CN100438773C - Method for reducing acrylamide in foods, foods having reduced levels of acrylamide - Google Patents

Abstract

A method for the reduction of acrylamide in food products, food products having reduced levels of acrylamide, and an article of commerce. In one aspect, the method comprises reducing the level of asparagine in a food material before heating. In one embodiment, the method comprises extracting at least a portion of the asparagine from the food material before heating. In yet another aspect, an article of commerce communicates to the consumer that a food product has reduced or low of acrylamide or asparagine.

Description

Reduce the method for acrylamide in the food, contain the food of reduced levels of acrylamide

Invention field

The present invention relates to reduce acrylamide content in the food, and relate to the food that contains reduced levels of acrylamide.The invention further relates to a kind of commodity.

Background of invention

Since civilization occurring, the food of carbohydrate containing has become the major part in people's diet.Nowadays, the food of carbohydrate containing (but as bread, breakfast cereal, dessert, biscuit, cookies, chips, ripe starchy vegetables, tower cake cake skin and snack) is eaten at large.Though available for countless years comes above-mentioned food to become the part of people's diet, the researcher just finds recently, comprises acrylamide in many these foods.

In April, 2002, the researcher of Sweden state food management board and Stockholm University has announced their discovery: produced acrylamide in the food that many kinds are cooked, a kind of compound that may be carcinogenic.Acrylamide in mouse body, have with food in the similar carciongenic potency of other carcinogenic substance, but for the mankind, its relative carciongenic potency in food is still unknown.For acrylamide, have only limited demographic data, and these data can not prove that occupational contact acrylamide has carcinogenic danger.(FAO/WHO is about the evaluation of acrylamide health problem in the food: summary report; Geneva, Switzerland, on June 25th to 27,2002)

Could determine people can produce what kind of health effect (if existence) with the edible acrylamide of content that is present in usually in the above-mentioned food, and many consumers pay close attention to this though need further research.Therefore, an object of the present invention is to provide the method that reduces acrylamide content in the food.The present invention also aims to provide the food that contains reduced levels of acrylamide.Say that further the present invention aims to provide a kind of commodity, these commodity inform that consumer's food contains decrement or a spot of acrylamide.

Summary of the invention

In one aspect, the invention provides a kind of method that reduces acrylamide content in the food.In one embodiment, reduce the asparagine concentration in the food material before this method is included in and heats.

In yet another aspect, the invention provides the method that reduces asparagine concentration in the food material.In one embodiment, this method is extracted into the small part asparagine before being included in heating from food material.

In yet another aspect, the invention provides food with reduced levels of acrylamide.

In yet another aspect, the invention provides food material with decrement asparagine.

In yet another aspect, the present invention also provides a kind of commodity, and these goods are informed the consumer, and food contains decrement or a spot of acrylamide or asparagine.

The relevant part of all references is incorporated herein by reference; Quoting of any document may not be interpreted as its approval as prior art of the present invention.

The accompanying drawing summary

Fig. 1. Fig. 1 has listed the reaction mechanism that is proposed, and according to this mechanism, acrylamide is formed at asparagine and carbonyl source (as glucose).R ₁And R ₂Can be H, CH ₃, CH ₂OH, CH ₂(CH ₂) _nCH ₃, or any other formed the component of reduced sugar; N can be any less than 10 integer.

Fig. 2. Fig. 2 has listed the reaction mechanism that is proposed, and according to this mechanism, asparaginase and asparagine react to stop the formation of acrylamide.

Fig. 3. Fig. 3 has listed the sample chromatogram of asparagine and aspartic acid LC analysis.The x axle is represented retention time, and the y axle is represented response.

Detailed Description Of The Invention

The applicant finds that when being heated, asparagine (being present in the naturally occurring amino acid in nearly all living things system) can produce acrylamide. Thereby when being heated, containing the more food of asparagine and be tending towards comprising the more acrylamide of high-load; When heating in the situation that has reduced sugar contains asparagine food, especially true. It has also been found that when food was cooked to lower final water content, the acrylamide of generation was more.

Without being limited by theory, it is believed that by reaction mechanism listed among Fig. 1 and can in food, produce acrylamide. The alpha-amido and the reaction of carbonyl source that it is believed that free asparagine form schiff bases. Under the heating condition, this schiff bases adduct decarboxylize forms product, this product can: (1) hydrolysis forms Beta-alanine acid amides (this compound can further be degraded and be formed acrylamide under the heating condition), or (2) are decomposed to form acrylamide and corresponding imines. (applicant finds that the atom of cyclic precursor comprises carbon atom and the nitrogen-atoms in the acrylamide. )

Therefore, the applicant further finds, removed the formation that at least part of asparagine can reduce the acrylamide in heat food before cooking. When heating contained the above-mentioned food of decrement asparagine, the amount of acrylamide of formation had also reduced. The method for optimizing of removing asparagine with solvent extraction. Preferred solvent is water.

A. reduce the method for Acrylamide in Foods

In one aspect, the invention provides the method that reduces Acrylamide in Foods, described method be included in final heating (as, cook) reduce before the asparagine concentration in the food material. In one embodiment, the method is extracted into the small part asparagine before being included in final heating from food material. In preferred embodiments, the method for reduction Acrylamide in Foods content comprises:

(1) provide food material, wherein said food material comprises asparagine;

(2) not necessarily reduce the particle diameter of food material;

(3) not necessarily increase the cell membrane permeability of food material;

(4) from food material, be removed to the small part asparagine; With

(5) the heated food material forms ultimate food.

In yet another aspect, the invention provides the method that reduces asparagine in the food material.In one embodiment, this method comprises be extracted into the small part asparagine from food material.In one embodiment, extraction is included in blanching food material in the solvent.Preferred solvent is a water.In preferred embodiments, the method for asparagine concentration comprises in the reduction food:

(1) provide food material, wherein said food material comprises asparagine;

(2) not necessarily reduce the particle diameter of food material;

(3) not necessarily increase the cell membrane permeability of food material; With

(4) from food material, extract asparagine.

1. food material is provided, and wherein said food material comprises asparagine

Term used herein " food material " comprises that used any kind contains the asparagine Edible material in the food making, comprises the mixture of two or more foods.

2. not necessarily reduce the particle diameter of food material

Nonessential but preferably, can reduce the particle diameter of food material, especially comprise when extracting when the removal of asparagus fern acid amides.Leaching process it is believed that it is a kind of DIFFUSION CONTROLLED phenomenon, and therefore shortening diffusion length can improve extraction efficiency.The particle diameter that reduces food material has reduced diffusion length, thereby has improved extraction efficiency.The reducing and can be finished by any suitable method of particle diameter, these methods comprise cutting, hack, macerate, pulverize, grind, tear up, push, smash to pieces, or the combination of these methods.

3. not necessarily increase the cell membrane permeability of food material

It is extremely important that important component is remained in the living cells pair cell viability.Many cells use the concentration that initiatively sends important component in the cell to remain on the higher level of concentration that is allowed than infiltration.Because this principle, it is difficult extracting some component from cell.Without being limited by theory, it is believed that asparagine is in the eucaryotic cell structure of food material; This makes asparagine be difficult for being applicable to extraction.The applicant has found to improve the extraction efficiency that permeability can improve asparagine greatly by changing membrane structure.

Available any suitable method changes the cell membrane of food material to improve the extraction of asparagine, described method comprises, but be not limited to, heating (as, convection current, radiation, microwave, infrared), osmotic pressure changes, changes the pH of cellular environment, with one or more enzymes processing (for example, cellulose degrading enzyme such as cellulase, hemicellulase, pectase or their mixture), freeze thawing circulation, other cell membrane breaking method (as, ultrasonication) or their combination.

In a preferred embodiment, use blanching to change cell membrane.In the blanching process, can influence the permeability of cell in various manners.For example, can enlarge cell content (as because starch gelatification), cause membranolysis.In addition, heating can make the cell membrane protein sex change, causes cell to leak.This can cause the extraction efficiency of asparagine to improve.

4. from food material, be removed to the small part asparagine

From food material, be removed to the small part asparagine.Preferably, the content of asparagine has reduced at least about 10% in the food material, preferably at least about 30%, and more preferably at least about 50%, also more preferably at least about 70%, even more preferably at least about 90%.

Can use the appropriate method of any removal asparagine.The method for optimizing of removing asparagine comprises extraction.Extraction used herein comprises and any food material being contacted with solvent to remove the method for part asparagine.The solvent of any solubilized asparagine can be used for extracting (for example, the food grade solvent of any solubilized asparagine, acid or alkali), yet preferred solvent is a water.Water is that the ideal solvent that asparagine extracts has several reasons: (1) asparagine very easily is dissolved in water; (2) water is dirt cheap; (3) water is considered to safe usually.

Can use any suitable extracting method.For example, extraction can comprise as methods such as immersion, lixiviate, washing, rinsing, main dipping baths, or the combination of these methods.

Preferably, leaching process solvent be liquid and food material physical attribute (as, to bigger those of ultimate food influence) can not carried out under the temperature of adverse effect.When water was used as solvent, lowest temperature was typically about 0 ℃; If when reducing freezing point, lower limit can be lower than 0 ℃ yet use salt or cosolvent (or some obtains the method that freezing point reduces).Temperature upper limit is typically the denaturation temperature that is lower than protein, for example, is lower than about 170 °F (77 ℃).In one embodiment, extracting temperature is about 5 ℃ to about 70 ℃, is preferably about 10 ℃ to about 60 ℃.

