah, A. Associations of serum aflatoxin B1-lysine adduct level with socio-demographic factors and aflatoxin intake from nuts and related nuts products in Malaysia. Int. J. Hyg. Environ. Health 2012, 215, 368–372. [Google Scholar] [CrossRef] [PubMed]
  22. Farah Nadira, A.; Rosita, J.; Norhaizan, S.; Mohd Redzwan, S. Screening of aflatoxin M1 occurrence in selected milk and dairy products in Terengganu, Malaysia. Food Control 2017, 73, 209–214. [Google Scholar] [CrossRef]
  23. Malaysian Food Regulations 1985. Food Safety Information System of Malaysia, Ministry of Health (MOH). Available online: (accessed on 22 December 2017).
  24. Sempere Ferre, F. Worldwide occurrence of mycotoxins in rice. Food Control 2016, 62, 291–298. [Google Scholar] [CrossRef]
  25. Shahzad, Z.I.; Muhammad Rafique, A.; Usman, H.; Muhammad, Z.; Jinap, S. The presence of aflatoxins and ochratoxin A in rice and rice products; and evaluation of dietary intake. Food Chem. 2016, 210, 135–140. [Google Scholar] [CrossRef]
  26. Shahzad, Z.I.; Sonia, N.; Muhammad Rafique, A.; Jinap, S. Natural incidence of aflatoxins, ochratoxins A and zearalenone in chicken meat and eggs. Food Control 2014, 43, 98–103. [Google Scholar] [CrossRef]
  27. Filazi, A.; Sireli, U.T. Occurrence of aflatoxins in food. In Aflatoxins-Recent Advancement and Future Prospects; Razzaghi-Abyane, M., Ed.; INTECH Open Access: Rijeka, Croatia, 2013; pp. 150–151. [Google Scholar]
  28. Mohd Redzwan, S.; Mohd Sokhini, A.M.; Jia-Sheng, W.; Zuraini, A.; Min-Su, K.; Nurul Aqilah, A.R.; Elham, N.N.; Rosita, J. Effect of supplementation of fermented milk drink containing probiotic Lactobacillus casei Shirota on the concentrations of aflatoxin biomarkers among employees of Universiti Putra Malaysia: A randomized, double-blind, cross-over, placebo-controlled study. Br. J. Nutr. 2016, 115, 39–54. [Google Scholar] [CrossRef] [PubMed]
  29. Schleicher, R.L.; McCoy, L.F.; Powers, C.D.; Sternberg, M.R.; Pfeiffer, C.M. Serum concentrations of an aflatoxin-albumin adduct in the National Health and Nutrition Examination Survey (NHANES) 1999–2000. Clin. Chim. Acta 2013, 423, 46–50. [Google Scholar] [CrossRef] [PubMed]
  30. International Agency for Research on Cancer (IARC). Mycotoxin Control in Low- and Middle-Income Countries; IARC Working Group Reports; IARC: Lyon, France, 2015; ISBN 978-9283225102. [Google Scholar]
  31. Ali, N.; Hossain, K.; Blaszkewicz, M.; Rahman, M.; Mohanto, N.C; Alim, A.; Degen, G.H. Occurrence of aflatoxin M1 in urines from rural and urban adult cohorts in Bangladesh. Arch. Toxicol. 2016, 90, 1749–1755. [Google Scholar] [CrossRef] [PubMed]
  32. Chin, C.K.; Abdullah, A.; Sugita-Konishi, Y. Dietary intake of aflatoxins in the adult Malaysian population—An assessment of risk. Food Addit. Contam. Part B Surveill. 2012, 5, 286–294. [Google Scholar] [CrossRef] [PubMed]
  33. Mohd-Redzwan, S.; Rosita, J.; Mohd Sokhini, A.M.; Nurul Aqilah, A.R.; Zuraini, A.; Karimi, G.; Parvaneh, K. Ultra-high performance liquid chromatographic determination o aflatoxin M1 in urine. World Mycotoxin J. 2015, 8, 405–413. [Google Scholar] [CrossRef]
