Prevalence of Aspergillus Spp. and Occurrence of Aflatoxins in Peanut Sauce Processing by Peanut Sauce Manufacturers

International Food Research Journal 27(1): 121 - 130 (February 2020) Journal homepage: http://www.ifrj.upm.edu.my

Prevalence of Aspergillus spp. and occurrence of aflatoxins in peanut sauce processing by peanut sauce manufacturers

1Farawahida, A. H., 1,2*Jinap, S., 1Nor-Khaizura, M. A. R., 1,2Samsudin, N. I. P., 3Chin, C. K. and 1,4Norlia, M.

1Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia 2Laboratory of Food Safety and Food Integrity (FOSFI), Institute of Tropical Agriculture and Food Security (ITAFoS), Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia 3Food Safety and Quality Division, Ministry of Health Malaysia, Precinct 3, 62675 Putrajaya, Malaysia 4Food Technology Division, School of Industrial Technology, Universiti Sains Malaysia, 11800 Minden, Penang, Malaysia

Article history Abstract

Received: 21 April 2019 The aims of the present work were to determine the prevalence of Aspergillus spp. and occur- Received in revised form: rence of aflatoxins (AFs) along the peanut sauce processing line from different peanut sauce 5 January 2020 companies in Malaysia, and to determine to which extent peanut sauce processing steps Accepted: 10 January 2020 employed by the peanut sauce industries could efficiently reduce AFs in peanut sauce. Peanut and chili samples were collected at each processing step along the peanut sauce production Keywords from three peanut sauce companies which were different in companies’ profile. Peanut fungi, samples from Companies B (87.5%) and C (100%) were contaminated with AFs. Of these, satay sauce, mycotoxins, 12.5% (Company B) and 75% (Company C) samples exceeded the Malaysian regulatory limit. High-performance Liquid None of the samples from Company A was contaminated. The steps efficient in reducing AFs Chromatography, in peanut sauce identified in the present work were (i) safety monitoring of raw materials, (ii) food safety sorting of raw materials, and (iii) heat treatment of raw materials. © All Rights Reserved

Introduction are allowed in chili (EC, 2010). In Malaysia, the implementation of Hazard Aflatoxins (AFs) are toxic, mutagenic, and Analysis and Critical Control Points (HACCP) carcinogenic compounds (Zain, 2011) that might principle by the food industries is optional (Standards occur in various types of foods and feedstuffs Malaysia, 2007). The Critical Control Points (CCPs) (Afsah-Hejri et al., 2013). The main producers of put in place by established peanut sauce companies AFs are Aspergillus flavus and A. parasiticus (Zain, associated with microbial hazards (bacteria, not 2011). Twenty AFs analogues have been identified Aspergillus spp.) are filling / sealing and retort thus far, but only AFB1, AFB2, AFG1, and AFG2 are processing. None of the peanut sauce companies in the primary contaminants of foods and feeds, with Malaysia has thus far established the CCPs to prevent

AFB1 being the most toxic (IARC, 1993). Aspergillus spp. and AFs contamination. In South East Asia, peanuts are used as the The aims of the present work were therefore main ingredient in popular foods such as peanut to determine the prevalence of Aspergillus spp. and sauce. Peanut sauce is usually consumed with sate occurrence of AFs along the peanut sauce processing (traditional skewered grilled meat; Jinap et al., line from different peanut sauce companies in Malay- 2013), nasi impit (compact rice), and yong tau foo sia; and to determine to which extent peanut sauce (tofu dish). Peanut sauce main ingredients, which are processing steps used by peanut sauce industries in peanuts and dried chili, have been shown to be Malaysia could efficiently reduce AFs in peanut contaminated with AFs (Kiran et al., 2005; Arzandeh sauce. and Jinap, 2011). According to the Malaysian Regu- lation (Food Act, 1983) and European Commission Materials and methods (EC, 2010), the maximum permitted level of total AFs in peanuts for further processing is 15 ng/g, Sampling while only 5 ng/g of AFB1 and 10 ng/g of total AFs Samples (1 kg each) comprising of peanut,

