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Current Nutrition & Food Science, 2019, 15, 678-684

RESEARCH ARTICLE

ISSN: 1573-4013 eISSN: 2212-3881

The Effect of Different Roasting Durations on the Acrylamide Concentra- tions in the Beverages

BENTHAM SCIENCE

Amal H. Alshawi1,*

1Nutrition and Food Sciences Department, College of Home Economic, Princess Nourah Bint Abdulrahman University, Riyadh,

Abstract: Background: Acrylamide is the most powerful carcinogenic and neurotoxic compound widely distributed in thermally processed foods. This compound is formed during early stages of roasting processes of coffee beans through Millard reaction between reducing and asparagine. Due to the huge consumption of coffee throughout the globe, it has been considered as the major die- tary source of this dangerous compound. Thus, this study aims to investigate the effect of roasting A R T I C L E H I S T O R Y conditions on sugars and acrylamide contents in the Arabic coffee beans and beverages.

Received: December 21, 2017 Methods: Arabic coffee beans were roasted at 125°C for 30 to 60 min, ground to powders, brewed in Revised: May 05, 2018 boiling water for 20 min and then analysed for sugars and acrylamide contents. Accepted: May 09, 2018

DOI: Results: Roasting conditions significantly (P ≤ 0.05) reduced the content of both reducing and non- 10.2174/1573401314666180515115709 reducing sugars compared to untreated samples. Increasing roasting duration to 40 min significantly (P ≤ 0.05) increased the acrylamide concentration values to the maximum, whereas extending the du- ration over 40 min (P ≤ 0.05) reduced the acrylamide concentration of coffee beans and beverages. Conclusion: The present study confirms that the process of roasting coffee beans is a significant fac- tor in the formation of acrylamide in Arabic coffee powders and beverages.

Keywords: Acrylamide, arabic coffee beans, HPLC, qahwa, roasting conditions, sugars composition.

1. INTRODUCTION foods, the ratio of the surface area to the volume of the heat- ed foods, and the thermal conditions [3]. During heat pro- Acrylamide (2-propenamide) is a hazardous compound cessing of foods, the carbohydrates and proteins are degrad- which has raised high health concerns in recent years due to ed to simple sugars (reducing and non-reducing sugars) and its potentially high carcinogenicity, neurotoxicity, mutageni- amino acids, which in turn, react together through Millard city, reproductive and developmental toxicity and genotoxi- reaction and form several compounds. Acrylamide is one of city [1]. the compounds that form through the interaction between In 2002, the Swedish State Agency on Food and re- reducing sugars, mainly glucose and fructose, with the amino searchers from Stockholm University announced that acry- acid asparagine at high processing temperatures and long lamide was widespread in thermally processed carbohydrate- durations [3]. Taking together the widespread of acrylamide rich foods. In addition, the use of polyacrylamide in the wa- in thermally processed foods and its harmful effect on human ter refining process causes an accidental occurrence of acry- health, JECFA and EFSA emphasize that the presence of lamide in the drinking water [2]. However, the compound acrylamide in foods is a major public health concern that is was found in numerous foods and that have been necessitating great scientific, governmental, and industrial treated at temperatures above 120°C during roasting, frying, efforts to reduce its exposure [5, 6]. In this regard, research boiling, toasting, grilling, and baking processes [3]. Among to alleviate acrylamide formation in foods by controlling the thermally treated foods, potato chips and crisps, coffee beans thermal treatment conditions is of high importance. and beverages and cereal-based foods such as pastries, cook- The tree belongs to the genus of the ies, bread, and biscuits are the major sources of acrylamide family Rubiaceae, and among the species of this genus, [4]. The formation and development of acrylamide in foods , and var. Robusta are the and drinks likely depend on the chemical composition of the most important commercial species [7]. Throughout the globe, coffee is the second most extensively consumed food *Address correspondence to this author at the Nutrition and Food Sciences preceded by water [7]; consequently, the research on coffee Department, College of Home Economic, Princess Nourah Bint Abdulrah- is of much importance for both national and international man University, Riyadh, Saudi Arabia; Tel: +966504135581; consumers and health professionals [8]. As coffee is one of E-mails: [email protected]; [email protected] Current Nutrition & Food Science the major sources of acrylamide in food, it contributes signif-