In the leaching process, not only can remove asparagine, but also can remove various other solvent solubilized (as, water-soluble) component.Because the soluble component of many these solvents can comprise flavor compounds, may negative effect be arranged to the local flavor of ultimate food so extract.The applicant finds, uses main dipping bath extracting method to minimize and remove other solvent solubilized component from food material, comprises flavor compounds.

Main dipping bath extracting method

Can use main dipping bath to come from food material, optionally to extract one or more components, and other component concentrations is had no adverse effect.Extract several food materials in succession with solvent-laden dipping bath liquid and set up a main dipping bath liquid with foundation or near one or more balances of extracting component in the food material with solvent extraction.These components can use any suitable method optionally to separate from solvent or remove.Remaining solubilized component and food material are set up or near balance.Successively, handle another batch or several food materials with the dipping bath liquid of determining.This causes the described component of selective removal, and the concentration of other combination is had no adverse effect.After every batch of food material is handled, can add additional solvent to keep the body lotion constant volume.

After as long as main dipping bath liquid is set up, several continuous food materials can with in batches, half in batches or continuation mode (as, adverse current continuation method, wherein food along a direction pump into and solvent pump in opposite direction) extract.

In the present invention, can use main dipping bath to come from food material, optionally to remove asparagine.Can implement main dipping bath method in various manners and come from solvent, to remove asparagine.For example, these modes can comprise asparagine conversion enzyme is added in the solvent of becoming owner of dipping bath, solvent is aspirated by being fixed with the post of asparagine conversion enzyme, or with solvent suction by comprise to asparagine have selectivity (as, comprise asparagine is had specific acceptor site) the adsorbent post.

In one embodiment, a kind of asparagine conversion enzyme is joined in the main dipping bath liquid optionally to remove asparagine." asparagine conversion enzyme " used herein or " enzyme " comprise any enzyme that can change the asparagine chemical constitution.For example, the deamidase with asparagine mapping function is included in these terms.Term used herein " asparagine conversion enzyme " and " enzyme " comprise one or more enzymes; For example, this term comprises the mixture of two or more enzymes.

In a preferred embodiment, asparagine conversion enzyme be can the hydrolysis amide group of free asparagine enzyme.The preferred enzyme that is used for this paper is an asparaginase.The preferred source of asparaginase is Sigma-Aldrich, and catalog number (Cat.No.) is #A2925.Without being limited by theory, it is believed that adding this kind of enzyme can make the side chain of asparagine degrade; When doing like this, amido link is hydrolyzed, and asparagine is transformed into aspartic acid.This reaction mechanism is listed among Fig. 2.

When the enzyme in the main dipping bath was transformed into aspartic acid with asparagine, this additional asparagine to several food materials of adding afterwards extracted and has produced driving force.Extractable matter and food material balance make other solubilized food material component except that asparagine not be extracted out, and asparagine continues reaction and by enzymatic conversion.The aspartic acid that is formed at asparagine soaks to be got back in the food material and reaches balance.Every batch of food material is handled the back and is added the solvent that replenishes and/or contain enzyme solutions, to compensate the solution of being taken away by the last consignment of food material; This makes winner's dipping bath keep constant volume.

Enzyme is sold according to active unit, rather than according to weight or volume.Therefore, the effective dose of the required enzyme of ultimate food acrylamide reduction of acquisition expectation will depend on the activity of used concrete enzyme product.

The amount that adds enzyme depends on the reduction of asparagine and the reduction of desired thus acrylamide.The amount that adds enzyme also depends on the asparagine amount that contains in the food material; The food material that contains more asparagine usually needs to increase the amount of enzyme or increases the reaction time, to reach the reduction of identical acrylamide.The amount that adds enzyme also depends on used concrete enzyme (for example, the ability of concrete enzyme degraded asparagine) and handled concrete food material.Those skilled in the art can be according to concrete food material, concrete enzyme, the concrete activity of enzyme and the result of expectation, the effective dose of decision enzyme.

When enzyme has reacted to required degree, not necessarily make its passivation or from food material, remove.When using enzyme safe to eat (as natural existence and be present in the common food), can select not with enzymatic inactivation or remove.Alternatively, can use any suitable inactive enzyme method to make enzyme deactivation.For example, regulate, use Protease Treatment by utilizing heat, pH, or their combination, can make enzyme deactivation.And, can use any suitable method that enzyme is removed from food material, this method includes but not limited to extract.The combination that enzyme can be passivated, remove or carry out passivation and remove.

In another embodiment, directly asparagine conversion enzyme is not joined in the main dipping bath liquid, but with the extract in dipping bath liquid suction bed or the post (described enzyme is adsorbable or be chemically bonded on the wall of substrate (being preferably inert substrate) or hollow membrane pipe) by being fixed with asparagine conversion enzyme as plastic sheet in the post or globule.The major advantage of this method is the not direct contact food material of resolvase, otherwise food material can be taken away the part enzyme from main dipping bath liquid, thereby needs to replenish enzyme.With the fixing expense that can reduce or eliminate the enzyme that replenishes costliness of enzyme.This embodiment makes other solvent solubilized component remove minimum advantage in addition from food material.

In another embodiment, be sent to from the solvent liquid stream in the main dipping bath liquid of determining and comprise hollow-fibre membrane (as United States Patent (USP) 5,869, those disclosed in 297), dialysis material or size exclusion material (as, zeolite) in the post, this size exclusion material allows asparagine molecule or other to equate or the molecule of smaller szie diffuses out from liquid stream.Final effect is a selective removal asparagine from food material, reaches required acrylamide reduction, and can reduce the local flavor of ultimate food sharply.

In another embodiment, the liquid stream of extract is sent in the post of the adsorbent that comprises selectivity absorption asparagine.Suitable adsorbent can include, but not limited to molecular sieve, zeolite, cyclodextrin, clay, diatomite, silica (as, such as

Magnesium silicate type), ion exchange resin (anion or cation or hybrid resin, as

) or their combination.Final effect is a selective removal asparagine from food material, reaches required acrylamide reduction, and can reduce the local flavor of ultimate food sharply.

In another embodiment, the liquid stream of extract is sent in the post that comprises asparagine specificity absorbent, and this absorbent comprises asparagine is had specific acceptor site.The final effect of this method is a selective removal asparagine from food material, reaches required acrylamide reduction, and can reduce the local flavor of ultimate food sharply.

Although each above-mentioned embodiment all uses the method for post to describe, but be to be understood that, these embodiments also can realize with any other suitable manner, as fluid bed, plant-scale continuous liquid chromatography (for example, such as being published in the sort of method described in the United States Patent (USP) 4,210,594 of authorizing people such as Logan on July 1st, 1980), or, wherein absorbent is added in the liquid stream and then absorbent is separated from liquid stream with batch mode.

In addition, although above-mentioned embodiment has been described with main dipping bath and has been handled the batch methods that food material is removed asparagine, but be to be understood that, food material and solvent can with half in batches or continuation mode (as the adverse current continuation method, wherein food material along a direction pump into and solvent pump in opposite direction) contact.In addition, food material can also be the bed in the post, and this post is passed through in the solvent pumping.

5. the heated food material forms final food

Heated food material in due form then, as baking, fried, extruding, oven dry (as, by vacuum drying oven or rotary-drum drier), expanded or microwave.In the embodiment of enzyme contact food material, may make enzymatic inactivation in the heating process, therefore nonessential passivation step and heating (as, cook) step can carry out simultaneously.Can make enzyme denaturation and passivation via cooking the heat treated of carrying out, make food material no longer have lasting enzymatic activity.And in heating steps, give at least a portion time, enzyme is reacted.

" ultimate food " used herein includes, but not limited to edible food and is used as the food of the composition of making other food.

Preferably, the content of acrylamide has reduced at least about 10% in the ultimate food, preferably at least about 30%, more preferably at least about 50%, also more preferably at least about 70% and even more preferably at least about 90%.

B. implement the mode of described method

Can implement the present invention in any suitable manner.For example, can be in batches, semi-batch or continuous mode implement method of the present invention.

C. the food that contains reduced levels of acrylamide

The method of this paper can be applied to producing any suitable food, carbohydrate containing food, especially the low moisture food of heating when including, but not limited to make (as, be less than about 10% moisture).For example, can use this method to reduce the acrylamide content that is present in mashed potato, potato block, fabricated snack, chips, breakfast cereal, bread, cookies, biscuit, cake, pizza skin, salty crisp volume cake, Hash Browns, potato pompon, tortilla and the Ta Ke cake cake skin.

In one embodiment, the acrylamide content of fried processing potato chips preferably less than about 300ppb, is more preferably less than about 200ppb less than about 400ppb, also is more preferably less than about 50ppb, even is more preferably less than about 10ppb.

In another embodiment, the acrylamide content of saratoga chip preferably less than about 30ppb, is more preferably less than about 20ppb less than about 40ppb, even is more preferably less than about 10ppb, most preferably less than about 5ppb.

In another embodiment, less than about 40ppb, preferably less than about 30ppb, be more preferably less than about 20ppb, by the acrylamide content of the chips made of cutting potato most preferably less than about 10ppb.

