MXPA01009069A - Ice confection containing no other proteins than natifreeze proteins - Google Patents

Abstract

The use of an antifreeze protein within an ice confection to restrict the flow of flavour or colour ions or molecules present as either solutes or a dispersion, wherein the ice confection contains no protein other than the antifreeze protein.

Description

ICE CREAM CONFECTION THAT DOES NOT CONTAIN PROTEINS OTHER THAN ANTIFREEZE PROTEINS TECHNICAL FIELD OF THE INVENTION The invention relates to the use of antifreeze proteins in ice cream jams. In particular, the invention relates to the use of antifreeze proteins in ice cream confections to restrict the flow of flavors and / or color. Background of the Invention It is a well-known problem that when an ice cream jam such as a sorbet is consumed, the taste and color is quickly sucked out of the product leaving essentially an ice block which is unappetising to eat. In addition, in ice cream jams such as sorbet which is comprised of a number of different components, each has a different color or flavor, the boundary between each component is not marked and is different because of the flow of color or flavor to a certain degree from one component to another. Consequently, to date it has not been possible to provide an ice cream jam that has REF: 132729 light components of a different flavor or color which remain different. WO 98/04146 (Unilever) discloses that AFPs can be incorporated into frozen food products such as ice cream jams to provide desirable product properties that provide that the product and processing conditions are varied so that the ice crystals provided in the product they have an aspect ratio of more than 1.9, preferably 1.9 to 3.0. The specific examples in WO 98/04146 are all cream ice cream compositions. WO 98/04146 does not teach that it is possible to restrict the flow of color and / or flavor in sorbet products by the inclusion of an antifreeze protein in the sorbet composition. WO 96/39878 describes a method for producing a frozen composition for storage, the method does not require a hardening step prior to storage. The frozen composition contains an antifreeze protein, in particular Type I AFP. The examples show the preparation of an aerated cream ice cream and an aerated frozen yogurt. WO 96/39878 does not teach that it is possible to restrict the flow of color and / or flavor in sorbet products by the inclusion of an antifreeze protein in the sorbet composition. US 5 118 792 (Warren et al) describes the addition of fusion proteins, and in particular protein A-Saf5 fusion protein in foods which are to be consumed frozen, for example, ice cream, frozen yogurt, Milk ice cream, snow, popsicles and frozen whipped cream. No examples are given where a final ice cream confection product containing such fusion proteins is provided. It is shown in Example 3B that when a paddle formulation is used within the "crush test", the growth of the ice crystals is restricted. US 118 792 does not teach that it is possible to restrict the flow of color and / or flavor in sorbet products by the inclusion of an antifreeze protein in the sorbet composition. It has now been found that the addition of antifreeze proteins to ice cream jams restricts the flow of flavor and / or color.

Description of the Invention Accordingly, the invention provides the use of an antifreeze protein within an ice cream confectionery to restrict the flow of ions or molecules of flavor or color present as either solutes or a dispersion wherein the ice cream jam does not contain proteins other than the antifreeze protein. By antifreeze protein (AFP) is meant a protein which has significant ice recrystallization inhibition properties when measured according to Example 1. AFP provides an ice particle size in the recrystallization of less than 20 μm, more. preferable from 5 to 15 μm. Preferably the ice cream jam comprises at least 0.0005% by weight of antifreeze protein, more preferably 0.0025% by weight of antifreeze protein. Typically the ice cream jam will comprise from 0.0005% by weight to 0.005% by weight of antifreeze protein. For some applications it may be advantageous to include a mixture of two or more different AFPs in the ice cream jam. The AFP for use in products of the invention can be any AFP suitable for use in food products. Examples of suitable sources of AFP are given, for example, in the article "Antifreeze proteins and their potential use in frozen food products", Marylin Griffith and K. Vanya Ewart, Biotechnology Advances, vol. 13, pp. 375-402, 1995 and in Patent applications WO 98/04699, WO 98/04146, WO 98/04147, WO 98/04148 and WO 98/22591. The AFPs can be obtained from their sources by any suitable process, for example the isolation processes as described in the documents mentioned above. A possible source of AFP materials is fish. Examples of fish AFP materials are antifreeze glycoproteins (for example, available from Atlantic cod, Greenland cod and Tomcod), AFP Type I (eg obtainable from Winter flounder, Yellowtail flounder, Shorthorn sculpin and Grubby sculpin), Type II AFP (for example, obtainable from Sea raven, Smelt and Atlantic herring) and Type III AFP (for example, obtainable from Ocean Pout, Atlantic olffish, Radiated Shannon, Rock gunnel and Laval 's eelpout). A preferred example of the latter type is described in WO 97/02343. Another possible source of AFP material is invertebrates. Bacterial AFPs can also be obtained.