2212-3881/19 $58.00+.00 © 2019 Bentham Science Publishers The Effect of Different Roasting Durations on the Acrylamide Concentrations Current Nutrition & Food Science, 2019, Vol. 15, No. 7 679 icantly to the daily dietary exposure. In some European 17 min in a glass-resistant kettle. The samples were filtered countries, the intake of acrylamide obtained from the coffee through Whatman No 1. filter paper. reaches close to 40% [9]. There are many factors affecting the acrylamide levels in the coffee beans such as coffee spe- 2.4. Reducing Analysis cies, beans maturity, post-harvest processing, roasting, stor- age, and beverage preparation conditions [10]. The Robusta The concentration of sugar was estimated by comparing species contain higher amounts of acrylamide than Coffea the concentration of sugar in the samples and the standard arabica. Moreover, during the early stage of roasting of both solution using HPLC (Agilent Technologies, Palo Alto, CA, types, the acrylamide formation increases. At the later stage USA) 1200 series [15]. of roasting, the concentration of acrylamide tends to decrease 2.4.1. Preparation of Standard Sugar Solution [10]. In addition, different preparation methods of coffee beverage could result in different concentrations of acryla- A mobile phase was composed as a reagent by stages at a mide in the final product [10]. Since roasting degree is the volume of 1 L, by 75%:25% acetonitrile (LC grade): ul- most influencing factor of acrylamide level in the coffee trapure water. Then, vacuum filtration was used to filter and beans, it is of utmost importance to optimize roasting condi- degas the mobile phase through a 0.45µm PTFE membrane tions to reduce acrylamides in the roasted coffee. In the Ara- filter. To prepare the standard stock solution of sucrose, bic coffee (Qahwa or Gahwa) preparation in some Arabian around 8g/L was used. To represent the vital concentration countries, the beans are lightly or heavily roasted at 165°C to of sucrose, aliquots of this solution were used. To compose 210°C, brewed for about 20 min in boiling water with added the standard collaboration mixture, concentration ratios of and other species like , cloves, and cinna- 1:0.2:1 g/L, 2:0.4:2 g/L and 5:1:5 g/L of fructose: sucrose: mon, and then served in small quantities together with dates glucose were used respectively, by adding accurate weighted several times per day [11]. The degree of roasting and masses of glucose and fructose to the pipetted volumes of the amounts of added coffee and cardamom make significant stock solution of sucrose. Standards were injected into the differences in the sensory attributes of Qahwa [12]. Despite HPLC at 0.6 ml/min flow rate and 0.9 ml/min flow rate to the massive studies on the acrylamide formation during observe the effect. Once optimum flow conditions had been thermal processing of several types of coffee beans, studies established, repeated injections of standards were performed on the acrylamide concentration in Qahwa are rare [13, 14] to obtain reproducible elution times and peak heights [15]. and no work has been reported on the effect of roasting con- ditions on acrylamide in Qahwa. Therefore, the present study 2.4.2. Preparation of Coffee Samples Solutions for Sugar aims to investigate the effect of roasting condition especially Analysis roasting durations on the acrylamide concentration in Arabic The coffee samples solutions were prepared with approx- coffee beans and beverage. imately 10 g/L of sugar content (fructose and glucose). For recovery check analysis, a solution of each coffee sample 2. MATERIALS AND METHODS was composed of approximately 5 g/L, by combining 2.1. Materials weighted mass of around 0.5 g of glucose and 2.5 g of the sample and made up to 500 ml. Vacuum filtration was used Green coffee beans were purchased from a local market to degas and filter the samples by using cellulose-acetate in Riyadh, Saudi Arabia. The samples were washed and membrane filters [15]. thoroughly dried in air. The beans were stored in polyeth- ylene bags at 4°C until used for roasting treatment, sugars 2.4.3. Determination of Sugars by HPLC and acrylamide analysis. To determine the content of sugars by operates HPLC column conditions to inject samples and standard as follows: 2.2. Treatment of Coffee Beans Column: Grace-Davison Prevail Carbohydate ES5µm. The beans were divided into five portions of (500 g 150mm x 4.6mm i.d. Acetonitrile: Water (75:25). RID detec- each), the first portion was kept as a control without roasting, tor. Flow rate: 0.9ml/min. The environmental conditions during the test were 25°C and 40% humidity. The average of the second portion was roasted at 125°C for 30 min, the third peak heights was used to construct calibration graphs for portion was roasted at 125°C for 40 min, the fourth portion fructose and glucose. Using the same conditions, samples was roasted at 125°C for 50 min and the last portion was and recovery check samples were injected; and calibration roasted at 125°C for 60 min. The treatments were replicated graphs were used to determine the fructose and glucose con- five times. All samples were coarsely ground (<1000 mm) tent of the coffee samples [15]. by using a coffee grinder. The powdered samples were di- vided into two portions: the first portion (250 g) was used to 2.5. Acrylamide Analysis analyse the acrylamide in the dry powders, while the second portion (250g) was used to prepare the coffee extracts. All The concentration of acrylamide was estimated by com- dried powders were sealed in sterile polyethylene bags and paring the concentration of acrylamide in samples and the stored at 4°C until used for further analysis. standard solution using HPLC (Agilent Technologies, Palo Alto, CA, USA) 1100 series [16]. 2.3. Preparation of Coffee Beverage Samples 2.5.1. Preparation of Standard Acrylamide Solution For each sample, about 50g of coffee powders from dif- Standard acrylamide was purchased from sigma corpora- ferent treatments were added to 100 mL of boiled water for tion with molecular formula C3H5NO and molecular weight 680 Current Nutrition & Food Science, 2019, Vol. 15, No. 7 Amal H. Alshawi