In another embodiment, the acrylamide content of corn snack preferably less than about 50ppb, is more preferably less than about 10ppb less than about 75ppb.

Although the method for this paper will be described with preferred dehydrated potato products, processing chips, potato block, chips and corn snack usually, it will be understood by those of skill in the art that this paper method can be applicable to any suitable product.Non-limiting example comprises biscuit, bread is (as rye, wheat, oat, potato, the full cereal products of white, mixed flour, big wheat cake, twisted shape bread, bun, web-like bread, than tower cake, matzo, Italy's vanilla bread, roasting crisp rusk, zwieback, crouton, soft muffin, soft pilot bread rod, heating instant type bread) the baked cake of, cookies, Danish pastry, croissant, tart, send skin, piecrust, muffin, chocolate cake, the rectangle cake, doughnut, dessert is (as pretzel, tortilla chips, cornflakes, potato chips, fabricated snack, the processing potato chips, extrude dessert, extrude filling dessert, carry-on solid food, granola, assorted dessert, the fried potato silk), flour, corn flour, compound is (as the cake compound, the biscuit compound, the brownie compound, the bread compound, the pancake mixes material, the pancake compound, cream batter compound, the pizza dough), frozen dough is (as biscuit, bread, bread stick, croissant, the meal bag, the pizza dough, cookies, Danish pastry, chocolate cake, send skin), frozen food is (as sending skin, pie, tart, the volume cake, pizza, the skin of food, cake, French fries, Hash Browns, be coated with food such as the chicken and the fish of cooking after the crumbs, be coated with the vegetables of cooking after the crumbs), bagel, cereal breakfast, dessert, French fries, vegetables are (as drying, bake, baking, fry in shallow oil, explode, vacuum drying), but tower cake cake skin, Hash Browns, mashed potato, zwieback, toastie, flour and tortilla, pancake, pancake, waffle, the cream batter, the pizza skin, rice, based on the food of nut (as peanut butter, contain the food that shreds nut), fruit is (as drying, bake, baking, fry in shallow oil, explode, vacuum drying, cure, fruit jelly, pie is sandwich, burnt, raisins, mossberry, cherry), fried frying in shallow oil really, alcoholic beverage (as beer and malt liquor), contain fry cocoa bean food (as dog feed, cat feed, the ferret feed, the cavy feed, the gerbil jird feed, the hamster feed, the bird feed, the camel feed, feed for ostrich, the emu feed, the ox feed, the deer feed, the elk feed, the buffalo feed, the rabbit feed, the mouse feed, the mouse feed, the chicken feed, the turkey feed, pig feed, the horse feed, feed for goat, the sheep feed, the monkey feed, fish meal).

1. the dehydrated potato products that contains decrement asparagine and acrylamide

Can use the present invention to make the dehydrated potato products that contains reduced levels of acrylamide by the asparagine concentration that reduces in the food material.To set forth the method for optimizing of making above-mentioned dehydrated potato products below, but the invention is not restricted to this specific embodiment.Though this embodiment that elaborates had below been described before the potato of boiling oven dry and extracted asparagine, should be appreciated that the extraction of enzyme asparagine can be finished in any any appropriate steps that is suitable for making the dehydrated potato products method.For example, can cook preceding, cook the back, pulverize before, pulverize after, or make the extraction of carrying out asparagine in preceding any other the suitable procedure of processing of final dewatering potato products.This paper method also can be implemented together in conjunction with any method that is suitable for making dehydrated potato products known in the art, as be set forth in " Potato Processing ", the 4th edition, Talburt and Smith edit, AVI Books, Van NostrandReinhold Co., New York, 1987, [hereinafter being " PotatoProcessing "]) those of the 535th page to 646 pages.

In preferred embodiments, can make dehydrated potato products, as potato block, latke or potato grain according to following method.Usually, this method comprises:

(1) provides potato;

(2) not necessarily reduce the particle diameter of potato;

(3) not necessarily increase the cell membrane permeability of potato;

(4) boiling potato;

(5) from potato, extract asparagine;

(6) make wet mashed potato with potato; With

(7) the dry dehydrated potato products that forms of mashed potato that will wet.

Be listed in step (5) in the above-described embodiment though should be appreciated that extraction step, extraction step can be finished in any other appropriate steps of this method.In another embodiment, the method for making dehydrated potato products comprises:

(1) provides potato;

(2) not necessarily reduce the particle diameter of potato;

(3) not necessarily increase the cell membrane permeability of potato;

(4) boiling potato;

(5) make wet mashed potato with potato;

(6) the dry dehydrated potato products that forms of mashed potato that will wet; With

(7) from potato, extract asparagine, before wherein said any one that is extracted in the above-mentioned steps 1 to 6, among or finish afterwards.

Should be appreciated that above-mentioned steps 1 to 6 can be undertaken by the order of any appropriate.

Can use any suitable potato (as be used to make conventional potato block, cake or grain those) to make the dehydrated potato products of this paper.Preferably, can adopt, for example, but be not limited to, the potato of kinds such as Norchip, Norgold, Russet Burbank, Lady Rosetta, Norkotah, Sebago, Bintje, Aurora, Saturna, Kinnebec, IdahoRusset, Altura, Russet Norkotah, Atlantic, Shepody, Asterix and Mentor is made dehydrated potato products.

Contain less than about 5% reduced sugar (is with the dehydrated potato benchmark calculate),, be more preferably less than about 2% potato for preferred preferably less than about 3%.For example, the potato that contains low content reduced sugar (promptly less than 1.5%) is especially preferred for the dehydrated potato products that making is used to make the fried potato snack.

Make it softening through boiling potato so that smash into mud.Potato can be peeled, peel in the part or do not peel.Potato can be complete before the boiling, maybe can be cut into the thin slice of any size.Boiling operation can be that potato is softening with any heating of it being smash into mud or the cooking methods of other type.For example, can come boiling by potato is immersed in water or the steam.

For example, typically, average thickness be about 0.95cm (3/8 inch) to the potato block usable temp of about 1.27cm (1/2 inch) be about 200 (93 ℃) extremely vapour cooking about 12 minutes to about 45 minutes of about 250 (121 ℃), more particularly about 14 minutes to about 18 minutes.Typically, being cut into thread potato block usable temp and being the vapour cooking 7 minutes to about 18 minutes of about 200 (93 ℃) to about 250 (121 ℃), more particularly is about 9 minutes to about 12 minutes, to reach required hardness.

Asparagine available water in the potato of boiling (as, the potato block that boils) is extracted.The size of the potato block that not necessarily, boils can reduce so that extract processing with above-mentioned the whole bag of tricks.The potato of boiling can reach predetermined a period of time and extracts by being immersed in the water.Extraction time can from a few minutes by several hours, this depends on required asparagine decrease.Typical extraction time is about 30 minutes to about 4 hours.After the extraction stage, Ma Lingzhu is separated from extract solution.This available any suitable method (as, filtration, centrifugal or decantation) finish.

After extract handling, the potato after the extraction is pulverized and can finish by any suitable method, and these methods are such as but not limited to pulverizing, smash to pieces, shred or their combination, to form wet mashed potato.

Nonessential composition can be added and sneaks in the wet mashed potato.Above-mentioned nonessential composition can comprise starch.Starch can include, but not limited to any suitable native starch or modified starch, comprises any dried potato goods that join or be added back in the mashed potato.Also not necessarily in wet mashed potato, add emulsifying agent as processing aid.

After forming mashed potato, can as described belowly further be dried and be processed to form dehydrated potato products.These dehydrated potato products can be Any shape, for example, but are not limited to flakelet, pie, particle, agglomerate, thin slice, fragment, small pieces, fine powder or particulate.Alternatively, wet mashed potato can be used for making these products: for example, but be not limited to mashed potato, potato patti, potato flapjack and potato snack (as pressing chips, potato slices and potato chips).For example, can use wet mashed potato to make the pressure chips, as described in the United States Patent (USP) of announcing on April 9th, 1,963 3,085,020 of authorizing people such as Backinger those.

Any appropriate method of utilizing mashed potato to make above-mentioned dehydrated potato products (those methods for example known in the art) can be adopted and any suitable device can be used.For example, can the mashed potato drying be made sheet according to known method, those methods described in for example following patent of this known method: the United States Patent (USP) of announcing on May 23rd, 2,000 6 of authorizing people such as Villagran, 066, the United States Patent (USP) of announcing on August 19th, 353 and 1,956 2,759,832 of authorizing people such as Cording and the United States Patent (USP) of announcing February 5 nineteen fifty-seven 2 of authorizing people such as Willard, 780,552.Can the mashed potato drying be made pie according to the method for being set forth in the United States Patent (USP) of announcing September 11 calendar year 2001 6,287,622 of authorizing people such as Villagran.According to the United States Patent (USP) of announcing on November 4th, 1,975 3 of authorizing people such as Purves, 917, method described in 866, or with other known method (as the United States Patent (USP) of announcing on December 6th, 1,949 2 of authorizing people such as Greene, 490, described in 431 those), can be made into granular by processing described mashed potato.Suitable drier can be selected from those drying devices of knowing, and includes but not limited to, fluidized bed dryer, scrapes wall type heat exchanger, rotary-drum drier, freeze-dryer and gas lift drier etc.