A third possible source of AFP material is plants. Examples of plants that contain AFPs are gralic mustard, blue wood aster, spring oats, winter cress, winter cañola, Brussels sprouts, carrots, Dutchman tails, spurge, yellow lily, winter barley, plant leaf Virginia aquatic plant, narrow-leaved plantain, plantain, spikelet, Kentucky blue-green silky grass, poplar of the Orient, white oats, winter rye, sweet-sour blackberry, potato, alfalfa, yellow, spring and winter wheat, triticalo , periwinkle, violet and yerba. Both naturally occurring species and species that have been obtained through genetic modification can be used. For example, microorganisms or plants can be genetically modified to express AFPs and then the AFPs can be used according to the present invention. Genetic manipulation techniques can be used to produce AFPs having at least 80%, more preferably more than 95%, more preferred 100% homology for AFPs directly obtained from natural sources. For the purpose of the invention these AFPs that possess this high level of homology are also comprised within the term "AFPs". The techniques of genetic manipulation can be used as follows: An appropriate host cell or organism could be transformed by a construct of genes containing the desired polypeptide. The nucleotide sequence encoding the polypeptide can be inserted into a suitable expression vector that encodes the elements necessary for transcription and translation and in a manner that they will be expressed under appropriate conditions (e.g. in the proper orientation and reading structure). correct and with appropriate target formation and expression sequences). The methods required to construct these expression vectors are well known to those skilled in the art. A number of expression systems can be used to express the sequence encoding the polypeptide. These include, but are not limited to, bacteria, yeast insect cell systems, plant and plant cell culture systems all transformed with the appropriate expression vectors. A wide variety of plants and plant cell systems can be transformed with the nucleic acid constructs of the desired polypeptides. Preferred embodiments could include, but are not limited to, corn, tomato, tobacco, carrots, strawberries, rapeseed and sugar beet. For some natural sources, the AFPs may consist of a mixture of two or more different AFPs. Preferably the antifreeze protein is chosen so as to provide an aspect ratio of more than 1.9 for the ice crystal, preferably from 1.9 to 3.0, more preferably from 2.0 to 2.9, even more preferably from 2.1 and 2.8 (see WO 98/04146). The aspect ratio is defined as the maximum diameter of a particle divided by its minimum diameter. The aspect ratio can be determined by any suitable method. A preferred method is illustrated in the Examples (Example 6). For the purpose of the invention, the preferred AFPs are derived from fish. Especially preferred is the use of type III fish proteins, HPLC 12 is more preferred as described in WO 97/02343. Suitable ice cream confections, which do not contain proteins other than AFP, include water ice, snow, granitas and frozen fruit purees.

Preferably the ice cream jam is a sorbet. Sorbet means a frozen solution made essentially of water, sugar, fruit acid or other acidifying agent, color, fruit or fruit flavoring. The sorbet will typically have an ice content of at least 30% by volume when measured at -18 ° C, more preferably at least 40% by volume when measured at -18 ° C, most preferably at least 50 ° C. % in volume when measured at -18 ° C. The ice content can be determined following the techniques described in the article by B of Cindio and S It will run in the Journal of Food Engineering, Volume 24, pages 405-415, 1995. The enthalpy data required for this technique is obtained using adiabatic calorimetry (Holometrix Adiabatic Calorimeter). The ice contents as expressed herein are measured on a sample of 80 g poured into the sample container of the calorimeter and cooled to -75 ° C by placing the assembly on dry ice prior to placement in the calorimeter (pre-cooled to between -70 ° C and -80 ° C). The enthalpy data obtained was analyzed to give the ice content as a function of the temperature following the method of Cindio and Carrera.

