' a may .~ 0'41; r} ,I;, n, flunk I.) be u. .fi. ' '1”: ,' ”Wilfifgifi: 14:2».- ‘f‘qm. mun-.5 ' n A: Law éwiil; ‘:—~‘r-., ¢ L'\ 5““? , I J11 2! - - v f ’ . ‘ ' > «add-:95?“ .- tax; ‘ . . ' . f“ v ‘ ‘» ~¢ ,a‘fiazfifi . ', ‘- "~91 S .‘ A; ‘ ‘. .. ' v’ 5‘." .1 -, ’;,. ‘\ : . . ‘sfigbp‘uguu 4-4;». m “ ":32, J-& .2 . z a V . ‘ . . ,. g” III >"""" .14" ”(if i". gfixiamsym y“ la: , ~ gig-$.33? "‘ Byrd .{v ‘ WE 'v ‘ ; : . . ‘1 ~‘ '\ 1.133% “‘ £ ‘ ‘ ' ‘ . , ' ' '1“ ‘w‘ ‘n‘ 1ficnv ~ 2 P . v , , , . . . . 1 ‘ A. . _ . . J r! '« ‘ ‘ ‘ ' 'nu.‘ :, ‘ ‘ , - > {"7 "Qf-v‘. "n R I .ling».gltx§ ‘ , ‘ m ' ‘ -' . -» " 4 ‘ ~ 'u . I; -. . a” " -w.v/4‘U ' 1.15.2.7” 1 _ l A A ' . ‘ , ‘ . . . ; 1.: , .' , 1" : , - , {X fizzy. I at, ‘Hv‘u‘fi 1' , I' - - 3w .‘ , ., ' ‘ ”my; . 9 ‘ W '9' my?“ ' x“ ' «1‘: ‘. . ‘ )1 :v V w _ q. ,\‘_ ‘ ‘ . - ‘4 , . v , ‘f‘éfly‘l “VIN“ h ‘ ¢ , r . I. . {v . ,, _ .. . ‘ "‘2. [5“.f. fir -_ _ ‘ - ~, . ‘ "I'L . 4-4. A .x- «W 3 . H“ 'giuflm... . . m » . f 1L0» 1‘ a, '. ‘ - h, h;- j. I‘ ‘ :- ni”.." '4" ~ A ‘ - ’ ‘ P" - "r?" '7" “ , f y A 1.: . . ‘ r ’- V‘H‘n' J30 )- . ‘ \ v . w H ‘4" , A M ' . A 4. ‘ «r ‘5 4 _; ‘ . ’1; .‘a \4 1. ‘ J -. . .r. . "*1 ,g‘ an Ivy-,5, .. , J‘Jn v ‘. . A ‘N' '4 I? 'I'“ fl . 1 5""; ' a.’ *3: fl I. I . ., ., .rmmm'uuuyu.omrm ‘ ,1, ,,,.... ‘ .Afiu: , Suu»r V' 0‘ . wins 173650 a 0 ‘t ‘{ '“llhll;lollLLJJlIW‘ um Michigan State University This is to certify that the thesis entitled THE EFFECT OF TEMPERATURE AND VIBRATION COMBINATIONS ON EMULSION STABILITY OF MAYONNAISE IN TWO DIFFERENT PACKAGE TYPES presented by Jeffrey Philip Mackson has been accepted towards fulfillment of the requirements for Master of Science degree in Packaging 6/454 Major prfl 0-7639 MS U is an Affirmative Action/Equal Opportunity Institution PLACE IN RETURN BOX to remove this checkout from your record. TO AVOID FINES return on or before die the. IDA DUE DATE DUE DATE DUE l ___J ____i fiV—l—j MSU Is An Affirmative Action/Equal Opportunity lnetltutlon THE EFFECT OF TEMPERATURE AND VIBRATION COMBINATIONS ON EMULSION STABILITY OF MAYONNAISE IN TWO DIFFERENT PACKAGE TYPES BY Jeffrey Philip Mackson A THESIS Submitted to Michigan State University in partial fulfillment of the reguirments for the degree of MASTER OF SCIENCE School of Packaging 1989 @64-l23i ABSTRACT THE EFFECT OF TWERATURE AND VIBRATION COMBINATIONS ON EMULSION STABILITY OF MAYONNAISE IN TWO DIFFERENT PACKAGE TYPES BY Jeffrey Philip Mackson Mayonnaise in two different packages was compared for its suseptability to vibration induced breakdown. The study investigated the effect of truck vibration on emulsion stability and compared two different package systems. The containers were 32 oz. glass and 32 oz. polyethylene terephthalate (PET) . Mayonnaise was conditioned at three different temperatures 40, 72, and 100 P, for three time periods 21, 26, and 36 days, and vibrated at normal and worst-case transportation 9- levels. Emulsion stability was quantified by specific gravity, percent of surface oil, and qualitatively rated by a visual pass- fail test. The specific gravity measure positively correlated to surface oiling. . A consistent difference in stability was found between plastic and glass only at the 1.09 level. Glass showed a higher rate of failure at that level. The typical transit combinations were established to be 0.75g at 40 F and 0.759 at 72 P. Mayonnaise contained in plastic showed a higher trend towards emulsion failure at those levels. ACHIOWLEDGMENTS I would like to acknowledge Dr. S.P. Singh for setting up the experimental design, guidance through the statistical evaluation, and in obtaining the product from the Kraft company. I would like to thank the Kraft company for the mayonnaise and the funding for this research. It‘ll}, l'll‘ i TABLE OF CONTENTS LIST OF TABLES LIST OF FIGURES 1.0 INTRODUCTION 2.0 REVIEW OF LITERATURE 2.1 Emulsifiers 2.2 Emulsion Stability 2.2.1 Interfacial Film 2.2.2 Kinetic Study 2.3 Emulsion Failure 2.3.1 Sedimentation 2.3.2 Phase Inversion 2.3.3 Demulsification 2.4 Temperature Influences 2.5 Vibration Influences 2.6 Testing Techniques 3.0 EXPERIMENTAL DESIGN 3.1 Product 3.2 Packages 3.2.1 Glass jar 3.2.2 PET jar 3.2.3 Case pack 3.3 Equipment 3.4 Temperature Selection 3.5 Vibration Selection 3.6 Time Periods iv Page vi vii 3.7 Methodology 3.8 Sampling RESULTS AND DISCUSSION Data Calculations