Preferred drying means comprises that those reduce the method for total amount of heat input.For example, when making the dehydrated potato sheet, the mobile drying of freeze drying, drum-type drying, resonance or pulsed, infra-red drying or their combination are preferred; When making dehydration Ma Lingzhu grain, gas lift drying, fluidized bed drying or their combination are preferred.

Though the dehydrated potato products of this paper is mainly described with potato block, it will be apparent to one skilled in the art that mashed potato of the present invention can dehydratedly make any required dehydrated potato products that is derived from mashed potato.

Drum-type drying (for example using the rotary-drum drier that is generally used for potato products industry) is with the dry method for optimizing that forms thin slice of mashed potato.Preferable methods is utilized single drum dryer, and the mashed potato that wherein will wet spreads upon on the cylinder with laminar, and the thickness of thin slice is about 0.005 " to about 0.1 ", preferred about 0.005 " to about 0.05 ", more preferably from about 0.01 ".Typically, when using rotary-drum drier, mashed potato is added on the upper surface of cylinder by the transmission means.The roll that minor diameter does not heat little by little is coated in fresh mashed potato on the part on the cylinder, forms sheet or layer with predetermined thickness thus.The peripheral speed of little roll is identical with the peripheral speed of cylinder.After the potato mud layer rotates along a part of circumference of cylinder, thereby scraping blade peels off the thin slice of drying and takes off dry thin slice from cylinder.In typical case, be pressurized to about 483kPa (70 pounds/square inch) and rotary-drum drier self be heated to about 250 (121 ℃) extremely about 375 (191 ℃) to about 965kPa (140 pounds/square inch) by being included in steam in the cylinder, preferred about 310 °F (154 ℃) to about 350 °F (177 ℃), more preferably from about 320 °F (160 ℃) are the temperature of about 333 (167 ℃) extremely.Be obtaining optimum, suitably control the rotating speed and the internal temperature thereof of dryer drum, is about 5% to about 14% so that obtain moisture, preferred about 5% to about 12% final products.Typically, about 9 seconds/go to about 25 seconds/change preferred about 11 seconds/to go to the about 20 seconds/rotating speed (second/commentaries on classics) that changes be enough.

In case after wet mashed potato is made into thin slice and drying, just the drying slice of gained potato block can be broken into less sheet (if necessary).These less sheets can be any required size.Can use anyly the infringement of starch and potato cell can be reduced to minimum broken thin slice method, for example crush, grind, prick broken, cutting or pulverize.For example, available UrschelLaboratories, (Valparaiso, Indiana) UrschelComitro of Sheng Chaning pulverizes thin slice so that the thin slice fragmentation to Inc..Alternatively, this sheet potato block can be kept perfectly.As used herein, a complete potato block and less sheet part all are included in the term " potato block ".

2. the food of making by the dehydrated potato products that contains reduced levels of acrylamide and asparagine Product

The dehydrated potato products that contains reduced levels of acrylamide and asparagine can be used for making any suitable food.A kind of particularly preferred purposes of dehydrated potato products is to make the processing potato chips of being made by dough/pasta.The embodiment of above-mentioned processing potato chips comprises: be described in the United States Patent (USP) of announcing on December 21st, 1976 of authorizing Liepa 3,998,975, the United States Patent (USP) of announcing November 7 nineteen ninety-five 5 of authorizing people such as Villagran, 464,642, the United States Patent (USP) of announcing November 7 nineteen ninety-five of authorizing Lodge 5,464,643 and people such as Dawes those in the PCT application PCT/US95/07610 that announced on January 25th, 1996 with WO 96/01572.

But dehydrated potato products is rehydration also, and be used to make food, and for example mashed potato, potato pie, potato pancake and other potato snack are for example pressed chips and potato slices.For example, can use dehydrated potato products make to press fried potato products, for example be described in the United States Patent (USP) of announcing on April 9th, 1,963 3 of authorizing people such as Backinger, 085, in the United States Patent (USP) of announcing on October 18th, 020 and 1976 of authorizing Cremer 3,987,210 those.Dehydrated potato products also can be used in bread, thicken soup meat soup, flavoring, baby food or any other suitable food.

3. the potato block that contains reduced levels of acrylamide

Can use the present invention to make the potato block that contains reduced levels of acrylamide.To set forth the method for optimizing of making above-mentioned potato block goods below, but the invention is not restricted to this specific embodiment.The typical method of making potato block is set forth in " Potato Processing ", the 371st page to 489 pages.

In a preferred embodiment, the invention provides the method that reduces acrylamide content in the potato block, this method comprises:

(1) not necessarily peels off jacket;

(2) not necessarily wash potato;

(3) potato slices is made potato flakes;

(4) rinsing potato flakes not necessarily;

(5) blanching potato flakes not necessarily;

(6) extract potato flakes to reduce asparagine concentration;

(7) not necessarily dry potato flakes;

(8) the fried potato thin slice is made potato block.

Most preferably, potato flakes blanching before carrying out the asparagine extraction.Though more than described in above-mentioned (6) step and extracted asparagine, should be appreciated that extraction can carry out in any suitable step of this method.In another embodiment, the method for acrylamide content comprises in the reduction potato block:

(1) not necessarily peels off jacket;

(2) not necessarily wash potato;

(3) potato slices is made potato flakes;

(4) rinsing potato flakes not necessarily;

(5) blanching potato flakes not necessarily;

(6) not necessarily dry potato flakes;

(7) the fried potato thin slice is made potato block.

(8) extract potato flakes reducing asparagine concentration, wherein said be extracted in above-mentioned

Before step 1 any one in 7, among or finish afterwards.

Should be appreciated that above-mentioned steps 1 to 7 can be undertaken by any suitable order.

Extraction step can be finished with any appropriate method.Preferable methods can comprise immersion, extract and rinsing with main dipping bath.

In one embodiment, thickness for about 0.5mm extremely the potato flakes of about 1.5mm be used to make potato block.The blanching thin slice was heated to about 54 ℃ (130 °F) to about 77 ℃ (170 °F) about 15 seconds to about 3 minutes in water.Thin slice after the blanching not necessarily cools off.Then the thin slice after the blanching is ducked in drink and extracted it in about 15 minutes to about 4 hours.Extraction can be finished in one or more extraction steps.The asparagine concentration of gained potato flakes reduces.Then fried potato flakes is not necessarily dry before making potato block.

The acrylamide content of the potato block of making according to the inventive method preferably less than about 30ppb, is more preferably less than about 20ppb, even is more preferably less than about 10ppb, most preferably less than about 5ppb less than about 40ppb.

4. chips

Can use the present invention to make the chips that contain reduced levels of acrylamide.To set forth the method for optimizing of making above-mentioned chips below, but the invention is not restricted to this specific embodiment.For example, can in any suitable procedure of processing of making chips method known in the art, carry out asparagine and extract, for example be set forth in " Potato Processing " the 491st page to 534 pages those, or be described in United States Patent (USP) 6,001,411 and 6, those methods in 013,296.

In a preferred embodiment, the invention provides the method that reduces acrylamide content in the chips, this method comprises:

(1) not necessarily peels off jacket;

(2) not necessarily wash potato;

(3) cut potato and make potato slices;

(4) rinsing potato slices not necessarily;

(5) blanching not necessarily or not necessarily half French fried potatoes;

(6) not necessarily cool off potato slices;

(7) from potato slices, extract asparagine;

(8) not necessarily dry potato slices;

(9) not necessarily wrap the face clothing to potato slices; With

(10) half French fried potatoes are made the semi-finished product chips.

Most preferably, potato slices blanching before carrying out the asparagine extraction.Though more than described in above-mentioned (7) step and extracted asparagine, should be appreciated that extraction can carry out in any suitable step of this method.In another embodiment, the method for acrylamide content comprises in the reduction chips:

(1) not necessarily peels off jacket;

(2) not necessarily wash potato;

(3) cut potato and make potato slices;

(4) rinsing potato slices not necessarily;

(5) blanching not necessarily or not necessarily half French fried potatoes;

(6) not necessarily cool off potato slices;

(7) not necessarily in dry Ma Lingzhu bar;

(8) not necessarily wrap the face clothing to potato slices;

(9) half French fried potatoes are made the semi-finished product chips; With

(10) from potato slices, extract asparagine, before wherein said any one that is extracted in the above-mentioned steps 1 to 9, among or carry out afterwards.

Be to be understood that above-mentioned steps 1 to 9 can be undertaken by any suitable order.

Extraction step can be finished with any appropriate method.Preferable methods can comprise immersion, extract and rinsing with main dipping bath.

Then, the semi-finished product chips are freezing, packing and storing so that fried is later on made the finished product chips.Term used herein " potato slices " is enough broad sense, comprises the potato of any suitable shape, as potato ball, magnificent husband's chips, the chips that curl, potato ball, Hash Browns, thick French fries, jacket or any other potato part.

Most preferably, potato slices blanching before asparagine is extracted.Wrap the face clothing to chips if desired, can use suitable wafer material (as starch or comprise the material blends of one or more starch) to be rolled on the potato slices before fried half.

The acrylamide content of the chips of being made by semi-finished product chips of the present invention preferably less than about 30ppb, is more preferably less than about 20ppb, most preferably less than about 10ppb less than about 40ppb.

