Comparing equivalent thermal, high pressure and pulsed electric field processes for mild pasteurization of orange juice. Part I: Impact on overall quality attributes

Mild heat pasteurization, high pressure processing (HP) and pulsed electric field (PEF) processing of freshly squeezed orange juice were comparatively evaluated examining their impact on microbial load and quality parameters immediately after processing and during two months of storage. Microbial counts for treated juices were reduced beyond detectable levels immediately after processing and up to 2 months of refrigerated storage. Quality parameters such as pH, dry matter content and brix were not significantly different when comparing juices immediately after treatment and were, for all treatments, constant during storage time. Quality parameters related to pectinmethylesterase (PME) inactivation, like cloud stability and viscosity, were dependent on the specific treatments that were applied. Mild heat pasteurization was found to result in the most stable orange juice. Results for HP are nearly comparable to PEF except on cloud degradation, where a lower degradation rate was found for HP. For PEF, residual enzyme activity was clearly responsible for changes in viscosity and cloud stability during storage.

Industrial relevance

Development of mild processing technologies with a minimal impact on fruit juice can be considered as a true alternative of fresh fruit. The present work presents a fair comparison of mild heat treated, high pressure (HP) and pulsed electric field (PEF) processed orange juice as an alternative for thermal pasteurization. Orange juices were monitored during two months of storage.     

Headspace fingerprinting as an untargeted approach to compare novel and traditional processing technologies: A case-study on orange juice pasteurisation

As a rule, previous studies have generally addressed the comparison of novel and traditional processing technologies by a targeted approach, in the sense that only the impact on specific quality attributes is investigated. By contrast, this work focused on an untargeted strategy, in order to take into account unexpected and unintended effects of (novel) processing, and to possibly uncover unknown compounds resulting from alternative processing. The potential of headspace GC–MS fingerprinting was explored as a tool to compare the impact of thermal, high pressure (HP) and pulsed electric field (PEF) processing for mild pasteurisation of orange juice. This study demonstrated that when processing conditions are selected based on equivalent microbial safety, the impact of heat, HP and PEF pasteurisation on the volatile profile of orange juice can be considered comparable. During refrigerated storage, however, indirect impact differences were revealed, which were attributed to differences in degree of enzyme inactivation.     

Carotenoid and flavanone content during refrigerated storage of orange juice processed by high-pressure, pulsed electric fields and low pasteurization

The impact of different processing technologies, including non-thermal technologies, on bioactive compounds of orange juice was investigated. Freshly squeezed orange juice was treated by high pressure (HP) (400 MPa/40 °C/1 min), pulsed electric fields (PEF) (35 kV cm⁻¹/750 μs) and low pasteurization (LPT) (70 °C/30 s). The stability of main carotenoids and flavanones was studied just after treatment and during 40 days of refrigerated storage at 4 °C. Just after treatment, HP juice showed a significant increase on total carotenoid and flavanone content extracted (45.19 and 15.46%, respectively) and on vitamin A value (30.89%) with regard untreated juice, whereas no significant changes were observed for PEF and LPT juices. For all treated orange juices, flavanone content decreased significantly (around 50%) during the first 20 days of storage at 4 °C while carotenoid content showed a moderate decrease (less than 11%) that took place during the last 20 days. In general, during refrigerated storage, carotenoids and flavanones remained higher in HP juice than in LPT and PEF juices. Hence, HP and PEF technologies were as effective o even more than LPT to preserve bioactive compounds in orange juice during refrigerated storage.     

ENZYME INACTIVATION ON APPLE JUICE TREATED BY ULTRAPASTEURIZATION AND PULSED ELECTRIC FIELDS TECHNOLOGY

Apple juice was pasteurized by an ultra-high temperature treatment (UHT) at 115, 125 and 135C for 3 and 5 s, and compared with a high-voltage pulsed electric field treatment (PEF) at ranges between 33 and 42 kV/cm with frequencies of 150, 200, 250 and 300 pulses per second (pps). Enzyme inactivation and physicochemical properties of the treated juices were compared using a nontreated sample as control. The UHT treatment was more efficient in enzyme inactivation, reducing 95% the residual activity of polyphenoloxidase at the maximum temperature and time. However, a PEF treatment at 38.5 kV/cm and 300 pps combined with a temperature of 50C achieved a 70% reduction of residual PFO activity. In terms of quality characteristics as a function of physicochemical properties, color, pH, acidity and soluble solids were all less affected by PEF than by UHT when compared with the untreated juice.

