CN104719610A - Hypoallergenic milk protein powder and preparation method thereof - Google Patents
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Abstract
The invention relates to hypoallergenic milk protein powder and a preparation method thereof and belongs to the field of processing of milk products. The preparation method provided by the invention comprises the following steps: by taking milk protein powder as a raw material, fully dissolving in water, then adding compound proteinase, obtaining a milk protein enzymolysis solution by using a biological enzymolysis technology, performing ultrafiltration on the milk protein enzymolysis solution through a ceramic membrane to obtain a protein solution with molecular weight of less than 10000KD, concentrating and drying to obtain the hypoallergenic milk protein powder provided by the invention. The hypoallergenic milk protein powder is whole milk protein, so that the sensitization of whey protein and casein protein is reduced, and the local flavor is good; and the hypoallergenic milk protein powder is safe to eat, has no additives and can provide the complete milk protein for people sensitive to the whey protein. The method for preparing the milk protein powder is simple and easier to realize industrial production and has huge application prospects and market values.
Description
Technical field
The present invention relates to milk product manufacture field, particularly, relating to a kind of is raw material with lactoprotein, utilizes biological enzymolysis technology to prepare the method for low sensitization lactoprotein powder, and the low sensitization lactoprotein powder obtained by the method.
Background technology
Milk allergy is the one of anaphylactia.Its mechanism can be divided into immediate hypersensitivity and delayed allergy.Clinical symptoms show: skin is allergic dermatitis, and respiratory system is rhinitis, asthma, and digestive organs are diarrhea, vomiting.These symptoms separately or may merge generation, also may show as systemic allergic, even anaphylactic shock time serious.A large amount of epidemiological studies shows, lactoprotein allergy is the most general concerning infant, and its incidence is approximately 2-6%.Milk allergy has become people's nutrition especially important Community health's problem in child nutrition.
The protein content of cow's milk is 3.0% ~ 3.5%, and its main component is casein and lactalbumin, and ratio is 80:20.Research shows, in cow's milk, all albumen is all potential anaphylactogen, but most of Milk allergy patient, usually easily by several lactoprotein sensitization, comprises beta lactoglobulin, ALA, casein (main alpha-casein, beta-casein), bovine serum albumin(BSA), lactoferrin.Wherein, beta lactoglobulin, casein and ALA are topmost anaphylactogens in cow's milk, their respectively can by 62% ~ 80%, the SERUM IgE identification of the Milk allergy patient of 60-80% and 50-60%, especially casein autopath showed a rising trend in recent years.
Proteolysis is a kind of conventional protein-modified method, and it can not affect the nutritive value of product.Zheng Hai (2009) adopts 8 kinds of protease to carry out enzymolysis to lactalbumin, and result shows that alkali protease and papain can reduce the antigenicity of beta lactoglobulin and ALA preferably.The results of study such as Wroblewska (2005) show: alkali protease-papain fractional hydrolysis is one of effective ways reducing lactalbumin allergenicity, and papain is added the effect can played and improve hydrolysate organoleptic properties as the second enzyme, especially bitter taste can be reduced.
The current report about low sensitization lactoprotein exploitation mainly about the method for lactalbumin anaphylactogen degraded, and has no the patent or bibliographical information that reduce casein sensitization method in lactoprotein.Lactalbumin and caseic sensibiligen synchronously can be reduced and simple method efficiently if can develop, the lactoprotein obtained both possessed low irritability, possess again the abundant nutrition composition of lactoprotein and good local flavor, certainly will will have huge application prospect and market value.
Summary of the invention
In order to solve the problems of the technologies described above, the object of the invention is to, a kind of preparation method of low sensitization lactoprotein powder is provided.
Another object of the present invention is to, a kind of low sensitization lactoprotein powder obtained by described method is provided.
Another object of the present invention is, provides described low sensitization lactoprotein powder in the application of the low sensitization food of preparation.
To achieve these goals, the preparation method of low sensitization lactoprotein powder provided by the invention, comprises the following steps:
(1) lactoprotein powder is mixed with water, fully dissolve and obtain milk protein solution, ultrasonic wave process milk protein solution;
(2) add compound protease and carry out enzyme digestion reaction, after enzyme digestion reaction terminates, insulation, then be cooled to room temperature, obtain lactoprotein enzymolysis liquid;
(3) lactoprotein enzymolysis liquid is passed through ceramic membrane ultrafitration, collect filtered fluid;
(4) filtered fluid is concentrated, dry, obtain the lactoprotein powder of low sensitization.