5. corn snack

In another embodiment, the acrylamide content of corn snack can preferably less than about 50ppb, be more preferably less than about 10ppb less than about 75ppb.Preferred corn snack comprises tortilla chips and cornflakes.Although the method for this paper is described with preferred tortilla chips usually, should be appreciated that this method can be used for making any suitable corn snack.

Tortilla chips is welcome especially consumption snack goods.Traditionally, tortilla chips is made by whole kernel corn, wherein this whole kernel corn in hot limewash boiling about 5 to about 50 minutes, then the dipping spend the night.This boiling-dipping method can soften shell, and can make the starch part gelatinize in the corn embryosperm.Then, wash this boiling-impregnated corn (being called " nixtamal ") and remove decapsidate, and grind and make plastic dough (being called " dough "), it comprises about 50% moisture.The dough that has just ground is made sheet, be cut into the dessert base, and to the temperature of 600 (302 ℃ to316 ℃), toasted about 15 to about 30 seconds, water content is reduced to about 20% to about 35% at about 302 ℃ (575 °F).The dessert base that will toast is then fried in deep fat and is made water content less than about 3% tortilla chips.Referring to, for example, the United States Patent (USP) of announcing on November 1st, 1,958 2 of authorizing people such as Anderson, 905, the United States Patent (USP) of announcing on September 12nd, 559 and 1,972 3,690,895 of authorizing people such as Amadon, and the 410th page of " Corn:Chemistryand Technology " (American Association of CerealChemists, people such as Stanley A.Watson compile) is to 420 pages (1987).

Also can make tortilla chips by the dried noodle cooking starch.In the process of the above-mentioned dried noodle cooking starch of typical making, the United States Patent (USP) of for example announcing March 1 nineteen fifty-five 2 of authorizing people such as de Sollano, 704,257 and the United States Patent (USP) 3 of authorizing people such as Gonzales announced February 20 nineteen sixty-eight, 369, described in 908 those will grind with the corn of lime treatment and dehydration forms stable state.Afterwards, available water is made the dough dough with dried noodle cooking starch rehydration, makes tortilla chips with this dough dough then, those described in the WO 01/91581 that announces in December 6 calendar year 2001 as people such as Zimmerman.

In one embodiment, can make the corn snack with following method, this method comprises:

(1) provide dough/pasta, wherein said dough/pasta comprises the dough with decrement asparagine;

(2) make the dessert base by dough/pasta; With

(3) cook the dessert base, make the corn snack.

In another embodiment, can make the corn snack with following method, this method comprises:

(1) blanching corn not necessarily;

(2) extract corn to form the corn of asparagine decrement;

(3) make nixtamal with corn;

(4) make the dessert base with nixtamal; With

(5) cook the dessert base, make the corn snack.

The corn snack that useful this paper method is made comprises tortilla chips, cornflakes or extruding corn snack.Suitable cooking method can comprise cure, fried, extruding and their combination.

Be to be understood that extraction can carry out in any appropriate steps of process.In one embodiment, the method for making corn snack comprises:

(1) blanching corn not necessarily;

(2) make nixtamal with corn;

(3) make the dessert base with nixtamal;

(4) cook the dessert base, make the corn snack; With

(5) extract corn obtaining the corn of asparagine decrement, before wherein said any one that is extracted in the above-mentioned steps 1 to 4, among or carry out afterwards.

D. commodity

Another embodiment of the invention is commodity, and it comprises:

(a) food, wherein said food contains the acrylamide of decrement;

(b) contain the container of food; With

(c) information relevant with container.

This information is told the consumer, and this food comprises the acrylamide of decrement.Suitable information comprises, but be not limited to, inform the information of " decrement " or " on a small quantity " acrylamide, inform and contain, and inform that food meets or exceeds the information of suggestion amount or standard volume (as the threshold value of regulation or the content surpassing) less than the ormal weight acrylamide information of (as less than 5ppb).In one embodiment, information is told the consumer, and food is to make with the composition that contains decrement or a small amount of asparagine, thereby hints that therefore this food contain decrement or a spot of acrylamide.

In another embodiment, these commodity comprise:

(a) food, wherein said food contains the asparagine of decrement;

(b) contain the container of food; With

(c) information relevant with container.

This information is told the consumer, and this food comprises decrement or a spot of asparagine.

This information can be directly or indirectly to be attached on the container, directly or indirectly to be attached near the printed article the container, or alternatively, can be printing, electronics or the broadcasting information relevant with container.

Can ration, present, any container of displaying or storage food all is suitable.Suitable containers includes, but not limited to bag, jar, box, bowl, dish, basin and tube.

Analytical method

Quantize to be used to characterize the parameter of key element of the present invention with specific analytical method.These methods are described in detail as follows.

1. acrylamide

Measure the method for acrylamide (AA) in the food

General introduction

With 1- ¹³The C-acrylamide ( ¹³C-AA) be incorporated in the food, use hot water extracting then.With the moisture supernatant of ethyl acetate extraction three times, combined ethyl acetate extract then, and concentrating, then with have special detection AA with ¹³The LC/MS of the selected ion monitoring of C-AA analyzes.

The extraction sample

1. weighing 6.00 ± 0.01g sample is put in the 125ml conical flask.Attention: sample is put in the food processor, and pulsed 30 seconds, make particle diameter reach about 0.3cm (1/8 inch) or littler.If sample is too little so that can not be pulverized effectively in food processor, then sample is placed a new plastic bag (as Whirl-Pak ^TMOr equivalent), pulverizes with rubber hammer then, reach about 0.3cm (1/8 inch) or littler until particle diameter.

2. use the 100ng/ μ L of adjustable 1000 μ L pipettes (calibrating) with 120 μ L ¹³C-AA deionized-distilled water solution (ISTD 2) directly is added on the sample.

3. the use distributor joins the 40mL deionized-distilled water in the flask, and covers with paper tinsel.

4. in 65 ℃ water-bath, placed 30 minutes.

With distributor with 10mL, the 2-dichloroethanes joins in the flask, uses Tekmar Tissumizer then ^TM(SDT-1810) or

(T18 Basic) homogenize 30 seconds, or until evenly.With deionized-distilled water flushing probe, washing lotion is connected in this flask.

6. the 25g homogeneous mixture is put in the bottle of 8 dram.

7. cover the tight mouth of pipe, centrifugal 30 minutes then with 2500 to 5200 rev/mins speed.

8. the 8g supernatant is transferred in the bottle of another 8 dram carefully, noted solid particle not being shifted in the past.

9. add 10mL ethyl acetate with distributor, close the lid, and vortex 10 seconds.

10. make all emulsion breakings; This can or vibrate once or twice by means of vortex, makes each layer separately then.

11. upper strata as much as possible (ethyl acetate) is transferred in the scintillation vial, and is not shifted any liquid (water) at the interface.Use the 5mL ethyl acetate extraction more than twice at every turn, and join in the same scintillation vial.Then, add about 2g anhydrous sodium sulfate.

12. in 60 ℃ to 65 ℃ water-bath, this extract is concentrated into about 1ml with gentle nitrogen stream.This extract is transferred to Pierce REACTI-VIAL ^TMOr in the cone-shaped glass bottle of equivalence, and further concentrate this extract, be about 100 μ L to 200 μ L until final volume.This extract is put in the automatic sampling bottle of band taper.

The preparation reference material

Storing solution and internal standard compound

Solution	Weight	Volumetric flask	Solvent	Concentration (ppm)
					Storing solution 1	0.1000g acrylamide (AA)	100mL	Ethyl acetate	1000
ISTD 1	0.0100g 13The C-acrylamide	100mL	Ethyl acetate							100
					Storing solution 2	0.1000g acrylamide (AA)	100mL	Deionized-distilled water	1000
ISTD 2	0.0100g 13The C-acrylamide	100mL	Deionized-distilled water	100

The intermediate standard thing

Solution	The volume of storing solution 1AA (μ L)	Volumetric flask (mL)	Solvent	Concentration (ppm)
					INT 1	100	10	Ethyl acetate	10
INT 2	1000	10	Ethyl acetate	100

Calibration standard

Standard	Volume INT 1 (μ L)	Volume INT 2 (μ L)	The volume of ISTD1 (μ L)	Volumetric flask (mL)	Solvent	Concentration AA (ppm)	Concentration ISTD 1 (ppm)
								0	0	0	450	10	Ethyl acetate	0	4.50
0.25	250	0	450	10	Ethyl acetate	0.250	4.50
								0.75	750	0	450	10	Ethyl acetate	0.750	4.50
1.5	0	150	450	10	Ethyl acetate	1.50	4.50
								3.0	0	300	450	10	Ethyl acetate	3.00	4.50
5.0	0	500	450	10	Ethyl acetate	5.00	4.50

The homogenizer clean method

At the interval of measuring each sample, with this clean method cleaning homogenizer.

1. hot tap-water is injected the conical flask (≈ 80% is full) of a 1L, add a Dawn then ^TMDishwashing detergent liquid is (available from Procter ﹠amp; Gamble Co.) or equivalent.

2. the probe with dispersive element is inserted in the water as far as possible.

3. with about 10 to 15 seconds of solution homogenize.

4. cleaning solution is poured out from conical flask; With hot tap-water flushing and this conical flask that reinjects.