*Corresponding author. Email: [email protected] ; [email protected] 122 Farawahida, A. H., et al./IFRJ 27(1) : 121 - 130

peanut sauce, dried chili, and chili powder were blender (Waring, Torrington, CT, USA) for 2 min. collected along the processing line from three differ- Next, 25 g of sub-samples and 5 g of sodium chloride ent peanut sauce manufacturers (Companies A, B, (NaCl; Merck) were added to 125 mL of metha- and C) from July 2016 to June 2017 in Selangor, nol/water (70:30, v/v), homogenized for 2 min, and Malaysia. The samples taken followed the ongoing filtered with fluted filter paper (24 cm Ø; VICAM, process from different batches randomly, and includ- Germany). Then, 15 mL of filtrate were diluted with ed at least three different locations of the container. 30 mL of purified water followed by second filtration The samples were packed in air-tight polyethylene with a microfiber filter (11 cm Ø; VICAM, USA). bags, labelled and stored at -18°C prior to analysis. Next, 15 mL of filtrate was passed through the AflaTest WB SR (Super Recovery) IAC (VICAM, Companies’ profile USA) at a rate of 1 - 2 drop/sec. The column was The selected companies were asked to fill up twice washed with 10 mL of purified water at similar a questionnaire regarding their background and rate. Finally, trapped AFs were eluted with 1 mL of processing activities. To disguise their identities, the methanol, followed by dilution with 1 mL of purified companies were coded A, B, and C (Table 1). water.

Materials Determination of aflatoxins by HPLC

Mixed AFs standards containing AFB1 and The method from Arzandeh et al. (2010) was

AFG1 at concentration of 1,000 ng/mL, and AFB2 followed in the determination of AFs by High Pres- and AFG2 at concentration of 300 ng/mL were sure Liquid Chromatography (Waters 600 Control- purchased from Supelco (Bellefonte, PA, USA). All ler; NY, USA) joined with a Multi λ Fluorescence solvents used in the present work were of Detector (HPLC-FLD) (Waters 2475; NY, USA) HPLC-grade and supplied by Merck (Darmstadt, with a post-column Photochemical Reactor for Germany). AflaTest WB SR (Super Recovery) Enhanced Detection (PHRED; Aura Industries, NY, Immunoaffinity Column (IAC) were purchased from USA) with a slight modification. A reverse-phase

VICAM (Watertown, MA, USA). symmetry XBridge C18 column (25 cm length × 4.6 mm width and 5 µm particle sizes) running on a Moisture content determination Waters 2475 HPLC system was used at an excitation The moisture content (%) of samples was wavelength of 365 nm and emission wavelength of determined by the oven-drying method (AOAC, 435 nm. Methanol (100%), acetonitrile (100%) 1990). Briefly, empty crucibles and lids were (Merck, Germany), and purified water (Elga Purelab labelled and dried in the oven (Memmert 854, Classic UV MK2, UK) were separately filtered Schwabach, Germany) for 30 min and transferred to through nylon membrane filter (0.45 µm Ø; Merck, desiccator to cool before being weighed. Next, 5 g of Germany), degassed by Microprocess Controlled samples were weighed into the crucibles and spread Bench-top Ultrasonic Cleaner (Powersonic 420, with spatula. The crucibles with samples and lids Hwashin Technology, Seoul, Korea) and used as were placed in the oven and heated to 105°C. After mobile phase for the HPLC-FLD in the ratio of drying, the crucibles with partially covered lids were water/methanol/acetonitrile (55:35:10, v/v/v) with a transferred to desiccator to cool. The crucibles and flow rate of 0.6 mL/min. The injection volume was their dried samples were weighed daily until constant 20 µL. The determination was done in triplicates. weight was achieved. Moisture content was deter- Empower 2 Chromatography Data Software (Waters, mined by using Eq. 1: Milford, MA, USA) was used for data processing. Linearity, limit of detection (LOD), limit of [(Fresh weight – Dry weight) / (Fresh weight)] × 100 quantification (LOQ), linear equation, and coefficient of regression (R2) of the analytical method were (Eq. 1) also determined. Mixed AFs standards at seven concentrations of 2, 4, 6, 10, 25, 50, 100 ng/mL for

Aflatoxin extraction and immunoaffinity clean-up AFG1 and AFB1, and 0.6, 1.2, 1.8, 3.0, 7.5, 15.0, and

AFs were extracted and determined follow- 30.0 ng/mL for AFG2 and AFB2 were injected to ing the AOAC Method 991.31 (AOAC, 2000) with estimate the linearity. The injection was done in slight modification. Samples in liquid or semi-liquid triplicates. To determine the recovery, AFs were form were first dried by using freeze dryer (Labcon- spiked in peanut, peanut sauce, and chili samples at co, Kansas City, Missouri) prior to aflatoxin extrac- concentrations of 0.50, 5.00, and 30.00 ng/mL for tion. Dried samples were then ground using a Waring AFB1 and AFG1, and 0.15, 1.50, and 9.00 ng/mL for Farawahida, A. H., et al./IFRJ 27(1) : 121 - 130 123

Table 1. Profiles of Companies A, B, and C.