Table 1. The effect of roasting conditions on the reducing and non-reducing sugars (%) in Arabic coffee beans.

Roasting Time (min) Fructose % Glucose % Sucrose % Total %

00.00 0.82 b ± 0.02 2.90 a ± 0.02 1.06 a ± 0.07 4.78 a ± 0.06

30.00 0.45 c ± 0.01 1.03 b ± 0.03 1.01 a ± 0.00 2.48 b ± 0.02

40.00 0.44 c ± 0.00 0.58 c ± 0.02 0.99 a ± 0.02 2.02 c ± 0.02

50.00 0.37 d ± 0.01 0.14 d ± 0.06 0.66 b ± 0.01 1.20 d ± 0.07

60.00 0.98 a ± 0.08 0.01 e ± 0.00 0.19 c ± 0.01 1.18 d ± 0.42

Values are the mean of 5 replicates and the results are expressed as the mean ± SD (P < 0.05). a-e Mean values with various letters are significantly different. = 70.8. The acrylamide standard solution was prepared by 3. RESULTS AND DISCUSSION dissolving 10 mg of standard acrylamide in 10 ml of HPLC 3.1. Sugars Contents of Coffee Beans water. Then, it was mixed by the vortex for 1 minute. After that, different concentrations of this solution were prepared. As roasting temperatures and durations are expected to Light absorption of the solutions was measured at wave- affect the sugar composition of roasted food materials, the length 230 nm by a spectrophotometer. The solution was sugar composition of Arabic coffee beans was studied. The then transferred to a 100-ml flask containing 25 ml of results showed that roasting temperature and roasting dura- HPLC-grade methanol. Next, the solution was filtered tion reduced both reducing and non-reducing sugars com- through a disposable 0.25 μl PTFE syringe filter [16]. pared to control samples (Table 1). However, the responses of sugars to the increased roast- 2.5.2. Preparation of Coffee Samples Solutions for Acryla- ing duration at 125°C were varied. Glucose and sucrose con- mide Analysis tents were concomitantly (P ≤ 0.05) reduced as the roasting 1g of each homogenous sample was diluted in 9 ml of duration increased reaching the minimum percentage at 60 HPLC water. Then mixed for 10 minutes and centrifuged at min, whereas fructose fluctuated between the minimum 900 cpm/30 min. The aqueous layer was removed and then amounts at 50 min and the maximum at 60 min. The sum of filtered while the filtrate was re-centrifuged at 400 cpm/ 10 the sugars (reducing and non-reducing) was also showed a min and filtered again. concurrent reduction with the roasting time. Overall, the re- sults demonstrated that roasting duration has a significant 2.5.3. Determination of Acrylamide by HPLC effect on the reducing and non-reducing sugars of the coffee beans. Thus, roasting coffee for a shorter time is better than a To analyse the acrylamide in the coffee samples, 2 mg of longer time. The reduction in sugars while roasting the cof- each sample was injected into the injector and operated un- fee beans could be attributed to the formation of the Millard der the following conditions: the acrylamide chromatograph- reaction between sugars and free amino acids mainly aspara- ic column length was 350 mm, and the internal diameter was gine [18]. Similarly, it has been reported that while roasting 25 mm filled with silica gel fine particles with reversed the Arabic and Robusta coffee, both reducing (glucose and phase column C18CLC/ODS. The injection volume was 30 fructose) and non-reducing sugars (sucrose) were rapidly μg with a flow rate of 0.5 ml/ min and wavelength of 230nm. degraded, reaching the minimal level in roasted beans com- The oven temperature was 26°C. Mobile phase: aqueous pared to the unroasted ones [19]. In addition, the findings are 0.1% acetic acid: 0.5% methanol 1:1 [16]. also well comparable to the fact that heating temperature and duration can increase the rate of the Millard reaction, in 2.6. Statistical Analysis which the free amino acid asparagine interacts with reducing The results were calculated using a Microsoft Excel sugars, primarily glucose and fructose [18]. The reduction of spreadsheet, and statistical analyses were carried out using sugars is consistent with the colour results of the roasted SPSS version [17]. The Kolmogorov-Smirnov Z test was coffee beans. A clear difference in the colour of the roasted used to ensure that data followed by a normal distribution. coffee beans was observed (Fig. 1); the development and the Kolmogorov-Smirnov Z coefficients for dry and liquid sam- intensity of the brown colour in the coffee beans increased ples were 1.06 at the significance of 0.210 and 0.995 at the concurrently with the increasing of the roasting durations, significance of 0.275 respectively. The data were reported as and the maximum intensity was observed in the sample that the mean ± SD. One-way ANOVA was performed to deter- roasted for 60 min. mine if any significant differences existed between the sam- It is also worth to note that after 30 min roasting the ples. Independent t-tests were used to identify significant brown colour is seen, whereas unroasted samples have no differences between the means. ANOVA with Scheffe’s test brown colour. This could explain the significant differences was used to determine the source of the significant differ- in the reducing sugars between the roasted and unroasted ences. coffee beans samples. A similar observation on the develop- The Effect of Different Roasting Durations on the Acrylamide Concentrations Current Nutrition & Food Science, 2019, Vol. 15, No. 7 681