5. about 10 to 15 seconds of homogenize once more.

6. with the conical flask turned letter, hot tap-water reinjects; Homogenize is about 10 to 15 seconds once more.

7., then continue on demand the repeatedly clean hot tap-water of homogenize, until reaching this condition if water is not clarified and particle is arranged.

8. when hot tap-water is clarified and is not had particle, with deionized-distilled water flushing probe.

Analyze with LC/MS

Use is connected Waters 2690 LC on the Micromass LCZ mass spectrograph, and sample is analyzed.

Phase flows	100％ H 2O，10mM NH 4Ac, the pH value is adjusted to 4.6w/ formic acid
		Chromatographic column	2.0mm * 150mm, YMC C18 AQ (available from Waters Corp.)
Flow velocity	0.2ml/ minute
		The interface	Directly (no layering)
Sampling volume	5μL
		MS ionization mode	Electron spray, positive ion mode
The MS detection mode	Selected ion monitoring: m/z 72 (AA), m/z 73 ( 13C-AA); The time of staying: 0.5s

Data analysis

For five standard samples in a series of ethyl acetate, with response ratio (area at AA peak/ ¹³The area at C-AA peak) to corresponding concentration ratio mapping.All standard samples comprise 4.5 μ g/mL's ¹³C-AA, and the concentration of AA is from 0 μ g/mL to 5 μ g/mL.Linear regression produces calibration curve, by this calibration curve, determines the concentration ratio in the extract from the response ratio of being measured.When multiply by step 2 at extracting process, this concentration ratio joins accurately knowing in the sample ¹³During C-AA content (nominal 2ppm), just can obtain the ppm content of AA.

The sample of LC/MS calculates:

By with the response ratio on the y axle (area of area/m/z 73 of m/z 72),, produce calibration curve to the concentration ratio on the x axle ([AA]/[13C-AA]) mapping.For present embodiment, the equation of this line is y=0.899x+0.0123.

4.0 minute the time AA peak (m/z 72) the mensuration area: 100,000

4.0 minute the time 13C-AA peak (m/z 73) the mensuration area: 500,000

Response ratio R _r=0.200.From the slope and the intercept of calibration curve, calculating concentration compares R _c: R _c=(0.200-0.0123)/0.899=0.209

The incorporation of 13C-AA (2ppm) in the known sample, then the mensuration content of AA is 0.209 * 2ppm=0.418ppm

Quality assurance/quality control (QA/QC)

1. all balances that use when preparing reference material and/or sample must be checked their calibration weekly with the series of standards counterweight.These balances should use at least three counterweights that cover testing sample/reference material weight range to check.

2. should make the calibration curve of six somes every day.

3. should use each group sample analysis work reference material (WRM).The running mean value of this material concentration should be within 2 σ.If not in this scope, then this instrument should be recalibrated, and recomputates WRM.

2. asparagine

Measure asparagine and aspartic acid in the F﹠B goods

Principle

To claim the sample of weight to mix, and heated homogenize then 30 minutes with 5%HCl.A part of homogeneous mixture is centrifugal, to handle then with a part of supernatant dilution, and with FMOC reagent (9-fluorene methyl chloro-formate), this reagent and asparagine and aspartic acid reaction form the hyperfluorescence derivative.With reversed-phase HPLC the FMOC-asparagine is separated from other sample substrate component then.Excite at the 260nm place, detect the fluorescent emission that 313 nanometers (nm) are located.The reference material analysis of concentration known can be carried out quantitatively.

The linearity

The correlation that the working stamndard curve of four standard samples (50-600ppm) provides is 0.998 or higher.The curve that is drawn by the sample of 2000ppm has also provided 0.998 correlation.

Precision

Potato products:

Mix the asparagine of four kinds of content and aspartic acid (40,200,400 and 600ppm) in the farina.Asparagine be recovered as 100% (relative standard deviation is less than 4%), and aspartic acid be recovered as 110% (relative standard deviation is less than 4%).

List of references

1.Herbert，P.；Santos，L；Alves，A.Journal of FoodScience(2001)，66(9)，1319-1325。

2.Heems，Dany；Luck，Geneviewe；Fraudeau，Chrisophe；Verette，Eric.Journal of Chromatography，A(1998)，798(1+2)，9-17。

The system repeatability

The work reference material of duplicate parallel determination potato chips in 5 days.The result is as follows:

The ug/g asparagine The ug/g aspartic acid

Mean value7832.07 1440.98

STD 625.59 195.80

％RSTD 7.99 13.59

Below be chemicals and the instrument that suggestion is used; Yet allow to replace with equivalent substance.

Chemicals

Water, HPLC level or Milli-Q ^TMLevel

(Millipore)

Acetonitrile, HPLC level Burdick ﹠amp; Jackson

#AH015-4

Methyl alcohol, HPLC level Fisher #A452-4

Ethyl acetate Baker #9280-3

Pentane Burdick ﹠amp; Jackson

#GC312-4

Asparagine monohydrate EM Science

Aspartic acid Sigma #A-8949

Aminoisobutyric acid Sigma #A-8379

9-fluorenyl chloro-formate (FMOC) ICN #150200

Boratex EM Science#SX 0355-

1

Boric acid Fisher #A-73

Sodium acid carbonate ICN #194847

Tetramethyl ammonium chloride Fisher #04640-500

Natrium citricum MCB #SX445

Anhydrous citric acid Baker #0122-01

Acetone Burdick ﹠amp; Jackson

#010-4

Hydrochloric acid, 0.1N Fisher #SA48-500

Calcium chloride dihydrate Aldrich #22,350-6

Equipment

Pipette, polyethylene (Samco #222)

Volumetric flask (25,100,250,1000ml)

Constant volume pipette (10ml)

Graduated cylinder (100-1000ml)

HPLC liquid reservoir (500ml, 1L or 2L)

Beaker

Magnetic stirring apparatus/stirrer

Assay balance (4)

Scintillation vial

Centrifuge tube, screw-cap (100 * 16mm) with cover

Automatic sampling bottle (8 * 30mm, 1ml), lid with lug

Security: this method need be used fume hood, and relates to the contact chemicals.Please refer to safety rule about fume hood uses and chemicals spills.

Instrument Model The manufacturer

Automatic instrument

SPE Hamilton

Pump/HPLC syringe HP 1100 Agilent

Detector RF10AXL Shimadzu

Data system Chemstation Agilent

Chromatographic column

(2) 3 microns # 00D-4251-EO of Phenomenex Luna 100 * 4.6mm C-18

Reagent preparation

(pH 8.3 to 8.5 for diluent; 1000ml)

1. weighing 3.0 gram Boratexes, 3.0 gram boric acid and 8.0 gram sodium acid carbonates have been removed the peel in the heavy dry beaker in one.

2. on magnetic stirring apparatus, place the 800ml beaker of a sky.Add about 500mlMilli-Q ^TMWater and stirrer.Vigorous stirring water, but splash is not come out.

3. quantitatively transfer to the reagent of step 1 in the water; Stirring is dissolved fully until it.

4. the solution of step 3 is quantitatively transferred in the volumetric flask of 1L, and used Milli-Q ^TMWater is diluted to volume; Mix.Can stablize to place and reach six (6) individual months.

Calcium chloride solution (100 gram)

1. weighing 40 grams two hydration calcium chloride are in removing the peel heavy 250ml beaker.

2. add 60 gram Milli-Q ^TMWater.Evenly mixed.Under environmental condition, be stored in the vial of lid.Can stablize to place and reach 1 year.

Extractant (pentane: ethyl acetate 80: 20; 500ml)

Security: pentane and ethyl acetate are volatile and inflammable.In fume hood, carry out following operation.

1. the 400ml pentane is transferred in the 500ml HPLC reservoir.

2. add 100ml ethyl acetate.Evenly mixed.In fume hood/preservation closes the lid down.

(buffer: methyl alcohol: acetonitrile 60: 5: 35, pH 3.2,2L) for the phase that flows

1. weighing 1.35 gram tetramethyl ammonium chlorides, 3.65 gram citric acids and 1.60 gram natrium citricums are in removing the peel heavy dry beaker.

2. on magnetic stirring apparatus, place the 800ml beaker of a sky.Add about 500mlMilli-Q ^TMWater and stirrer.Vigorous stirring water, but splash is not come out.

3. quantitatively transfer to the reagent of step 1 in the water; Stirring is dissolved fully until it.

4. the solution of step 3 is quantitatively transferred in the 1L graduated cylinder, and used Milli-Q ^TMWater is diluted to 1000ml; Mix.

5. it being transferred to 2 liters HPLC flows in the phase liquid reservoir.

6. add 200ml Milli-Q ^TMWater, 100ml methyl alcohol and 700ml acetonitrile.Under vigorous stirring, add back two kinds of solvents lentamente.In fume hood, carry out this operation, and dress individual protective equipment.Detail sees also relevant material data of safety table (MSDS).

7. when stirring, outgas mutually to flowing with the vacuum aspiration.

FMOC reagent solution (in the acetone)

1. weighing 0.10 gram FMOC reagent is in removing the peel heavy 100ml volumetric flask.

2. the adding acetone solution is diluted to volume with it equally.Evenly mixed.In fume hood, carry out this operation.The PPE that stipulates among the MSDS of chemicals on the wearing.

