WO2017086784A1 - Procédé de conservation d'aliment liquide utilisant un traitement de champ électrique pulsé - Google Patents

Procédé de conservation d'aliment liquide utilisant un traitement de champ électrique pulsé Download PDF

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Publication number
WO2017086784A1
WO2017086784A1 PCT/NL2016/050799 NL2016050799W WO2017086784A1 WO 2017086784 A1 WO2017086784 A1 WO 2017086784A1 NL 2016050799 W NL2016050799 W NL 2016050799W WO 2017086784 A1 WO2017086784 A1 WO 2017086784A1
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Prior art keywords
liquid product
product
liquid
process according
pef
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PCT/NL2016/050799
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English (en)
Inventor
Rian Adriana Hendrika Timmermans
Ricardo Ermirio De Moraes
Hendrikus Cornelis Mastwijk
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Stichting Wageningen Research
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Priority to EP16810096.4A priority Critical patent/EP3376878A1/fr
Priority to US15/776,164 priority patent/US11903400B2/en
Priority to JP2018544752A priority patent/JP6921838B2/ja
Priority to MYPI2018701823A priority patent/MY189799A/en
Priority to BR112018009870-1A priority patent/BR112018009870B1/pt
Priority to SG11201803585PA priority patent/SG11201803585PA/en
Priority to CN201680066218.4A priority patent/CN108471787A/zh
Priority to EA201890901A priority patent/EA037900B1/ru
Priority to AU2016357683A priority patent/AU2016357683B2/en
Priority to MX2018006003A priority patent/MX2018006003A/es
Priority to NZ742053A priority patent/NZ742053B2/en
Publication of WO2017086784A1 publication Critical patent/WO2017086784A1/fr

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Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/005—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by heating using irradiation or electric treatment
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
    • A23B5/00—Preservation of eggs or egg products
    • A23B5/005—Preserving by heating
    • A23B5/01—Preserving by heating by irradiation or electric treatment
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C3/00—Preservation of milk or milk preparations
    • A23C3/02—Preservation of milk or milk preparations by heating
    • A23C3/03—Preservation of milk or milk preparations by heating the materials being loose unpacked
    • A23C3/033—Preservation of milk or milk preparations by heating the materials being loose unpacked and progressively transported through the apparatus
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C3/00—Preservation of milk or milk preparations
    • A23C3/02—Preservation of milk or milk preparations by heating
    • A23C3/03—Preservation of milk or milk preparations by heating the materials being loose unpacked
    • A23C3/033—Preservation of milk or milk preparations by heating the materials being loose unpacked and progressively transported through the apparatus
    • A23C3/0335—Preservation of milk or milk preparations by heating the materials being loose unpacked and progressively transported through the apparatus the milk being heated by electrical or mechanical means, e.g. by friction
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/42—Preservation of non-alcoholic beverages
    • A23L2/46—Preservation of non-alcoholic beverages by heating
    • A23L2/48—Preservation of non-alcoholic beverages by heating by irradiation or electric treatment
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/16—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by heating loose unpacked materials
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/16—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by heating loose unpacked materials
    • A23L3/18—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by heating loose unpacked materials while they are progressively transported through the apparatus
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F1/00—Treatment of water, waste water, or sewage
    • C02F1/48—Treatment of water, waste water, or sewage with magnetic or electric fields

Definitions

  • the present invention relates to a process for fast and homogeneously heating a liquid product to a predetermined temperature by means of resistive heating.
  • the present invention further relates to said process wherein the liquid product is pre-heated prior to subjecting the liquid product to the process.
  • Pulsed electrical fields is used as a technology to induce electroporation of a cell membrane by the application of pulses of a short period of time by an external electrical field of high intensity.
  • the most widely accepted theory for this phenomena is that by application of an external electrical field on a biological membrane, local instabilities in the lipid bilayer are induced, eventually leading to pore formation.
  • the formation of pores enhances permeability across the membrane (electro-permeabilization) that, depending on the intensity of the applied electric field, is either a reversible process or when applied at high voltages, is irreversible, leading to cell death.
  • Irreversible electroporation is effective on vegetative micro-organism at field strengths in the range of 10-20 kV/cm when using pulses of 2 microsecond duration for a total treatment time of 100-400 microseconds (Fig. 1).
  • Reynard and co-workers (1998) have investigated the critical effect on the pulse duration of a single pulse for gene transfer. They found that a minimum pulse time is needed for orientation of ⁇ 1 millisecond and indicated critical response times for permeabilization from 3 to 5 milliseconds using pulses of 24 milliseconds duration at an electrical field strength of 1-2.7 kV/cm.
  • AC currents are used to invoke high electrical field conditions in a liquid.
  • AC currents with a fixed frequency (f) can be viewed as pulses with a duration of 1/f, the characteristic pulse shape considered here is rectangular meaning that the repetition frequency of pulses is less than the bandwidth (1/pulse duration).
  • AC currents at frequencies greater than 1 MHz (or pulse durations less than 1 microsecond) has been considered in US 2010/0297313.
  • Reversible electroporation is a procedure regularly used in molecular biology and clinical biotechnology to introduce small or large molecules into the cell, i.e. drugs, oligonucleotides, antibodies and plasmids into the cytoplasm, aiming at keeping cells to stay alive.
  • Irreversible electroporation can be used to extract molecules from the cell or to inactivate cells.
