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7 Macromolecular Chemistry, Cadi Ayyad University, Abdelkrim Khattabi st, Marrakech 4000,
8 Morocco
9 Abstract: The equilibrium moisture content of green beans was investigated in order to
10 determine their hygroscopic stability, the optimal storage condition and the net isosteric heat.
11 Furthermore, how different drying methods affect the nutritional content and the quality of
12 some foods was checked up. Thereby, the effect of freeze-drying, forced convection, and
13 direct sunlight drying on nutritional value of green beans was evaluated. To assess the
14 suitability of each drying process, sugars, proteins, vitamin C, titratable acidity, polyphenols,
15 polyphenoloxydase and peroxydase activities, antioxidant activity and color change were
16 investigated. The obtained results favored the freeze-drying technique for nutritional
17 considerations. In fact, the amounts of sugars, proteins, polyphenols and vitamin C were
18 highly conserved, and the titratable acidity was low (0.54±0.05 mg equivalent malic acid /
19 100g DM). The forced convection solar drying at 40°C led to sufficient quality given that the
20 proteins and vitamin C contents were quite high (respectively 6.21±0.05 mg protein / g DM
21 and 1.94±0.09 mg / g DM). Contrariwise, the direct sunlight drying led to a final product with
22 the lowest quality. Ultimately, solar drying by convection at 40°C should be a potential
23 method since it retained the nutritional value and color for low energy costs.
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Corresponding author: Phone: +212 6 6694 6021; Fax: +212 5 2443 3170; E-mail: n.abdenouri@uca.ma
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24 Keywords: Food conservation, Convection drying, Solar drying, Freeze-drying, Nutritional
25 quality.
26 1. Introduction
29 sunlight drying is the most common drying method of fruits and vegetables in the developing
30 countries. The indirect solar drying is faster, leads to edible product, therefore, it can be
32 In some cases, at industry scale, freeze drying is more suitable for drying products at low
34 Despite many advantages, freeze-drying has always been recognized for its high costs of
35 investment, maintenance and for the required energy (Karam, Petit, Zimmer, Djantou, &
36 Scher, 2016).
37 The heat and mass transfers occurring during the drying process affect strongly the
38 structure and biological contents. For instance, decomposition and evaporation of some
39 essential compounds occur at high temperatures. Besides, the drying temperature is the most
40 determining factor since an inadequate temperature treatment leads to the loss of volatile
41 compounds (Bernhard, 1968) and thermal denaturation of proteins (Bull et al., 2017). Several
42 studies had shown that drying causes significant losses of ascorbic acid in different fruits such
43 as pear, and kiwi (Djendoubi, Boudhrioua, Kechaou, Courtois, & Bonazzi, 2012; Orikasa et
44 al., 2014). In other studies on banana (Guiné et al., 2015) and plums (Turturicə, Stənciuc,
45 Bahrim, & Râpeanu, 2016), the drying process at high temperatures led to a high drop of
47 Green beans (Phaseolus vulgaris) are originally from South and Central America (Chacón
48 S, Pickersgill, & Debouck, 2005). In Morocco, the production of green beans has increased to
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49 more than 100 000 tons in 2004. It is the second more exported vegetable after tomato, its
50 exports reached 73 000 tons in 2016. Green beans is a traditional food in the human diet, it is
51 rich in proteins, vitamin C, complex carbohydrates, polyphenols and fiber. In addition, the
52 consumption of dry beans has been linked to reduced risk of obesity (Geil & Anderson, 1994)
53 and cancer (Azevedo et al., 2003). Polyphenols have positive effects on human health and it
54 has been reported that they have antioxidant properties (Gamez et al., 1998). Vitamin C is a
55 usual marker of drying's impact on the nutritional value of plant foods. Furthermore,
56 polyphenoloxidase and peroxidase are heat and pressure stable enzymes in green beans
57 (Guenes & Bayindirli, 1993). In addition, the color evolution has to be examined during
58 drying. In fact, color is a visual index of the dried products’ quality, and it sorely influences
60 The properties and quality of dried green beans depends to a great extent on their
61 hygroscopic stability. This stability is determined by the relationship between the equilibrium
62 moisture content and its corresponding water activity at a given temperature which is called
63 sorption isotherm. The sorption isotherms are a good way to describe how active water is
64 bound to a wet product (Abdenouri, Idlimam, & Kouhila, 2010). They are also an extremely
65 valuable tool because it provides precious information about the optimal storage humidity and
67 The present work focuses on the hygroscopic behavior of green beans, and on the effect of
68 freeze-drying, forced convection drying (at 40, 60 and 80°C), and direct sunlight drying on its
69 quality parameters. To assess the quality of the dried samples, a list of quality parameters
72 change.