Description Company A Company B Company C Status of Medium Enterprise* Small Enterprise** Small Enterprise** company Machine / manual Machine Manual Manual processing a. Chili paste (L) a. Dried chili (D) a. Chili powder (D) b. Peanut crush b. Storage 1 (peanut, b. Cooked chili paste (D) (rough) (D) after receiving) (D) c. Peanut crush (fine) c. Storage 1 (peanut, after c. Storage 2 (peanut, (D) receiving) (D) after sorting) (D) d. Cooking and stirring d. Storage 2 (peanut, after Raw d. Frying (peanut) (D) (L) sorting) (D) materials / e. Filling and sealing e. Storage 3 (peanut, after e. Grinding (peanut and processing (S) oil-less frying) (D) chili powder) (D) steps taken f. Mixing (peanut, chili f. Storage 4 (peanut, after f. Sterilization (S) powder , and other grinding) (D) ingredie nts) (D) g. During cooking (peanut g. Holding (peanut g. Delivery (S) sauce) (S) sauce) (D) h. After cooking (peanut h. Packaging (peanut

sauce) (S) sauce) (D) Ready -to-eat peanut End products RTE peanut sauce Pre -mix peanut sauce sauce (RTE) Form o f end Semi -liquid Semi -liquid Dry products Destination of end Local + export Local Local products GMP, MS ISO 9001, Quality ISO 22000, HACCP None 1Malaysia Best certification (MS 1480), BRC Global Standard *Medium Enterprise generates annual sales of MYR 15 - 50 mil, or 75 - 200 employees. **Small Enterprise generates annual sales of MYR 0.3 to < 15 mil, or 5 - 74 employees. L = liquid; S = semi-liquid; D = dry.

AFB2 and AFG2. To estimate the LOD and LOQ, grams of samples were added to 90 mL of 0.1% three times standard deviation (SD) and ten times SD peptone water and homogenized by BagMixer 400 were used, respectively. For quantification of AFs in (Interscience, France) for 2 min. Next, 100 µL of the samples, a calibration curve with seven points homogenate was inoculated onto Aspergillus flavus was constructed for AFG2, AFG1, AFB2, and AFB1, and parasiticus agar (AFPA). Inoculated plates were respectively. incubated at 30°C for 48 h. Aspergillus flavus and A. parasiticus produced orange-yellow reverse colony Enumeration of Aspergillus flavus and A. parasiticus pigmentation. A. ochraceus also exhibits orange-yel- Enumeration of A. flavus and A. parasiticus low reverse colony but it requires longer incubation was performed following the method of Pitt et al. period. Light yellow reverse colony indicates the (1983) with slight modification. Samples were presence of A. niger. However, after prolonged incu- collected from each step of peanut sauce processing bation, A. niger will produce black conidial heads and used for Aspergillus spp. enumeration. Ten and is easily differentiated from A. flavus (Pitt et al., 124 Farawahida, A. H., et al./IFRJ 27(1) : 121 - 130

1983). The results were presented as log Colo- Table 3 lists the recovery of AFs on spiked ny-Forming Unit per gram (log CFU/g). samples using the immunoaffinity columns (IAC). The recovery rates obtained were within the regula- Statistical analysis tion limits for AFs determination (Codex Alimentari-

All experiments were performed in tripli- us, 1995; EC, 2006) except for AFG2 in chili. The cates, and readings were reported as mean ± SD. The levels of AFG2 in the spiked samples were very low individual significance probability for each inde- with no detection in most of the tested samples. Low pendent variable was shown by p-value. p < 0.05 was detection of AFG2 might be due to its low affinity accepted as significant difference. All data were with the antibodies in the IAC, depending on the subjected to univariate one way analysis of variance sample matrices and diluents used for samples (ANOVA) using Minitab® (version 16.0, Pennsylva- extraction (Ali et al., 2005; Zhao et al., 2016). nia, USA). Considering that AFG2 is not as carcinogenic as