  • irreversible electroporation we aim for irreversible electroporation as a non-thermal preservation method, where the maximum temperature obtained by PEF processing and the holding time is lower than by conventional heat pasteurisation. This results for example amongst other beneficial aspects in a better preservation of the fresh taste and nutritional values of the product.
  • the processing conditions that are selected for pulsed electrical treatment when aimed at microbial inactivation is dependent on several factors but can be classified into three groups: processing parameters, microbial characteristics and treatment medium characteristics.
  • the temperature is considered as critical to the effectiveness of microbial inactivation by PEF (Raso et al, 2014). Increase of electrical field strength and treatment time will lead to an increased PEF lethality. As a result of these conditions, more energy will be applied per mass unit, leading to more heating up of the product.
  • Typical process conditions used for irreversible electroporation are in the range of short pulses of micro-seconds at a high voltage (5-80 kV/cm).
  • the extent of microbial inactivation by PEF is enhanced by increasing the temperature of the medium, e.g. the liquid food product, prior to PEF treatment, even in the range of temperatures that are not lethal for micro-organisms. Without wishing to be bound by theory, this pre-heating effect has influence on the phospholipid bilayer structure of the cell membrane, making the cells more vulnerable for the PEF-process (Wouters et al, 1999).
  • Characteristics of the micro-organism have influence on the effectiveness of microbial inactivation by PEF. Generally, it has been reported that relatively large micro-organisms are more sensitive towards PEF than smaller micro-organisms, and Gram-negative microorganisms are more sensitive towards PEF than Gram-positive micro-organisms.
  • PEF PEF treatment is often studied in liquid media suspended with microorganisms. Characteristics of this treatment medium have been investigated, and pH has been reported to be of major importance for the efficacy of the treatment. That is to say, PEF is much more effective in media at low pH than in media at neutral pH.
  • Applied process conditions are suitable for liquid food products with low pH, i.e. high-acid fruit juice, with a pH below about 4.6. These process conditions appear in several applications to be suitable for inactivating larger sized micro-organisms in liquid food products. Furthermore Gram-negative micro-organisms can be inactivated more effective than Gram-positive micro- organisms. Especially inactivating small size Gram-positive bacteria is in most cases cumbersome with the currently known PEF process conditions. In addition, current low-pH process conditions are not applicable in an effective way for food products having a pH higher than about 4.6. Current PEF processes for liquid food products encompasses electrical field strengths that are relatively high, i.e. 5 kV/cm and higher, typically 10-30 kV/cm.
  • the current invention relates to a process for fast and homogeneously heating a liquid product to a predetermined temperature by means of resistive heating, to obtain heated liquid product, comprising:
  • the pulse duration of a single pulse is a critical factor rather than the total effective treatment time.
  • pulse duration of 2 microseconds ( ⁇ ) and electrical field strengths (E) of 10 kV/cm the inventors found that inactivation was not efficient, despite that the total effective treatment time calculated as E 2 ⁇ was 4 times higher than at 20 kV/cm where conventional PEF treatment is employed.
  • E electrical field strengths
  • the process of the invention is applicable to liquid food products and liquid feed products, and the PEF processing conditions of the invention are equally effective in inactivating Gram- negative bacteria as well as Gram-positive bacteria.
  • the PEF processing conditions are applicable to liquid food products and liquid feed products, which conditions are effective in both inactivating relatively large microbes and inactivating relatively small microbes.
  • the inventors surprisingly found PEF processing conditions now applicable under currently applied conditions of relatively low pH, as well as applicable under conditions at higher pH.
  • a second aspect of the current invention relates to a liquid product obtainable by the process according to the invention.
  • Figure 1A Figure IB. Reduction of viable counts of Escherichia coli, Listeria monocytogenes, Lactobacillus plantarum, Salmonella Senftenberg, Saccharomyces cerevisiae in orange juice at pH 3.8 after various PEF-treatment conditions. Panels on the left represent PEF conditions currently used, and panels on the right show PEF conditions of the invention. Reference to the various PEF-treatment conditions related to each panel is made below the panels of Figure IB.
  • Solid black triangles 10 kV/cm, 2 microseconds; solid gray diamonds: 15 kV/cm, 2 microseconds; open white circles: 20 kV/cm, 2 microseconds; solid gray circles: 0.9 kV/cm, 1000 microseconds; solid black diamonds: 2.7 kV/cm, 1000 microseconds; open white diamonds: 2.7 kV/cm, 100 microseconds; dashed line: detection limit.
  • Figure 3 Reduction of viable counts of E. coli and L. monocytogenes in orange juice, coconut water and watermelon juice after PEF-treatment at 2.7 kV/cm, 1000 microseconds.
  • Figure 6 Amount of soluble solids (°Brix) in orange juice before PEF treatment and after PEF treatment during 3 months of storage at 7°C and at ambient temperature.
  • Figure 8. pH of orange juice before PEF treatment and after PEF treatment during 3 months of storage at 7°C and at ambient temperature.
  • Figure 10 Vitamin C content of orange juice before PEF treatment and after PEF treatment during 3 months of storage at 7°C and at ambient temperature.
  • the inventors now found PEF processing conditions applicable to liquid food products and liquid feed products, which conditions are equally effective in inactivating Gram-negative bacteria as well as Gram-positive bacteria.
  • the inventors also found PEF processing conditions applicable to liquid food products and liquid feed products, which conditions are effective in both inactivating relatively large microbes and inactivating relatively small microbes.
  • the inventors surprisingly found PEF processing conditions now applicable under conditions of relatively low pH, as well as applicable under conditions at higher pH.