  34. National Health and Morbidity Survey 2014: Malaysia Adults Nutrition Survey (MANS). Volume III, Food Consumption Statistics of Malaysia by Ministry of Health (MOH, Putrajaya). Available online: (accessed on 1 December 2017).
  35. Omar, S.S. Incidence of aflatoxin M1 in human and animal milk in Jordan. J. Toxicol. Environ. Health Part A 2012, 75, 1404–1409. [Google Scholar] [CrossRef] [PubMed]
  36. Nurshad, A.; Meinolf, B.; Khaled, H.; Gisela, H.D. Determination of aflatoxin M1 in urine samples indicates frequent dietary exposure to aflatoxin B1 in the Bangladeshi population. Int. J. Nyg. Environ. Health 2017, 220, 271–281. [Google Scholar] [CrossRef]
  37. Herzallah, S.M. Determination of aflatoxins in eggs, milk, meat and meat products using HPLC fluorescent and UV detectors. Food Chem. 2009, 114, 1141–1146. [Google Scholar] [CrossRef]
  38. Kensler, T.W.; Roebuck, B.D.; Wogan, G.N.; Groopman, J.D. Aflatoxin: A 50 year odyssey of mechanistic and translation toxicology. Toxicol. Sci. 2011, 120, S28–S48. [Google Scholar] [CrossRef] [PubMed]
  39. General Standard for Contaminantsand Toxins in Food and Feed (CODEX STAN 193–1995). Codex Alimentarius Commission (CAC). Available online: (accessed on 22 December 2017).
  40. Wagacha, J.M.; Muthomi, J.W. Mycotoxin problem in Africa: Current status, implications to food safety and health and possible management strategies. Int. J. Food Microbiol. 2008, 124, 1–12. [Google Scholar] [CrossRef] [PubMed]
  41. Dors, G.; Pinto, E.; Badiale-Furlong, E. Migration of mycotoxins into rice starchy endosperm during the parboiling process. Food Sci. 2009, 42, 433–437. [Google Scholar] [CrossRef]
  42. Ephrem, G. Implication of aflatoxin contamination in agricultural products. Am. J. Food Nutr. 2015, 3, 12–20. [Google Scholar] [CrossRef]
  43. Leong, Y.-H.; Ismail, N.; Latif, A.A.; Ahmad, R. Aflatoxin occurrence in nuts and commercial nutty products in Malaysia. Food Contr. 2010, 21, 334–338. [Google Scholar] [CrossRef]
  44. Iqbal, S.Z.; Rafique Asi, M.; Zuber, M.; Akram, N.; Batool, N. Aflatoxins contamination in peanut and peanut products commercially available in retail markets of Punjab, Pakistan. Food Contr. 2013, 32, 83–86. [Google Scholar] [CrossRef]
  45. Amiri, M.J.; Karami, M.; Sadeghi, E. Determination of afb sub 1 in peanut, almond, walnut, and hazelnut in Kermanshah markets, Iran. Int. J. Agric. Crop Sci. 2013, 6, 1199–1202. [Google Scholar]
  46. Karlovsky, P.; Suman, M.; Berthiller, F.; De Meester, J.; Eisenbrand, G.; Perrin, I.; Oswald, I.P.; Speijers, G.; Chiodini, A.; Recker, T.; et al. Impact of food processing and detoxification treatments on mycotoxin contamination. Mycotoxin Res. 2016, 32, 179–205. [Google Scholar] [CrossRef] [PubMed]
  47. Sana, J.; Farah, N.T.; Shafi, M.N.; Hassan, I.A. Contamination profile of aflatoxin M1 residues in milk supply chain of Sindh, Pakistan. Toxicol. Rep. 2015, 2, 1418–1422. [Google Scholar] [CrossRef]
  48. Bakirci, I. A study on the occurrence of aflatoxin M1 in milk and milk products produced in Van province of Turkey. Food Control 2001, 12, 47–51. [Google Scholar] [CrossRef]