In general, the sorbet has a total content of soluble solids, less than 40% by weight, preferably less than 25% by weight, more preferably less than 15% by weight. For low calorie water snows, the content of soluble solids can be as low as about 5% by weight. The total soluble solids content is measured at 4 ° C and is the% by weight of the total composition that dissolves at this temperature. The ice cream jam can be aerated or not aerated, preferably the ice cream jam is not aerated. By non-aerated is meant an ice cream confection having a surplus of less than 25% (equivalent to 0.2 volume fraction of air), preferably less than 10% (equivalent to 0.09 volume fraction of air). During the processing of the ice cream jam, steps such as shake are not deliberated which are assumed to increase the gas content of the product. However, it should be done during normal methods for the preparation of non-aerated ice cream jams, low levels of gas or air can be incorporated into the product, for example due to the use of mixing conditions.

Typical color materials used in ice cream jams could include, for example, carmoisine, carotene, anthocyanin, chlorophyll, chlorophyllins, chlorophyll and chlorophyllin copper complexes, riboflavin, riboflavin-5'-phosphate, candy, charcoal black , hot pepper extract, capsanthin, capsorubin, red beet, calcium carbonate, titanium dioxide, iron oxides and hydroxides, achiote extract, curcumin, tartrazine, quinoline yellow, sun yellow FCF, cochineal, ladybird 4R , AC alura red, blue patented V, carmine indigo, bright blue FCF, green S, bright black BN, coffee HT, lycopene, beta-apo-8'-carotenal (C30), ethyl ester of acid Beta-apo-8 ' -carotenic (C30) and lutein. Typical flavor materials used in ice cream jams could include, for example, naturally flavored compounds, identical to natural or synthetic, examples of which include; cherry, strawberry, raspberry, orange, banana, lemon, lime, lychee, guava, passion fruit, mango, grape, kiwi, melon, pineapple, papaya, apple, plum, apricot, peach, pear, mint, marshmallow, caramel, caramels licorice, coffee, cotton candy and pump gum.

The inclusion of antifreeze proteins in ice cream jams results in the formation of a dense, strong continuous network of ice crystals within the ice cream jam. By dense continuous network of ice crystals it is understood that any given ice crystal is connected to at least one ice crystal. In non-aerated ice cream jams which have been frozen with agitation, the degree of network formation can be measured as contiguity. Contiguity is defined as the ratio of the particle-to-particle interface area divided by the total interface area. It is therefore a measure of the degree of network formation of the particle phase. Example 2 shows a method for contiguity measurement. The non-aerated ice cream jams according to the invention have a contiguity of at least 0.2, when measured by the test given in Example 2, for an ice content of 50-90%, preferably 54-85% by weight when it is measured at -18 ° C. In non-aerated ice cream jams which have been frozen by any means, the degree of network formation can be measured as the Euler-Poincare characteristic of the ice phase. The Euler-Poincare characteristic is a measure of the degree of network formation of a particular phase. The lower and more negative is the value of the Euler-Poincare characteristic, the greater the continuity of the phase in question. Example 4 shows a method for measuring the Euler-Poincare characteristic. The non-aerated ice cream jams according to the invention have a Euler-Poincare feature of ice phase of less than -150mm "2, when measured by the test given in Example 4, for an ice content of 50. -90%, preferably 54-85% by weight when measured at -18 ° C. The use of an antifreeze protein within the ice cream jam to restrict the flow of ions or molecules of flavor or color present as either solutes or A dispersion provides a number of advantages, In particular, ice cream jam products are provided for which the taste and / or color is not significantly sucked during consumption.The flavor and / or color is retained throughout the ice cream jam. during the time of total consumption.

A further advantage of the use of AFP to restrict the flow of flavor and / or color is that multi-component products can be provided, each component has a different taste and / or color and the distinction between each component remains very marked. In particular, this allows products that have light components of different color and / or taste to be provided. Each light component remains distinct from each other, these are not substantially combined of different taste and / or color over time. The ice cream jam according to the invention may comprise the entire product or may be included within a composite product. For example, a product having a conventional ice cream core coated with 2 or more thin layers of sorbet containing AFP can be provided, each sorbet layer being a different flavor and / or color.