  • the inventors found PEF processing conditions applicable to larger volumes of throughput of liquid food products and liquid feed products than the volumes that were previously possible with currently available processes.
  • the inventors provide for a process that addresses many of the shortcomings related to currently known processes for heating a liquid product to obtain liquid product with a diminished microbial load.
  • the current invention relates to a process for fast and homogeneously heating a liquid product to a predetermined temperature by means of resistive heating, to obtain heated liquid product, comprising:
  • the process for fast and homogeneously heating a liquid product to a heating temperature by means of resistive heating provides a heated liquid product with a diminished microbial load.
  • Heating a liquid product to a temperature above a certain maximum temperature may cause unwanted reduction of fresh flavours, vitamins and nutrients and denaturation of proteins present in the fresh (untreated) product.
  • the degree of reduction and denaturation of the components is related to the temperature and time the product is exposed to the treatment. Varying liquid products are exposed to varying temperature-time combinations to obtain the desired degree of enzymatic and microbial inactivation.
  • Alternative (non)-thermal processes with a reduced temperature and/or time exposure to the product are therefore gaining a lot of interest, as they can better retain the fresh characteristics of the product. When either the temperature or the exposure time can be reduced, a better product quality can be expected.
  • the exposure time to the heat is reduced tremendously, and due to the chosen process conditions the maximum temperature of the liquid product autonomously remains below about 92°C during the resistive heating.
  • the maximum temperature of the liquid product autonomously remains below a critical temperature during the resistive heating according to the process of the invention, at which temperature of the liquid product does not suffer from reduction of the heat- sensitive components or denaturation of the proteins, if present in the product, while at the same time the microbial load in the liquid product is reduced to an acceptable level aimed for. It is now due to the process of the current invention that processing conditions have become applicable which both prevent overheating of the liquid product while still effectively and efficiently lowering the microbial load of the liquid product.
  • the pulse duration of a single pulse is a critical factor rather than the total effective treatment time.
  • pulse duration times of 2 microseconds and electric field strengths of 10 kV/cm it was determined that inactivation of microbes was not efficient, despite that the total effective treatment time was four times more than at 20 kV/cm, which is the electric field strength at which conventional PEF treatment is employed.
  • the explanation is that at the reduced electrical field strength of 10 kV/cm the electroporation effect is compromised.
  • an external electrical field applied to a product has an influence on the protein channels in the cell membrane, and/or the lipid domain of the cell membrane of a micro-organism present in the product, resulting in conformational changes in the channels and/or the domain.
  • Membrane protein channels open at 50 mV membrane potential, which is considerable lower than the 150-400 mV required for pore formation in the lipid double layer (Tsong, 1992).
  • protein channels may be affected at electrical field strengths 3 to 8 times smaller than the electrical field strengths at which lipid double layers are affected; that is to say, an electrical field strength of between 2.5 kV/cm and 7 kV/cm for inactivation of the protein channel compared to the required 20 kV/cm for irreversible damage by electroporation of the lipid double layer (i.e. conventional PEF conditions).
  • an electrical field strength of between 0.1 kV/cm and 5 kV/cm in combination with a pulse duration of 10- 1000 microseconds is sufficient and efficacious with regard to establishing efficient inactivation of microbes in a liquid product.
  • the process of the invention is applicable for a liquid product in a 1 L/h PEF apparatus ('PEF system').
  • the pulse duration was set to either 100 microseconds or 1000 microseconds and the selected electrical field strength, or 'electric field strength', was either 0.9 kV/cm or 2.7 kV/cm.
  • the number of pulses applied to the liquid product varied between 0 and 35, the time lapse between two consecutive pulses varied between 0.6 milliseconds and 199 milliseconds, and as a result the maximum temperatures obtained varied between 36°C and 92°C. See for a more detailed outline of examples demonstrating the efficiency of the process of the invention the Example 1, below.
  • the process according to the invention provides for an efficient mechanism of inactivating spores as well.
  • Spores contain essential proteins for germination in the inner membrane and in the spore cortex that are targets for external electrical stimuli.
  • a batch of liquid product is processed in the process of the invention, applying a 1200 L/h PEF apparatus for fast and homogeneously heating a liquid product to a predetermined temperature by means of resistive heating.
  • the pulse duration is 1000 microseconds and the electrical field strength is 2.0 kV/cm.
  • the number of pulses applied to the liquid product is about 5 pulses and the time lapse between two consecutive pulses is 3.8 milliseconds. See also Example 3, below, for the detailed embodiment of the invention.
  • One embodiment of the invention is the process according to the invention, wherein the pH of the liquid product is between pH 1.5 and 9.0, preferably above 4.6, preferably between 4.8 and 9.0, more preferably between 5.5 and 8.0, more preferably between 6.0 and 7.5.
  • One embodiment of the invention is a process of the invention, wherein the pH is above about 5.0, preferably about 6.0.
  • the invention relates to the process according to the invention, wherein the pH of the liquid product is lower than 4.6, preferably between 1.5 and 4.6, more preferably between about 1.5 and about 3.8.
  • the pH of the liquid product is higher than 4.6, preferably between 4.6 and 9.0.