AFB1, and that the mean recovery for AFB1 were Results and discussion within the specified limits, the IAC procedures were accepted for subsequent quantification of AFs. Method validation The standard curve constructed for all AFs Companies’ profiles analogues (AFG2, AFG1, AFB2, AFB1) yielded very The profiles of companies A, B, and C are good correlation coefficient (R2 > 0.999) as shown in listed in Table 1. For Malaysia, the definition of Table 2. This indicated that the quantification proce- Small and Medium Enterprises (SMEs) is based on dures were accurate and reliable. Low LOD and LOQ employment and sales (Ndubisi and Nwankwo, also signified the sensitivity and accuracy of the 2013). quantification instruments (Shrivastava and Gupta, 2011). Company A Company A produced 120 kg peanut sauce Table 2. LOD, LOQ, and R2 for aflatoxin quantification. daily. The process of making peanut sauce started with receiving rough and fine roasted crushed LOD LOQ Aflatoxins R2 peanut, dried chilies, shrimp paste, tamarind, water, (ng/mL) (ng/mL) onion, lemon grass, granulated sugar, salt, and palm AFG2 0.06 0.20 0.9994 olein. Next, the raw materials except crushed peanuts were mixed in a tank and cooked at 90 - 95°C for 2 h AFG1 0.09 0.30 0.9996 into gravy. Then, rough and fine crushed peanut were AFB2 0.01 0.05 0.9994 added to the gravy, passed through the piping line, and placed in a holding tank at 65°C for 4 h. The AFB1 0.04 0.14 0.9994 product was filled into an aluminium pouch and

Table 3. Recoveries of aflatoxins in spiked samples. Concentration of spiked Samples Aflatoxins Mean recovery ± SD (%) aflatoxins (ng/mL)

AFG2 0.15 - 9.00 71.63 ± 2.00

AFG1 0.50 - 30.00 94.79 ± 3.7 Peanut AFB2 0.15 - 9.00 96.36 ± 8.2

AFB1 0.50 - 30.00 83.56 ± 3.2

AFG2 0.15 - 9.00 73.70 ± 1. 2

AFG1 0.50 - 30.00 82.76 ± 5.5 Peanut sauce AFB2 0.15 - 9.00 102.21 ± 11.7

AFB1 0.50 - 30.00 97.83 ± 8.3

AFG2 0.15 - 9.00 44.71 ± 1.0

AFG1 0.50 - 30.00 102.12 ± 2.6 Chili AFB2 0.15 - 9.00 86.15 ± 2.5

AFB1 0.50 - 30.00 85.84 ± 8.4 Farawahida, A. H., et al./IFRJ 27(1) : 121 - 130 125 sterilized at 121°C for 26 min before being cooled after, the sorted peanuts were fried at 120°C for 20 (90°C) and dried (70°C, 15 min). The product was min. After the removal of oil by machine, the fried then passed through the X-ray machine to detect peanuts were left to cool down for 10 min and mixed metal contaminant, and stored in the warehouse at with chili powder and ground together. Onion and 4°C before being delivered to customers. garlic were sliced, fried, blended, and separately stored in containers. Other ingredients were added to Company B the mixture of ground peanut and chili powder, and Company B produced 100 kg peanut sauce mixed by machine. After holding for approximately daily. The process of making peanut sauce started 4 h, peanut sauce was packed manually by sealer in with receiving peanut kernels, dried chilies, palm aluminium pouch (primary packaging) and delivered olein, water, granulated sugar, and salt. Raw peanuts to regular customers or sold at retail markets. were manually sorted to remove defect peanuts. The sorted peanut kernels were then oil-less fried and Moisture contents, aflatoxin levels, and A. flavus and ground. Dried chilies were blanched at 90°C for 3 A. parasiticus loads in samples from the companies min, sieved and then ground into chili paste. Then, Company A the chili paste was cooked with palm oil at 60°C for No contamination of AFs, A. flavus, and A. 10 min. All the raw ingredients including cooked parasiticus was detected in any of the sample (Table chili paste were mixed in a big pot and cooked manu- 4). This might be due to the fact that the company has ally for 3 h at 99°C. After cooking, the peanut sauce given a specification to the supplier of less than 4 was left to cool down for 5 min before packing and ng/g AFs, and during receiving the concentration was delivery. verified again by using RIDA AFs test kit (R-Biop- harm AG, Darmstadt, Germany). This company also Company C controlled the temperature during storage and Company C produced 20 kg peanut sauce processing (4°C). Since Company A practiced proper daily. The process of making peanut sauce started storage, it had one time higher in the implementation with receiving palm olein, onion, garlic, shrimp of hygiene practices as compared to Companies B paste, tamarind, peanut kernels, chili powder, salt, and C (Mohd Azaman et al., 2016). Other than that, monosodium glutamate (MSG), and sugar. Then, the the peanut sauce processing was found to be a contin- peanut kernels were sorted and stored. Several days uous process utilizing fully automated machine

