Volume 20
`
`Issue 4
`
`Case No.: IPR2023-00206
`U.S. Patent No. 9,943,096
`
`Motif Exhibit 1134, Page 1 of 30
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`

`

`Critical ReviewsrM
`• Ill
`Food Science
`and
`Nutrition
`
`Editor
`
`Thomas E. Furia
`President
`lntechmark Corporation
`Palo Alto, California
`
`CRC Press. Inc.
`Boca Raton. Florida
`
`Case No.: IPR2023-00206
`U.S. Patent No. 9,943,096
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`Motif Exhibit 1134, Page 2 of 30
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`This journal represents information obtained fro m authentic and highly regarded sources. Reprinted ma(cid:173)
`terial is quo ted with permission, and sources are indicated . A wide variety of references ·are listed . Every
`reasonable e ffort has been made to give reliable data and information, but the a utho r and the publisher
`canno t assume responsibility for the validity o f all materials o r for the consequences o f their use.
`
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`ISSN 0099-0248
`
`© 1984 by C RC Press, Inc.
`
`Case No.: IPR2023-00206
`U.S. Patent No. 9,943,096
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`Motif Exhibit 1134, Page 3 of 30
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`CRC CRITICAL REVIEWS in
`FOOD SCIENCE AND NUTRITION
`
`Volume 20 Issue 4
`
`TABLE OF CONTENTS
`Influence of Time, Temperature, Moisture, Ingredients, and Processing Conditions
`on Starch Gelatinization ................ .............. ..... . ................. . ...... .. 249
`Daryl Lund, author. B .S .. M.S .. Ph.D. , University of Wisconsin, Madison. Professor of Food Engineering,
`Depanments of Food Science and Agricultural Engineering, University of Wisconsin, Madison, Wisconsin.
`Klau s J. Loren z. referee. Ph. B .. Nonhwestem Uni versity. Chicago, Illinois: M .S .. Ph. D . , Kan sas State University,
`Manhallan. Professor. Depanment of Food Science and Nutrition, Colorado State University, Fon Collins, Colorado.
`Starch gelatinization phenomena is extremely imponant in many food systems. This review focuses on factors
`affecting gelatinization characteristics of starc h . lm ponant variables which must be considered in design of processes
`in which starch undergoes gelatinization are heat of gelatinizatio n and temperature of gelatinization. Major inter(cid:173)
`actions are reviewed for the effects of li pids. moisture content, nonionic constituents and electrolytes on these
`c haracteristics. Funhe rmorc, treatment of starch-containing systems prior to heating into the gelatinization tem(cid:173)
`perature range can ha ve a s ignificant effect o n ultimate gelatinization characteristics .
`Optimization Methods and Available Software. Part I •.••••••.•.••.............••.. 275
`Israel Saguy, co-author. B.Sc .. M.Sc .. D .Sc .. Tec hnion. Haifa. Israel. Senior Scientist. Dcpanment of Food
`Technology. Agricultural Research Organization . The Volcani Center . Bet Dagan. Israel.
`Martin A. Mishkin. co-author. B.S ., Re nsselae r Polytechnic Institute. Troy. New York: Ph. D .. Massachuseus
`Ins titute of Technology. Cambridge . Dcpanment of Food Produc t Development. Procter & Gamble Co mpany,
`Cincinnati. Ohio.
`Marcus Karel. co-author. A . B .. Bosto n University. Bosto n. Massachuscus: Ph.D .. Massachusells Institute of
`Tec hno logy. Cambridge . Professor of Food Engineering. Dcpanment of Nutrition and Food Science . Massachusetts
`Institute of Technology. Cambridge.
`Anhur A. T eixeir-J. referee. B.S .. M .S .. Ph. D .. University of Massachuseus. Amherst. Associate Professor of
`Food Engineering. Institute of Food and Agric ultural Sciences. De panment of Agricultural Engineering. University
`of Florida. Gainesville. Florida.