  • One embodiment of the invention is a process according to the invention, wherein the pH of the liquid product is between 5.0 and 9.0, preferably between 6.0 and 9.0. It is now due to the applicability of the process of the invention, that liquid products having a wide range of pH are processed in one and the same process according to the invention. Since the process of the invention is applicable for processing liquid products with such widely varying pH, the diversity of liquid products for which PEF processing is desirable and which are selectable for processing in the process of the invention, is very large. Virtually any liquid product, ingredient or semi-finished product applied in for example food processing is now suitable for fast and homogenously heating by the process of the invention. Due to the surprising finding of the inventors that their PEF process is effective and efficient in such a wide pH range, processing food products with a process incorporating PEF according to the invention has now become wider accessible than before.
  • one embodiment of the invention is the liquid product according to the invention, wherein the liquid product has an electrical conductivity between 0.01 and 10 S/m measured at 20°C, more preferably between 0.1 and 3 S/m measured at 20°C, most preferably between 0.2 S/m and 0.8 S/m measured at 20°C.
  • electrical conductivity at 20°C has been measured for several batches of liquid food products and was for example 0.1 S/m for a cranberry juice, 0.15 S/m for a beer, 0.2 S/m for an apple juice, 0.4 S/m for a chocolate milk, 0.45 S/m for a whole milk, 0.4 S/m for soy milk, 0.25 S/m for almond milk, 1.0 S/m for a carrot juice, and 1.8 S/m for tomato sauce.
  • the process of the invention is applicable for a wide array of widely varying liquid food products.
  • liquid food products are effectively pasteurized, i.e. microbes were inactivated effectively and efficiently, when applying PEF processing conditions of the invention comprising a surprisingly low electrical field strength of 0.1-5 kV/cm, preferably 4 kV/cm or lower, more preferably 3 kV/cm or lower, in combination with a pulse duration of 10-1000 microseconds, preferably about 1000 microseconds, more preferably about 100 microseconds, and at a maximum temperature of the liquid food product of between 40°C and 92°C, preferably between 50°C and 92°C, more preferably between about 60°C and 85°C.
  • PEF processing conditions of the invention comprising a surprisingly low electrical field strength of 0.1-5 kV/cm, preferably 4 kV/cm or lower, more preferably 3 kV/cm or lower, in combination with a pulse duration of 10-1000 microseconds, preferably about 1000 microseconds, more preferably about 100 microseconds, and at a maximum temperature of the liquid food
  • pasteurization upon applying the process of the invention is particularly efficient and effective when the number of pulses applied to the continuously flowing liquid product is at least 1, preferably 1 to 100, more preferably 5-50 for each fluid element during passage within the treatment zone.
  • the number of pulses is given by equation 1, where n, is the number of pulses, Vis the volume of the treatment chamber (L), is the pulse frequency used (Hz), and ⁇ is the flow rate (L/h): v -f
  • the number of pulses is not a critical step in designing the process, as long as at least one pulse is applied to every fluid element flowing through the treatment chambers of the apparatus for fast and homogeneously heating a liquid product, according to the invention.
  • the system has to be designed with the purpose that the residence time in the treatment chamber is larger than l/f.
  • the number of pulses applied will be the result of the process design, based on desired throughput of liquid product ( ⁇ , L/h), the conductivity of the liquid product ( ⁇ , S/m), specific heat capacity of the product (c p , kJ/kg-K), density of the product (p, kg/m 3 ), applied electrical field strength (E, V/m), pulse duration (x pu ise, s) and temperature gradient ( ⁇ , °C) obtained with the process (difference between inlet temperature of the liquid product and outlet temperature). Relationship between these parameters is given in equation 2.
  • the pulse duration is critical for the effectiveness of the PEF treatment according to the process of the invention.
  • application of one relatively long pulse can be more effective than application of more short pulses with a similar total effective treatment time.
  • the pulse duration is about 100-1000 microseconds, and the electrical field strength is about 2.7 kV/cm.
  • the number of pulses is then about 1-25, and the time laps between two consecutive pulses is about 0.6-39 milliseconds, dependent on the desired temperature increase across the treatment chambers, being the difference between the inlet temperature and maximum temperature.
  • the pulse duration is 1000 microseconds, and the electrical field strength is about 2.0 kV/cm, in the process according to the invention.
  • the number of pulses is then about 5, and the time laps between two consecutive pulses is about 3.8 milliseconds.
  • the process of the invention is applicable for processing a liquid product in a PEF apparatus having a throughput of between 30 L/h and 200 L/h, according to the invention.
  • the process of the invention is applicable for processing a liquid product in a PEF apparatus having a throughput of about 30.000 L/h, according to the invention.
  • One embodiment of the invention is the process of the invention, wherein the apparatus for fast and homogeneously heating a liquid product to a predetermined temperature has a throughput of between about 1 L/h and about 30.000 L/h, preferably about 1 L/h or about 30 L/h, or about 200 L/h, or about 1200 L/h, or about 30.000 L/h.
  • these PEF processing conditions of the invention result in inactivation of microbes following the theoretical mechanism of inactivating the protein channels of the membrane of cells.
  • These new PEF processing conditions of the invention provides new opportunities in microbial inactivation compared to the currently used treatment conditions for PEF.
  • Gram-positive micro-organisms such as for example Listeria monocytogenes , Lactobacillus plantarum, Leuconostoc strains, and Streptococcus species are also inactivated in liquid products. Based on the expected theoretical mechanism, it is expected that also spore-forming bacteria can be inactivated, as their cell membrane also contains voltage-gated channels, like Alicyclobacillus bacteria and Clostridium bacteria. Furthermore, spores itself contain essential proteins for germination in the inner membrane and the spore cortex that may be targeted by the external applied pulses.