  49. Choudhary, P.L.; Sharma, R.S.; Borkhartria, V.N. Effect of chilling and heaing on aflatoxin M1 content of contaminated Indian cow’s milk. Egypt. J. Dairy Sci. 1998, 26, 223–229. [Google Scholar]
  50. Reddy, U.M.; Rani, P.C. Effect of processing on detoxification of aflatoxins in maize. Indian J. Nutr. Diet. 2000, 37, 59–63. [Google Scholar]
  51. Innocent, M.; Christo, D.I.; Leshweni, J.S.; David, R.K. Aflatoxin bioamarkers in hair may facilitate long-term exposure studies. J. Appl. Toxicol. 2017, 37, 395–399. [Google Scholar] [CrossRef]
Table 1. Socio-demographic characteristics of respondents (n = 444).
Table 1. Socio-demographic characteristics of respondents (n = 444).
Marital statusSingle32072.1
Educational levelNo education30.7
Personal income≤RM150030067.6
Household income≤RM150013029.3
Age 129.25 ± 11.87
1 Presented as mean ± standard deviation.
Table 2. Median of food intake for each group.
Table 2. Median of food intake for each group.
Food GroupsMedian
Cereal products452.68
Nuts and legumes54.19
Dairy products77.68
Table 3. Aflatoxin occurrences in Malaysians’ foodstuff and the Chi-Square analyses on the association between dietary intake level and AFM1 biomarker presence among respondents a.
Table 3. Aflatoxin occurrences in Malaysians’ foodstuff and the Chi-Square analyses on the association between dietary intake level and AFM1 biomarker presence among respondents a.
GroupAflatoxin Occurrence in Malaysian’s FoodstuffMedian of Total Intake (g/day)AFM1Pearson Chi-Square
Phi-Value Association
Positive, n (%)Negative, n (%)
Cereal products-Aflatoxin ranged from 0.19 to 3.96 ng/g in rice sold in the Malaysian General Market in Kuala Lumpur [19]
-AFM1 levels in a maize and two rice samples from the selected Malaysian markets exceeded the European regulatory limits [20]
≤452.68 (low)91 (41.2)130 (58.8)2.36−0.070.12
>452.68 (high)108 (48.4)115 (51.6)
Eggs-No recent cases were reported in Malaysia≤22.71 (low)93 (39.9)140 (60.1)4.77−0.100.03 *
>22.71 (high)106 (50.2)105 (49.8)
Nuts and legumes-Aflatoxin existed in 32 from 196 nuts and its products samples in Penang, ranging from 16 µg/kg to 711 µg/kg [21]≤54.19 (low)99 (44.4)124 (55.6)0.03−0.010.86
>54.19 (high)100 (45.2)121 (54.8)
Dairy products-19 out of 53 dairy products positive AFM1, ranging from 3.5 to 100.5 ng/L [22]≤77.68 (low)76 (34.4)145 (65.6)19.360.210.00 **
>77.68 (high)123 (55.2)100 (44.8)
* p-value < 0.05, ** p-value < 0.01, a Computed for 2 × 2 table.

Share and Cite

MDPI and ACS Style

Sulaiman, S.H.; Jamaluddin, R.; Sabran, M.R. Association between Urinary Aflatoxin (AFM1) and Dietary Intake among Adults in Hulu Langat District, Selangor, Malaysia. Nutrients 2018, 10, 460.

AMA Style

Sulaiman SH, Jamaluddin R, Sabran MR. Association between Urinary Aflatoxin (AFM1) and Dietary Intake among Adults in Hulu Langat District, Selangor, Malaysia. Nutrients. 2018; 10(4):460.

Chicago/Turabian Style

Sulaiman, Siti Husna, Rosita Jamaluddin, and Mohd Redzwan Sabran. 2018. "Association between Urinary Aflatoxin (AFM1) and Dietary Intake among Adults in Hulu Langat District, Selangor, Malaysia" Nutrients 10, no. 4: 460.

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop