Changes in viscosity and pectolytic enzymes of tomato and strawberry juices processed by high-intensity pulsed electric fields

The effects pulse frequency (50–250 Hz), pulse width (1.0–7.0 μs) and polarity (monopolar or bipolar) of high-intensity pulsed electric field treatments (35 kV cm⁻¹ and 1000 μs) on viscosity and the pectin methylesterase (PME) and polygalacturonase (PG) activities were evaluated using a response surface methodology. Second-order expressions were accurate enough to fit experimental results. Tomato juice apparent viscosity increased within the range of the assayed conditions, achieving the highest values at 250 Hz and 7.0 μs in bipolar mode. At the same conditions the lowest residual PME (RA_PME = 10%) and PG (RA_PG = 45%) activities were observed in the juice. Apparent viscosity of strawberry juices slightly rose when frequencies higher than 100 Hz and 1-μs monopolar pulses were applied to the juice. Treatments causing the greatest increase in strawberry juice apparent viscosity also led to the lowest RA_PME (10%) and RA_PG (75%) values. In contrast, viscosity loss was promoted under the rest of assayed HIPEF conditions despite the low RA_PME values (<20%) achieved. Moreover, RA_PG did not decrease below 75% throughout the range of studied conditions.     

Inactivation of Salmonella enterica Ser. Enteritidis in tomato juice by combining of high-intensity pulsed electric fields with natural antimicrobials

ABSTRACT:  The effect of high-intensity pulsed electric field (HIPEF) treatment (35kV/cm, 4 μs pulse length in bipolar mode without exceeding 38 °C) as influenced by treatment time (200, 600, and 1000 μs) and pulse frequency (100, 150, and 200 Hz) for inactivating Salmonella enterica ser. Enteritidis inoculated in tomato juice was evaluated. Similarly, the effect of combining HIPEF treatment with citric acid (0.5%, 1.0%, 1.5%, and 2.0%[wt/vol]) or cinnamon bark oil (0.05%, 0.10%, 0.2%, and 0.3%[vol/vol]) as natural antimicrobials against S. Enteritidis in tomato juice was also studied. Higher treatment time and lower pulse frequency produced the greater microbial inactivation. Maximum inactivation of S. Enteritidis (4.184 log₁₀ units) in tomato juice by HIPEF was achieved when 1000 μs and 100 Hz of treatment time and pulse frequency, respectively, were applied. However, a greater microbial inactivation was found when S. Enteritidis was previously exposed to citric acid or cinnamon bark oil for 1 h in tomato juice. Synergistic effects were observed in HIPEF and natural antimicrobials. Nevertheless, combinations of HIPEF treatment with 2.0% of citric acid or 0.1% of cinnamon bark oil were needed for inactivating S. Enteritidis by more than 5.0 log₁₀ units (5.08 and 6.04 log₁₀ reductions, respectively). Therefore, combinations of HIPEF with organic acids or essential oils seem to be a promising method to achieve the pasteurization in these kinds of products.     

Impact of high-intensity pulsed electric field variables affecting peroxidase and lipoxygenase activities of watermelon juice

The effects of pulse frequency (50-250 Hz), pulse width (1.0-7.0 μs) and polarity (monopolar or bipolar) of high-intensity pulsed electric field (HIPEF) treatments (35 kV/cm and 1000 μs total treatment time) on peroxidase (POD) and lipoxygenase (LOX) activities were evaluated using a response surface methodology. Second-order expressions were accurate enough to fit experimental results. HIPEF bipolar treatments resulted to be more effective than monopolar treatments in reducing POD and LOX activities. Watermelon juice LOX was more resistant to HIPEF than POD within the range of assayed conditions. HIPEF treatments set at 50 Hz for 1.0 μs pulse width could attain minimum residual POD activity values up to 10%, whereas the highest POD activity reduction was reached by combining high frequencies and pulse widths. Thus, POD could be totally inactivated by applying 7.0-μs bipolar pulses at frequencies higher than 114 Hz. In addition, the effect of frequency on the LOX activity was highly affected by the pulse width of the treatment. Thus, treatments conducted at 220 Hz with bipolar pulses of 1.0 μs led to the lowest residual LOX activity (50%).     