Quantitative Results 1 Series A and B Series C Correlations Comparisons Stability Trends 2 3 .4 .5 Qualitative Results .1 Comparing qualitative results CONCLUSIONS APPENDICES A. Calculation method for specific gravity and percent oiling B. Cross weight of sample in pycnometer C. Weight of surface oil versus weight of centrifuged sample D. Analysis of variance test results E. Minimum significant difference calculation for specific gravity and surface oiling F. Standard deviation for specific gravity and surface oiling measurements G. Gross weights of empty pycnometer and pycnometer with distilled water LIST or REFERENCES 21 22 56 S7 66 73 77 79 81 82 LIST OF TABLES Table 1 Number of days of conditioning and number of vibration sessions given to the series. 2 Total number of containers tested at each temperature, g-level, and conditioning time. 3 Resonant frequencies of the test samples. 4 Averaged specific gravity of mayonnaise for all plastic and glass test containers evaluated. 5 Averaged percent surface oil of mayonnaise for all plastic and glass test containers evaluated. 6 Comparison of Series A-B containers for significant differences. 7 Comparison of Series C containers for significant differences. 8 Visual pass-fail rating for all plastic and glass test containers. vi Page 20 27 30 31 32 35 37 49 LIST OF FIGURES Figure Page 1 Pynometer................ ....... . ................... 24 2 Fresh and demulsified mayonnaise....... ............. 25 3 Trends in specific gravity for Series A............. 40 4 Trends in specific gravity for Series B ............. 41 5 Trends in specific gravity for Series C....... ...... 42 6 Trends in percent oiling for Series A... ............ 43 7 Trends in percent oiling for Series B............... 44 8 Trends in percent oiling for Series C............... 45 9 Percent containers passing visual test for Series A. 50 10 Percent containers passing visual test for Series B. 51 11 Percent containers passing visual test for Series C. 52 vii 1.0 INTRODUCTION The trend of moving from glass to plastic in consumer grocery items is increasing. Ketchup, peanut butter, and pickles are popular examples of this transition from glass to plastics and plastic laminates. This benefits the producer through decreased shipping weight and easier stocking, and the consumer by offering a "shatterproof" feature and easier handling. It is hypothesized that the physical differences between glass and plastic containers could cause variation in emulsion stability of mayonnaise ‘when the two types of jpackages are subjected to identical vibration and storage combinations. Temperature and vibration have an influence on mayonnaise stability, but it is the visual implications seen on the surface of the product which directly effect the consumer and make the product acceptable. We are concerned with discovering if these influences have discernable differences when applied to identical mayonnaise product in glass versus plastic packages. The objectives of this research were: i) to compare differences in mayonnaise emulsion stability by measuring specific gravity on the top surface between identically l vibrated and temperature conditioned polyethylene terephthalate (PET) and glass containers. ii) to compare .the’ percent of visible surface oiling between identically vibrated and temperature conditioned glass and PET containers. iii) to qualitatively compare the mayonnaises' visual acceptability between identically vibrated and temperature conditioned glass and PET containers. 2.0 REVIEW OF LITERATURE The following section is a brief overview of oil in water emulsions: the forces that hold them together, classification of separation mechanisms, and the influence of temperature and vibration upon emulsion stability. It must be recognized that many variations exist in emulsion science terminology. Sutheim (1946) gives one of the most appropriate definitions: An emulsion is a heterogeneous system, consisting of at least one immiscible liquid intimately dispersed in another in the form of droplets, whose diameters, in general, exceed one micrometer. Such systems possess a minimal stability, which may be accentuated by such additives as surface-active agents, finely-divided solids, etc. . Emulsions are classified as being oil in water (herein referred to as O/WI or water in oil (herein