Wherein, in step (1), the protein content of lactoprotein powder is 65%-70%, lactose is 16%-20%, fat is 1%-2%, moisture is 3%-5%, and described % is mass percent.
Above-mentioned lactoprotein powder is preferably concentrated milk albumen powder.
In the step (1) of said method of the present invention, lactoprotein powder and the mixed mass ratio of water are 6%-10%; 35-45 DEG C of aquation 60-120min, makes lactoprotein powder fully dissolve, prepares milk protein solution.
Preferably, lactoprotein powder and the mixed mass percent of water are 8%; 40 DEG C of aquation 120min, make lactoprotein powder fully dissolve, prepare milk protein solution.
In step (1), the time of ultrasonic wave process is 15 ~ 30 minutes; Ultrasonic frequency is 20 ~ 25KHz.Preferably, the time of ultrasonic wave process is 30 minutes; Ultrasonic frequency is 25KHz.
In step (2), the consumption of compound protease is the 0.5%-2% of lactoprotein grain weight amount; Preferably 1%; Described compound protease comprises flavor protease, alkali protease and neutral proteinase, and the mass ratio of three is 2-4:2:1.Preferably, three's mass ratio is 3:2:1.
In step (2), the condition of enzyme digestion reaction comprises: temperature is 45-55 DEG C, and pH value is 7.0 ~ 8.0, and the reaction time is 30-120min.
Preferably, in step (2), the condition of enzyme digestion reaction comprises: temperature is 50 DEG C, and pH value is 7.5, and the reaction time is 120min.
Further, in enzyme digestion reaction process, NaOH or HCl of pH value 1mol/L regulates.
In step (2), at 90 ~ 95 DEG C, be incubated 3 ~ 5 minutes after enzyme digestion reaction terminates, then be cooled to room temperature.
In step (3), ceramic membrane aperture is 0.001 μm, can retain and be greater than 10000 daltonian protein.
The invention provides the purposes of above-mentioned any one method in the low sensitization lactoprotein powder of preparation.
The invention provides the low sensitization lactoprotein powder that said method prepares, its protein molecular weight < 10000 dalton.
Present invention also offers the application of low sensitization lactoprotein powder in the low sensitization food of preparation that said method obtains.
Low sensitization lactoprotein powder, preparation method thereof provided by the invention, preferably, comprises the following steps:
(1) concentrated milk albumen powder mixes its mass percent rear with the clean water meeting sanitary standard for drinking water is 6%-10%, 35-45 DEG C of aquation 60-120min, makes it fully dissolve, prepares milk protein solution; By milk protein solution in supersonic generator, be the ultrasonic wave process 15-30 minute of 20-25kH through frequency; It is the institutional framework changing lactoprotein by the object of ultrasonic wave process;
(2) according to lactoprotein grain weight amount in milk protein solution 0.5 ~ 2% percentage by weight ratio in the milk protein solution through ultrasonic wave process, add compound protease (flavor protease: alkali protease: the mass ratio of neutral proteinase composition is 2-4:2:1), 45 ~ 55 DEG C, pH carries out enzyme digestion reaction 30 ~ 120min under being 7.0 ~ 8.0 (regulating with NaOH or HCl of 1mol/L) condition; Then at 90 ~ 95 DEG C, be incubated 3 ~ 5 minutes, be cooled to room temperature, obtain lactoprotein enzymolysis liquid;
(3) by the lactoprotein enzymolysis liquid of step (2) gained by aperture be the ceramic membrane ultrafitration of 0.001 μm, proteins and peptides molecular weight being less than 10000 is separated, and gets the filtrate after ultrafiltration;
(4) get the protein liquid that molecular weight is less than 10000, by concentrated, spraying dry, obtain the total milk protein powder of low sensitization.
The advantage that the present invention has is as follows:
(1) low sensitization lactoprotein preparation method of the present invention is simple, more easily realizes industrialization and produces, and solution current reduction total milk protein sensitization realizes the problem that industrialization is produced effectively.