PRACTICAL APPLICATIONS

Apple juice is a popular beverage worldwide and it is consumed nearly as much as orange juice. Consumers prefer fresh-squeezed fruit juices with high nutrient value and fresh-like sensory attributes. Enzymatic browning negatively impacts appearance, nutritive value and flavor of fruit juices. The use of ultra-high temperature processing is efficient in microbial control, as well as in enzyme inactivation. Any thermal processing may, however, decrease the overall quality of the treated juices. Pulsed electric field processing provides a potential alternative to thermal pasteurization of fruit juices.     

Cost Analysis and Environmental Impact of Pulsed Electric Fields and High Pressure Processing in Comparison with Thermal Pasteurization

The cost of high pressure processing (HPP) and the environmental impact of pulsed electric fields (PEF), HPP and thermal pasteurization of orange juice were estimated in the US. The cost analysis was based on commercial processing conditions that were validated for a 2-month shelf-life of orange juice under refrigeration conditions. Total electricity consumption was estimated to be 38,100 and 1,000,000 k Wh/year for thermal and HPP processing, respectively. Total pasteurization cost of HPP was estimated to be 10.7 ¢/l for processing 16,500,000 l/year (3,000 l/h). Of this, capital costs accounted for 59 % (6.3 ¢/l), labor costs accounted for 37 % (4.0 ¢/l) and utility charges, mainly electricity, accounted for 4 % (0.4 ¢/l). The total HPP cost was 7-folds higher than that of conventional thermal processing (1.5 ¢/l). The equivalent CO₂ emission was 90,000 kg for thermal processing and 700,000 and 773,000 kg for PEF and HPP, respectively. This corresponds to an increase between 7- and 8-folds in comparison to the thermal processing. Increasing the production output by 2- to 6-folds reduced the total production costs of nonthermal processing by 50–75 %. A deeper knowledge of the processing costs and environmental impact of nonthermal technologies will afford companies a better understanding of the benefits and limitations of these novel systems.     

Effects of Pulsed Electric Fields on the Activities of Microorganisms and Pectin Methyl Esterase in Orange Juice

Orange juice was treated with pulsed electric fields (PEF) in a pilot‐plant system to optimize PEF‐processing conditions for maximum microbial inactivation and to investigate the effects of PEF on pectin methyl esterase (PME) activity. Electric‐field strengths of 20,25,30, and 35 kV/cm and total treatment times of 39, 49, and 59 μs were used. Higher electric‐field strengths and longer total treatment times were more effective to inactivate microorganisms and PME (p < 0.05). PEF treatment of orange juice at 35 kV/cm for 59 μs caused 7–log reductions in total aerobic plate count and yeast and mold counts. About 90% of PME activity was inactivated by PEF treatment at 35 kV/cm for 59 μs. PEF‐treated orange juice at 35 kV/cm for 59 ms did not allow growth of microorganisms and recovery of PME at 4, 22, and 37 °C for 112 d.     

Effect of refrigerated storage on vitamin C and antioxidant activity of orange juice processed by high-pressure or pulsed electric fields with regard to low pasteurization

A better knowledge of the effect of refrigerated storage on the nutritional and antioxidant characteristics of foods processed by emerging technologies with regard to thermal traditional technology is necessary. Thus, freshly squeezed orange juice was processed by high-pressure (HP) (400 MPa/40 °C/1 min), pulsed electric fields (PEF) (35 kV/cm/750 μs) and low pasteurization (LPT) (70 °C/30 s). The stability of vitamin C and antioxidant activity was studied just after treatment and during 40 days of refrigerated storage at 4 °C. The determination of total vitamin C (ascorbic acid plus dehydroascorbic acid) was achieved by HPLC whereas the antioxidant activity was assessed by the measurement of the DPPH• radical scavenging. Just after treatment, all treated orange juices showed a decrease lower than 8% in vitamin C content compared with the untreated one. At the end of refrigerated storage, HP and LPT juices showed similar vitamin C losses (14 and 18%, respectively) in relation to untreated juice, although HP juices maintained better the vitamin C content during more days than LPT juices. Regarding antioxidant activity, after 40 days at 4 °C, differences among treated juices were no significant in terms of antiradical efficiency (AE=1/EC₅₀T_EC50). HP and PEF may be technologies as effective as LPT to retain antioxidant characteristics of orange juice during refrigerated storage.     