Optimizing critical high-intensity pulsed electric fields treatments for reducing pectolytic activity and viscosity changes in watermelon juice

A response surface methodology was used to determine the combined effect of high-intensity pulsed electric fields (HIPEF) variables such as frequency, pulse width and polarity on the inactivation of pectolytic enzymes involved in viscosity changes of juices. Pectin methylesterase (PME) and polygalacturonase (PG) activities as well as viscosity were determined in watermelon juices processed at pulse frequencies from 50 to 250 Hz and pulse widths ranging from 1.0 to 7.0 μs in monopolar or bipolar mode. Electric field strength and total treatment time were maintained constant in all treatments at 35 kV/cm and 1,000 μs. Second-order expressions were accurate enough to fit the experimental results. The great PME reduction contrasted with the low impact of HIPEF on the PG activity of watermelon juice within the range of assayed conditions. Minimal residual PME activity values (15%) were obtained by selecting pulse widths higher than 5.5 μs at 250 Hz in bipolar mode, whereas the lowest PG residual activities (60%) were achieved after applying 7.0-μs bipolar pulses at 250 Hz. Moreover, watermelon juice viscosity increased throughout the range of studied conditions. The highest viscosity observed in the juice after applying 7.0-μs bipolar pulses at 250 Hz was related to the lowest PME activities obtained in the product treated under those conditions. Hence, the HIPEF processing optimization through frequency, pulse width and polarity could contribute to assure enzymatic inactivation while keeping valuable attributes of juices.     

Volatile compounds and changes in flavour‐related enzymes during cold storage of high‐intensity pulsed electric field‐ and heat‐processed tomato juices

BACKGROUND: The effects of high‐intensity pulsed electric field (HIPEF) processing (35 kV cm^?1 for 1500 µs, using 4 µs bipolar pulses at 100 Hz) on the production of volatile compounds and flavour‐related enzymes in tomato juice were investigated and compared with those of thermal processing (90 °C for 30 or 60 s). RESULTS: Tomato juice treated by HIPEF showed lower residual lipoxygenase (LOX) activity (70.2%) than juice heated at 90 °C for 60 s (80.1%) or 30 s (93.2%). In contrast, hydroperoxide lyase (HPL) was almost completely inactivated when the juice was subjected to 90 °C for 60 s, whereas roughly 50% of the control tomato juice was depleted after HIPEF treatment or thermal processing at 90 °C for 30 s. A slight decrease was observed in the initial LOX activity of treated and untreated samples during storage, whereas initial HPL activity was strongly affected over time. CONCLUSION: HIPEF‐treated juice exhibited higher levels of compounds contributing to tomato aroma than untreated and heat‐treated juices throughout storage. Thus HIPEF processing can preserve flavour quality and stability of tomato juice compared with conventional thermal treatments. Copyright © 2010 Society of Chemical Industry     

Inactivation of Oxidative Enzymes by High-Intensity Pulsed Electric Field for Retention of Color in Carrot Juice

The effects of high-intensity pulsed electric fields (HIPEF) on oxidative enzymes and color of fresh carrot juice were studied. A response surface methodology (RSM) was used to evaluate the effect of pulse polarity (mono or bipolar mode), pulse width (from 1 to 7 μs), and pulse frequency (from 50 to 250 Hz) on color and peroxidase (POD) inactivation of carrot juice treated by HIPEF. The total treatment time and the electric field strength were set at 1,000 μs and 35 kV/cm, respectively, at a temperature below 35°C. The physicochemical characteristics of carrot juice were measured. There was a linear relationship between electrical conductivity and temperature of the carrot juice. The results showed that HIPEF-treated carrot juice at 35 kV/cm for 1,000 μs applying 6 μs pulse width at 200 Hz in bipolar mode led to 73.0% inactivation of POD. The color coordinates did not change significantly. Therefore, HIPEF was effective in POD inactivation and carrot juice color preservation.     