Fig. (1). The effect of roasting durations on the brown colour development in the Arabic coffee beans and beverages. ment of the brown colour during roasting the coffee beans, beans at 125°C for less than 30 min to keep the concentra- wheat flours and potato chips has been reported [20]. During tion of acrylamide at the minimum. The initial increase in the roasting the coffee beans, the reactive breakdown the prod- concentration of the acrylamide could be due to the en- ucts of the carbohydrates, such as reducing sugars, are react- hancement of the acrylamide formation from its precursor by ed with the side chain of basic amino acids, leading to the the roasting temperature and time, whereas the reduction of formation of melanoidins [19]. The brown colour developed the acrylamide when the roasting time increased over 40 min during roasting the coffee beans resulted from the Millard could be due to the consumption of the precursor or the deg- reaction, where melanoidins are known as the main end radation of the acrylamide. Generally, the major precursors products of the reaction [20]. Moreover, the formation of the of the acrylamide in coffee beans are sugars (sucrose, glu- brown colour in the coffee beans during the roasting is a cose, and fructose) and the amino acid asparagine, and the significant indicator of the acrylamide formation and devel- reaction between these components initiate Millard reaction opment [20]. The Acrylamide formation occurs via the inter- and the acrylamide formation [10]. action of glucose and asparagine at temperatures above During Millard reaction, melanoidin is formed, and this 120°C for 60 min or more [1]. compound could control the acrylamide formation and elim- ination pathway during the , and it thus serves 3.2. Acrylamide Concentration in the Coffee Beans Pow- as acrylamide mitigating substance [2]. Similar trends of the der acrylamide formation during roasting of coffee beans have The acrylamide concentration in the coffee beans pow- been stated previously [21]. In addition, the acrylamide con- ders significantly affected by roasting temperature and dura- tent was analysed in both Arabica and Robusta coffee beans tions (Fig. 2). during roasting; it was found that the acrylamide level was at Roasting at 125°C for 30 to 60 min significantly (P ≤ its highest in lightly roasted beans, while the elongated roast- 0.05) increased the acrylamide concentration of the coffee ing time significantly reduced the acrylamide levels in both beans compared to unroasted samples. Increasing the roast- types of coffee beans [22]. The roasting conditions, particu- ing time up to 40 min greatly (P ≤ 0.05) increased the larly temperature and durations, are the main factors that acrylamide concentration from 112.36 ± 4.43 ppb in the un- contribute to the initial increase of the acrylamide concentra- roasted samples to 330.03 ± 8.71 ppb, whereas elongating tion and its later reduction in the coffee beans [21]. During the time over 40 min progressively (P ≤ 0.05) decreased the the whole roasting process, the range of acrylamide (112.36- acrylamide concentration to177.08 ± 5.40 and 122.33 ± 9.35 330.03 ppb) in the current study fall within the range (40-400 ppb at 50 and 60 min, respectively, suggesting that the deg- ppb), which has been determined for various types of coffee radation rate of acrylamide exceeded the formation rate. beans [23]. However, other reports showed high levels of However, the acrylamide concentration at higher roasting acrylamide in the coffee beans [22]. The variation between time (50 and 60 min) is higher (P ≤ 0.05) than that at 30 min. these studies could be due to the differences in the genetic It could thus be recommended to roast the Arabic coffee makeup, agronomical practices, environmental conditions, 682 Current Nutrition & Food Science, 2019, Vol. 15, No. 7 Amal H. Alshawi