(2) the low sensitization lactoprotein of the present invention's exploitation is total milk protein, synchronously reduce the anaphylaxis of lactalbumin (beta lactoglobulin, ALA) and casein (alpha-casein, beta-casein), current low sensitization lactoprotein is mainly low sensitization lactalbumin, is not total milk protein.
(3) the present invention utilizes ultrasonic technology process lactoprotein, improves the reduction degree of lactoprotein sensitization.
(4) the present invention utilizes ultrasonic technology process milk protein solution, ultrasonic wave produces vibration and impact due to the effect of high radiation pressure and superb sound pressure to medium, makes protein denaturation, cell metaplasia, some chemical bond rupture, biomolecule cracking; Thus obviously can improve the structure of lactoprotein, improve the yield of low sensitization lactoprotein powder product.
(5) the present invention adopts food-grade compound protease (flavor protease, neutral proteinase and alkali protease), through in a mild condition, obtains specific total milk protein through appropriate enzymolysis, does not add any additive, edible safety.
(6) the low sensitization lactoprotein of the present invention's exploitation, has good local flavor, for the exploitation of lactoprotein allergic human population breast group food provides milk protein raw material.
In a word, the present invention is directed to the present situation of current low sensitization milk product exploitation, with lactoprotein powder for raw material, through a large amount of experiments, develop a kind of preparation method of simple efficient low sensitization lactoprotein powder; The antigenicity of the ALA of the lactoprotein powder obtained, beta lactoglobulin, alpha-casein and beta-casein all reduces more than 70%; In allergenicity, the allergenicity of ALA, beta lactoglobulin, alpha-casein reduces more than 50%, the allergenicity of beta-casein reduces more than 25%, show that method provided by the invention can prepare the product of low sensitization lactoprotein, more easily realize industrialization to produce, there is huge application prospect and market value.
Detailed description of the invention
Following examples for illustration of the present invention, but are not used for limiting the scope of the invention.
Room temperature in the present invention is 25 DEG C.Protein content unit " % " is mass percent.
In embodiments of the invention, concentrated milk albumen powder (MPCs, Fonterra, milky way road, Beijing Trade Co., Ltd. provides).Protein content determination adopts Kjeldahl's method (GB5009.5-2010).Described flavor protease (purchased from Novozymes Company of Denmark), alkali protease and neutral proteinase (purchased from the magnificent prosperous and powerful Bioisystech Co., Ltd in east, Beijing), ceramic membrane is purchased from Jiangsu Jiuwu High-Tech Co., Ltd..
In the present invention, water used is the clean water meeting sanitary standard for drinking water.
PH value in the present invention all regulates with NaOH or HCl of 1mol/L.
The unclassified stores used in the present invention is the conventional material that can commercially obtain.The equipment used also is the conventional equipment of this area.In the present invention, NM operation is the routine operation of this area.
The preparation method (1) of embodiment 1 low sensitization lactoprotein powder
(1) concentrated milk albumen powder 10 grams (protein is 67.2%, lactose is 17.8%, fat is 1.6%, moisture be 4.8%) is got, add water 115 grams, the mass percent making cow's milk protein powder account for mixed liquor is 8%, 40 DEG C of Water Under 120min, make its abundant stirring and dissolving, obtained milk protein solution.By milk protein solution in supersonic generator, process 30 minutes through ultrasonic wave (frequency 25kH);
(2) according to albumen powder weight in milk protein solution 1% percentage by weight in the milk protein solution of the ultrasonic wave process in step (1), add compound protease 0.1 gram (flavor protease: alkali protease: the mass ratio of neutral proteinase composition is 3:2:1), 50 DEG C, pH carries out enzymolysis 120min under being 7.5 conditions; Then at 95 DEG C, be incubated 3 minutes, be cooled to room temperature, obtain lactoprotein enzymolysis liquid;
(3) by the lactoprotein enzymolysis liquid of step (2) gained by aperture be the ceramic membrane ultrafitration of 0.001 μm, get filtered fluid, obtain the protein liquid that molecular weight is less than 10000;
(4) protein liquid that obtains of step (3), by concentrated, spraying dry, obtains the albumen powder 8.8 grams of low sensitization.The protein content of low allergic protein powder is 69.0%.