3. freezing preservation is no more than six (6) individual months.

The acid solution (5% HCl) that is used for sample extraction

1. with 100ml Milli-Q ^TMWater adds in the 200ml volumetric flask.

2. the HCl with 4ml 1N adds in the volumetric flask.

Use Milli-Q ^TMWater is diluted to volume.

Preparation internal standard compound (aminoisobutyric acid)

ISTD A-internal standard compound storing solution A

1. weighing 0.5 gram aminoisobutyric acid is in removing the peel heavy 250ml volumetric flask.

2. add 25ml 1.0N HCl and about 100ml Milli-Q ^TMWater.Vortex mixed is until dissolving.

Use Milli-Q ^TMWater is diluted to volume, and mixes.Freezing preservation is no more than six (6) individual months.

ISTD B-work internal standard substance solution B(this solution is joined in the calibration standard)

1. inhale and move in 1ml internal standard substance solution A to the 100ml volumetric flask.

2. use Milli-Q ^TMWater is diluted to volume.Can stablize and preserve 1 month.

The preparation calibration standard

The deposit calibration solution

In removing the peel heavy 50ml volumetric flask, weighing 0.100g (+/-0.001g) asparagine and 0.100g (+/-0.001g) aspartic acid.Add 25ml Milli-Q ^TMThe HCl of water and 1ml 1N.Be placed in the sound wave bath,, use Milli-Q then until dissolving ^TMH2O is diluted to volume.Freezing solution down can be remained valid 6 months.

The working stamndard thing

Prepare following work calibration standard:

Std #	The mL storing solution	Final volume (mL)	ppm
				1	5	200	50
2	5	100	100
				3	1	10	200
4	3	10	600

Freezing solution down can be remained valid 1 month.

The preparation sample

1. weighing 1g sample is in the 125ml conical flask.

2. the HCl solution that adds 48.0ml 5% in each sample.

3. add 2ml ISTD A in each sample.

4. with aluminium foil each flask is covered, and in the water-bath of 60C, placed 30 minutes.

5. 10mL dichloroethanes in each sample.

6. with sample homogenize 60 seconds.

7. a part of sample is poured in the 30ml centrifuge tube.

8. under 5 ℃ centrifugal 32 minutes with 10000rpm.Supernatant is used for " sample-dilution " step 1.

Preparation reference material and sample

Use three kinds

Method is come dilute sample/reference material, adds internal standard compound and is formed the FMOC derivative.They are summarized as follows.

Operation Used Microlab method

Dilution TRANSDIL

Add internal standard compound ADDISTD

Form FMOC derivative ADDFMOC

Use

Automatically instrument is prepared sample and reference material

Step 1: reference material-adding ISTD and dilution step

1. for each reference material, prepare two set of tubes.In a sleeve pipe, put into about 2mL reference material, these pipes that reference material is housed are placed on

On the leftmost position.

2. the pipe support that will be placed with empty pipe is placed on

On the rightmost chord position.

3. the internal standard substance solution B that will work annotates in 20ml glass (flicker) pipe, and is placed on

The workspace on.

4. system of selection ADDISTD.(mix 200 μ l ISTD B, 50 μ l reference material solution, use Milli-Q ^TMWater is diluted to 4000 μ l with total measurement (volume)).

5. implement this method.

6. take off the pipe group from left position, abandon on one side.

From

The work internal standard substance solution is taken out in the workspace, and freezing.

The pipe that stays the right is used for step 3.

Step 2: sample-dilution step (in the sample process for preparation, having added ISTD)

1. for each sample, prepare two set of tubes.In a set of tubes, put into about 2mL sample, these pipes that sample is housed are placed on

On the leftmost position.

2. the pipe support that will be placed with empty pipe is placed on

On the rightmost chord position.

3. system of selection TRANSDIL.(sample number is set, and sample size is 50 μ l, and uses Milli-Q ^TMWater-reducible final amount of dilution is 4000 μ l.)

4. implement this method.

5. take off the pipe group from left position, abandon on one side.

The pipe that stays the right is used for step 3.

Step 3: add FMOC reagent-preparation fluorescent derivative

1. prepare the nut pipe of a 100 * 16mm.

2. pipe support is placed on

On the rightmost chord position.

3. the standard property management and the sample cell that will derive from top dilution step are positioned over

On the leftmost chord position.

4. the FMOC reagent solution of five equilibrium (22ml) is transferred in the glass scintillation pipe.Add about 100 μ L, 40% calcium chloride solution; Mix.(add calcium chloride and make that FMOC reagent " charged "-this is to use

Measure necessary).

5. scintillation vial is positioned over

On the workspace.

6. system of selection ADDFMOC.

7. syringe 1 and 2 is switched to diluent (pH 8.3 to 8.5) from water.

8. with diluent (pH 8.3 to 8.5) syringe 1 and 2 is washed at least 5 circulations.

9. implementation method ADDFMOC.(450 μ l FMOC solution, 250 μ l are derived from the sample mix of top ADDISTD, and being diluted to final volume with diluent solution is 1300 μ l).

10. take off the pipe group from the sample chord position, and abandon.

11. from

The FMOC reagent solution is taken out in the workspace, and freezing.

13. take off the pipe group from rightmost position, and place fume hood.Left standstill at least 10 minutes, or until solution clarification (but being no more than 20 minutes).

14. add the 2ml extractant in each pipe.Close the lid, and high speed vortex 2 minutes, to extract unreacted FMOC reagent.

15. prepare the pipe of another pipe support 55 * 16mm.Add the mobile phase solution of 1ml in each pipe.

16. (lower floor) transfers to from centrifuge tube in the pipe of 55 * 16mm with the 1.0mL water layer.

17. discard upper strata (organic layer).

18. sample is transferred in the automatic sampling bottle, and sealing.

Chromatography

Operating condition

The HP 1100 of Chem Station software is housed

Detector: Waters 474 scanning fluorescence detectors

Model: Norm

Signal: 0.0000

Wavelength: excite 260

Emission 313

Gain: 10

Decay: 1

Response: FST

Chromatographic column: Phenomex Luna C18 (2) 100 * 4.6mm 3u

The LC method

Flow: 1.000ml/ minute

Deng degree operation (referring to the reagent preparation-phase flows)

Volume injected: 10.0ul

Temperature is provided with: temperature control not

Calculate

The usable floor area value, with respect to the calibration curve calculation sample solution of known quantity:

y＝mx+b

Y (asparagine/ISTD ratio)=m (slope) x (acrylamide concentration)+b (y y-intercept)

(y-b)/m＝x

Asparagine (ppm)=(asparagine area/ISTD area-intercept)/slope

[ppm＝ug/mL]

Embodiment

Asparagine (ppm)=(215.45436/551.828--0.0165)/0.0023=176.93ppm

Correction for dilution/homogenizing in the sample preparation steps.

Asparagine (ppm) X of asparagine (ug/g)=record The sample diluting liquid body Amass (50) (ml)

The gram number of sample

[ppm＝ug/mL]

Embodiment

Asparagine (ug/g)=176.93ppm X 50ml=8773.65 (ug/g)

1.0083g

Carry out the standard of qualified judgement:

● the degree of accuracy of working standard control sample must be within 10% scope of asparagine known results.

● the linearity (r of calibration curve ²) be necessary for 0.995 or bigger.

The LC chromatography of sample

Fig. 3 has listed the LC chromatogram analysis of sample.

Retention time	Compound
		4.5 minute	Asparagine
6.6 minute	Aspartic acid
		11.5 minute	FMOC reagent
20.7 minute	ISTD

3. the minimizing percentage of acrylamide

The content of acrylamide in the minimizing %=[(control sample of acrylamide-handled the content of acrylamide in the sample)/control sample in the content of acrylamide] * 100.

Handled the identical mode of sample and make control sample to be used for making, difference is that control sample do not remove processing through asparagine.

4. the minimizing percentage of asparagine

The minimizing percentage of asparagine=[content of asparagine in (content of asparagine in the control sample-handled the content of asparagine in the sample)/control sample] * 100.

Handled the identical mode of sample and make control sample to be used for making, difference is that control sample do not remove processing through asparagine.

Embodiment

Following examples illustrate the present invention, but are not intended to limit it.

Embodiment 1-contains the dehydrated potato products of decrement acid amides

The Russet potato peeled and be cut into 0.64cm ( ¹/ ₄" in) and sheet.Boiling is to deliquescing (～93 ℃ (200 °F) continue 30 minutes) in water.Dry and use water rinse.The sheet that uses the rubber scraper firmly to make to boil is made mashed potato (centre-to-centre spacing of mesh wire is 1.67mm) by silk screen.This mashed potato of part is reserved is used as contrasting mashed potato.

With 453g (1 pound) mud shape potato and 1,361g (3 pounds) water adds the Kitchen-that has whipper

In the food processor (model is #K45), the speed of whipper per minute=2.With scalping (twine centre-to-centre spacing is 1.67mm) filtering mixt.With dusting cover (twine centre-to-centre spacing is 1.11mm) filter liquide part (after filtration).Two solid portions are mixed.Repeat to wash five times for the first time.

Together partially mixed with behind all once washings.To the mashed potato repeated washing process behind the once washing to obtain～1, the mashed potato after the secondary washing of 905g (4.2lbs).