EXAMPLES The invention will now be illustrated by means of the following examples.

Example 1 Method for determining whether an AFP possesses ice recrystallization inhibition properties The recrystallization inhibition properties can be measured using a modified "crush test" (Knight et al, 1988). 2.5 μl of the solution under investigation in 30% (w / w) of sucrose is transferred on a 16 mm circular coverslip, appropriately labeled, clean. A second coverslip is placed on top of the drop of solution and the sandwich is pressed together between the index finger and the thumb. The sandwich is dripped in a hexane bath maintained at -80 ° C in a dry ice box. When all the sandwiches have been prepared, the sandwiches are transferred from the hexane bath at -80 ° C to the observation chamber containing hexane held at -6 ° C using forceps pre-cooled in the dry ice. During the transfer at -6 ° C, the sandwiches can be observed to change from a transparent to an opaque appearance. Images are recorded by video camera and recorded in an image analysis system (LUCIA, Nikon) using a 20x objective. The images of each crush are recorded at a time = 0 and again after 60 minutes. The size of the ice crystals in both tests is compared by placing the slides inside a temperature controlled cryostat showcase (Bright Instrument Co. Ltd. Huntingdon, UK). The images of the samples are transferred to a Quantimet 520 MC image analysis system (Leica, Cambridge UK) by means of a Sony monochrome CCD camcorder. The sizing of the ice crystals is done by hand designing around the ice crystal. At least 400 crystals are sized for each sample. The size of the ice crystal was taken as being the largest dimension of the 2D projection of each crystal. The average crystal size was determined as the numerical average of the individual crystal sizes. If the size at 30-60 minutes is similar or only moderately (less than 10%) increased compared to the size at t = 0, and / or the size of the crystal is less than 20 micrometers, preferably from 5 to 15 micrometers this is an indication of the good ice recrystallization inhibition properties.

Example 2 Measurement of Contiguity Contiguity is measured using microstructural images of ice cream jam using a cryogenic scanning electron microscope (SEM). The structures are formed in images using the technique described in "A low temperature scanning electron microscopy study of ice cream I. Techniques and general microstructure" Food Structure Vol. 11 (1992), pp 1-9. In a particulate composite, the Contiguity of the particulate material phase is defined as the ratio of the area of particle to particle interconnection divided by the total internal interconnection area. It is a measure of the degree of network formation of the particulate material phase. In ice cream jams the particles are ice crystals inside the matrix and thus the contiguity of the ice is defined as; where Cu is the contiguity, Aa is the total interfacial surface area of ice-ice interfaces and Aim is the interfacial surface area of ice-matrix interfaces. The contiguity can be measured from microstructural images of flat surfaces randomly cut through the material. The Cryo-SEM images of the flat fracture surfaces of the non-aerated ice cream jam are sufficient for this. By placing an array of lines in the image of the microstructure, the number of intersections or crystallographic parameters of these lines with ice-ice and ice-matrix interfaces are counted and combined in the following equation, to give the contiguity; C ,, = - 2N, m? Nim) where Na = number per unit length of intersections or ice-ice crystallographic parameters and Nim = number per unit length of intersections or crystallographic parameters of ice-matrix. Ideally, there are approximately 800 interfaces out of a total of 5 images that are representative of each sample structure. To determine the contiguity, two sets of measurements were taken from each image. After placing a regular set of lines in the image, the number of intersections of the ice-matrix and ice-ice interfaces were counted with these lines, only including all the obvious ice-ice interfaces. The count was repeated then, but this time with all possible ice-ice interfaces included. Such as, a maximum ice contiguity measurement and a minimum ice contiguity measurement were made for each image. The average of these figures is then taken as the contiguity value.

