J. Agr. Sci. Tech. (2015) Vol.

17: 837-846

Modeling the Simultaneous Effects of Microwave and
Ultrasound Treatments on Sour Cherry Juice Using
Response Surface Methodology

B. Hosseinzadeh Samani1, M. H. Khoshtaghaza1∗, S. Minaee2, and S. Abbasi2


This study aimed to examine the effectiveness of combined microwave-ultrasonic
pasteurization system on Escherichia coli and vitamin C content in sour cherry juice
(SCJ). Based on the findings, microwave output power, ultrasound power, and ultrasonic
exposure time as well as the microwave-induced temperature were the most effective
factors in reducing E. coli and vitamin C content. In addition, the microwave-induced
temperature and ultrasonic exposure time, as independent variables, were both effective
on E. coli removal. At higher temperatures, the effectiveness of ultrasonic waves as well as
cavitation intensity declined. However, their combined effect (ultrasound and
temperature) was more significant than their individual effect. It was also found that any
increase in ultrasound power, ultrasonic exposure time, and microwave output power led
to a significant reduction in vitamin C content, while the ultrasound power was the most
effective. On the basis of RSM modeling, the optimum processing condition was: 352.21W
microwave output power, 49.94˚C temperature, 475.13W ultrasound power and 6 minutes
of exposure time. On the basis of response surface methodology (RSM) modelling, the
maximum vitamin C content was 142.5 mg per 100 mL with no remaining E. coli.

Keywords: Cavitation, Microwave output power, Pasteurization, Ultrasound power.

INTRODUCTION 2004; Valdramidis et al., 2010).. However,
the main challenge facing the non-thermal
In order to reduce the adverse effects (loss technologies in food processing is the
of vitamins, flavor, and non-enzymatic inactivation of pathogenic microorganisms
browning) of the thermal pasteurization and food spoilage agents, which can be
method, other methods capable of achieved by various methods. According to
inactivation of microorganisms can be the literature, the non-thermal methods not
applied. In doing so, non-thermal methods only totally eliminate the microorganisms,
are of interest, including pasteurization but also decrease or, in some cases,
using high hydrostatic pressure processing completely remove the heat required for the
(HPP), electric fields, and ultrasound waves destruction of microorganisms (Piyasena et
(Toepfl et al., 2007; Aronsson et al., 2001; al., 2003). Moreover, different studies have
Mertens , 1992). The ultrasound technology reported on the capability of ultrasound in
has been the main focus of studies in recent deactivating microorganisms and enzymes
years, which mainly uses the frequency (Hosseinzadeh Samani et al., 2013;
range of 20 kHz to 10 MHz (Knorr et al.,
Biosystems Engineering Department, Tarbiat Modares University, P. O. Box: 14115-139, Tehran, Islamic
Republic of Iran.

Corresponding author, e-mail: khoshtag@modares.ac.ir
Department of Food Science and Technology, Tarbiat Modares University, P. O. Box: 14115-139,
Tehran, Islamic Republic of Iran.