referred to as W/O). Mayonnaise is a O/W emulsion. This means that the dispersed phase is oil and the continuous phase is water. Through mechanical input and chemical emulsifiers the two immiscible liquids in mayonnaise may exist as one system. Any emulsion is inherently unstable and will eventually break down to form two layers. Mayonnaise is a particularly delicate food emulsion because for a one system it contains a very large amount of oil. High amounts of oil do not favor O/W emulsions (Becher,1985). Considerable research has been devoted to theoretical considerations of emulsion stability in general, but 4 in comparison little has been published concerning practical aspects of mayonnaise stability. One of the few papers of this type was published by Corran (1948), who reported on the effect of product formulation on the stability of mayonnaise. The review is broken into six segments: 2.1) Emulsifiers 2.2) Emulsion stability 2.3) Emulsion failure 2.4) Temperature influences 2.5) Vibration influences 2.6) Measurement techniques 2.1 Emulsifiers The FDA uses the following definition for mayonnaise: Mayonnaise is a semisolid emulsified food prepared from edible vegetable oil, water, acidifying ingredients and yolk-containing ingredients. It must contain not less than 65 percent‘by'weight of oil. It may contain salt, sweetening ingredients and suitable harmless food seasonings or flavorings that do not impart color simulation the color of egg yolk. The acidifying agents may include vinegar, lemon juice, or lime juice. The egg-yolk containing ingredients may be liquid, frozen or dried egg yolk or whole eggs or any of the foregoing mixed with liquid or frozen egg white. It may contain calcium disodium EDTA or disodium EDTA or both. (0.5. FDA,1964) In mayonnaise the emulsifying agent is a complex of lecithin and protein from egg yolk (Chang and Fennema,1972). Due to the large positive free energy at the interface of the two liquids, emulsions are thermodynamically unstable. They require the help of emulsifiers and sometimes other compounds to remain stable. 5 To produce stable emulsions, the tendency to minimize interfacial area through coalescence must be counteracted and this is usually accomplished by adding emulsifiers (King,1941) . Most emulsifying agents are amphiphilic compounds. They concentrate at the oil- water interface causing a significant. lowering of interfacial tension, providing a physical resistance to coalescence, and sometimes increase surface charge (Becher,1977). Emulsifiers in an O/W system surround the dispersed oil droplets allowing them to remain dispersed throughout the continuous water phase. Lecithin is a'common name for 3-sn-phosphatidylcholine. Instead of being a simple triglyceride, one position on the glycerine is occupied by the substituted phosphoric acid. Only a few fatty acids are found in lecithin: those fatty acids are adsorbed at the oil droplet surface leaving the carboxylic heads penetrating through the interface into the water phase. The carboxylic ends are ionized forming at the surface of the droplet a coating of negative charges. This coating sets up what is referred to as the electrical double layer. Emulsion stability is often attributable to the presence of repulsive electrical charges on the surfaces of emulsion droplets (Becher,l977). Dispersed particles are subject to two independent forces: the van der Waals force of attraction and the electrostatic force of repulsion arising from the presence of electrical double-layers at the particle surfaces. If the repulsion potential exceeds the attraction potential an energy barrier opposing collisions results; if the magnitude of this 6 energy barrier exceeds the kinetic energy of the particles, the suspension is stable (Friberg,1976). That is the case in mayonnaise. More detail on this subject is discussed by Friberg using the (Derj aguin, Landau, Verwey, and Overbeek) DLVO theory. Also contained in mayonnaise are finely ground particulates of onion. Finely ground particulates like sugar, salt, onion and other spices function as O/W stabilizers by lowering the surface tension (Chang et a1,1972). Solid particles of very small size, as compared to the size of the dispersed oil droplet, can stabilize an emulsion by adsorbing at the interface to form a physical barrier around the droplets (Fennema,1985) . 