Inactivation of tomato juice peroxidase by high-intensity pulsed electric fields as affected by process conditions

A response surface was used to establish the high-intensity pulsed electric fields (HIPEF) conditions in processing tomato juice to obtain the greatest peroxidase (POD) inactivation. Keeping constant the electric field strength at 35 kV/cm and the temperature below 35 °C, the treatments were set at pulse frequency from 50 to 250 Hz, pulse width from 1 to 7 μs and treatment time from 1000 to 2000 μs, using monopolar or bipolar mode. The effect of these parameters on POD inactivation was evaluated through a second order model that adequately fitted the experimental data (p = 0.0001), with a determination coefficient (R²) of 0.85. HIPEF treatment resulted to be more effective in bipolar than monopolar mode to reduce POD activity and the longer the treatment time, the greater the reduction on the enzyme activity. A pulse frequency of 200 Hz was enough to reach a minimum value of residual POD activity. The significant interaction term pulse frequency and treatment time was included in the model, showing that different combinations of both variables can lead to the same level of residual POD activity. The effect of pulse width was enhanced by using a bipolar mode, being feasible to maximize POD inactivation selecting pulse width higher than 5.5 μs in bipolar mode.     

Inactivation of orange juice peroxidase by high‐intensity pulsed electric fields as influenced by process parameters

The inactivation of orange juice peroxidase (POD) under high‐intensity pulsed electric fields (HIPEF) was studied. The effects of HIPEF parameters (electric field strength, treatment time, pulse polarity, frequency and pulse width) were evaluated and compared with conventional heat pasteurization. Samples were exposed to electric field strengths from 5 to 35 kV cm^?1 for up to 1500 µs using square wave pulses in mono‐ and bipolar mode. Effect of pulse frequency (50–450 Hz), pulse width (1–10 µs) and electric energy on POD inactivation by HIPEF were also studied. Temperature was always below 40 °C. POD was totally inactivated by HIPEF and the treatment was more effective than thermal processing in inactivating orange juice POD. The extent of POD inactivation depended on HIPEF processing parameters. Orange juice POD inhibition was greater when the electric field strength, the treatment time, the pulse frequency and the pulse width increased. Monopolar pulses were more effective than bipolar pulses. Orange juice POD activity decreased with electric energy density input. The Weibull distribution function adequately described orange juice POD inactivation as a function of the majority of HIPEF parameters. Moreover, reduction of POD activity related to the electric field strength could be well described by the Fermi model. Copyright © 2005 Society of Chemical Industry     

High-intensity pulsed electric fields processing parameters affecting polyphenoloxidase activity of strawberry juice

High-intensity pulsed electric fields (HIPEF) were applied to strawberry juice to study the feasibility of inactivating polyphenoloxidase (PPO). Response surface methodology was used to evaluate the effect of HIPEF processing, in which total treatment time (1000 to 2000 μs), pulse frequency (50 to 250 Hz), pulse width (1.0 to 7.0 μs), and polarity (monopolar or bipolar) were the controlled variables at a constant electric field of 35 kV/cm. The proposed 2nd-order response functions were accurate enough to fit experimental results. Strawberry juice PPO was strongly reduced within the range of assayed conditions. HIPEF treatments were more effective in bipolar than in monopolar mode in inactivating PPO. Treatments of longer duration resulted in reductions of the enzyme activity. Moreover, it was feasible to minimize residual PPO activity (down to 2.5%) by selecting bipolar treatments at frequencies higher than 229 Hz and pulse widths between 3.23 and 4.23 μs for a constant total treatment time of 2000 μs.     