the 1999). De Gennaro et al. The qualitative index. remaining suspended particles. Microwave and ultrasound processing MATERIALS AND METHODS The effect of microwave-ultrasound combination on pasteurization of SCJ was Preparation of SCJ Samples studied. 1998). Baumann et al.. 2002. A total of 3 strains was observed. In order to separate pulp 838 . effectiveness on E. In doing so. product uniformity and yields. so that the resulting cellular mass while no such interaction with the other two was suspended in sterile SCJ. Huang et al. to date no studies have been mL of SCJ. This was kept vegetative cells in laboratory medium. _____________________________________________________________ Hosseinzadeh Samani et al. cleaned.. improving 2013). total phenol content. Acierno mention that the dimensions of the reactor et al.000 rpm between microwave and ultrasonic treatment (7. It should be noted that the cherry juice (SCJ) pasteurization. Hungary). The et al. proliferation included nutrient solid/liquid ultrasonic system and examine its cultures (Kuldiloke. 2004..656 g). It is necessary to (Hosseinzadeh Samani et al. 2010). the strain was microwaves improved levels of ascorbic cultivated on an agar nutrient culture acid. the selected prepared fruits were then dewatered using an variables included microwave power.307 g) for 20 a heating mechanism other than the direct minutes. coli and vitamin C content in SCJ. coli was considered as an indicator for the effectiveness of these waves on Sour cherry fruit of Mashhad Champa microorganisms.. Using coli vial was opened. Moreover. a loop activities compared to the conventional of grown microbial strain on agar was pasteurization method (Igual et al. and antioxidant (Micromedia.. To this effect. 2005. and protecting food into a reactor with diameter and height of 80 from surface browning and crusting and 50 mm. coli in vinegar and apple juice compared with conventional pasteurization practices (Cañumir et al. Wang inside an oven for 18−24 hours at 37°C. Therefore. Afterwards. (2010) showed that there was a liquid culture containing the grown cells was synergetic interaction in killing DSM 4006 centrifuged for 5 minutes at 8. the amount of variety was purchased from local markets remaining vitamin C was selected as a and washed. inoculated to a 25 mL nutrient liquid culture Utilization of microwave and ultrasonic is under sterile conditions in order to prepare useful for inactivating Alicyclobacillus the microbial suspension. the destruction level of E. it was reported in the literature on simultaneous held for 15−30 minutes before the effect of microwave and ultrasound on sour deactivation. Once the lyophilized standard strain of E. suspensions and tissue components. electric juicer. extracted juice was poured into a centrifuge A novel thermal processing method using with the speed of 6. 2007). Many were optimized during pretests. This study cultures used for primary activation and aimed to develop a combined microwave. the samples were poured unique micro-structure. 2013. Tajchakavit et al... mL of this suspension was inoculated to 300 However. 2002). Lactobacillus Preparation of Microbial Suspensions plantarum and E. For complete separation of the thermal method is called microwave.. studies have reported a faster eradication of Saccharomyces cerevisiae.. enhancing vacuum pump (Hosseinzadeh Samani et al. and cored. respectively. For adaptation purpose. the Microwave heating is widely used in the transparent portion of the extract was passed food industry because of its reduced through a Whatman filter paper using a processing time and costs.000 rpm (4.

800 watt. Korea) was placed in they were superficially cultivated on line. a E. Switzerland) working at 20±1 kHz In order to determine the cells survival frequency was used. Adjusting a level meter. In this and exposure time of samples to ultrasound. consumption was continuously recorded Germany).000W electric generator (Model MPI. Finally. 2009.. In order to obtain the reactor where they were exposed to optimum value using the RSM. In order to supply uniform ultrasonic waves. Range of level Independent variable -α -1 0 1 α Microwave Power (W) 200 350 500 650 800 Temperature (°C) 20 30 40 50 60 Ultrasound power (W) 200 400 600 800 1000 Ultrasonic exposure time (Min) 3 6 9 12 15 Figure 1. Iran) was used to al. were counted after 48 hours (Kuldiloke. selected as the size of each sample. E. (Figure 1) A schematic diagram of the set up used for the experiment is shown in Figure 1. Ultrasound and Sour Cherry Juice ___________________________________ sample temperature.coli Survival Counts 1. The experimental treatments were set according to Table 1. Laboratory circulating was used (Halim et al. 200 mL of SCJ was Vitamin C Assay Table 1. the treated and untreated samples processing system. the inoculated SCJ was poured into 2002). the treated samples were removed from the system for the intended (1) experiments. 839 . First. Various levels of independent variables with regards to the employed analysis method.coli plates were placed inside using a power analyzer (Lutron. It is noteworthy that the power MacConkey sorbitol agar (Merck. DW-6090) the oven at 35˚C and the emerged colonies throughout the experiment. Equation (1) ultrasonic waves. To complete the count. the ultrasound power. Mostafaei et water (Sahand 1000i.. 2013): maintain the samples’ temperature during Yi = β 0 + ∑ βi X i + ∑ βij X i X j + ∑ βij X i2 + ε sonication. the system’s tank. a microwave oven were diluted using physiological serum and (Samsung. system. the samples were first heated in a The RSM using the central composite design microwave oven to a desired temperature (CCD) was employed for data analysis and and were then placed inside the ultrasonic optimization (Table 1).Microwave. Schematic diagram of the designed microwave-ultrasound processing system.