`Optimi1.ation theories and generally applied optimization techniques are reviewed. The versatility and the complexity
`anticipated in actual problems are simplified to enable the food practitioners interested in the subject to overcome
`some of the barriers whic h prevented full utilization of optimization. The paper summerizes the various mathematical
`methods avai lable for solving problems of product and process optimization and provides informatio n and advice
`concerning the advantages and limitations of each tec hnique . A compiled list of optimization subroutines. guideli11C$
`and criteria for choosing the proper software are furnished .
`
`Case No.: IPR2023-00206
`U.S. Patent No. 9,943,096
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`Motif Exhibit 1134, Page 4 of 30
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`The Role of Collagen in the Quality and Processing of Fish .................. . ..... 30 I
`
`Zdzistaw E. Sikorski, co-author. II .Sc .. M.Sc .. Ph.D .. D.Sc . . Politechnika Gda,bka. Gda,bk. Poland. Professor
`of Food S1..·icncc. Depart men I of Food Prl·,l.'rvalilHl and Tcchni1..·al Mir rohioltlgy. Pt1lill.'t·hnika Gdaltska. Gdaflsk.
`Poland.
`
`Dorian N. Scott. co-authll T. B. Tl.'ch . . Ma:-.,t:y Univl.'r-,it~ o f Marrnwatu. Pahm.·rston North. Nrw Zealand. Food
`Technolofist. Oi,·i,ion of Horticulture: and Processing . D.S .I.R .. Mt. /\ ll'N:rt Rc,c~irch Center. Auckland . New
`Zealand.
`
`Da,•id H. Buisson. t.'lH.1uthor. ll. Se .. M .S .. : .. Ph .D .. Univl.'r,ity nf Au...:kland. Au('kland. New Zealand . S1..:icnti..,t.
`Divi,iun uf lfonicullurc :,ml Pr,,ce"i ng. D.S.I.R . . Mt. All><,rt Rc,earch Center. Auc kl,111d. New Zealand.
`
`R. Malcolm Love. referee. B.Sc.. Ph.D .. D.Sc. Univer,it) nf Liverpool. Lanca,hire. England. Section Head.
`Section of Applied Biology. Torry Re,earch Station. Mini,tr) of Agri,·ulturc. Fi, herie, & Food . Aberdeen. Scotland .
`
`Collagen in the mw,r k :-. of fish cun~tituh.~, the m.1in r11mpnnl'nl 111' the ,·,,nnccti vc: ti:-.:-.uc mcmbanc:-. joining individual
`myotomt':,,. and i:-. rC~J)(.Hhihk for lht.~ in1cgri1y of the filki-.. Thl' conh:nt of l'ollagcn in lh h mu'.'--cle:,,. i'.'-- fmm abo ut
`0.2 to 1.-Vk and in :,.quid mantel about 2.6'/4. Fi:-.h and lnh.'rtt.'brata collagen:-. nmtain :-.ligh1ly more e:-.:-.enlial amino
`acids 1han in1rnmuscular hovin1..· connt.'ctivc tb:-.ut.' l'oll;.1gc11. The invertebra1;.1 collagen!'- arc exceptionally rich in
`,ugars linked ,m,inly O-glyrn,idk ally Ill hydrnxylysinc rc,iduc,. Durinf ma1uration of lish 1he proportion of wllafc n
`10 total protein in the mu:,,,clc:,,. innca:-.l'"i whik the cxtcnl of 1..To,:,,.lin~ing doe:,. not change :-i ignificantly. The thermal
`propcnics of fi:-.h collagt.•n~ dl'f>t.'!H.I :,,.ignilkantly 011 the content or hydrox)pruline and proline re:,,.idues which in
`turn j:,,. corrdatl'd to thl' tl'mrcratun: t>f the hahi1at. Gt..'na;.11ly tht..' :-.hrinkagc temperature of fish skin collagen, is
`about ~0°C lowt.·r than that of mammalian hick~ collagen" . In -.,cvi:ral :,,,pccii::,,. of fish the weakening of the connective
`1b:-.ucs pthl mortem may l~ad to ~t.·rit>m, (JUality dctl'ritlration thal manife ~t:,,. ibelf by J isin1egration o f the fillch.