  • the process of the invention is suitable for inactivation in liquid products of relatively large micro-organisms, such as for example yeasts and moulds.
  • the process of the invention is also suitable for inactivation in liquid products of relatively small micro-organisms, such as for example L.monocytogenes.
  • the process of the invention is equally suitable for inactivation in liquid products of micro-organisms with sizes in between the sizes of these exemplified micro-organisms, shown in example 1
  • the process of the invention is especially suitable for liquid food products subjected to the process of the invention in an apparatus for fast and homogeneously heating a liquid product to a heating temperature by means of resistive heating, when the residence time of the fluid in the high field region is n about 17 milliseconds to 2 seconds. Frequency of the pulses is restricted between 1 kHz and 50 kHz, to avoid metal release of the electrodes (Mastwijk, 2006).
  • the flow rate of the liquid product is then between about 1 L/h and 5000 L/h, preferably between about 1000 L/h and 30.000 L/h .
  • a further embodiment of the invention is a process according to any of the previous embodiments of the invention, wherein the liquid product is a liquid food product or a liquid feed product.
  • liquid product is an ingredient, semi-finished product, or final liquid product, like fruit juice, vegetable juice, infant food, jam, spread or smoothie, an alcoholic or non-alcoholic beverage , dairy product, plant milk product, liquid egg, a soup or a sauce.
  • One embodiment of the invention is a process for fast and homogeneously heating a liquid product to a predetermined temperature by means of resistive heating according to the invention, wherein the dairy product is selected from milk, a milk product or a liquid composition comprising a milk component or a milk fraction.
  • an embodiment of the invention is a process according to the invention, wherein the liquid product is a dairy product comprising milk, a milk product, a milk component or a milk fraction.
  • An important aspect of the invention is the finding that during the PEF processing, no cooling section between the treatment chambers of an apparatus applied in the process according to the invention is required in order to keep the temperature of the liquid product below about 92°C, or below about 85°C, or below about 70°C, or below about 60°C, according to the invention.
  • cooling sections are added between treatment chambers to avoid overheating of the liquid product.
  • the liquid product is preheated to a temperature in the range of from 20°C to 70°C before being supplied to the apparatus, preferably from 35°C to 65°C, more preferably from 40°C to 60°C.
  • the liquid product is preheated before being supplied to the apparatus to a temperature in the range of from 20°C to 70°C, preferably from 35°C to 65°C, more preferably from 40°C to 60°C.
  • liquid products subjected to the process according to the invention are cooled to ambient temperature or below immediately, e.g. cooled to 2-8°C. Since no holding time is required, cooling of a liquid product proceeds directly (preferably within 3 seconds) after the liquid product leaves the high field region.
  • cooling tubes may be installed 0.5 m downstream from the high field region.
  • autonomous has its regular meaning, and here refers to the temperature of the liquid product that reaches a certain value in the process of the invention unaided by external cooling (or heating) during the treatment.
  • the liquid product such as for example a liquid food product selected from orange juice, a dairy product, coconut water, watermelon juice, is for example pre-heated to about 40°C, about 50°C or about 60°C.
  • the liquid product is preheated to between about 30°C to 65°C, preferably between 36°C and 59°C.
  • the maximum temperature of the liquid product autonomously remains below about 85°C during the resistive heating, more preferably, below about 70°C, or below about 63°C, or below about 60°C, according to the invention.
  • process parameters of the process of the invention are selected such that the maximum temperature of the liquid product autonomously remains below a selected temperature. At or below the selected temperature efficient and effective killing of the micro-organisms present in the liquid product is ensured, whereas unwanted reduction of fresh flavours, vitamins and nutrients and denaturation of proteins present in the fresh (untreated) liquid product is prevented, or at least prevented to a large extent, when applying the process of the invention.
  • the inventors By applying the electrical field strength according to the invention, with a pulse duration according to the invention, the inventors surprisingly found that the temperature of the liquid product remains below a maximum temperature of about 92°C, or about 85°, or about 70°C, or about 60°C, according to the invention, making the process of the invention particularly suitable for implementation in a large scale setting, e.g. a commercial setting.
  • An example of a commercial application of the process of the invention is the processing of a liquid food product in an apparatus for fast and homogeneously heating a liquid product to a predetermined temperature by means of resistive heating, wherein the flow of liquid food product through the apparatus is between about 500 L/h to 30.000 L/h, for example about 1200 L/h, according to the invention.
  • one embodiment of the invention is a process according to the invention, wherein the temperature of the liquid product autonomously remains below 85°C during the resistive heating, preferably below 75°C, more preferably below 60°C.
  • the invention relates to the process according to the invention, wherein the electrical field strength is lower than about 5 kV/cm.
  • the electrical field strength is 0.5 to 5 kV/cm, more preferably 2.5 to 4 kV/cm.
  • the invention relates to the process of the invention, wherein the electrical field strength is below about 3 kV/cm, preferably about 2.7 kV/cm or lower, more preferably between about 0.9 and about 2.5 kV/cm.
  • One embodiment of the invention is a process according to the invention, wherein the pulse duration is at least 10 microseconds, more preferably 10 to 2000 microseconds, even more preferably 50 to 500 microseconds, most preferably 50 to 100 microseconds. In one embodiment the invention relates to the process according to the invention, wherein the pulse duration is between about 100 microseconds and about 1000 microseconds. In a further embodiment the invention relates to a process according to the invention, wherein the pulse duration is 1000 microseconds or lower, preferably about 100 microseconds.