95015 1 11. the interaction effect of microwave power and temperature.0002 MP×UP 0.1025 10 0. Vitamin C= 219.41697×t+4.062371 5.159693 1 0.3599 Residual 0.039442 3.0121 Lack of Fit 0.424234 117. coli count was simplification purposes.00458×Temp×t +0.0001 Microwave Power (MP) 0.003533×MP×t+0.36588+0.302212 1 0.6-dichlorophenol-indophenol ultrasonic exposure time were not significant colorimetry using a Cecil spectrophotometer in the fitted model (P< 0.0025 Temp×UP 0.01025 0.2E- temperature. _____________________________________________________________ Hosseinzadeh Samani et al. As shown in Tables 2 and 3.021954 1.141173 1 0. the final statistically significant.0001 Time (t) 0. 840 .70303 0. 2006).10 denotes significant effect. respectively.1980 Temp2 0.010794 1 0.0911 Temp×t 0.302149 1 0.010794 0. it could be asserted that the accuracy of the RESULTS AND DISCUSSION developed model is acceptable.600587 1 5.06348×UP-2.600587 462.062371 1 0.0384 MP×t 0. -4. the non. 0. (2) significant (P< 0.6167 < 0.79E-06×UP2 (CV) of the model were 0. equations were as follows: significance of the “lack of fit” indicates the Log(N/N0)= effectiveness of the resulting model (Table 2).00022×MP×t-2.302149 24.259703 0.7383 Pure Error 0. as well as Table 2.97104 0.073639 6.0063×UP+0.10).096503 7.05621×Temp- respectively.98 and 5.0158 Total 20.202106 16.10).171088×Temp- Except for the square of microwave power and 0.08426×t2 of microwave power.0128 Temperature (Temp) 11.000187×UP×t- 0.096503 1 0.021954 1 0. some of the As the analysis of variance (ANOVA) model coefficients were not significant and results for the second order model show.0262 MP×Temp 0.3%.08E-06×MP*UP.5E-05×Temp×UP- of determination and coefficient of variation 0.6%.00175×Temp2+ 1.0001 Ultrasonic Power (UP) 5.892069 0.6E-05×UP2+0.94 and 7.859 < 0.203052×t-9. Therefore.93928 14 1.073639 1 0. coli values.648734 0. Since the “lack (λ= 500 nm) and reported as mg 100 mL-1 of fit” is non-significant and also the values for SCJ (Burdurlu et al.1815 15 0. The adjusted coefficient 0. The first and second order variables 1.0010 MP2 0.97624 0.0002 UP×t 0.66721 0.085841 0.079 5 0.141173 11. Sum of Mean df F value P-value a Source squares square Model 19.002734×MP+0.814393 0. all the other factors were 05×MP×Temp+2.0038 t2 0.302212 24.001054×Te (3) amount of vitamin C remaining after the mp×UP-0.1429 < 0. Also. Vitamin C content of SCJ was determined the interaction effect of temperature and the by 2.7054 < 0.51E- Table 3 shows the ANOVA results for the 05×MP×UP+0.975457 0. the coefficient of determination and coefficient of variation were 0.95015 987.00698×Temp2- process. the were excluded from the fitted model for data from the surviving E.154606 0.836628 69.836628 1 0.202106 1 0.039442 1 0.159693 13.4945-1.0001 UP2 0. ANOVA results for first and second order coefficients using response surface method for E.04605×Temp×t+0..12078 29 a P-value< 0.19781 0.