73 Nomenclature
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74 Xeq Equilibrium moisture content (% d.b)
75 aw Water activity
83 The used green beans are fine green filet beans “Rolande” with slender pods of 15±1cm,
84 and they were collected from Souss region (Morocco). Samples were cleaned and edges-
85 trimmed before the experiments. Three drying techniques were conducted on fresh samples
86 with initial water content of 89.1±0.5%. The samples were dried until a final water content of
87 14±1%.
88 For the direct sunlight drying, samples were placed directly under solar radiation (about
89 850W/m²) for three days (discontinuous drying). The ambient temperature was almost 36°C,
90 and the mean relative humidity and the atmospheric pressure were respectively 39% and
91 1018hPa.
92 For the forced convection drying, the experimental apparatus consists of an indirect forced
93 convection solar dryer (Fig. 1), with a solar air collector, an electric auxiliary heater, a
94 circulation fan and a drying chamber. The dryer was described in detail by EL Ferouali et al.
95 (2016). Samples were dried at three temperatures (40, 60 and 80°C), at an air flow rate of
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97 For the freeze-drying technique, samples were frozen at -20°C and then lyophilized during
98 20 hours using a lyophilizer (Alpha 2-4 LP+) at a temperature of -80°C and a pressure of
99 0.001mbar.
101 The hygroscopic equilibrium was achieved by the static gravimetric technique, it consists
102 on using six saturated salt solutions (KOH, MgCl 2 6H2O, K2CO3, NaNO3, KCl, BaCl2 2H2O).
103 These salts provide a range of relative humidity of 5–90 % (Greenspan, 1977).
105 The applied biochemical protocols for the determination of the studied quality parameters
107 The adopted method for the sugars content was described by Dubois, Gilles, Hamilton,
108 Rebers, & Smith (1956). 200mg of samples were ground in 5ml of ethanol at 80%. Then, the
109 solution was centrifuged for 10min at 5000rpm. After that, 0.1ml of the supernatant was
110 added to 1ml of phenol at 5% and to 5ml of concentrated H 2SO4. The mixture was put into a
111 boiling water bath for 20 minutes, and then cooled in the dark. The absorbance was measured
112 at 495nm by using a spectrophotometer (VWR UV-3100PC) and sugars content was
114 Total proteins determination was performed according to Bradford (1976). Thereby,
115 100mg of samples were ground in 2ml of cold phosphate buffer (pH = 6.8). The extract was
116 centrifuged at 4°C for 20 min at 12500×g. Then, 50µl of the supernatant was added to 100µl
117 of distilled water and 2ml of Bradford reagent. The mixture was stirred and the homogenate
118 was incubated for 5min at 27°C. The absorbance was measured at 595nm, and the results
119 were expressed in Bovine Serum Albumin equivalent by reference to a standard range
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121 The direct titration method of vitamin C with diiodine was used for the determination of
122 vitamin C content Suntornsuk, Gritsanapun, Nilkamhank, & Paochom (2002). To achieve
123 that, 1g of the product was homogenized with 20ml of distilled water. After filtering, 10ml of
124 the filtrate was added to 10ml of distilled water and a few drops of starch. Titration was
126 The measurement of titratable acidity was carried out according to Türkmen & Eksi
127 (2011). The titration of the free acid was accomplished with NaOH solution (0.1N) in the
129 For the determination of total phenols; 100mg of samples were ground in 2ml of methanol
130 at 80%. Then, the mixture was sonicated for 10min in an ultrasonic bath containing distilled
131 water. The homogenates were centrifuged at 15 000×g during 10min. Supernatants were
132 tested by a differential assay in the presence and absence of PVPP according to Bridi et al.
133 (2014). Samples were passed through three cycles of pre-treatment. At first, 50mg of PVPP
134 was added to 500µl of the supernatant. Then, the solution was centrifuged at 10000×g for
135 5min. Then, 250µl of the supernatant was recovered, and 25mg of PVPP was added to it and
136 the obtained mixture was stirred and centrifuged again. Finally, 125µl of the supernatant was
137 treated by 12.5mg of PVPP, and the obtained mixture was stirred and centrifuged again in the
138 same experimental conditions as mentioned before. The total polyphenols concentration was
139 determined using the Folin-Ciocalteu reagent according to Singleton & Rossi (1965).