Changes in Quality and Nutritional Parameters During Refrigerated Storage of an Orange Juice–Milk Beverage Treated by Equivalent Thermal and Non-thermal Processes for Mild Pasteurization

The effects of high-pressure (HP) treatment (400?MPa at 42 °C for 5 min) and pulsed electric field (PEF) processing (25 kV/cm at 57 °C for 280 μs) on ascorbic acid, total carotenoids, total phenolic compounds and total antioxidant capacity (TEAC and ORAC) of an orange juice–milk (OJ-M) beverage along the storage time at 4 °C were compared with a conventional heat preservation technology used in industry (90 °C for 15 s). During storage, ascorbic acid, total carotenoids and antioxidant capacity (TEAC) depleted with time regardless of the treatment applied. Instead, total phenolic content and antioxidant capacity measured by the ORAC method increased at the end of the storage. Non-thermal-treated beverage had less non-enzymatic browning than the thermally pasteurized one. There were no significant variations in the hidroxymethylfurfural (HMF) content of the HP- and PEF-treated OJ-M, whilst a significant increase was obtained after thermal treatment. During refrigerated storage, HMF was always below the maximum values established. The HP treatments reduced the L* value of the treated beverages immediately after processing and during refrigerated storage and induced an increase in total colour differences of beverages treated by HP compared with PEF and thermally processed orange juice–milk. Hence, alternative methods such as HP and PEF may give new opportunities to develop orange juice–milk with an equivalent shelf life to that of thermally treated orange juice mixed with milk in terms of microbial, physicochemical and nutritional characteristics.     

Comparing the impact of high pressure,pulsed electric field and thermal pasteurization on quality attributes of cloudy apple juice using targeted and untargeted analyses

The impact of low-oxygen spiral-filter press technology combined with thermal pasteurization (TP), pulsed electric field (PEF) and high pressure processing (HPP) on cloudy apple juice quality was investigated immediately after the treatments and after 3 weeks of storage at 4 °C. Based on equivalent levels of microbial safety and desired shelf-life, low and high processing intensities were selected: TP (72 °C/15 s; 85 °C/30 s), PEF (12.5 kV/cm, 76.4 kJ/L; 12.3 kV/cm, 132.5 kJ/L), and HPP (400 MPa/3 min; 600 MPa/3 min). High intensity thermal treatment resulted in a bright, yellowish color which was maintained during storage. PPO and POD activities were largely reduced by high intensity PEF and TP yet showed high resistance to HPP. The highest vitamin C content was provided by fresh juice followed by PEF-treated juices. Due to oxidative degradation reactions, vitamin C of all treated samples significantly decreased during storage. Immediately after processing, high cloud stability values were obtained in all samples; however, cloud stability decreased during storage particularly for HPP juices with high residual PME. No significant changes were observed in pH, titratable acidity, organic acid and sugar content which also corresponded to sweet and sour taste. Results from untargeted volatile profiles showed that esters increased after PEF and were better retained after HPP. Contrary to TP treatment where ester degradation reactions occurred together with the formation of off-flavors. Most of the volatiles decreased during storage which could be linked to oxidation and ester hydrolysis reactions.Industrial relevanceBeing one of the most popular fruit juices consumed worldwide, cloudy apple juice can still undergo quality changes such as color degradation, cloud loss (fast sedimentation) and flavor changes during processing and storage. This study evaluates the potential of low-oxygen spiral-filter press in combination with different preservation technologies to obtain a maximal quality of cloudy apple juice. Results showed that high intensity thermal pasteurization can effectively inactivate quality-degrading enzymes, therefore it is useful to obtain an optimal cloudy apple juice product in terms of color and cloud stability. Although HPP has minimal impact on aroma of the juice, shelf-life of the juice may be limited due to incomplete enzyme inactivation. In the case of PEF treatment, thermal effects may contribute to maintain apple juice quality.     

Commercial-Scale Pulsed Electric Field Processing of Orange Juice

Effects of commercial‐scale pulsed electric field (PEF) processing on the microbial stability, ascorbic acid, flavor compounds, color, Brix, pH, and sensory properties of orange juice were studied and compared with those of thermal processing. Freshly squeezed orange juice was thermally processed at 90 °C for 90 s or processed by PEF at 40 kV/cm for 97 ms. Both thermally processed and PEF‐processed juices showed microbial shelf life at 4 °C for 196 d. PEF‐processed juice retained more ascorbic acid, flavor, and color than thermally processed juice (P<0.05). Sensory evaluation of texture, flavor, and overall acceptability were ranked highest for control juice, followed by PEF‐processed juice and then by thermally processed juice (P<0.01).     