The preparation method (2) of embodiment 2 low sensitization lactoprotein powder
(1) concentrated milk albumen powder 50 grams (protein is 67.2%, lactose is 17.8%, fat is 1.6%, moisture be 4.8%) is got, add water 450 grams, the mass percent making cow's milk protein powder account for mixed liquor is 10%, 40 DEG C of Water Under 120min, make its abundant stirring and dissolving, obtained milk protein solution.By milk protein solution in supersonic generator, process 20 minutes through ultrasonic wave (frequency 20kH);
(2) according to albumen powder weight in milk protein solution 1.2% percentage by weight in the milk protein solution of the ultrasonic wave process in step (1), add compound protease 0.6 gram (flavor protease: alkali protease: the mass ratio of neutral proteinase composition is 3:2:1), 45 DEG C, pH carries out enzymolysis 80min under being 7.5 conditions; Then at 95 DEG C, be incubated 3 minutes, be cooled to room temperature, obtain lactoprotein enzymolysis liquid;
(3) by the lactoprotein enzymolysis liquid of step (2) gained by aperture be the ceramic membrane ultrafitration of 0.001 μm, get filtered fluid, obtain the protein liquid that molecular weight is less than 10000;
(4) protein liquid that obtains of step (3), by concentrated, spraying dry, obtains the albumen powder 43.8 grams of low sensitization.The protein content of low allergic protein powder is 68.9%.
The preparation method (3) of embodiment 3 low sensitization lactoprotein powder
(1) concentrated milk albumen powder 100 grams (protein is 67.2%, lactose is 17.8%, fat is 1.6%, moisture be 4.8%) is got, add water 900 grams, the mass percent making cow's milk protein powder account for mixed liquor is 10%, 35 DEG C of Water Under 90min, make its abundant stirring and dissolving, obtained milk protein solution.By milk protein solution in supersonic generator, process 30 minutes through ultrasonic wave (frequency 25kH);
(2) according to albumen powder weight in milk protein solution 1% percentage by weight in the milk protein solution of the ultrasonic wave process in step (1), add compound protease 1.0 grams (flavor protease: alkali protease: the mass ratio of neutral proteinase composition is 3:2:1), 55 DEG C, pH carries out enzymolysis 100min under being 7.0 conditions; Then at 95 DEG C, be incubated 3 minutes, be cooled to room temperature, obtain lactoprotein enzymolysis liquid;
(3) by the lactoprotein enzymolysis liquid of step (2) gained by aperture be the ceramic membrane ultrafitration of 0.001 μm, get filtered fluid, obtain the protein liquid that molecular weight is less than 10000;
(4) protein liquid that obtains of step (3), by concentrated, spraying dry, obtains the albumen powder 87.8 grams of low sensitization.The protein content of low allergic protein powder is 68.5%.
The low antigenicity of sensitization lactoprotein of embodiment 4 and the mensuration of allergenicity
The low sensitization lactoprotein powder prepared with embodiment 1-3 6% the aqueous solution for subjects, carry out the antigenicity of lactoprotein and the detection of allergenicity.Measurement result is in table 1.
1, the mensuration of sample antigen
Adopt the antigenicity of ALA (α-LA), beta lactoglobulin (β-LG), alpha-casein (α-CN) and beta-casein (β-CN) in indirect competitive ELISA method working sample, concrete steps are as follows:
(1) antigen coated: using α-LA, β-LG, α-CN or β-CN (being all purchased from Sigma company) as antigen, after being diluted to finite concentration with coating buffer, to be coated in 96 hole ELISA Plates, every hole 100 μ L, 4 DEG C of refrigerator overnight.
(2) antigen and antibody response: the antigenicity measuring α-LA, β-LG, α-CN and β-CN, sample is respectively 1000 through PBS extension rate, and 2000,1000,2000.The extension rate measuring rabbit anti-serum corresponding to α-LA, β-LG, α-CN and β-CN antigenicity is respectively 1:30000,1:60000,1:100000 and 1:40000.The two equal-volume is mixed, does not add the reaction tube of sample as uncontested reaction system.The reaction tube prepared is in 4 DEG C of refrigerator overnight.
(3) wash: liquid in 96 ELISA Plate holes, hole of inclining next day (after 14h), wash plate 4 times (first time leaves standstill 1min, rear 3 concussion 3min) with cleaning fluid, every hole 250 μ L, pats dry on clean blotting paper.