  • the applied pulses are bipolar pulses.
  • one embodiment of the invention is a process according to the invention, wherein the minimal one pulse applied to the liquid product is a pulse applied in bipolar pulse form. It is advantageous to apply a bipolar pulse to the liquid product, to avoid electrode damage (Loeffler, 1996). Of course, it is part of the invention that also other types of pulses are equally applicable in the process of the invention.
  • the process of the invention is particularly suitable for inactivation of micro-organisms in liquid food products such as juices, sauces, dairy products.
  • liquid food products such as juices, sauces, dairy products.
  • examples of such liquid food products are an ingredient, a semi-finished product, or a final liquid product, like a fruit juice, a vegetable juice, an infant food, a jam, a spread or a smoothie, an alcoholic or nonalcoholic beverage, a dairy product, a plant milk product, a liquid egg, a soup or a sauce.
  • One embodiment of the invention is a process according to the invention, wherein the process is a process for inactivation of micro-organisms in the liquid product.
  • the process of the invention has many advantages over current processes for heating a liquid product by means of resistive heating.
  • One of the major advantages achievable with the process of the invention is the pasteurization of a liquid product, e.g. a liquid food product, wherein the micro-organism is small or large, and wherein the micro-organism is a Gram-negative microbe or a Gram-positive microbe.
  • liquid products subjected to the process according to the invention such as a liquid food product, are cooled to ambient temperature or below, e.g. cooled to 2-8°C, immediately after the process of the invention is applied to the liquid product.
  • one embodiment of the invention is a process, wherein the heated liquid product is cooled immediately after flowing through the apparatus for fast and homogeneously heating a liquid product to a predetermined temperature by means of resistive heating.
  • the cooling is suitably applied immediately from the moment the liquid product is flown through the apparatus, e.g. as fast as possible, preferably within 3 seconds.
  • an embodiment of the invention is a process according to the invention, wherein the heated liquid product is cooled after being transferred through the apparatus for fast and homogeneously heating a liquid product to a heating temperature by means of resistive heating.
  • the process according to the invention is suitable for application with an apparatus for fast and homogeneously heating a liquid product to a heating temperature by means of resistive heating, which apparatus is run at a flow rate of liquid product of between about 0.5 L/h to about 2000 L/h, preferably at about 0.5 L/h to about 2 L/h, more preferably at about 1 L/h, or equally preferably at about 100 L/h to about 2000 L/h, preferably at about 1000 to 1500 L/h, more preferably at about 1200 L/h.
  • the flow rate is about 30.000 L/h.
  • the liquid product is adequately pre-heated before subjecting the liquid product to the process for fast and homogeneously heating a liquid product to a predetermined temperature by means of resistive heating, without the necessity for cooling during the process.
  • the liquid product is preheated to a temperature in the range of from 20°C to 70°C before being supplied to the apparatus, preferably from 35°C to 65°C, more preferably from 40°C to 60°C.
  • the process according to the invention is very effective and efficient in inactivating microorganisms present in the liquid product subjected to the process for fast and homogeneously heating a liquid product to a predetermined temperature by means of resistive heating according to the invention. Applying the process of the invention to a liquid product comprising microorganisms, the microbial count is reduced with at least 2 log cfu/mL, most preferably 6 log cfu/mL or more.
  • the invention is a process, wherein the microbial count (colony forming unit; cfu) in the liquid product is reduced with at least 2 log cfu/mL, preferably at least 5 log cfu/mL, most preferably 6 log cfu/mL or more.
  • One embodiment of the invention is a process according to the invention, wherein the microbial count in the liquid product is reduced with at least 4 log cfu/mL, preferably with at least 7 log cfu/mL.
  • taste and smell of the liquid food product orange juice is preserved for about 60 days or more, upon application of the process of the invention to the orange juice, when the juice is stored at about 7°C.
  • Equally beneficial is the consolidation of quality of the orange juice for about 23 days, when kept at ambient temperature, after subjecting the orange juice to the process of the invention.
  • the process of the invention turns out to be equally applicable for inactivating Gram-positive micro-organisms in a liquid product, and for inactivating Gram-negative micro-organisms in a liquid product.
  • the size of the micro-organism does not play a limiting role, meaning that either small sized or larger sized micro-organisms are inactivated by the process according to the invention.
  • the various aspects and embodiments of the invention are thus an important contribution to the art, since up to the present invention, small Gram-positive micro- organisms could not be efficiently inactivated with known processes for fast and homogeneously heating a liquid product to a heating temperature by means of resistive heating.
  • One embodiment of the invention is therefore a process according to the invention, wherein the microbe optionally comprises a Gram-positive micro-organism.
  • the pH of the liquid product is minimally altered during the course of the process, if changed at all.
  • the invention thus relates to a process, wherein the pH of the liquid product at the end of the process is within 0.5 pH unit from the pH at the start of the process, preferably within 0.2 pH unit, more preferably within 0.1 pH unit, most preferably within 0.05 pH unit.
  • the process according to the invention is suitable for liquid products, in particular liquid food products.
  • a second aspect of the invention relates to a liquid product obtainable by the process according to the invention as has been described above.
  • the invention is further illustrated by the following non-limiting examples, provided below.
  • Pathogenic and spoilage micro-organisms were selected based on their morphology and their association with and prevalence in fruit juice. Furthermore, heat-resistance or PEF resistance of the strains was used as a criteria for selection of the strains. Selected micro-organisms are listed in Table 1.