Example 3 Measurement of Aspect Ratio Samples were equilibrated at -18 ° C in a Prolan environmental cabinet for approximately 12 hours. The microscopic slides were prepared by staining a thin layer of the ice cream jam from the center of the thin glass plates. Each slide was transferred to a temperature-controlled microscopic stage (at -18 ° C) where images of the ice crystals (about 400 individual ice crystals) were collected and re-emitted through a video camera to a storage of image and analysis system. The images of the stored ice crystal were visually enhanced manually by drawing around its perimeter which then visually enhances the entire crystal. The images of the visually enhanced crystals were then measured using the set of software (image analysis software) which counts the number of pixels required to complete the longest diameter, (length), the shortest diameter (width), the aspect ratio (length / width). The average aspect ratio for the crystals was calculated.

Example 4 Measurement of the Euler-Poincare characteristic The Euler-Poincare characteristic is measured using microstructural images of ice cream jam using a cryogenic scanning electron microscope (SEM). The structures are formed in images using the technique described in "A low temperature scanning electron microscopy study of ice cream I. Techniques and general microstructure" Food Structure Vol. 11 (1992), pp 1-9.

In a composite structure of two phases, the degree of continuity of a phase can be measured using the Euler-Poincare characteristic. The lower the value of the Euler-Poincare characteristic for a phase, the more continuous or connected that phase will be within the microstructure. The Euler-Poincare characteristic can be a positive or negative number. The definition of the Euler-Poincare characteristic is given in "Unbiased estimation of the Euler-Poincare characteristic" by B.P. Pinnamaneni, C. Lantuejoul, J.P. Jernot and J.L. Chermant, Acta Sterelogica, 1989, 8/2, p 101-106. To measure the Euler-Poincare characteristic for ice in ice cream jams, the identification of the ice and matrix phases in the microstructural images is performed and an image analysis system is used; the Euler-Poincare characteristic of the ice phase was determined using a specifically written analysis program. When the contrast in the images was insufficient for the image analysis system to automatically distinguish ice and matrix separately, the interface between the two was manually identified, thus making it possible to ensure the determination of the Euler-Poincare characteristic.

The Euler-Poincare feature can be measured for ice in an ice cream jam produced by any processing route.

Example 5, Comparative Example A A sorbet solution having the following composition was prepared as follows; % by weight Sucrose 20.0 Locust bean gum 0.2 Water at 100 Total Soluble Solids; 20.2% by weight Ice content at -18 ° C; 70% by weight The sorbet solution was prepared as follows; All sorbet ingredients were mixed together, except AFP, using a high-cut mixer for approximately 3 minutes. The water was added at a temperature of 80 ° C. The temperature of the sorbet mixture is about 55-65 ° C after mixing. The mixture was then passed through a plate heat exchanger for pasteurization at 81 ° C for 25 seconds. The mixture was then cooled to approximately 4 ° C in the plate heat exchanger prior to use. After pasteurization AFP Type III (as described in WO 97/02343) was added to the sorbet solution in the following concentrations; Example 5 - 0.005% by weight Comparative Example A - without AFP The sorbet solution was frozen at rest without surplus which is introduced as follows: The sorbet solution was poured into divided metal molds producing bars having the dimensions 25 x 25 x 200mm They were then placed in the cold room overnight to freeze at rest at a temperature of -25 ° C. The next day, the molds were removed from the test bars, placed in polyethylene bags and stored at -25 ° C. The Euler-Poincare characteristic was measured as in Example 4. The results are shown in Table 1.

Table 1 Examples 6 and 7, Comparative Example B A sorbet solution having the following composition was prepared as follows; % by weight Sucrose 20.0 Locust bean gum 0.2 Water at 100 Total soluble solids; 20.2% by weight Ice content at -18 ° C; 70% by weight The sorbet solution was prepared as in the Example 5. After the pasteurization, AFP Type was added III (as described in WO 97/02343) to the sorbet solution in the following concentrations; Example 6 - 0.0005% by weight Example 7 - 0.005% by weight Comparative Example B - without AFP The sorbet solution was frozen in a Technohoy MF 75 striped surface heat exchanger with no surplus being introduced. The sorbet was extruded at a temperature of -3.9 ° C to -5.6 ° C. The product was then hardened in a forced air freezer at -35 ° C, then stored at -25 ° C. The contiguity was measured as in Example 2. The results are shown in Table 2 Table 2 Example 8 The production of a "big caramel" product having different colored sorbet and flavor layers sequentially processed around a core of ice cream.