Impact of PEF treatment inhomogeneity such as electric field distribution, flow characteristics and temperature effects on the inactivation of E. coli and milk alkaline phosphatase

High intensity pulsed electric field (PEF) treatment was investigated focusing on the alteration of electric field distribution, flow characteristics and temperature distribution due to the modification of the treatment chamber. The aim was the improvement of the effectiveness of microbial inactivation of E. coli and to reduce the PEF impact on alkaline phosphatase (ALP) activity in raw milk. Mathematical simulation of the PEF process conditions considering different treatment chamber setups was performed prior to experimental verification. Finally the impact of the treatment chamber modifications on microbial inactivation and enzyme activity was determined experimentally. Using a continuous flow-through PEF system and a co-linear treatment chamber configuration the insertion of stainless steel and polypropylene grids was performed to alter the field strength distribution, increase the turbulence kinetic energy and improve the temperature homogeneity. The Finite Element Method (FEM) analysis showed an improved electric field strength distribution with increased average electric field strength and a reduced standard deviation along the center line of the treatment zone indicating a more homogenous electric field. The velocity profile was improved resulting in an increase of turbulence kinetic energy due to the insertion of the grids. As revealed by mathematical modeling, the temperature of the liquid was decreased, and formation of temperature peaks was avoided. Measured inactivation of heat sensitive alkaline phosphatase (ALP) was reduced from 78% residual activity to 92% after PEF treatment and it could be shown that thermal effects and temperature peaks have been the main reason for enzyme inactivation due to PEF. At the same time, an increase of microbial inactivation of 0.6 log–cycles could be determined experimentally due to the modification of the treatment chamber design.

Industrial relevance

The application of pulsed electric field as a non-thermal pasteurization technology requires an accurately defined treatment intensity followed by a predictable microbial inactivation. Unavoidable thermal effects occurring during PEF treatment due to ohmic heating have to be minimized to assure the retention of heat-sensitive nutrients and bioactive compounds. The presented investigations contribute to the fulfilment of these requirements for further successful industrial implementation of the PEF technology such as the selective inactivation or retention of enzyme activity in liquid food systems.     

The Impact of Thermosonication and Pulsed Electric Fields on Staphylococcus aureus Inactivation and Selected Quality Parameters in Orange Juice

The effect of thermosonication (TS) and pulsed electric fields (PEF) on inactivation of Staphylococcus aureus (SST 2.4) and selected quality aspects in orange juice was investigated. Conventional pasteurization (HTST, 94 °C for 26 s) was used as a control. TS (10 min at 55 °C) applied in combination with PEF (40 kV/cm for 150 μs) resulted in a comparable inactivation of S. aureus to that achieved by conventional HTST. TS/PEF did not affect the pH, conductivity, or °Brix and had a milder impact on the juice color than thermal treatment. Furthermore, the non-enzymatic browning index was significantly affected by HTST (P < 0.05) but not by TS and PEF. Ascorbic acid retention was almost complete after TS and PEF (96.0%), but it was substantially lower (P < 0.05) after HTST (80.5%). Residual activity of pectin methyl esterase (PME) decreased as PEF field strength and treatment time increased; however, applying TS and PEF in combination left a greater residual PME activity than HTST (12.9 vs 5.0%, respectively).     

DEVELOPMENT OF A PROTEIN FORTIFIED FRUIT BEVERAGE AND ITS QUALITY WHEN PROCESSED WITH PULSED ELECTRIC FIELD TREATMENT

Pulsed electric field treatment (PEF), was explored to process a beverage at 30C. A protein fortified fruit based beverage was developed using proteins, orange juice, guar gum, sucrose, calcium lactate, citric acid, natural flavor and color. It was processed at low pH (3.75) using both a heat pasteurization and a PEF treatment. PEF treated beverage had less protein denaturation and lower loss of vitamin C compared with that of the heat treated beverage. The heat treated beverage had a slightly higher apparent viscosity than the PEF treated beverage and developed sedimentation in the container during storage. The PEF processed beverage maintained its natural orange juice like color better than the heat treated beverage, which developed a slightly whitish color. However, the PEF treated product was less microbiologically stable (5 months) at refrigeration temperature compared with the heat treated product which was stable for more than 12 months. It was concluded that more technological improvements are needed to process food safely by PEF technology.     