2.2 Emulsion Stability Emulsion formation and stability is much too complex to be explained by a single coherent theory. The literature reviewed, by in large, has been limited to specialized systems and what has been true for certain O/W systems under certain conditions is not necessarily applicable to other 0/w systems. This is especially true with food emulsions where ingredient choice is limited (by regulation) yet varies by taste preferences throughout different regions of the country. Hence, results from studies on similar products, while interesting for comparison, must be looked at cautiously in their relationship to mayonnaise. The primary concern to the manufacturer is the end effect of a distribution cycle upon the surface integrity of the mayonnaise. 7 To understand the end effects on the stability of mayonnaise, and hence its surface, it is necessary to understand emulsion microstructure. The investigation of emulsion microstructure basically falls into two categories (Tomoyoshi,1983). These are: -investigation of the thinning of the liquid film between the globules and -kinetic study of coagulation of the globules. 2.2.1 Interfacial Film Emulsifiers, as they orient at the oil-water interface, will form an interfacial film. The nature of this film is important in determining the stability of the emulsion. In order to understand the behavior of these films one needs to consider two aspects of their physics: (1) the nature of the forces acting across the film - these determine whether the film is thermodynamically stable, metastable, or unstable and (2) the kinetic aspects associated with local (thermal or mechanical) fluctions in the film thickness. Details on the physics of film behavior is addressed by Tadros and Vincent, (Becher,1983.) Influences affecting the film are important because before two droplets unite the film separating them must rupture. Film thinning is a precursor to rupturing (Brown,1968). Becher (1977) pointed out that the rupture of the film is most likely to occur at the thinnest portion and that at the interface the thickness is likely to vary as a result of vibrations or thermal effects. 8 Some of the variables affecting film thinning are summarized by Woods (1972). These are: - the viscosity and density of the two phases - the interfacial tension and its gradient - the interfacial shear, dilational viscosities and elasticities - the droplet sizes - electrostatic (van der Waals) forces - electrical double layer interaction forces and other forces acting between the phases. 2.2.2 Kinetic Study The second category describes the kinetic study of coagulation of globules. Understanding globule kinetics and the forces influencing them is essential for determining the likelihood of temperature or vibration having an effect on stability. The motion and influences upon globule motion leading to aggregation and interaction between droplets is another popular method of examining O/W microstructure. When viewed under a microscope the particles which make up a_fine suspension are seen to undergo a continuous random motion. This Brownian motion increases the probability of interparticle collisions and hence of coagulation/coalescence (Becher,1977). This effect is not significant for emulsions with larger sized particles (greater than four micrometers). However, this could have as effect on mayonnaise which has an average globule diameter of about 2.2 micrometers (Chang et al.1972). A popular method of researching coagulation kinetics, and hence O/W 9 emulsion stability, is the examination of globule size and distribution within a system and their affects on the rate of coagulation (Tomoyoshi,1983). Understanding globule kinetics and the forces influencing them is essential for determining the likelihood of temperature or vibration having an effect on stability. 