140 Polyphenoloxidase and peroxidase activities were determined by the following method:
141 50mg of samples were ground in 1ml of phosphate buffer (0.1M, pH 6) containing the
142 insoluble PVP (5%), and then centrifuged for 30min at 12000×g. After that, the protocol
143 depends on the nature of the sought activity. For the polyphenoloxidase activity, 100μl of
144 supernatant were added to 2ml of catechol (10mM) and then the absorbance was picked up
145 after 3min at 410nm. For the peroxidase activity, 100μl of supernatant were added to 1ml of
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146 guaiacol (20mM). The reaction was started up by the addition of 0.5ml of hydrogen peroxide
147 (0.1%) and the absorbance was measured after 3min at 470nm.
149 radical (DPPH) as described by Scherer & Godoy (2009) with some modifications. For this
150 purpose, 100μl of different concentrations of each phenolic extract were added to 0.9ml of the
151 freshly prepared DPPH (0.004%) methanol solution. Negative control was prepared in parallel
152 by mixing 100μl of methanol with 0.9ml of the methanol solution of DPPH. The absorbance
153 was measured at 517nm, after incubation in a dark place for 30min.
155 All measurements were performed in triplicate and were reported as mean and standard
156 deviation. All data were analysed using the SPSS 21.0 statistical package. An analysis of
157 variance (ANOVA) followed by the Student Newman–Keuls post hoc test were used to
158 compare differences in the means. The level of significance was defined as P<0.05.
160 The CIE Lab color space was used for color analyses based on imaging process of
161 samples. L* represents lightness, ranging from 0 (black) to 100 (white). The parameter a*
162 ranges between -128 (green) and 127 (red), and b* ranges between -128 (blue) and 127
163 (yellow). The L*, a* and b* color parameters were determined by using a Minolta Chroma
164 meter. Values of different parameters were expressed as the mean ± standard deviation.
168 Fig. 2 shows the effect of temperature on desorption and adsorption of green beans. Xeq
169 increases by decreasing temperature at constant water activity. This result may be explained
170 by the higher excitation state of water molecules at higher temperature thus decreasing the
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171 attractive forces between them. The hysteresis appears between desorption and adsorption.
172 This phenomenon is explained by thermo dynamical irreversible processes that occur during
175 All experimental data of desorption and adsorption at 30, 40, 50 and 60°C were gathered
176 on the same graph (Fig. 3). The sorption isotherm may particularly be described by a 3 rd
177 degree polynomial. The polynomial fitting is presented in the same figure and its relative
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179 X eq =1.8561+100.49951 a w -325.44754 a w +318.05885 a w (1)
180 This curve allowed to determine the value at which the second derivative of Xeq equals to
181 zero (inflection point) and consequently the optimum value of water activity for storage. The
182 found value for green beans (awop=0.34±0.005) is in agreement with the general stability
183 domain of biological products that ranges between 0.2 and 0.4 (Le Meste, Roudaut, Chiotelli,
186 The net isosteric heat of sorption (∆hd) or differential enthalpy stands for the binding
187 energy of the water to the substrate. This energy must be added to the heat vaporization
188 energy (∆hvap) for dehydration (Eq. (2)). The net isosteric heat of sorption was calculated from
189 the experimental data using the Clausius Clapeyron equation given by Eq. (3) (Beristain,
[( ) ]
d ( ln a w ) - ∆hd
=
192 1 R (3)
d
T Xeq
193 This equation involves determining the isotherms at different temperatures in order to
194 calculate the logarithmic variation of the water activity as a function of inverse temperature
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195 for fixed equilibrium moisture content. It is assumed that isosteric heat does not vary with the
196 temperature. Hence, ∆hd is determined from the slope -∆hd/R (Eq. (4)).
- ∆hd 1
197 ln( a w )= +cst (4)
R T
198 Fig. 4 presents the net isosteric heat of sorption of green beans at temperatures ranging
199 between 30°C and 60°C. This curves show that the isosteric heat is higher (59.97±1.01
200 kJ.mol−1) for small values of the moisture content (Xeq=12% d.b) indicating the highest
201 binding energy for water removal, and it decreased along with the increase of Xeq.