Impact of high-intensity pulsed electric fields variables on vitamin C, anthocyanins and antioxidant capacity of strawberry juice

A response surface methodology was used to determine the combined effect of HIPEF critical processing conditions on vitamin C, anthocyanins and antioxidant capacity of strawberry juice. Keeping constant the electric field strength at 35 kV/cm and the treatment time at 1000 μs, the treatments were set at frequencies from 50 to 250 Hz, pulse width from 1 to 7 μs using monopolar or bipolar mode. A second order response function covering the whole range of experimental conditions was obtained for each health-related compound. Strawberry juice antioxidant potential was affected linearly by frequency, pulse width and pulse polarity. The quadratic term of frequency and the combined effect of frequency and pulse width were also significant. HIPEF treatments conducted at 232 Hz with bipolar pulses of 1 μs led to strawberry juices with the greatest presence of health-related compounds. The evaluation of the HIPEF critical parameters influence on health-related compounds can contribute to achieve optimal processing conditions to obtain strawberry juices with high antioxidant potential.     

Influence of high-intensity pulsed electric field processing on lipoxygenase and β-glucosidase activities in strawberry juice

The influence of high-intensity pulsed electric field (HIPEF) parameters, pulse frequency, pulse width and pulse polarity in strawberry juice lipoxygenase (LOX) and β-glucosidase (β-GLUC) was studied using a response surface methodology. The studied parameters affected on both residual enzymatic activities at unchanging electric field strength of 35 kV/cm and treatment time for 1000 μs. The contour plots showed a minimum defined space where residual activity of LOX remained at 65% and 70% in monopolar and bipolar mode, respectively. Low pulse frequencies (up to 61.6 Hz) in monopolar treatments as well as pulse frequencies and widths higher than 218 Hz and 5.4 μs in bipolar treatments did not have any effect on LOX inactivation. On the other hand, the higher the pulse frequency and pulse width, the higher the β-GLUC inactivation obtained. Moreover, when the HIPEF treatment was applied in monopolar mode, an enhancement in β-GLUC activity was observed in most of the experimental range. HIPEF treatments have demonstrated adequately that can reduce activity of enzymes that are involved in the formation of desirable flavor compounds, helping processors to obtain juices that keep their fresh flavor.

Industrial relevance

High-intensity pulsed electric fields (HIPEF) have proved to be effective in the interaction of microorganisms and enzymes in juices, maintaining their quality and freshness.HIPEF juice processing has demonstrated to have some advantages with regard to conventional thermal treatment. HIPEF treatments can reduce adequately enzymes that are involved in the formation of desirable flavor or color compounds. Thus, HIPEF technology can help processors to obtain juices that keep their fresh flavor by achieving optimal inactivation of related enzymes. This would prevent the product from undesirable off-flavor formation, which in turn would result in greater acceptability by consumers.     

Comparative study on color,viscosity and related enzymes of tomato juice treated by high-intensity pulsed electric fields or heat

The effects of high intensity pulsed electric fields (HIPEF) processing (35 kV/cm for 1,500 μs using bipolar 4-μs pulses at 100 Hz) on color parameters and viscosity, as well as peroxidase (POD), pectin methylesterase (PME) and polygalacturonase (PG), were evaluated during 77 days of storage at 4 °C and compared to thermal treatments at 90 °C for 1 min or 30 s for unprocessed tomato juice. HIPEF-treated tomato juice showed higher values of lightness than the thermally processed and the untreated juice throughout storage time (P < 0.05). Viscosity of HIPEF-treated tomato juice was also greater than both thermally treated and untreated for the first 35 days of storage. POD of HIPEF-treated tomato juice was inactivated by 97% whereas in the case of the thermally treated, 90 and 79% inactivation was achieved after 1 min and 30 s, respectively. The highest PME inactivation in tomato juice was obtained by PEF (82%) and heat treatment at 90 °C for 1 min (96%). PG of PEF-treated tomato juice was inactivated by 12% whereas thermal treatments at 90 °C for 1 min or 30 s achieved 44 and 22%, respectively. Despite the low rates of PG inactivation obtained, the pattern followed in the residual activity along the storage time was similar in the tomato juice treated by HIPEF than the thermally processed.     