Table 4. Moisture content, prevalence of Aspergillus spp. (log CFU/g), and occurrence of aflatoxins (ng/g) in samples from Company A. Total Raw materials / Moisture AFG2 AFG1 AFB2 AFB1 Company A Log CFU/g AFs processing steps content (ng/g) (ng/g) (ng/g) (ng/g) (ng/g)

82.38 ± < 2.00 ± Chili paste (L) NDa NDa NDa NDa NDa 0.40a 0.00a

Peanut crush 4.74 ± < 2.00 ± Raw m aterials* NDa NDa NDa NDa NDa (rough) (D) 0.64b 0.00a

Peanut crush (fine) 4.19 ± < 2.00 ± NDa NDa NDa NDa NDa (D) 0.71b 0.00a

Cooking and 83.19 ± < 2.00 ± NDa NDa NDa NDa NDa stirring (L) 0.93a 0.00a

Filling and sealing 61.86 ± < 2.00 ± NDa NDa NDa NDa NDa (S) 0.87b 0.00a Processing steps* 62.59 ± < 2.00 ± Sterilization (S) NDa NDa NDa NDa NDa 0.75b 0.00a

62.59 ± < 2.00 ± Delivery (S) NDa NDa NDa NDa NDa 0.75b 0.00a

ND: not detected. Different letters within the same column indicate significant difference (p < 0.05). *Separate statistical analyses were conducted on raw material and processing steps. L = liquid; S = semi-liquid; D = dry. 126 Farawahida, A. H., et al./IFRJ 27(1) : 121 - 130 which further reduced the risk of cross-contamina- another factor contributing to increasing AFs tion usually brought about by manual labour. The contamination. After cooking process significantly owner and production manager of the company reducing 41% of AFs, the peanut sauce was fully possess the knowledge about AFs and food safety. cooked at temperature 99°C for 3 h. Since Company A had quality certifications such as Good Manufacturing Practices (GMP) and HACCP, Company C it had four times likelihood towards minimizing AFs 37.5% of the samples were contaminated contamination as compared to Companies B and C with A. flavus and A. parasiticus which exceeded the (Mohd Azaman et al., 2016). regulatory limit (log 3.44 - 5.05 CFU/g; Table 6). Similar pattern of Aspergillus spp. prevalence has Company B been observed. Aspergillus spp. was detected in chili 12.5% samples were contaminated with A. powder, thus their total AFs increased. Besides, flavus and A. parasiticus, and exceeded the regulato- Aspergillus spp. was found in Storage 1 (after peanut ry limit (Table 5). During Storage 1, peanuts received receiving) and Storage 2 (after peanut sorting). had exceeded log CFU/g of Aspergillus spp. and total Meanwhile, all the samples were contaminated with AFs. However, during Storage 2, sorting of peanuts total AFs (1.71 - 537.09 ng/g), with 75% of them reduced Aspergillus spp. and AFs below the regula- exceeding the regulatory limit. This company did not tory limits. 87.5% of the samples were contaminated have a Standard Operating Procedure (SOP) for with total AFs (0.58 - 32.91 ng/g), with 12.5% of incoming raw materials, no record-keeping, and did them exceeding the regulatory limit. The total AFs in not hire Production Supervisor or Quality Control dried chili far exceeded the regulatory limit of 5 ng/g Manager to oversee the processing line. The owner (Food Act, 1983) due to the favourable condition for and operators of the company also did not possess mould growth when the moisture content was above any knowledge about AFs. Therefore, they were 11% (Toontom et al., 2012). However, after cooked three times less likely to implement high level of into chili paste, the total AFs was reduced by 88% to hygienic practices (Mohd Azaman et al., 2016). an acceptable level. This might be due to the heat Similar to Company B, the chili powder treatment applied during cooking and lower moisture from Company C too had high level of AFs. It could content of cooked chili paste. According