`esprci,1lly unde r thl' strai n of rough handling and of rigor morti :-. at ambient tcmpcrnturc . Thermal change:,. in
`collagen ar1..• the necc~:-.ary ri:~ult of the cooking nf fi sh. :,,.4ui<l. and minced li:-.h products and contribute to the
`dcsirahlt: tl'xtun.: 111' tht.· mc:.it. However. lhl'Y may lead It) !'teriou~ los~c:-. during hot smoking due to a reduction in
`lhc hrc-aking :-.tn:ngth of the-
`tis:,,.uc..; when heating. is condul'tl'd at high relative humidity. Bccau:,,.e of th1..• high
`vi:,£0:,,.i1y of gdatinitt.!d collagl' n. it i, not po:,,.sihk to (onccntrntc 1hc fi:,h stickwaters. a protcinaccou:,,. bypro<ltu.:t
`of thl· fi,h mt..'al indu~try. 1t1 more than )O'h dry mailer. Better knowledge of the contents and prupertie:-. of fish
`collagcn:-. could he helpful in rationali;,ing many a:-.pccl:-. of fbh prrn:e~sing .
`
`Subject Index .............. . . .... ......... .. . ............ .................... . . . .. ..... 344
`
`Author Index .......... ... .................. ............. .............................. 345
`
`Case No.: IPR2023-00206
`U.S. Patent No. 9,943,096
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`Motif Exhibit 1134, Page 5 of 30
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`.INFLUENCE OF TIME, TEMPERATURE. MOISTURE. INGREDIENTS. AND
`PROCESSING CONDITIONS ON STARCH GELATINIZATION
`
`Volume 20. Issue 4
`
`249
`
`A utho r:
`
`Daryl Lund
`Departments of Food Science and
`Agricultural Engineering
`University of Wiscons in
`Madison. Wisconsin
`
`Referee:
`
`Klaus J. Lorenz
`Dcpanmcnt of Food Science and Nutrition
`Coluratlo Stale University
`Fon Collins. Colorado
`
`I. INTRODUCTION
`
`Cereal grains comprise the larges t s ing le food group in the human die t in the world. Cereal
`grain~ a re used in developed and underde ve loped countries as a primary caloric source and
`contribute s ignificant and often sole sources of v itam ins. minerals . and protein . Processing
`cereal g rain into more des irable food forms (e.g .. bread) and improving upon their consumer
`4uality charac terist ics (flavor. tex ture. and co lor) has been practiced for mille nia. and ref(cid:173)
`erences are made to cereal grai n processing in the earliest known records.
`Curre ntly the food process industry uses sta rch. the primary const ituent of cereal grains.
`in a variety of ways ,md a varil:ty of products. With such a long history of use and its
`importance in foo<l processing. it wou ld he expected that changes in cereal grains and their
`starches as a func tion of' processing parame ters would be we ll established and characteri zed.
`However. this is not the case. Physical and chemical changes in starch as a func tion o f
`processing conditions have been described and some 4uantitative information is avai lable
`for e ng ineering des ig n of processes (e.g .. pasting temperatures). The physicochemical fun(cid:173)
`damentals for these c ha nges and the effect of other const itue nts on sta rch cook ing phenomena
`have no t been dctcm1incd .
`The purpose of this paper is to rev iew the s tate of knowledge on starc h cooking phenomena.
`A brief introduct ion to starc h gelati ni zation w ill be followed by a review of methods to
`study gela tin izat ion phenomena . Special emphasis will be g iven to differential scanning
`calorimetry (DSC) because of its increased use in s tudying a nd monitoring sta rc h gela tini(cid:173)
`zat ion. The major part of the paper wi ll the n focus on the influence of time. temperature,
`moisture conte nt. ing redients. and processing conditions on starch gelatinization .
`
`II. GELATINIZATION OF STA RCH
`
`Starch granules are insoluble in cold water but swell when heated in an aqueous medium .
`Initially . the swelling is reversible and the optical prope rties of the granule arc retained (e.g ..