(4) close: add confining liquid and close, every hole 100 μ L, is placed in wet box, incubation 1h in 37 DEG C of incubators.Wash plate 4 times by step (3) cleaning fluid, pat dry.
(5) application of sample: add in ELISA Plate by liquid in reaction tube in step (2), every hole 100 μ L, is placed in wet box, incubation 1h in 37 DEG C of incubators.Wash plate 4 times by step (3) cleaning fluid, pat dry.
(6) ELIAS secondary antibody is added: add in 96 hole ELISA Plates with after confining liquid dilution HRP-goat anti-rabbit igg to 10000 times, every hole 100 μ L, is placed in wet box, incubation 1h in 37 DEG C of incubators.Wash plate 4 times by step (3) cleaning fluid, pat dry.
(7) develop the color: preparation tmb substrate solution, every hole 100 μ L, is placed in wet box, incubation 10min in 37 DEG C of incubators.
(8) cessation reaction: add 2mol/L H immediately
2sO
4, every hole 50 μ L.
Measure light absorption value: the light absorption value OD (dual wavelength: 450nm and 630nm) measuring each hole with ELIASA.The actual OD=OD in each hole
450-OD
630.
(9) data processing: the antigenicity size that measured object remains can be evaluated with its antigen inhibiting rate, is calculated as follows: antigen inhibiting rate=(1-B/B
0) × l00%
In formula: wherein B represents the light absorption value of each detected sample, B
0for the light absorption value of uncontested system.
The size of the ability that in inhibiting rate characterizing sample, α-LA (β-LG, α-CN or β-CN) suppresses antiserum and ELISA Plate wrap the α-LA of quilt, (β-LG, α-CN or β-CN) combines, is designated as α-LA inhibiting rate (β-LG inhibiting rate, α-CN inhibiting rate or β-CN inhibiting rate).The allergenicity of inhibiting rate and sample is proportional, and inhibiting rate is higher, and allergenicity is higher.Measurement result is in table 1.
2, the mensuration of sample allergenicity
Using α-LA, β-LG, α-CN or β-CN as anaphylactogen, indirect competitive ELISA method is adopted to measure the allergenicity of lactoprotein, method and step are as (1) in the mensuration of sample antigen-(8), measure α-LA, β-LG, α-CN with β-CN allergenicity process, sample is respectively 1000 through PBS extension rate, 2000,1000,2000.Wherein antibody is newborn allergic patients sera, and extension rate is 1:5, and ELIAS secondary antibody is HRP-sheep anti human IgE, and extension rate is 500 times.
Data processing: the allergenicity size that sample remains represents with anaphylactogen inhibiting rate, and computing formula is as follows: anaphylactogen inhibiting rate (%)=(1-B/B0) × 100%
Wherein B represents the light absorption value of each detected sample, and B0 is the light absorption value of uncontested system.
The size of the ability that in inhibiting rate characterizing sample, α-LA (β-LG, α-CN or β-CN) suppresses antiserum IgE and ELISA Plate wrap the α-LA of quilt, (β-LG, α-CN or β-CN) combines, is designated as α-LA inhibiting rate (β-LG inhibiting rate, α-CN inhibiting rate or β-CN inhibiting rate).The allergenicity of inhibiting rate and sample is proportional, and inhibiting rate is higher, and allergenicity is higher.Measurement result is in table 1.
Table 1 low sensitization lactoprotein antigenicity inhibiting rate and allergenicity inhibiting rate measuring result
| Testing index | Concentrated milk albumen powder | Embodiment 1 | Embodiment 2 | Embodiment 3 |
| α-LA antigenicity inhibiting rate (%) | 98.6％ | 25.7％ | 29.5％ | 29.0％ |
| β-LG antigenicity inhibiting rate (%) | 94.7％ | 22.3％ | 24.4％ | 22.1％ |
| α-CN antigenicity inhibiting rate (%) | 91.2％ | 14.8％ | 14.2％ | 16.4％ |
| β-CN antigenicity inhibiting rate (%) | 82.6％ | 18.9％ | 17.7％ | 20.3％ |
| α-LA allergenicity inhibiting rate (%) | 90.23％ | 42.56％ | 40.86％ | 38.78％ |
| β-LG allergenicity inhibiting rate (%) | 89.16％ | 30.64％ | 31.43％ | 29.76％ |
| α-CN allergenicity inhibiting rate (%) | 86.38％ | 40.67％ | 41.88％ | 42.89％ |
| β-CN allergenicity inhibiting rate (%) | 62.65％ | 40.68％ | 39.86％ | 41.44％ |
As can be seen from Table 1, embodiment of the present invention 1-3 obtain 6% the low sensitization concentrated milk albumen powder aqueous solution, in antigenicity, the antigenicity of ALA, beta lactoglobulin, alpha-casein and beta-casein all reduces more than 70%; In allergenicity, the allergenicity of ALA, beta lactoglobulin, alpha-casein reduces more than 50%, and the allergenicity of beta-casein reduces more than 25%.This result shows that the lactoprotein powder developed can be used for developing the product of low sensitization lactoprotein.