204 Fig. 5 shows the sugars content of fresh and dried samples. In all studied cases, sugars
205 content drop after drying. The reduction of sugars content can be explained by the Maillard
206 reaction occurred in samples (Mota, Luciano, Dias, Barroca, & Guiné, 2010). The maximum
207 value was obtained by freeze-dried samples (529.44±3.01 mg glucose equivalent / g DM).
208 The minimum value was recorded in the case of direct sunlight drying (205.37±9.11 mg
209 glucose equivalent / g DM). Furthermore, under forced convection, the sugars content in the
210 dried samples increased gradually with temperature, which is in accordance with previous
213 All dried samples were characterized by low proteins content compared to fresh ones (Fig.
214 6). The maximum value was obtained by the freeze-drying technique (8.69±0.23 mg protein /
215 g DM). On the other hand, convective drying at 80°C led to the lowest value (4.01±0.13 mg
216 protein / g DM). In addition, it was noticed that proteins' loss are substantial at higher
217 temperatures in convection drying, which is in agreement with previous results on maize
218 (Odjo, Malumba, Dossou, Janas, & Béra, 2012). This decrease in proteins content at high
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219 temperatures is due to breakage of the hydrophobic bonds, which causes the proteins
222 Significant loss of Vitamin C was noticed by comparing dried samples to fresh ones (Fig.
223 7). The freeze dried samples recorded the highest value (2.98±0.10 mg / g DM) followed by
224 the convective dried samples at 40°C. Direct sunlight drying and convective drying at 60°C
225 led to final products with almost the same vitamin C contents. Finally, the lowest value
226 (1.17±0.09 mg / g DM) was obtained by convective drying at 80°C. The same trend was
227 revealed during convective drying of pears in which it is shown that vitamin C degradation is
228 accelerated at higher temperatures (Djendoubi et al., 2012). This decrease of vitamin C is
230 non-enzymatic decomposition caused by the presence of oxygen in the atmosphere (Orikasa
233 The highest titratable acidity was recorded by the forced convection drying at 40°C (Fig.
234 8). Besides, it decreased inversely with temperature; its value is 4.02±0.05, 3.48±0.05 and
235 2.40±0.17 mg equivalent malic acid / 100g DM for 40°C, 60°C and 80°C, respectively. A
236 high titratable acidity was also perceived for direct sunlight drying (3.62±0.16 mg equivalent
237 malic acid / 100g DM). On the other side, the minimum value (0.54±0.05 mg equivalent malic
239 These results showed that titratable acidity varied according to both the drying technique
240 and the drying temperature. The same trend was perceived for other products. For instance,
241 titratable acidity of dried onion decreased gradually by increasing the drying temperatures
242 from 30°C to 70°C (Mota et al., 2010). This phenomenon could be related to the drying time
243 that is extended at low temperatures; leading to partial fermentation of samples. Furthermore,
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244 it could be related to evaporation of volatile acids at high temperatures (Mota et al., 2010).
245 Ultimately, results showed that freeze-drying technique is the most suitable for lowering free
246 acids.
248 The highest polyphenols content is assigned to the freeze-dried samples (1.98±0.30 mg
249 equivalent gallic acid / g DM) (Fig. 9). Otherwise, the polyphenols content increased
250 correspondingly with temperature for the forced convection technique; it increased from
253 Other authors have also noticed the decrease of the polyphenols content by convective
254 drying at low temperatures for banana and aronia (Guiné et al., 2015; Samoticha, Wojdyło, &
255 Lech, 2016). In fact, extended drying time at low temperature leads to irreversible chemical
256 changes (Guiné et al., 2015). This decrease could also be attributed to the exposure of the raw
257 material close to the optimal temperature of the polyphenoloxidase (40°C). Therefore, fast
258 and short drying at high temperature can lead to the inactivation of oxidative enzymes and
261 According to Fig. 10, the highest PPO activity was obtained for forced convection dried
262 samples at 40°C (44.03±1.22 EU / mg protein / min), whereas, lowest values were recorded
263 for freeze-dried samples and convective dried ones at 80°C (respectively 22.32±1.91 and
264 20.34±1.17 EU / mg protein / min). Similarly, the highest peroxidase activity was sighted for
265 forced convection dried samples at 40°C (1226.03±4.12 EU / mg protein / min) (Fig. 11). And
266 the lowest value was obtained for freeze-dried samples (366.25±3.58 EU / mg protein / min).