to Farawa- be deduced that either the dried chili used to prepare hida et al. (2017), oil-less frying of chili powder the chili powder was contaminated with the fungi, or could reduce 33 - 41% of AFs, with higher reduction the fungi colonized the chili powder during storage could be achieved with moist samples (Rustom, prior to arrival at the company. The levels of AFs 1997). Reduction of Aspergillus spp. and AFs (95%) during storage of peanut kernels (Storage 1) were were also observed in peanuts after manual sorting as also very high (537.09 ± 2.39 ng/g), which was 36 compared to the received raw materials. Since very times higher than the permitted level. This could be low concentrations of AFs (0.58 ± 0.20 ng/g) were due to several factors. The peanut sauce was manual- detected after sorting, oil-less frying of peanuts ly processed in batches and in the traditional way. further reduced AFs to a non-detectable level. Arzan- The temperature in the storage and production areas deh and Jinap (2011) reported that roasting peanut (32.8°C) provided optimal growth condition for kernels at 130 - 150°C for 120 min reduced 57 - 70% Aspergillus spp. (Hocking, 1997). For warehouses of AFB2 while 78 - 80% reduction of AFB1 was dealing with food commodities, they are recom- achieved when peanuts were roasted at 150°C for mended to have a refrigeration temperature (0 - 120 min. Studies from Yazdanpanah et al. (2005) 10°C) to inhibit any microbial contamination (Codex showed that roasting peanut kernels at similar dura- Alimentarius, 2004). During manual sorting, it was tion and temperature reduced > 95% of AFB1. How- found that approximately 1.5% of peanuts were ever, storage of roasted peanut after grinding saw a rejected (based on the physical appearance), with significant increase in AFs. This might be due to reduction of AFs by 21%. This low rate of rejection improper storage practices which subsequently led to could have been translated to the high carryover of cross-contamination and increase in AFs production AFs after sorting (426.92 ± 7.93 ng/g). Based on (Udomkun et al., 2018). preliminary study, about 7.4% of peanuts could be The five-fold increase in AFs noted during removed by manual sorting. Other studies stated that cooking of peanut sauce mixture (as compared to 29 - 38% of total AFs could be reduced by electronic during storage 4) might be explained by the addition colour sorting (Whitaker et al., 2005), while manual of AFs-contaminated chili paste. Besides, unsani- sorting could reduce 40 - 80% of total AFs (Park, tised big pot used (from previous cooking) can be 2002). Farawahida, A. H., et al./IFRJ 27(1) : 121 - 130 127

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a l d s s r r i x h k r s i e o p i n l i t e t t ( s c c i m d p f

t o t o t e

M e u u a h e H o a f g G r d S S

c r n n l i c P w a n i n

a o a g t , i a h t , e r e t , u y i n t u R a p p r u p C n ( ( u n a n F n a e a a e e p e ( p p Table 6. Moisture content, prevalence of Aspergillus spp. (log CFU/g), and occurrence of aflatoxins (ng/g) in samples from Company C. Aspergillus 6. Moisture content, prevalence of Table ( ( p (

C g

l s i n * n s s w i a

s a p r a e p c t e t e R s o a m r m P C o ND: not detected. Different letters within the same column indicate significant difference (p < 0.05). letters within the same column indicate significant difference ND: not detected. Different *Separate statistical analyses were conducted on raw material and processing steps. D = dry. Farawahida, A. H., et al./IFRJ 27(1) : 121 - 130 129