32336 1 29.54719 < 0. Sum of Mean df F value P-valuea Source squares square Model 897. At lower temperatures. 841 .395741 0.44025 24. coli.31507 15 1. Ultrasound and Sour Cherry Juice ___________________________________ Table 3.7016 1 235.77374 9.0002 Temp×UP 71.8017 29 a P-value< 0.15057 1 71.556341 0.coli account.3266 1 115.002369 0.0070 Residual 24. removal gradient caused by the increased N0 is initial E.525669 0.Microwave. increasing power (W). Where.15057 43.1697 1 326.83832 0.24585 2.61319 6. coli survival. coli and its resulting heat.0001 Time (t) 235. By temperature on E.7016 145.075683 1 0. N is E.36353 1 13.53701 1 30. Comparison of experimental and model predicted data: (a) E.56341 10 1.94765 0.77374 1 15.4866 14 64. ANOVA results for first and second order coefficients using response surface method for remaining vitamin C.53701 18.0007 MP×t 40.08962 0. incremental gradient than the microwave- Figure 2 presents the goodness of fit for induced temperature.262503 1.8318 MP2 2.750331 Total 921.0001 Microwave Power (MP) 3. However.0001 Temp×t 30. it is likely due to the steeper microbial the faster reaching to the distinct higher (a) (b) Figure 2. the experimental and model predicted data.2146 < 0. increasing the microwave power had no It was found that the effect of final SCJ effect on the reduction of E.24585 1 3.0218 t2 15.3266 71. t is time (s).1697 201.243977 0.1775 Temperature (Temp) 115.0001 MP×Temp 0.coli count after treatment. (b) Vitamin C.61319 1 10.046689 0. Temp is temperature (°C).525669 1 0.4047 < 0. UP is microwave output power had a milder ultrasound power (W).10619 39.36353 8.621005 Lack of Fit 15.0006 UP×t 0. MP is microwave temperature.5775 MP×UP 29.262503 1 2.10 denotes significant effect.0001 Ultrasound Power (UP) 326.14517 < 0.324286 0.32336 18.44025 1 40. According to Figure reduction was enhanced possibly owing to 3a.88917 0.075683 0.730842 0.5929 Pure Error 8.751657 5 1.89289 < 0. the E. coli reduction was more increasing the temperature to 50 or 60˚C and significant than that of the microwave power raising the microwave power.0117 UP2 10.2558 Temp2 13.547291 0.

These findings could be Equation (1). temperatures and having insufficient any increase in the independent variable chance for the microorganisms to adapt with would result in a larger negative value in the new conditions. coli derived from Equation (1) as the product of removal. This shows that removal of microorganisms (Tajchakavit et 842 . Various researches have already microwave power multiplied by temperature indicated the effect of microwaves on had a negative coefficient. _____________________________________________________________ Hosseinzadeh Samani et al. (a) (b) (d) (c) (e) Figure 3. Influence of treatment variables on E. coli reduction. as well as more E.