The use of the sorbet composition according to the invention provides a product which is very hard and forces the consumer to lick, before biting, the product and then each layer of sorbet is gradually discovered. In addition, the sorbet layers remain separated and little or no "running" color is observed between the layers. A non-aerated sphere of ice cream (20-30 mm in diameter) having the following formulation was molded into a bar as follows; Cream ice cream composition% (weight) Double cream 26.5 Skimmed milk powder 9.2 Sucrose 16.0 Water at 100 The premixed cream ice cream was poured into an aluminum mold and the mold was cooled in a freezer by forced air at -35 ° C . When the premixed cream ice cream was partially frozen a bar was inserted. When the cream ice cream was completely frozen, it was removed from the mold by spraying the outside of the mold with water at 50 ° C.

The cream ice cream core was pre-cooled by immersion in solid C02 (dry ice) for approximately 2 minutes then immersed in a sorbet mixture having the following formulation; Sorbet composition % (weight) Sucrose 15.0 Dextrose 5.0 Locust bean gum 0.25 Citric acid '0.5 Flavor / Color 0.2 AFP * of Type III 0.005 Water at 100 * as described in WO 97/02343 Total solids; 20.5% Ice content at -18 ° C; 68.0% by weight The product was then submerged sequentially in the sorbet mixture to make a number of layers (typically 12 to 15) of different colors and flavors.

Between each submerged in the sorbet mixture the product was cooled in dry ice to facilitate the absorption of the next layer of sorbet. The resulting product was a sphere of approximately 3-5 cm in diameter.

Example 9 A sorbet solution having the following composition was prepared; % in weigh Sucrose 10.0 Glucose 5.0 Locust bean gum 0.2 Citric acid 0.5 Water at 100 The composition was divided into four and the following color and / or AFP was added. (i) flavor / color of cherry 0.5% p / p AFP * 0.005% p / p (ii) AFP * 0.005% p / p (iii) cherry flavor / color 0.5% w / w (iv) No additions * AFP as in WO 97/02343 Two color sorbet monobites were made either out of (i) and (ii) above (ie, containing AFP) or out of (iii) and (iv) above (without AFP) as follows: 5 ml aliquots of unbleached sorbet solution ((ii) or (iv)) were formed in latex bucket molds for ice. These were frozen for 1 hour in a forced air freezer at -35 ° C. 5 ml red sorbet mixture ((i) or (iii)) were then used to fill the remaining mold volume and the molds were frozen by forced air to -35 ° C for an additional hour. The molds were then transferred to a refrigerator at -25 ° C overnight before removal of the molds. Once the molds were removed, the monobits were transferred into individual plastic containers with sealed lids and stored at -10 ° C per 1, 2, 3, 5 and 7 weeks. The photographs were taken at time 0 and after the specified period of time. As long as the sample containing AFP shows little or no combination of colors even after 7 weeks at -10 ° C, the sample that does not contain AFP shows the color combination after only 1 week at -10 ° C and the complete sample was almost a single mixed color after 7 weeks at -10 ° C.

Example 10 Sorbet samples were produced and tested in three flavors, with and without AFP, by Time Sensitive Sensitive methodology, as detailed below.

Formulation of the sorbet % (P / p) Sucrose 13.7 Glucose 5.9 Stabilizer 0.15 Citric acid 0.3 AFP * 0.005 Taste / Color see later Water at 100 Taste / color of orange: 1.0% Flavor / strawberry color: 0.8% Flavor / cherry color: 0.3% * as described in WO 97/02343 All the ingredients of the sorbet except the AFP were mixed together using a high mixer Cut for approximately 3 minutes. The water was added at a temperature of 80 ° C. The temperature of the sorbet mixture is about 55-65 ° C after mixing. The mixture was then homogenized (2000 psi) and passed through a plate heat exchanger for pasteurization at 81 ° C for 25 seconds. The mixture was then cooled to approximately 4 ° C in the plate heat exchanger prior to use. All the products were prepared in the same way. The liquid mixtures were distributed in smaller plastic tubes (approximately 100 ml) at cooling temperature (4 ° C). Then these tubes were frozen by forced air for 3 hours at -35 ° C before they were transferred to the refrigerator at -25 ° C. Prior to the evaluation of these sorbet blocks, they were cut into pieces of uniform size (approximately 2 cm x 2 cm x 1 cm) and equilibrated at -18 ° C overnight.