Inactivation of Saccharomyces cerevisiae suspended in orange juice using high-intensity pulsed electric fields

Saccharomyces cerevisiae is often associated with the spoilage of fruit juices. The purpose of this study was to evaluate the effect of high-intensity pulsed electric field (HIPEF) treatment on the survival of S. cerevisiae suspended in orange juice. Commercial heat-sterilized orange juice was inoculated with S. cerevisiae (CECT 1319) (10(8) CFU/ml) and then treated by HIPEFs. The effects of HIPEF parameters (electric field strength, treatment time, pulse polarity, frequency, and pulse width) were evaluated and compared to those of heat pasteurization (90 degrees C/min). In all of the HIPEF experiments, the temperature was kept below 39 degrees C. S. cerevisiae cell damage induced by HIPEF treatment was observed by electron microscopy. HIPEF treatment was effective for the inactivation of S. cerevisiae in orange juice at pasteurization levels. A maximum inactivation of a 5.1-log (CFU per milliliter) reduction was achieved after exposure of S. cerevisiae to HIPEFs for 1,000 micros (4-micros pulse width) at 35 kV/cm and 200 Hz in bipolar mode. Inactivation increased as both the field strength and treatment time increased. For the same electric field strength and treatment time, inactivation decreased when the frequency and pulse width were increased. Electric pulses applied in the bipolar mode were more effective than those in the monopolar mode for destroying S. cerevisiae. HIPEF processing inactivated S. cerevisiae in orange juice, and the extent of inactivation was similar to that obtained during thermal pasteurization. HIPEF treatments caused membrane damage and had a profound effect on the intracellular organization of S. cerevisiae.     

Flavour retention and related enzyme activities during storage of strawberry juices processed by high-intensity pulsed electric fields or heat

The effects of high-intensity pulsed electric fields (HIPEF) processing (35 kV/cm for 1700 μs using pulses of 4 μs at 100 Hz in bipolar mode) and thermal treatments (90 °C for 30 s or 60 s) on lipoxygenase (LOX) and β-glucosidase (β-GLUC) activities as well as on the production of volatile compounds were assessed in strawberry juice for 56 days of storage. HIPEF-treated juice kept higher residual LOX activity than heat-treated juices during the first 28 days of storage. Moreover, β-GLUC increased its initial activity just after HIPEF processing. The concentration of DMHF in HIPEF-processed strawberry juice was above those of untreated and heat-treated juices during the first 14 days of storage. On the other hand, concentrations of ethyl butanoate and 1-butanol obtained after HIPEF processing were better maintained than after thermal processing. However, thermally-treated samples showed an increase in the amount of 1-butanol beyond day 35, causing an unpleasant flavour to the product. Thus, flavour stability in HIPEF-processed strawberry juice was greater than in thermally-treated samples during storage.     

Changes of health-related compounds throughout cold storage of tomato juice stabilized by thermal or high intensity pulsed electric field treatments

The effect of high intensity pulsed electric fields (HIPEF) processing (35 kV/cm for 1500 μs in bipolar 4-μs pulses at 100 Hz, with an energy density of 8269 kJ/L) on the main bioactive compounds and antioxidant capacity of tomato juice was investigated and compared to heat pasteurization (90 °C for 1 min or 30 s) having the fresh juice as a reference. HIPEF and heat treated tomato juices showed higher lycopene and lower vitamin C levels than the untreated juice. However, no significant changes in the total phenolic content and antioxidant capacity were observed between treated and fresh juices just after processing. Lycopene, vitamin C and antioxidant capacity of both treated and untreated juices decreased exponentially during storage following a first order kinetics (R² = 0.763–0.987), whereas tomato juices maintained their initial phenolic content. HIPEF-treated tomato juice maintained higher lycopene and vitamin C content than the thermally treated juices during the storage time. Hence, the application of HIPEF may be appropriate to achieve nutritious and fresh like tomato juice.

Industrial relevance

HIPEF processing can lead to tomato juice with higher nutritional value than that thermally processed. HIPEF-treated (35 kV/cm for 1500 μs with 4-μs bipolar pulses at 100 Hz, energy input of 8269 kJ/L) tomato juice shows greater lycopene, vitamin C and antioxidant capacity just after the treatment and during the storage time than heat treated (90 °C­30 s and 90 °C­60 s) tomato juice. Therefore, HIPEF technology is a feasible alternative to thermal treatment to obtain tomato juice with a high presence of health-related compounds.     