The mensuration of the bitterness value of the low sensitization lactoprotein of embodiment 5
Subjective appreciation group is made up of (being non-smoker) 10 people through screening, and after assessment officer gargles with distilled water, get 10% enzymolysis liquid about 2.0mL and be placed in mouth, 5-10 spues after second.Then according to the scoring benchmark of table 2 to 6% of the low sensitization lactoprotein powder that embodiment 1-3 obtains the aqueous solution mark, score value is exactly bitterness value, and the mean value drawn represents the bitterness of milk peptide.The concentrated milk albumen powder solution of 6% in contrast.
Table 2 hardship hides the standards of grading of value
Table 3 hardship hides the appraisal result of value
Carry out bitter taste estimate to total milk protein powder and low sensitization lactoprotein powder, result is as table 3.The low sensitization lactoprotein powder bitterness value of the present invention's exploitation is very low as can be seen from Table 3, and consumer is complete acceptable.
Although above the present invention is described in detail with a general description of the specific embodiments, on basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, all belong to the scope of protection of present invention.
Claims (10)
1. a preparation method for low sensitization lactoprotein powder, comprises the following steps:
(1) lactoprotein powder is mixed with water, fully dissolve and obtain milk protein solution, ultrasonic wave process milk protein solution;
(2) add compound protease and carry out enzyme digestion reaction, after enzyme digestion reaction terminates, insulation, then be cooled to room temperature, obtain lactoprotein enzymolysis liquid;
(3) lactoprotein enzymolysis liquid is passed through ceramic membrane ultrafitration, collect filtered fluid;
(4) filtered fluid is concentrated, dry, obtain the lactoprotein powder of low sensitization.
2. preparation method according to claim 1, is characterized in that, in step (1), the protein content of lactoprotein powder is 65%-70%, lactose is 16%-20%, fat is 1%-2%, moisture is 3%-5%, and described % is mass percent.
3. preparation method according to claim 1, is characterized in that, in step (1), mass percent shared after lactoprotein powder mixes with water is 6%-10%; 35-45 DEG C of aquation 60-120min, makes lactoprotein powder fully dissolve, prepares milk protein solution.
4. preparation method according to claim 1, is characterized in that, in step (1), the time of ultrasonic wave process is 15 ~ 30 minutes; Ultrasonic frequency is 20 ~ 25KHz.
5. preparation method according to claim 1, is characterized in that, in step (2), the quality of compound protease is the 0.5%-2% of lactoprotein opaque amount; Described compound protease comprises flavor protease, alkali protease and neutral proteinase, and the mass ratio of three is 2-4:2:1.
6. the preparation method according to claim 1-5 any one, is characterized in that, in step (2), the condition of enzyme digestion reaction comprises: temperature is 45-55 DEG C, and pH value is 7.0 ~ 8.0, and the reaction time is 30-120min.
7. the preparation method according to Claims 1 to 5 any one, is characterized in that, in step (2), is incubated 3 ~ 5 minutes, then is cooled to room temperature after enzyme digestion reaction terminates at 90 ~ 95 DEG C.
8. the preparation method according to Claims 1 to 5 any one, is characterized in that, in step (3), ceramic membrane aperture is 0.001 μm.
9. the lactoprotein powder for preparing of method described in claim 1-8 any one, its protein molecular weight < 10000 dalton.
10. method described in claim 1-8 any one or the application of lactoprotein powder according to claim 9 in the low sensitization food of preparation.
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