  • MRS medium containing 52.2 g MRS (De Man, Rosoga and Sharp broth, Merck) and 12 g agar per 1 L distilled water. Plates were overnight incubated at 30°C.
  • a single colony was used to inoculate a 100 mL flask with 10 mL MRS broth and cultivated for 24 h at 20°C in a shaking incubator (180 rpm). From this culture, 200 microliter was used to inoculate 19.8 mL fresh MRS broth, supplemented with 1 % glucose (100 mL flask) and incubated for 24 h at 20°C and 180 rpm. After culturing, cells were washed and suspension of the selected micro-organism was added to orange juice (Minute Maid®), to a final concentration of about 1.0E+8 - 1.0E+9 cfu/mL.
  • the inoculated suspension was pumped through a 1 L/h PEF system at a flowrate of 13.0 ⁇ 0.5 mL/min and preheated to 36°C prior to the electrical treatment.
  • suspension entered two vertically positioned co-linear treatment chambers, wherein the electrical treatment was given.
  • the intensity of the treatment conditions electric field strength, pulse duration and number of pulses applied
  • juice heated up to variable maximum temperatures autonomously No holding section was added, so directly after leaving the treatment chambers (within 3 seconds), juice was cooled down via a heating spiral that was immersed in a water bath.
  • samples were collected aseptically. Due to the variation in chosen frequency, the number of pulses applied and consequential the temperature increase leading to maximum temperature varied (Table 2). Samples were collected at different maximum temperatures, and kinetics for the fixed electrical field strength and pulse duration were determined.
  • the number of viable microbial cells was determined by plating 100 [iL of serially diluted PEF- treated juice in sterile peptone physiological salt diluent (PSDF) on suitable-agar plates supplemented with 0.1% sodium pyruvate to enhance growth of sub-lethally damaged cells (Timmermans et al, 2014). Surviving cells were enumerated after 5 days incubation at 25°C (S. cerevisiae), 30°C (L.monocytogenes, L. plantarum) or 37°C (S. Senftenberg, E.coli).
  • PSDF sterile peptone physiological salt diluent
  • Tested conditions are provided in Table 2. Dimensions of the treatment chamber varied to obtain a variable electrical field strength. As a result of this variable dimension, residence times within the treatment chambers were not the same for every tested condition. To obtain the desired maximum temperature, frequency was adjusted. Finally, the number of pulses was calculated by taking the product of residence time and frequency used. Time between two pulses was calculated by dividing the residence time by number of pulses, minus the pulse duration.
  • Inactivation is shown as the logarithm of number of surviving micro-organism at tested condition, N, divided by start concentration of micro-organisms, No, being log 10 (N/N 0 ).
  • the inactivation is shown as a function of the maximum temperature at the outlet of the treatment chamber for the tested micro-organisms.
  • On the left panel inactivation at various electrical field strength for short pulses (2 microseconds) are shown, demonstrating the current state of art, and on the right panel, inactivation at various electrical field strength for long pulses (i.e. 100 or 1000 microseconds) are shown demonstrating the invention described in this application.
  • an electrical field strength of 2.7 kV/cm with a pulse duration of 100 microseconds or of 1000 microseconds provides inactivation of micro-organisms to a higher degree, compared to higher electrical field strengths and shorter pulse duration commonly applied in the field.
  • the pulse duration plays a critical role in inactivating the micro-organism, hinting at another mechanism for the conditions described in the invention than today used. It is hypothesized that when the pulse duration is long enough, voltage-sensitive protein channels will be opened, and conduct a higher current than they are designed for. As a result, the channels will become irreversibly denaturated, and cells will lose their viability.
  • E.coli inactivation data shows no difference in degree of inactivation at 2.7 kV/cm when pulses of 100 or 1000 microseconds are used, hinting that the critical duration of a the pulse is lower than 100 microseconds.
  • both Gram-positive as Gram-negative micro-organisms can be inactivated up to 1.0E-7 (N/No) using this new PEF conditions.
  • size of the micro- organisms does not play a prominent in the degree of inactivation (right panel) as they do with the current state of the art conditions (left panel).
  • yeasts can be inactivated at lower maximum temperatures at 2.7 kV/cm than L. plantarum and S. Senftenberg, no big differences are found between E.coli and L.monocytogenes, at the new PEF conditions, while larger differences are found with current used PEF conditions (left panels).
  • Example 2 Microbial inactivation of E.coli and Listeria monocytogenes in products with variable characteristics ( 1 L/h scale) Escherichia coli (ATCC 35218) and Listeria monocytogenes NV8 were prepared from frozen stock and cultured in Tryptone Soy Broth (E. coli) or Brain Heart Infusion Broth (L. monocytogenes), according to the method described in (Timmermans et al, 2014).
  • Inactivation is shown as the logarithm of number of surviving micro-organism at tested condition, N, divided by start concentration of micro-organisms, No, being log 10 (N/N 0 ).
  • Example 3 microbial validation at 1200 L/h scale
  • Orange juice was pumped at a flow of 1.200 ⁇ 100 L/h and preheated to 59°C. Pulses were delivered in three vertically positioned treatment chambers to reach a maximum temperature of 70°C.