`birefringence) . But when a certain tempe rature is reac hed. the swe lling becomes irreversible
`and the struc ture o f the granule is altered significantly. The process is called ' · gc la tinization · ·
`and the te mperature at which gelatinization occurs is railed the· ' gclati nization temperature "'.
`At this te mpe rature the granule loses its bire fringence and material fro m the granu le diffuses
`into the water. For a population of granules. the range for gclatinization te mperature is
`usually between 5 to I 0°C. indicating that fract ions of granules exhib it different gclat inization
`temperatures.
`Aside from swelling during gclatinization . the viscosity of the medium also increases.
`Both the molec ula r and g ranular structures contribute to the increase in viscosity. In itially.
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`gclatin ization occurs in the more accessible and amorphous regions. As the temperature is
`raised above that for initiation of gelatinization. intermolecular hydrogen bonds which main(cid:173)
`tain the structural integrity of the granule continue to be disrupted. Water molecules solvate
`the liberated hydroxyl groups and the granule continues to swell. As a consequence of severe
`disruption of hydrogen bonds, the granu le will be fu lly hydrated and finally the micellar
`network separates and diffuses into the aqueous medium . After disruption of the granules,
`the viscosity decreases. The increase and decrease of viscosity during gelatinization can be
`followed by a Brabender Amylograph. The increase in viscosity in the early heating stages
`is due mainly to the release of amylose while, in later stages. the continued viscosity increase
`is due to interaction of extragranular material and swelling of the granules.
`Marchant and Blanshard" postulated three constituent processes for starch gelatinization
`based on nonequilibrium thermodynamics: ( I) diffu sion of water into the starch granules,
`(2) a hydration-facilitated helix-coil trans ition which is a melting process, and (3) swelling
`as a result of c rystallite disintegration (melting). Blanchard" reported that total exchange of
`water between a starch granule and the environment at ambient temperature occurs in about
`I sec. Based on this observation and the temperature dependence of gelatinization , the
`diffusion process per se is not responsible for starch gelatinization. Using a light-scattering
`method to further study melting and swelling processes during gelatinization shows that
`gelatinization may be described as a "semicooperative process". Each starch granule has
`its own degree of crystallinity with its unique energy characteristics. The imposition of a
`2°C temperature rise may result in certain granu les being totally gelatinized but, in others,
`only some of the crystallites will have their gelatinization threshold exceeded. If the tem(cid:173)
`perature increase is applied continuously. eventually the whole population of granules will
`be gelatinized. Si nce the amorphous regions hydrate initially. French 22 has proposed that
`swelling of the amorphous phase which occurs when starch is heated in excess water con(cid:173)
`tributes to the disruption of the crystallite reg ions by tearing molecules from the crystallites.
`Based on changes in characteristics of starch granules (using a Brabender Amylograph)
`during and after heating in aqueous medium. Olkku and Rha49 summarized the steps of
`gelatinization as follows:
`
`Granules hydrate and swell to several times their original size.
`I .
`Granules lose their birefri ngence .
`2.
`Clarity of the mixture increases.
`3.
`4. Marked , rapid increase in consistency occurs and reaches a maximum.
`Linear molecules dissolve and diffuse from ruptured granules.
`5.
`Upon cooling. uni formly dispersed matrix forms a gel or paste-like mass.
`6.
`
`III . DETERMINATION OF DEGREE OF GELATINIZATION
`
`Degree of gelatinization can be determined qualitatively and/or quantitatively by physical,
`c hemical. and biochemical methods ."'·"'·"
`
`A. Birefringence End Point Method
`As was pointed out earlier, loss of birefringence is characteristic of starch that undergoes
`thermal gelatinization. Watson"' determined the degree of gelatinizat ion by measuring the
`percent loss of birefringence with a Koeller electrically heated microscopic hot stage and a
`polarizing microscope. In the procedure. 0. 1 to 0.2% aqueous starch suspension is prepared
`and a small drop of this suspension is spotted on a microscope slide and surrounded by a
`continuous ring of high-viscosity mineral oil. A cover slip is placed on the drop in such a
`way that air bubbles are not present under the cover g lass. A uniformly distributed field of
`about 100 to 200 granules is usually observed during the examination . The stage is heated
`uniformly at the rate of about 2°C/min through a variable transformer and the field is watched
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`to record the temperatures corresponding with loss of birefingence by 2. 10. 25. 50. 75.