However, these high levels of AFs were method to determine the natural occurrence of greatly reduced more than 99% to permissible limits aflatoxins in commercial traditional herbal medi- by frying the peanuts. Peanut frying and roasting cines from Malaysia and Indonesia. Food and have been shown to be very effective in reducing AFs Chemical Toxicology 43(12): 1763–1772. (Razzazi-Fazeli et al., 2004; Mutegi et al., 2013). Arzandeh, S. and Jinap, S. 2011. Effect of initial Grinding peanuts with chili powder increased the aflatoxin concentration, heating time and roast- level of AFs due to the addition of the AF-contami- ing temperature on aflatoxin reduction in nated chili powder, and this increasing trend was contaminated peanuts and process optimization continuously observed in the subsequent steps (i.e., using response surface modelling. International mixing of ingredients, holding, and packaging). Journal of Food Science and Technology 46(3): Several factors might have contributed to this such as 485–491. the use of contaminated container during grinding; Arzandeh, S., Selamat, J. And Lioe, H. 2010. Afla- the use of stand fan during holding; and the increase toxin in raw peanut kernels marketed in Malay- of humidity during packaging (Abou-Arab et al., sia. Journal of Food and Drug Analysis 18(1): 1999). 44–50. Association of Official Analytical Chemists Conclusion (AOAC). 1990. Official methods of analysis of the AOAC International. 15th ed. Washington In the present work, the prevalence of Asper- (DC): AOAC. gillus spp. has been detected during raw materials Association of Official Analytical Chemists (peanuts and chili) receiving and sorting. Manual (AOAC). 2000. Official methods of analysis of sorting and heat treatment of peanut kernels (oil the AOAC International. 17th ed. Washington frying or oil-less frying) and cooking of chili paste or (DC): AOAC. peanut sauce have been shown to significantly reduce Codex Alimentarius. 1995. Codex general standard the AFs levels. Moreover, it is also important to for contaminants and toxins in food and feed - ensure that the peanuts and chili powder used in CXS 193-1995. Retrieved on December 25, 2018 peanut sauce manufacturing meet the AFs guidelines from FAO Website: http://www.fao.org/filead- before entering the manufacturing process. min/user_upload/livestockgov/docu- ments/1_CXS_193e.pdf Acknowledgement Codex Alimentarius. 2004. Code of practice for the prevention and reduction of aflatoxin contamina- Authors acknowledge Universiti Putra Malay- tion in peanuts - CAC/RCP 55-2004. Retrieved sia for financially supporting the present work (GP-IP- on February 1, 2019 from FAO Website: www.- B/2013/9425400 and UPM/800-3/3/1/GP- fao.org/input/download/standards/10084/CX- B/2018/9658100); the Malaysian Ministry of Educa- P_055e.pdf tion for the HICoE grant (HICoE-ITAFoS- European Commission (EC). 2006. Commission /2017/FS6/6369114); the Malaysian Ministry of Regulation (EC) No 401/2006 of 23 February Health and Federal Agricultural Marketing Authority 2006 laying down the methods of sampling and for assistance in sampling and questionnaire collection; analysis for the official control of the levels of and the companies involved for providing the samples. mycotoxins in foodstuffs. Official Journal of the European Union: L70/12 – L70/34. References European Commission (EC). 2010. Commission Regulation (EU) No 165/2010 of 26 February Abou-Arab, A. A. K., Soliman Kawther, M., El 2010 amending Regulation (EC) No 1881/2006 Tantawy, M. E., Badeaa, R. I. and Khayria, N. setting maximum levels for certain contaminants 1999. Quantity estimation of some contaminants in foodstuffs as regards aflatoxins. Official Jour- in commonly used medicinal plants in the Egyp- nal of the European Union: L 50/8 – L 50/12. tian market. Food Chemistry 67(4): 357–363. Farawahida, A. H., Jinap, S., Nor-Khaizura, M. A. R. Afsah-Hejri, L., Jinap, S. and Radu, S. 2013. Occur- and Samsudin, N. I. P. 2017. Reduction of Asper- rence of aflatoxins and aflatoxigenic Aspergillus gillus spp. and aflatoxins in peanut sauce in peanuts. Journal of Food, Agriculture and processing by oil-less frying of chili powder and Environment 11(3): 228–234. retort processing. Food Additives and Contami- Ali, N., Hashim, N. H., Saad, B., Safan, K., Nakaji- nants Part A 34(12): 2242–2250. ma, M. and Yoshizawa, T. 2005. Evaluation of a Food Act. 1983. Laws Malaysia, Act 281, Food 130 Farawahida, A. H., et al./IFRJ 27(1) : 121 - 130

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