400 a 350

300

250 b 200

150 c d 100 e

Acrylamide concentration (ppb) 50

0 0 30 40 50 60 Roasting time (min)

Fig. (2). The effect of roasting conditions on the acrylamide concentrations (ppb) in the dry coffee powders. Error bars indicate the standard error of five independent replicates. a-e Mean values with various letters are significantly different at P ≤ 05.

250 a

200

b 150

c 100 d

e 50 Acrylamide concentration (ppb)

0 0 30 40 50 60

Roasting time (min) Fig. (3). The effect of roasting conditions on the acrylamide concentrations (ppb) in the coffee beverage (Qahwa). Error bars indicate the standard error of five independent replicates. a-e Mean values with various letters are significantly different at P ≤ 0.05. and roasting conditions of the coffee beans. Overall, this mide concentration of the coffee beverage. As the roasting study suggested that roasting Arabic coffee beans at a tem- duration increased, the concentration of acrylamide progres- perature of 125°C for a duration shorter than 30 min or long- sively (P ≤ 0.05) increased to the maximum of 208.28±1.31 er than 60 min could potentially result in low levels of the ppb at 40 min and then gradually (P ≤ 0.05) decreased to acrylamide in the coffee beans powder. 151.88±1.47 and 101.56±2.07 ppb at 50 and 60 min, respec- tively. With the exception of the unroasted samples, the cof- 3.3. Acrylamide Concentration in the Coffee Beans Bev- fee beverage that prepared from short time roasted beans (30 erage min) showed the least (P ≤ 0.05) acrylamide concentration compared to other roasted samples. In the current study, a Considering that coffee is usually consumed in a range of acrylamide (76.61-208.28 ppb) in Arabic coffee beverage like form, the acrylamide concentration in the cof- beverage was higher than those that have been reported for fee beverage was assessed. The results showed a similar various coffee types from coffee vendor machines in Spain trend to that of dry coffee samples (Fig. 3). [21]. In addition, the extraction percentages of 68-86% were The acrylamide concentration of the coffee beverage that observed for the Arabic coffee in the current study, and this was prepared from unroasted beans was significantly lower fall within the ranges of 59-98% and 62-99% that reported than those prepared from roasted beans. Roasting durations for coffee which prepared respectively from Ro- again showed a significant (P ≤ 0.05) effect of the acryla- busta and Arabica coffee beans [21]. Acrylamide is a polar The Effect of Different Roasting Durations on the Acrylamide Concentrations Current Nutrition & Food Science, 2019, Vol. 15, No. 7 683