  • the volume of the treatment zone in the middle treatment chamber was smaller (dimensions length: 14 mm, diameter 8 mm) than the treatment zone of the first and third (outer) treatment chambers (dimensions length 14 mm, diameter 12 mm). Due to the connections of the power sources, this resulted in a double electrical field strength in the middle treatment chamber compared to the outer treatment chambers, being 1.9 kV/cm in the middle and 1.0 kV/cm in the outer treatment chamber. Pulses had a fixed duration of 1000 microseconds. Residence time in the middle treatment chamber was 5.4 ⁇ 0.5 milliseconds and in the outer treatment chambers 9.5 ⁇ 0.9 milliseconds the total number of pulses delivered during the treatment were 5.1 ⁇ 0.01 at a repetition rate of 207 ⁇ 18 Hz.
  • Juice was cooled directly after leaving the treatment chambers within 3 seconds, so holding time was minimised to reduce the heat load on the product. After cooling the juice was packed and aseptically and stored. Microbial samples of untreated and PEF -treated juice were analysed in duplicate in three different laboratories, having a total of 6 untreated and PEF samples to be analysed. Total mesophilic plate count, total coliforms and number of yeasts and moulds were analysed according to method described by Dowes and ITO (2001).
  • Acidothermophilic Spore-forming Bacteria was analysed according to the method of Eguchi et al, (1999), Salmonella was analysed according to the method of AO AC (2000), Listeria monocytogenes was analysed according to the method of ISO 11290-1 (1996) and lactic acid bacteria was analysed according to the method of Silva et al. (2007).
  • Results of the microbial inactivation in untreated orange juice and in PEF-treated orange juice is depicted in Figure 4A and B.
  • the initial microbial load in the untreated orange juice is relatively high, since about l,0E+5 cfu/mL in untreated orange juice were present (Figure 4A).
  • Example 4 Impact of PEF conditions of the invention to quality aspects and microbial shelf life
  • Activity of this enzymes is expressed as the release of acid per mL (multiplied by 1.0E+4) during pectin hydrolysis as a function of time at pH 7.8 and 20°C.
  • most juice pasteurisers provide juice having pectinesterase activity expressed in pectinesterase units (PEU) with values of between 1.0E-6to 1.0E-4.
  • Acidothermophilic sporeforming bacteria in orange juices: detection methods, ecology, and involvement in the deterioration of fruit juices. Report of the research project ABE Citrus, Campinas (SP), Brasil.
  • Escherichia coli electropulsation orientation, permeabilization and gene transfer. Biophysical Journal, 75, 2587-2596

Abstract

La présente invention concerne un procédé de chauffage rapide et homogène d'un produit liquide à une température prédéterminée au moyen de chauffage résistif. Selon l'invention, l'inactivation microbienne suffisante et efficace est obtenue lors de l' application d'une intensité de champ électrique entre 0,1 et 5,0 kV/cm pendant une durée prolongée, en sélectionnant une intensité de champ électrique relativement faible et une durée d'impulsion d'au moins 10 microsecondes tandis que la température maximale du produit liquide reste de manière autonome inférieure à 92 °C pendant le chauffage résistif. Le procédé de l'invention est efficace à un pH neutre et à un pH inférieur à 7. En outre, le procédé de l'invention est efficace pour inactiver une large plage de micro-organismes pertinents. La présente invention concerne en outre ledit procédé dans lequel le produit liquide est préchauffé avant de soumettre le produit liquide au procédé. La présente invention concerne également le produit liquide pouvant être obtenu par le procédé selon l'invention.
PCT/NL2016/050799 2015-11-17 2016-11-17 Procédé de conservation d'aliment liquide utilisant un traitement de champ électrique pulsé WO2017086784A1 (fr)

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EP16810096.4A EP3376878A1 (fr) 2015-11-17 2016-11-17 Procédé de conservation d'aliment liquide utilisant un traitement de champ électrique pulsé
US15/776,164 US11903400B2 (en) 2015-11-17 2016-11-17 Process for liquid food preservation using pulsed electrical field treatment
JP2018544752A JP6921838B2 (ja) 2015-11-17 2016-11-17 パルス電界処理を用いた液体食品保存のための方法
MYPI2018701823A MY189799A (en) 2015-11-17 2016-11-17 Process for liquid food preservation using pulsed electrical field treatment
BR112018009870-1A BR112018009870B1 (pt) 2015-11-17 2016-11-17 Processo para aquecimento rápido e homogêneo de um produto líquido
SG11201803585PA SG11201803585PA (en) 2015-11-17 2016-11-17 Process for liquid food preservation using pulsed electrical field treatment
CN201680066218.4A CN108471787A (zh) 2015-11-17 2016-11-17 利用脉冲电场处理保存液体食物的方法
EA201890901A EA037900B1 (ru) 2015-11-17 2016-11-17 Способ сохранения жидких пищевых продуктов c использованием обработки импульсным электрическим полем
AU2016357683A AU2016357683B2 (en) 2015-11-17 2016-11-17 Process for liquid food preservation using pulsed electrical field treatment
MX2018006003A MX2018006003A (es) 2015-11-17 2016-11-17 Proceso para la preservacion de alimentos liquidos usando el tratamiento de campo electrico pulsado.
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WO2020173644A1 (fr) * 2019-02-26 2020-09-03 Krones Ag Dispositif et procédé pour fournir un aliment liquide stérilisé ou pasteurisé au moyen de champs électriques pulsés
WO2021007553A1 (fr) * 2019-07-10 2021-01-14 Kemin Industries, Inc. Nouvelles applications de technologie à champ électrique pulsé et à faisceau électronique
CN112438359A (zh) * 2019-08-27 2021-03-05 湖南农业大学 一种真空冷冻干燥牛肉干及其加快干燥速率的方法
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