The reason the bubble wall. can lead to a loss of pigments and bubbles. the combined effect of Figures 3-e and -c... Ugarte-Romero et al. coli sensitive to heat.500˚C and 500 bar. 2006). Wider probing range would result in more According to Figures 4-a and -b. Thus. It is believed that increasing increased. and the microwave power waves remove the microorganisms by would lead to a reduction in vitamin C thinning the cell walls. causing a drastically for this reduction could be the wider reduced cavitation effect (Wu et al. and microwave power. concentrating heat. 1998. coli Cañumir et al. independent variables effective on E. contraction process occurs. During the ultrasonic treatment. as the temperature 843 .. C breakdown at high temperature The SCJ temperature and ultrasonic (Odriozola-Serrano et al. 2008.. Ultrasound and Sour Cherry Juice ___________________________________ al. This. therefore.. The main bacterial al. coli reduction temperature and ultrasonic exposure time gradient was greater in regions where the was more significant than their individual ultrasound power and exposure time were effects. the cavitation-induced regions with extreme temperature and thermolysis could be another reason for the pressure which. Similar findings breakdown of these compounds. any bubbles inside the fluid giving rise to increase in the ultrasound power. Gentry and Roberts. and the process due to providing higher sonic flow periods temperature is directly correlated with the in the reactor chamber. In addition.. but they are too effects (Vikram et al. 2002). Ugarte-Romero et al. 2008. effectiveness of ultrasonic reduction was greater than that of the waves and the cavitation intensity were both microwave power (Figure 3-b). the E. As shown in weakened. 2006). in turn. 2002). content of the SCJ. have been reported by other researchers (Wu thermal processing adversely affects vitamin et al. However. This. in some cases. increasing temperature (Figures 4-c and -d). Increasing the ultrasonic exposure time This clearly indicates that vitamin C is can decrease the survived number of E. Oxidative reactions can be the energy inside the bubble is not sufficient for reason for anthocyanins and ascorbic acid turning into the gas phase and. 2009. ultrasonic cavitation (Vichare et al. The impact of ultrasound and developing free radicals inside the fruit power was more effective than the juice. The hot points vitamin C of orange juice. movement range of the probe in the fluid.. in to the formation of gas and development of turn. 2008). 2005). contributing compounds such as anthocyanin. ultrasound discoloration. Torregrosa exposure time are both important et al. reach breakdown of these compounds (Tiwari et 5. It was reported that microwave effect of ultrasound is the pressure variations processing could increase the breakdown of induced by these conditions. removal.. at high The effect of ultrasound power on E. the breakdown during the ultrasound treatment. The contracted This process can be enhanced by the free molecules collide with each other and create radicals created through the ultrasound shock waves. coli temperatures. longitudinal waves are as the ultrasound power increased. The vapor pressure increase inside the collapsing removal capability of ultrasonic waves was bubbles which cushion the inward motion of enhanced as the power increased.. 2001). 2005. Ultrasonic exposure time.. than in the regions where only the temperature of the fruit juice can result in microwave output power was increased. Moreover. 2006). due to its thermal can kill some bacteria.Microwave. This created which set the stage for pressure ultrasound-induced loss in vitamin C content variation.. As shown in Figure 3-d. At the critical point. This is clearly shown by when ultrasonic waves collide with the the increased vitamin C reduction gradient liquid medium. concentrated and their influence area is not Vitamin C content of SCJ decreased with large (Kuldiloke. The shock waves develop treatment. The bubbles expand and breakdown or interaction with other attain larger cross-section area. produces bubbles was likely the result of ascorbic acid inside the medium.

94 kJ of energy. and reduction of E. and The objectives for the process optimization ultrasonic exposure time as well as sample were to obtain: minimum energy consumption. coli reduction. Ultrasound power. vitamin would become the leading factor in decreasing C content would be 142. This negatively affected the vitamin 844 . Effect of process variables on vitamin C content of SCJ. 49. with 475.94˚C temperature. 2011). Based on these values. (a) (b) (c) (d) Figure 4.21W waves was enhanced owing to the increased microwave power. vitamin C content. This weakens the impact of consumption energy. coli and retention of vitamin C maximum vitamin C retention. coli remaining. it was was increased.5 mg per 100 mL with the vitamin C content since cavitation intensity zero E. microwave power. coli count and 1. temperature exposure time. The optimum cavitation. optimal values on the device.13W ultrasound power and 6 minutes of further temperature increase. When the ultrasound power the priority of E.425. This increased duration of oxidative reactions and confirms the precision of the optimum occurrence of thermolysis.87 kJ of decreases. and temperature (Rawson et al. coli was weighted 5 in the final optimization while the decreased due to the increased intensity of rest were weighted 3. longer vitamin C content was obtained to be 144. _____________________________________________________________ Hosseinzadeh Samani et al. the actual On the basis of our findings. research showed that the breakdown level of ascorbic acid in watermelon juice increased CONCLUSIONS significantly with the ultrasonic amplitude.. the effectiveness of ultrasonic combination was obtained as 352. reached 50˚C. However. Moreover.2 ultrasonic exposure times decreased the mg per 100 mL with no E. By implementing the ultrasonic waves. the amount of E. internal pressure of bubbles.407. temperature are the influential factors in the reduce the E. Considering contents of SCJ. coli content to zero. more likely because of the while consuming 1. exposure time. another condition determined by the developed model.

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