267 In similar works, according to Baltacıoğlu, Bayındırlı, Severcan, & Severcan, 2015, PPO is
268 fully active at 40°C, but this activity decreases after heating treatment between 50°C and
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269 70°C. Moreover, Amiour & Hambaba (2016) reported that maximum PPO and PO activities
270 of two date varieties were obtained at 35°C after heating treatment at various temperatures
271 (ranging from 10 to 80°C). Thereby, the high PPO and PO activities at 40°C indicates that this
274 Freeze-dried samples exhibited the highest antioxidant activity (83.41±0.19%) (Fig. 12).
275 Besides, previous studies on bananas had also shown that freeze-drying keeps this activity
276 better than convective drying (Guiné et al., 2015). In addition, the drying temperature of 60°C
277 retained the antioxidant activity (73.11±0.19%) better than 40°C (72.16±0.09%), these results
278 suggest that the exposure time has strong impact on the degradation of antioxidants than
279 temperature agreeing with the finding of Sadilova, Carle, & Stintzing (2007). However,
280 higher drying temperature (80°C) led to a high drop of antioxidant activity (66.62±0.09%). In
281 fact, this temperature changed the physical structure of the material which increase its
282 exposure to oxygen and its contact with components that degrade the bioactives responsible
285 Table 1 exhibits the (L*, a*, b*) parameters of fresh and dried samples. Convection dried
286 samples at 40°C almost retained the lightness of fresh samples, it was also noticed that by
287 increasing the drying temperature from 40°C to 80°C, L* decreased from 39.13±1.96 to
288 35.95±1.27. Contrariwise, freeze-dried and direct sunlight dried samples showed respectively
289 the extreme values (59.52±1.13, 28.11±1.63). In addition, convective dried samples at 40°C
290 retained the green color of tomatoes since a* recorded the lowest value among other
291 techniques (-9.92±0.73). Furthermore, forced convection dried samples at 40°C held the
292 parameter b* closer to fresh ones, and the lowest value was got by the freeze drying
293 technique. As a result, freeze-drying caused complete loss of color followed by the convection
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294 drying at 80°C and direct sunlight drying. On the other hand, convective dried samples at
296 4. Conclusions
297 The present paper links between the hygroscopic behavior, the drying process and the
298 green beans’ quality. The optimal water activity for the storage of the product was estimated
299 at awop=0.34±0.005. The net isosteric heat of sorption was evaluated, it is in the range of
300 59.97±1.01 kJ.mol−1 for small values of the moisture content (Xeq=12% d.b), and it decreased
301 along with the increase of the Xeq; this thermodynamic quantity estimates the required energy
302 for dehydration processes. Furthermore, the present work investigated the effect of freeze-
303 drying, forced convection (at 40, 60 and 80°C), and direct sunlight drying on the quality
304 parameters of green beans. Biochemical analyses showed that freeze dried samples recorded
305 the lowest value of titratable acidity (0.54±0.05 mg equivalent malic acid / 100g DM).
306 Furthermore, sugars, proteins, total polyphenols and vitamin C contents showed the highest
307 values compared to the other drying techniques. In addition, a strong antioxidant activity,
308 close to that of the fresh material, was obtained. Besides, regarding the forced convection
309 drying technique, the proteins and vitamin C levels, and polyphenoloxydase and peroxydase
310 activities decreased strongly by increasing temperature from 40 to 80°C. It is due to protein
312 activation at high temperatures. The convection technique retained partially the color of fresh
313 samples especially at 40°C. However, at this drying temperature, low sugars and polyphenols
314 contents were noticed. Finally, the direct sunlight drying led to a final product with the lowest
315 quality. In fact, the titratable acidity was high (3.62±0.16 mg equivalent malic acid / 100g
316 DM). Also, low levels were noticed for vitamin C (1.53±0.09 mg / g DM), proteins
317 (5.67±0.23 mg / g DM), polyphenols (1.08±0.05 mg equivalent gallic acid / g DM), sugars
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319 direct sunlight drying caused a low quality dried product. The freeze-drying method
320 engendered the best nutritional quality of the final product. But, this method requires
321 advanced technology and it consumes a lot of energy. Contrariwise, forced convection drying
322 technique especially at 40°C reached acceptable quality dried product with significant energy
323 saving, and it is more suitable for most crop producing countries.
324 Acknowlegement
325 This work was supported by the research institute for solar energy and new energies
326 (IRESEN) as part of the project SSH and all of the authors are grateful to the IRESEN
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