With mashed potato and the contrast mashed potato drying of freeze drier after with secondary washing.

The asparagine concentration (based on dry weight) of the mashed potato after contrast mashed potato and the secondary washing is respectively 1.2% and less than 0.2%.The asparagine decrease of this method is greater than 84%.

Embodiment 2-contains the processing potato snack goods of decrement acid amides

Use the contrast and the dehydrated potato products after the secondary washing of the foregoing description 1 to be produced as follows the described processing potato snack of table:

	Tester	After the secondary washing
			The dehydrated potato products tester	100g
Dehydrated potato products after the secondary washing		100g
			Emulsifying agent	0.6g	0.6g
Water	50.5g	50.5g
			Frying temperature	191℃(375°F)	191℃(375°F)
The fried time	8 seconds	8 seconds

Dried potato products is worn into powder.With water and emulsifier, mix the formation dough/pasta fully with powder then.Then dough/pasta is ground in flakes by mixing roll, then sheeted dough is cut into oval-shaped dessert base.Then with dessert base fried formation processing potato snack in deep fat.

During the moisture of two samples and acrylamide content are listed in the table below:

Final products	Tester	After the 2X washing
			Moisture (%)	1.8	1.1
Acrylamide (ppb)	3219	60

The acrylamide decrease of the processing potato snack of use embodiment 1 extracting method is greater than 98%.

Embodiment 3-potato block

Only blanching: use unprocessed potato block, can make the potato block that contains reduced levels of acrylamide.With Atlantic Ocean peeling potatoes, and be cut into～sheet that 1.1mm is thick.Clean, and press dry.In 74 ℃ of (165) water, with potato block blanching 15 seconds.Cooling, the water in the potato block that scalded that reduces phlegm and internal heat side by side.The potato block of fried processing in the deep fryer that is set to 191 ℃ (375) 60 seconds.(embodiment 2A) as above makes two batches of potato chips again, and different is that blanching treatment time is become 60 seconds (embodiment 2B) and 180 seconds (embodiment 2C).

With be used to make the foregoing description 2A, the B mode identical and make a control sample with C, difference is not blanching of potato flakes.

The method of using this paper to set forth, the acrylamide content in analytic sample and the tester.

Only blanching	2A	2B	2C
				Blanching treatment time (second)	15	60	180
Control sample *In acrylamide (ppb)	1079	1079	1079
				Acrylamide in the blanching sample (ppb)	940	696	679
Reduce percentage (%)	13	35	37

^*Annotate-only prepared a control sample

Blanching and extraction: use unprocessed potato block, can make the potato chips that contain reduced levels of acrylamide.With Atlantic Ocean peeling potatoes, and be cut into～sheet that 1.1mm is thick.Clean, and press dry.In 74 ℃ of (165) water, with potato block blanching 15 seconds.Cooling, the water in the potato block that scalded that reduces phlegm and internal heat side by side.The potato flakes of 100 gram blanchings are immersed in the 250ml distillation/deionized water one hour extract it.Every 8 minutes vortex stirred samples 1 minute.Be set to high fire heating 2 minutes with micro-wave oven (Panasonic micro-wave oven, model NN-S5488A), then with about 800ml cold running water washing three times, each 10 seconds.The potato block of fried processing in the deep fryer that is set to 191 ℃ (375) 60 seconds.(embodiment 2D) as above makes two batches of potato blocks again, and different is that blanching treatment time is become 60 seconds (embodiment 2E) and 180 seconds (embodiment 2F).

With be used to make the foregoing description 2D, the E mode identical and make a control sample with F, difference be potato flakes neither blanching do not extract yet.

The method of using this paper to set forth, the acrylamide content in analytic sample and the tester.

Blanching and extraction:	2D	2E	2F
				Blanching treatment time (second)	15	60	180
Control sample *In acrylamide (ppb)	1079	1079	1079
				Acrylamide (ppb) in the sample of blanching and extraction	281	156	50
Reduce percentage (%)	74	86	95

^*Annotate-only prepared a control sample

Conventional potato chips do not carry out the blanching step, thereby the potato chips with blanching step are this area the unknowns.The foregoing description shows, by use the novel combination of blanching and extraction in the potato block preparation method, can obtain excellent result.

Embodiment 4-chips

Use unprocessed potato slices, can make the chips that contain reduced levels of acrylamide.

With Atlantic Ocean peeling potatoes, and be cut into the potato slices of cross-sectional area for about 8mm * 8mm.Clean, and press dry.In 74 ℃ of (165) water, with potato slices blanching 1 minute.Cooling, the water in the potato slices that scalded that reduce phlegm and internal heat side by side.The potato slices of 100 gram blanchings are immersed in the 250ml distillation/deionized water one hour extract it.Every 8 minutes vortex stirred samples 1 minute.Be set to high fire heating 2 minutes with micro-wave oven (Panasonic micro-wave oven, model NN-S5488A), then with about 800ml cold running water washing three times, each 10 seconds.The potato slices of fried processing in the deep fryer that is set to 191 ℃ (375) 3 minutes.(embodiment 3A) as above makes two batches of chips again, and different is that blanching treatment time is become 3.5 minutes (embodiment 3B) and 7 minutes (embodiment 3C).

To be used to make the foregoing description 3A, the B mode identical with C, make control sample, difference is that they are not extracted.

The method of using this paper to set forth, the acrylamide content in analytic sample and the tester.

	3a	3b	3C
				Blanching treatment time (minute)	1	3.5	7
Control sample *In acrylamide (ppb)	216	216	216
				Acrylamide (ppb) in the sample of blanching and extraction	130	84	5.7
Reduce percentage	40％	61％	97.4％

^*Annotate-only prepared a control sample

Embodiment 5-commodity

The potato block of embodiment 3 is packaged in the bag, with sales to consumers.Be printed on information on the bag, statement " no acrylamide product! "

Embodiment 6-commodity

The potato block of embodiment 3 is packaged in the bag, with sales to consumers.Be printed on information on the bag, " acrylamide content is low in statement! "

Embodiment 7-commodity

The potato block of embodiment 3 is packaged in the bag, with sales to consumers.Be printed on information on the bag, " acrylamide has reduced more than 90% in statement! " information of television advertising transmission of these potato chips is that " our potato chips acrylamide content is lower! "

Embodiment 8-commodity

The processing potato chips of acrylamide content less than the unified shape of the embodiment 2 of 100ppb are packaged in the cylinder tank, with sales to consumers.The information of the television advertising transmission of these potato chips is " to have reduced acrylamide content! "

Embodiment 9-commodity

The chips of embodiment 4 are packaged in the paper cover of an end opening (from then on French fries are held and stretched out), with sales to consumers.The billboard that posts up in the retail division of selling chips is write, and " our chips contain the acrylamide of decrement! "

Embodiment 10-commodity

The chips of embodiment 4 are packaged in the paper cover of an end opening (from then on French fries are held and stretched out), with sales to consumers.The billboard that posts up in the retail division of selling chips is write, and " our chips acrylamide content is low! "

Embodiment 11-commodity

The potato block of embodiment 3 is packaged in the bag, with sales to consumers.Be printed on information on the bag, " composition with low asparagine concentration is made in statement! "

Embodiment 12-dehydrated potato products

Water cleans the potato that yellowish-brown is baked and banked up with earth, and places one pot of boiling water then.Potato was boiled (submergence) 1 hour.Take out the potato of boiling from water, peeling is smashed pulp to pieces then.In the 15g mashed potato, add 45g water, and this mixture of homogenize, until evenly, make it not have agglomerate.

The glutaminase that in the solution of this homogenize, adds 200 units, and rocked this sample every 5 minutes, cultivated altogether 30 minutes.

After carrying out cultivating in 30 minutes, with high fiery microwave (Panasonic micro-wave oven, the model NN-S5488A) heating products that increased progressively in per 2 minutes 10 minutes, until drying (and becoming brown).When the methods analyst acrylamide content of setting forth with this paper, compare with using the dehydrated potato (control sample) that identical method is prepared except that not adding enzyme, the dehydrated potato products of handling with enzyme has been obtained the acrylamide reduction greater than 10%.

Although illustrated and described the present invention with specific embodiments, it is evident that those skilled in the art can make many other change and modifications under the situation that does not deviate from the spirit and scope of the present invention.Therefore in additional claims, comprise all such changes and modifications in the scope of the invention consciously.

Claims (4)

1. a reduction is selected from the method for asparagine concentration in the food material of potato, corn and composition thereof, and described method comprises:

(1) provides the food material that is selected from potato, corn and composition thereof that comprises asparagine;

(2) not necessarily reduce the particle diameter of described food material;

(3) cell membrane permeability of the described food material of increase; With

(4) from described food material, extract asparagine.

2. the method for claim 1 wherein increases the cell membrane of described food material by the described food material of blanching.

3. a reduction is selected from the method for acrylamide content in the food of potato, corn and composition thereof, and described method comprises:

(1) provides the food material that is selected from potato, corn and composition thereof that comprises asparagine;

(2) not necessarily reduce the particle diameter of described food material;

(3) cell membrane permeability of the described food material of increase;

(4) extraction unit is divided described asparagine from described food material; With

(5) the described food material of heating forms ultimate food.

4. method as claimed in claim 3 wherein increases the cell membrane of described food material by the described food material of blanching.

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