400 Dry coffee beans *** Coffee beverage 350

300

250 *** 200 ** 150 ***

100 *

Acrylamide concentration (ppb) 50

0 0 30 40 50 60 Roasting time (min) Fig. (4). The difference in acrylamide concentration between coffee beans powders and beverage as affected by roasting time. Error bars indicate the standard error of five independent replicates. *Significant at P ≤ 0.05, **significant at P ≤ 0.01, ***significant at P ≤ 0.001. compound with a high solubility in water; therefore, it could formation and leaching into beverages. In the Arabic coffee easily extract in water [2]. The chemical composition of the drinking traditions, the coffee beverage is served in small coffee beverage depends on various factors, such as coffee cups of approximatly15 mL a maximum volume for at the beans type, growing conditions and environment, roasting rate of 3 cups/ serving time/ day. The average of acrylamide temperature and duration, grinding processes and degree, in the Arabic coffee is about 142.45 ppb equalling 142.45 coffee/water ratio, water temperature and extraction times µg/L (minimum amount was 76.61ppb and maximum was [21]. The variation in the acrylamide concentration and the 208.28 ppb). Consequently, one cup of coffee (15 mL vol- extraction percentage could be due to the preparation method ume) might contain, on average, about 2.14 µg acrylamide. of the coffee. As in the current study, 20 minutes boiling was Assuming the average of the Arabic coffee consumption used, and the ratio of coffee powder/water was adjusted to among Arabian peoples is 5 cups/person/day, then the mean the Arabic traditional preparation methods. However, in es- daily intake of acrylamide could be about 10.7 presso coffee, hot water and high pressure were applied and µg/person/day. In this sense, considering the average of the the ratio of coffee powder/water is high resulting in high adult body weight of 70 kg, the mean acrylamide intake from extraction rate of acrylamide [21]. Generally, the concentra- coffee among the Arabian people could be 0.15 µg/kg body tion of acrylamide in the Arabic coffee beverages is high. weight/day. These values are similar to the estimated acrylamide intake of about 0.106, 0.168, and 0.171µg/kg 3.4. Comparing Acrylamide Concentration in the Coffee body weight/day for people in Denmark, France and Swe- Beans Powder and Beverages den, respectively [24]. However, it is higher than the people The statistical analysis of the acrylamide concentration in consuming other coffee types from vendor machines in the dry coffee beans powder and beverages demonstrated Spain [21]. Indeed, several factors affected the extractability significant differences between them (Fig. 4). of the acrylamide in the coffee beverages, such as the coffee beans type, roasting temperature and durations, grinding The acrylamide concentration in the coffee beverage was process, powder types, powder/water ratio and beverage lower than dry powder. For the unroasted samples, the preparation processes [10]. Furthermore, the intake of acrylamide concentration in the dry powder was higher (P ≤ acrylamide from the coffee could also depend on various 0.05) than the beverage. Significant differences (P ≤ 0.001) cultural consumption habits. Controlling these factors could in the mean of the acrylamide concentration were observed potentially reduce the acrylamide concentration in the coffee between the roasted dry beans and the beverages at 30, 40, beverage. Managing the daily consumption of the Arabic and 50 min. However, at the longest roasting duration, the coffee could also reduce the daily exposure and intake of the differences between dry powders and beverages were highly acrylamide. significant (P ≤ 0.001). It is clear that increasing the roasting duration has affected the acrylamide extractability, as elon- CONCLUSION gated time resulted in higher extractability (83-86%) com- pared to the shorter duration (68-69%). Overall, using a short The findings of this study concluded that the Arabic cof- time (30 min or less) to roast the coffee beans to prepare the fee (Qahwa) could be a significant source of dietary acryla- Arabic coffee beverage is highly recommended, due to the mide to its consumers, and preparation processes should be potentially lower concentration of acrylamide in coffee beans considered. The roasting conditions and particularly dura- roasted at 125°C for 30 min. In addition, the roasting tem- tions significantly affected the acrylamide concentration in perature might also be reduced to minimize the acrylamide Arabic coffee beans powder and beverage. Short roasting 684 Current Nutrition & Food Science, 2019, Vol. 15, No. 7 Amal H. Alshawi durations (up to 40 min) had a higher mean acrylamide con- Health and Disease Prevention. Academic Press: San Diego 2015; centration than those with long roasting durations. Therefore, pp. 575-84. [2] Morales FJ, Mesias M. Analysis of Acrylamide in Coffee. In: it could be suggested that preparing Arabic coffee (Qahwa) Preedy VR, Ed. Coffee in Health and Disease Prevention. Academ- by roasting the coffee beans at a temperature of 125°C for a ic Press: San Diego 2015; pp. 1013-21. time shorter than 30 min or longer than 60 min could poten- [3] Ingo L, Ternité Ruediger, Jochen W, Katrin H, Helmut G, van der tially reduce the level and exposure of acrylamide in Qahwa. Stegen GH. 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