`90. and 98% of the granules in the field . The 98% point is taken as the gelatinization
`temperature e nd-point (BEPT). In practice. usually on ly initial gelati ni zation and 98% loss(cid:173)
`temperature are recorded . Watson 's method was modified by Berry and White' to follow
`the progress of gelatinization by recording the light o utput on a photocell as a funct ion of
`hot stage temperature .
`
`B. Viscosity Method
`the most widely used method
`e.g .. in industry or commercially -
`In common practice -
`for determining the degree of gelatinization is based on changes in viscosity during gclat in(cid:173)
`ization. A viscoamylograph (or amy lograph) records the shear force induced o n a spindle
`as a func tio n of time/temperature when a starch slurry is rotated. The slurry is heated at a
`rate of I .5°C/min while the sample holder is rotated at speeds from 30 to 150 rpm: the
`granules swell and impinge on each other thus increasing the viscosity of the paste. However.
`when the integrity of the g ranule is lost. the viscosity decreases. In addition to measuring
`gclatinization temperatu re range. the instrument also provides information on temperatures
`of initial viscosity rise and maximum viscosity. time of cooking. and even viscosity changes
`during cooling if it is des ired.
`
`C. X-Ray Diffraction Method
`In certain cases. X-ray patterns can be used to differentiate between cereal and root starches
`and to detec t changes in crystallinity brought about by physical or chemical treatment of
`starch granules. The method has been used as a tool to measure the extent of gelatiniza(cid:173)
`tion.40·"·"' In principle. there arc two me thods of recording patterns in an X-ray diffraction
`unit: ( I) the diffractomete r method. in which rays scattered from the sample are recei ved
`by a Geiger- Muller counte r tube. the output then amplified and plotted by a chart recorder:
`(2) the photographic method. in which the pattern is recorded on a photographic film .
`X-rays arc a form of e lectromagnetic radiation with a wave length typically between 0. I
`and 1.0 nm ( I to 10 A). which is comparable to the molecular spacing in a crystal. When
`an X-ray beam impi nges o n a crystal which is held in a special mount that allows the crystal
`to be rotated with respect to the inc ident beam. diffract ion occurs. Diffraction is the phe(cid:173)
`nomenon that occurs whenever a wave motion interacts with an obstacle. The diffracted
`beams are recorded to obtain information on the structure of the crystal and the molecules
`within the crystal.
`In X-ray analysis of starch granules. satisfacto ry results are obtained if the specimens are
`properly prepared and mounted. Coarse powder gives less intense patterns. Therefore. the
`samples have to be less than 200 mesh (74 µm) in size and packed as densely as possible
`into a sample holder. The finished surface must be smooth and nush with the face of the
`sample holder. Upon recording the diffracted beams. the patterns are analyzed based on
`their interplanar spacings and relative intensities of the diffraction lines.
`
`D. Amylose/lodine Blue Value Method
`Amylose complexes with iodine to yield a brilliant blue color and this characteristic has
`been used as an analytical tool to measure amylose content. A quantitative method to
`determine the amount of amylase present in solution base has been developed by McCready
`' The absorbance of the blue color is
`and Hassid46 and modified by Gilbert and Spragg. 21
`measured with a spectrophotometer at 600 nm. However, it is s uspected that the blue colo r
`may not fully develop if retrogradation or incomplete solubilization of amylose is encoun(cid:173)
`tered. Under these conditio ns a more vigorous method of. mixing is required . Thus, the
`method that gives the maximum absorbance sho uld be chosen. In spite of these weaknesses,
`the iodine blue value method provides a rapid determination of amylose content that is
`adequate for most purposes.