Sensitive Methodology The products were valued by a group of people who appreciate the highly prepared smell and flavor. He used a methodology of descriptive analysis by which prepared group members identify and quantify the main sensory properties of water snow with and without AFP. The basic aspects which this method involves are given in Sensory Evaluation Techniques, 2nd Edition (1991) M Meilgaard, G.V. Civille and B. T. Carr, CRC Press, and include: • The development of the sensitive descriptors by the group, referred to later as the attribute profile. • Consensus agreement of common results on each sensitive attribute of a 'control' product. • Evaluation of some experimental and commercial samples to review the performance of the group prior to the selection of the jury of all the experimental samples. • All test sessions are conducted in individual boxes, in a controlled environment to eliminate the prejudice of external variables.

• Analysis of the data via the Analysis of Variance (ANOVA) with the Duncan Multiple Range Comparison test to verify the statistically significant differences (p = < 0.05) between the samples.

Choice of Jury for Time Intensity Time intensity (TI) is a sensitive profiling method that measures how an individual attribute changes over time, giving a quantifiable measure of the "dynamic" aspects of sensory perception. It differs from other sensitive techniques in that all panelists are treated as individuals and therefore it is important that they are reproducible within themselves rather than against any means of the jury.

Background for Data Analysis The approach to IT data analysis is to calculate a number of parameters (eg, Maximum height, area under the curve) that characterizes each curve, and then analyze how the test factors (such as the used product) affect the values of these parameters. The emphasis of this approach is, therefore, to find the significant differences between products rather than differences of the individual panelists.

Experimental Design The test was performed using a statistical design.

The products were presented in a random order, a hidden control was included and an open control was used at the beginning of each session. Panelist number: 10 Session number: 3 for each test Periods of time allowed for evaluation: 30 seconds Valuated Samples (formulations as detailed above); 1. Cherry Sherbet 2. Cherry Sherbet + AFP 3. Strawberry Sorbet 4. Strawberry Sorbet + AFP 5. Orange Sorbet 6. Orange Sorbet + AFP Results a. Strawberry Flavor The average duration of the maximum flavor resistance was significantly longer for the sample containing AFP than the control. b. Cherry Flavor The average duration of the maximum flavor resistance was significantly prolonged for the sample containing AFP than the control. c. Orange flavor The average duration of the maximum flavor resistance was significantly prolonged for the sample containing AFP than the control.

It is noted that in relation to this date the best method known by the applicant to carry out the aforementioned invention, is the conventional one for the manufacture of the objects or products to which it refers.

Claims (6)

1. CLAIMS Having described the invention as above, the contents of the following claims are claimed as property: 1. The use of an antifreeze protein within an ice cream jam to restrict the flow of ions or molecules of flavor or color present in either solutes or a dispersion, where the ice cream jam does not contain proteins other than the antifreeze protein.
2. 2. The use of an antifreeze protein according to claim 1, wherein the ice cream jam is a sorbet.
3. 3. The use of an antifreeze protein according to claim 2, wherein the sorbet is not aerated.
4. The use of an antifreeze protein according to any of the preceding claims, wherein the antifreeze protein is chosen so as to provide an aspect ratio of more than 1.9, preferably from 1.9 to 3.0, more preferably from 2.0 to 2.9, more preferably from 2.1 to 2.8, to the ice crystal.
5. 5. The use of an antifreeze protein according to any preceding claim, wherein the antifreeze protein is AFP Type III HPLC12. The use of an antifreeze protein according to any preceding claim, wherein the antifreeze protein is present at a concentration of at least 0.0005% by weight, preferably at least 0.0025% by weight.