Kinetics of Peroxidase Inactivation in Carrot Juice Treated with Pulsed Electric Fields

Peroxidase activity accounts for quality losses in many plant‐based foods. The paper provides insight into the inactivation kinetics of peroxidase (POD) in carrot juice treated with pulsed electric fields (PEF). Juice samples were subjected to electric field intensities of 20 to 35 kV/cm for 300 to 2000 μs. Up to 93% of the initial activity was inactivated after treating at 35 kV/cm for 1500 μs. POD activity inactivation correlated well with the increase in energy density input. A first‐order fractional conversion model best fitted the experimental results. Other kinetic approaches such as the Fermi's model can be used to estimate residual POD activity values in treated juices as a function of electric field strength. Practical Application : Pulsed electric fields (PEF) are a processing technology that can be used for the pasteurization of liquid food products. Peroxidase activity inhibition is required in carrot juices to prevent undesirable quality losses, such as discoloration, flavor changes, and loss of nutrients. The most significant processing parameters ruling POD inactivation in PEF‐treated carrot juice are identified and mathematical modeling of experimental data is conducted.     

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.     

Pasteurization of carrot juice by combining UV-C and mild heat: Impact on shelf-life and quality compared to conventional thermal treatment

The combination of UV-C radiation and mild heat (UV-H) treatment is a promising strategy for synergistically increasing microbial inactivation in low UV-transmitting juices. In this research, we explored the suitability of UV-H treatment in carrot juice pasteurization and its impact on juice quality during shelf-life compared to that of thermal pasteurization. UV-H treatment at 60 °C (3.92 J/mL, 3.6 min) enabled reductions of over 5 log₁₀ cycles in the reference pathogens and a significant reduction in spoilage yeasts, bacteria, and bacterial spores. The activity of pectin methylesterase and polyphenol oxidase was reduced by UV-H treatment to levels close to those of low-temperature pasteurization (60 °C/18.1 min). The native population of total aerobic bacteria, lactic acid bacteria, and yeasts and molds of UV-H-treated juice remained undetectable during 29 days of cold storage. Furthermore, viscosity, cloud stability, and the color of fresh juice were better preserved by UV-H treatment than by thermal pasteurization throughout storage.Industrial relevanceThis study demonstrates that UV-H treatment is a beneficial alternative to conventional thermal processing in carrot juice pasteurization, since appropriate inactivation levels of pathogenic and spoilage microorganisms can be reached while better preserving the quality attributes of fresh juice throughout its shelf-life.     

Inactivation Kinetics of Pectin Methylesterase,Polyphenol Oxidase,and Peroxidase in Cloudy Apple Juice under Microwave and Conventional Heating to Evaluate Non-Thermal Microwave Effects

Continuous-flow microwave pasteurization provides important advantages over conventional heat exchangers such as fast volumetric heating, lower tube surface temperature, and possible non-thermal effects that enhance enzymatic and bacterial inactivation. Conventional and microwave-assisted inactivation of pectin methylesterase (PME), polyphenol oxidase (PPO), and peroxidase (POD) in cloudy apple juice were investigated to evaluate non-thermal effects. Experiments were conducted to provide uniform heating with accurate temperature acquisition and similar temperature profiles for conventional and microwave treatments. A two-fraction first-order kinetic model was successfully fitted to the data in a procedure that took into account the whole time-temperature profile instead of assuming isothermal conditions. Predicted inactivation curves for pasteurization at 70 and 80 °C of the cloudy apple juice showed that PME has the highest thermal resistance (residual activity of 30% after 250 s at 80 °C) and that there was no evidence of non-thermal microwave effects on the inactivation of these enzymes.     

Thermal, high pressure, and electric field processing effects on plant cell membrane integrity and relevance to fruit and vegetable quality

Advanced food processing methods that accomplish inactivation of microorganisms but minimize adverse thermal exposure are of great interest to the food industry. High pressure (HP) and pulsed electric field (PEF) processing are commercially applied to produce high quality fruit and vegetable products in the United States, Europe, and Japan. Both microbial and plant cell membranes are significantly altered following exposure to heat, HP, or PEF. Our research group sought to quantify the degree of damage to plant cell membranes that occurs as a result of exposure to heat, HP, or PEF, using the same analytical methods. In order to evaluate whether new advanced processing methods are superior to traditional thermal processing methods, it is necessary to compare them. In this review, we describe the existing state of knowledge related to effects of heat, HP, and PEF on both microbial and plant cells. The importance and relevance of compartmentalization in plant cells as it relates to fruit and vegetable quality is described and various methods for quantification of plant cell membrane integrity are discussed. These include electrolyte leakage, cell viability, and proton nuclear magnetic resonance (1H-NMR).