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`Application of the amylase/ iodine blue complex method has been reported by Roberts et
`al. 57 as an index of parboiling in rice. Wootton et al .•• also utilized this method to detennine
`the degree of gelatinization of biscuits. However, the method was not applicable to parboiled
`rice since gelatinized rice flour was insoluble in water. Birch and Priestly7 modified this
`method by dissol ving the amylose in alkali solution and then subsequently neutralizing it
`with acid solution. The alkali treatment dissolves amylose in an aqueous solution . A critical
`concentration of alkali was found for gelatinized starch (0.2 N KOH) while for raw starch
`it was greater (0.5 N KOH) . This optimum concentration could be used to distinguish between
`raw and gelatinized starches. After addition of iodine reagents, the absorbance is measured
`with a spectrophotometer at 600 nm. The ratio of the absorbance of these two different
`mixtures (0.2 N vs. 0 .5 N KOH) is proportional to the degree of gelatinization.
`
`E. Other Methods
`Other methods in addition to those previously described are available, e.g., enzymatic
`digestability, 59 nuclear magnetic resonance ,37 light-extinction, 12 solubility or sedimentation
`of swollen granules,34 and absorption of congo red measurements." However, these methods
`are less popular due to the difficulties in their procedures or degree of accuracy in measuring
`the degree of gelatinization.
`
`IV . DIFFERENTIAL SCANNING CALORIMETRY (DSC) METHOD
`
`4 2
`•
`
`Stevens and Elton60 applied DSC to measure heat of gelatinization of starches, and since
`56 Banks and Greenwood3
`then many groups have applied DSC in the study of gelatinization.20•·
`suggested this method was valid by considering gelatinization to be the analog of a melting
`process for a crystal. The DSC method uses a small sample (IO to 20 mg at IO to 20% total
`solids) which minimizes the thennal lag within the system. Since the pans containing the
`sample are hennetically sealed, water will not be lost from the system under normal operating
`temperatures.
`The genn "differential scanning calorimetry" was initially a source of some confusion
`in thermal analysis. The parent technique to DSC is " differential thennal analysis (DTA)"
`and the indiscriminate use of the terms DSC and DTA resulted in IUPAC proposing defi(cid:173)
`nitions of these processes. Several excellent books on thermal analysis provide detailed
`infonnation on DTA and DSC, including McNaughton and Mortimer, 47 Wendlandt,"" Pope
`and Judd (Eds.)," and Mackenzie (Ed. ,). 41
`The purpose of differential thennal systems is to record the difference between an enthalpy
`change which occurs in a sample and that in some inert reference material when both are
`heated. Systems to accomplish this may be classified into three types: (I) classical OTA,
`(2) "Boersma" DTA , and (3) DSC.
`In the classical and " Boersma" DT A systems, both sample and reference are heated by
`a single heat source. Temperatures are measured by thennocouples embedded in the sample
`and reference material (classical), or attached to the pans (Boersma). The instrument measures
`the temperature difference between the sample and reference as a function of temperature
`and presents the data as a plot of AT vs. T . The magnitude of AT. at any given T (or time
`since the instrument is programmed to heat at a constant rate of temperature change, dT/dt)
`is a function of: (I) the enthalpy change, (2) the heat capacities. and (3) the total thermal
`resistance to heat flow. The thermal resistance to heat flow is dependent upon the nature of
`the sample, the way it is packed into the sample pan, and the extent of thermal contact
`between the sample pan and holder. In ''Boersma' ' DTA. the temperature sensors are attached
`directly to the pans in an attempt to reduce variations in thennal resistance caused by the
`sample itself. In either classical or Boersma OTA, the thennal resistance, and hence the
`calibration constant for the instrument, is a function of temperature. Although data from
`
`Case No.: IPR2023-00206
`U.S. Patent No. 9,943,096
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`DTA provide quantitative information on temperatures of transition. it is very difficult to
`transform the data to entha lpy changes. To calculate enthalpy changes from DTA data it is
`necessary to know heat capacities and the variation of the calibration constant w ith te m(cid:173)
`perature. Consequentl y OTA systems are not very suitable for calo rimetric measure me nts.