Comparative study on shelf life of orange juice processed by high intensity pulsed electric fields or heat treatment

The effects of high intensity pulsed electric fields (HIPEF) processing (35 kV/cm for 1,000 μs; bipolar 4-μs pulses at 200 Hz) on the microbial shelf life and quality-related parameters of orange juice were investigated during storage at 4 and 22 °C and compared to traditional heat pasteurization (90 °C for 1 min) and an unprocessed juice. HIPEF treatment ensured the microbiological stability of orange juice stored for 56 days under refrigeration but spoilage by naturally occurring microorganisms was detected within 30 days of storage at 22 °C. Pectin methyl esterase (PME) of HIPEF-treated orange juice was inactivated by 81.6% whereas heat pasteurization achieved a 100% inactivation. Peroxidase (POD) was destroyed more efficiently with HIPEF processing (100%) than with the thermal treatment (96%). HIPEF-treated orange juice retained better color than heat-pasteurized juice throughout storage but no differences (p<0.05) were found between treatments in pH, acidity and °Brix. Vitamin C retention was outstandingly higher in orange juice processed by HIPEF fitting recommended daily intake standards throughout 56 days storage at 4 °C, whereas heat-processed juice exhibited a poor vitamin C retention beyond 14 days storage (25.2–42.8%). The antioxidant capacity of both treated and untreated orange juice decreased slightly during storage. Heat treatments resulted in lower free-radical scavenging values but no differences (p<0.05) were found between HIPEF-processed and unprocessed orange juice.     

Effects of High-Intensity Pulsed Electric Fields Processing Parameters on the Chlorophyll Content and Its Degradation Compounds in Broccoli Juice

The influence of high-intensity pulsed electric fields (HIPEF) parameters including electric field strength (15–35 kV/cm), treatment time (500–2,000 μs), and polarity (monopolar or bipolar mode) on the content of chlorophylls (Chls), pheophytin (Phe), chlorophyllide (Chlide), and pheophorbide (Phb) and chlorophyllase activity (Chlase) in broccoli juice were assessed. A significant effect of HIPEF parameters on Chlase, Chls, and Chls degradation compounds was observed through a response surface methodology design. However, polarity did not exert influence neither on Chl a nor on Chl b. The optimum HIPEF treatment was found to be 35 kV/cm for 1,980 μs in bipolar mode, where the highest content of Chls was kept, the lowest Chlase residual activity was reached, and the minimal quantities of Chls degradation compounds content were formed. Additionally, at these HIPEF conditions, broccoli juice exhibited greater content of Chls than thermally treated or untreated juice. These outcomes demonstrated that HIPEF processing could be a suitable technology to maintain the Chls content in broccoli juice.     

Phenolic acids, flavonoids, vitamin C and antioxidant capacity of strawberry juices processed by high-intensity pulsed electric fields or heat treatments

The effect of high-intensity pulsed electric fields (HIPEF) processing (35 kV/cm for 1,700 μs in bipolar 4-μs pulses at 100 Hz) on individual phenolic compounds (phenolic acids and flavonoids), vitamin C and antioxidant capacity of strawberry juice was evaluated and compared to heat (90 °C for 60 or 30 s) and fresh juice as a reference. Although strawberry juice underwent a substantial depletion of health-related compounds with storage time irrespective of the treatment conducted, ellagic acid was enhanced. HIPEF-treated strawberry juice maintained higher amounts of phenolic acids (ellagic and p-coumaric acid) and total anthocyanins than the thermally treated juices during the storage period. Regarding the antioxidant capacity, similar DPPH and ABTS values were obtained so that differences among pasteurized juices were non significant. HIPEF processing may be a technology as effective as thermal treatments not only to achieve safe and stable juices, but also to obtain juices with a high content of antioxidant compounds.