`The important difference between OTA and DSC is that in DSC the sample and reference
`arc each provided with individual heaters which a llows the determination to be conducted
`with no temperature difference between sample and reference. Running in this mode allows
`two important simplificatio ns compared to DTA. First. because the sample and rcfcrcm:c
`pans arc maintained at the same temperature. the calibration constant for the instrument is
`independent of te mperature. Obviously this greatly simplifies experimental techn ique sini:c
`the calibrat ion constant need only be determined for one standard material. Second. since
`the sample and reference pan have independe nt heaters. the differcni:c in heat flow to the
`sample and reference in order to maintai n equal temperature can be measured directl y. Thus
`data arc obtained in the form of differential heat in put (dH/dt) vs temperature (or time . since
`constant heating rates arc used). These data arc readily used to obtain temperatures and
`enthalpy of trans itions or reactions.
`Like any instrumental technique . there arc aspects o f the operatio n which must be i:arcfully
`attended to in o rder to obtain data with the desired precision and accuracy. DSC is no
`exception. especially since a calibration procedure must be used to obtain the calibratio n
`constant and fix the te mpe rature scale accurate ly.
`Most of the DSC units curre ntly in use have small sample pans which generally hold 10
`to 20 µ t' of sample. Obviously. for pure materials. obtaining a representative sample of this
`size does not present any great difficulty. For optimum peak sharpness and resolutio n. the
`contac t surface between pan and sample sho uld be maximized. which is usually accomplished
`by having the sample as thin discs, or films. or fine granules. Frequently. in applications
`in foods. the sample will be dispersed o r soluble in wate r. which obviates any proble m w ith
`contact surface. For heterogeneous materials, sampling can present a problem because of
`the sma ll quantity required. In these cases homogenizing the sample may be required. with
`care being exercised not to heat the sample during homogenizing (causing some irreversible
`transitions which arc being looked for in DSC).
`Generally, the samples are encapsulated in a luminum pans with lids that are crimped into
`positio n. For samples containing water. the pan must be hermetically scaled to prevent
`evaporation of water (with its large e nthalpy of vapori zation). Generall y the pans and
`crimping devices currently in use will withstand internal pressures to 2 to 3 atm . For higher
`pressures, there arc high-pressure DSC cells available from instrument manufacturers. and
`flint glass ampules can be used which withstand internal pressures up to 30 atm.
`Calibratio n of the instrume nt is generally carried o ut with a high-purity metal with ac(cid:173)
`curate ly know n enthalpy of fusion and me lting point. The most commonly used calibrant
`is indium (AH,.'"""' = 6 .80 cal/g; mp I 56.4°C). Mc Naughto n and Mortimer" discussed the
`procedure which should be used to obtain te mperature data from the cndo/cxothcrm to within
`± 0.2°C and ±: 0. 1°C.
`Determination of enthalpy for the process under study requires measure ment of the area
`of the endo/exotherm . This can present some difficulty because the baseline for the endo/
`exothe rm may not be horizontal (a functio n of the match of heat capacities of the sample
`and reference pans) and the peak is generally not symmetrical. An accepted procedure for
`determining the baseline for the peak is illustrated in Figure I . First. the baseline o n each
`side o f the peak is extrapolated across the peak . The linear portion of each side of the peak
`is then extrapo lated to intercept the extrapolated baseline o n its respective side (i.e . . right
`of peak extrapolated to inte rcept right baseline). The intersections (points T,. and T, o n
`Figure I) represent the initial and final temperature of the transition. The baseline for
`determinatio n of the peak area starts at the deviation of the pen from the left-hand baseline
`
`Case No.: IPR2023-00206
`U.S. Patent No. 9,943,096
`
`Motif Exhibit 1134, Page 10 of 30
`
`

`

`254
`
`CRC Critical Reviews in Food Science and Nutrition
`
`-
`
`TEtvffRATl.f£ or Tt..£ - - -
`
`FIGURE I . Measurement of endotherm area and temperatures of gelatinization on
`a DSC thermogram.
`
`and terminates at the return of the pen to the right-hand baseline. The area is indicated by
`the shaded area in Figure I .
`The three basic approaches to determination of the peak area are "cut and weigh",
`planimeter, and integrator. Each method has some disadvantages and all require experience
`by the experimenter. For "cut and weight'', it may be necessary to retrace the data onto
`paper with a uniform density and to allow the paper to reach constant moisture content
`before weighing. With a planimeter. it may be necessary to enlarge the peak to improve the
`accuracy of measuring the area. Enlarging can conveniently be done by Xeroxing the original
`tracing onto a transparency , showing the transparency onto a screen, and tracing the enlarged
`peak. The magnification factor can be easily determined from the grid on the DSC paper
`or by marking a known length on the transparency. Electronic integrators would certainly
`have the required accuracy for use with DSC data; however, it may be necessary to correct
`the data for a nonhorizontal baseline.
`In the last 10 years. DSC has become the analytical method of choice for studying starch
`gelatinization phenonema. The method can only be applied to separated starch or ground
`cereal grains because of the nature of the samples. Thus in situ starch gelatinization phe(cid:173)
`nomena must be studied by other techniques such as staining and enzymatic digestion
`susceptibility.
`
`Case No.: IPR2023-00206
`U.S. Patent No. 9,943,096
`
`Motif Exhibit 1134, Page 11 of 30
`
`

`

`Table I
`SUMMARY OF GELATINIZATION CHARACTERISTICS OBTAINED
`FROM VARIOUS STARCHES AND METHODS
`
`Volume 20, Issue 4
`
`255
`
`Temperature (' Cl
`
`Type
`starch Method
`
`Wheal Hol stage
`DSC
`Amylograph
`Turbidi1y
`POlalO HOI slage
`DSC
`Com Hol siage
`DSC
`Rice Hot s1agc
`DSC
`DSC
`
`Onset
`(T. )
`
`55
`52
`54
`55
`59
`57-59
`65
`65
`72
`70
`68
`
`Peak
`(T,)
`
`6 1
`59
`
`63
`71
`69
`70.6
`75
`76.3
`74
`
`Conclusion
`(T, )
`
`~H
`(cal/g)
`
`Rer.
`
`66
`65
`67
`95- 100
`68
`94-95
`76
`77
`79
`82
`79
`
`2.4
`
`5. 1- 5.5
`
`3.3
`
`3. 1
`3.0
`
`16
`16
`27
`3
`58
`60
`16
`16
`16
`16
`66
`
`V. COMPARISON OF METHODS TO DETERMINE GELATINIZATION
`CHARACTERISTICS
`
`Several attempts have been made to compare methods for determination of gelatinization
`characteristics -
`i.e., temperature and heat of gelatinization . Data from several investigators
`are presented in Table I for gelatinization of starch in excess water. The data show that, in
`general. the initial temperature (T.,,,,.,,) obtained by different methods for the same type of
`starch are about the same. However, T re•• and Tcondu,;00 obtained from DSC are slightly
`higher than those obtained by other methods. This could be due to a high heating rate that
`caused overshooting. The end point temperature for wheat starch gelatinization based on
`turbidity measurement was also much higher than that reported for other methods. This is
`understandable since the method measures the intensity of light transmitted through the starch
`solution as it is gradually heated. The largest light transmission was obtained when the starch
`granules were completely destroyed and dispersed into the medium .
`An important observation based on Table I is that starches are different in their gelatin(cid:173)
`ization characteristics. Wheat starch has a low temperature of gelatinization (54 to 65°C)
`and for rice. it is higher (7 1 to 80°C). Thus the cooking characteristics of the starches differ
`and will affect the design of industrial processes. Furthermore. the heat of gelatinization
`(AH in Table I) is dependent on starch source. Wheat has the lowest (2.4 cal/g) and potato
`the highest (5.3 cal/g) of the four sources in the table. Thus for every gram of potato starch
`gelatinizcd in a process. 5 .3 cal of heat are required. Since the specific heat of starches is
`only on the order of 0.2