2.3 Emulsion Failure A brief explanation of separation mechanisms (Becher,1985). It has been said that an emulsion becomes stable only when it is broken. Thermodynamically this is true since this is the state of least energy. Broken emulsions, however, result in consumer rejection based on low visual acceptance. Over time any emulsion will deteriorate. Temperature changes, transportation and packaging all play a role in how this process accelerates. The phenomenon of emulsion separation can be broadly categorized into three parts. These include, -Sedimentation -Inversion -Demulsification These three categories do not always occur exclusively from each other. Demulsification, for instance, often occurs after sedimentation or inversion. They are discussed independently in this text. 10 2.3.1 Sedimentation Sedimentation, or creaming, is a process where one emulsion is separated into two emulsions. One is rich in dispersed phase and the other lacking in the dispersed phase. There is no change in the basic droplet size, similar sized droplets simply move to their own level forming a droplet concentration gradient. This phenomenon is not a breaking of the emulsion but it can result in consumer rejection due to aesthetics resulting from phase variation. 2.3.2 Phase Inversion Phase inversion occurs when an O/W system changes, or inverts, to a W/o system. Similarly, this occurrence is not a breaking of the emulsion but it can result in visual and or functional problems. 2.3.3 Demulsification Demulsification is a true breaking of an emulsion. It occurs in two stages: flocculation and coalescence. Flocculation is the build up of aggregates of droplets within the emulsion, the individual droplets still retaining their identity. In the second stage, two or more droplets combine forming a larger droplet. The decreased number of droplets (resulting from coalescence) leads to coagulation of the dispersed phase (in mayonnaise this is oil) and hence separated (surface) oil. Another type of demulsification phenomenon is Ostwald ripening. Ostwald ripening is the result of the system. trying to equilibrate, allowing all the droplets to attain the same size. 11 It should be noted that some authors make a distinction between complete breaking and a slower form of breaking called oiling. Whereas breaking is generally clear cut, oiling is more subtle and it may slowly become worse. This evidence of instability can generally be recognized by a small accumulation of oil at the interface of the container, air, and liquid (Carroll,1976). To conclude this discussion of emulsion failure it must be mentioned that the process of defining the exact nature of breakdown or separation mechanism in food emulsions is difficult. One major difficulty in the experimental study of emulsion breakdown is the isolation of the phenomenon described above. In practical systems, one or more processes may occur simultaneously or sequentially depending on the rate constants for the individual processes (Becher,1977). Even in fundamental studies, the isolation of the four processes has been experimentally difficult to achieve. 2.4 Temperature Influences The effect of low temperature on O/W emulsions is well researched. Freezing breaks emulsions by destroying the membrane of emulsifying agent surrounding each globule (Rochow,1936) . Whereas simply lowering temperature has been shown to increase stability in an O/W . emulsion (Reddy,1981). In experiments conducted by Gillespie and Rideal,(l956) the 12 probability of interfacial film rupture was measured for various temperatures. They found that small temperature fluctuations markedly affected the lifetimes of drops due to its affect on the continuous phase film. Increases in temperature can affect the behavior of emulsions considerably. In most cases an increase in temperature results in a decrease in emulsion stability. An increase in temperature increases the rate of adsorption, the van der Waals forces, Brownian motion, and decreases the viscosity of the continuous phase film all of which cause instability in the emulsion (Carroll,1976). 2.5 Vibration Influences The literature review indicates that no previous studies have been done describing the effect of vibration on mayonnaise in glass or PET packages. No studies were found relating differences in a materials coefficient of friction to the stability of O/W emulsions. Owing to the complexity of typical food systems and the relatively low level of basic research in this area, little information exists in literature about mayonnaise, salad dressing, and low calorie dressings (Becher,1985). Similarly, little information exists in literature about the effect of transit level vibration on food emulsions. Richmond (1984), addressed yogurt stability to vibration. Detailed research on the effect of vibration upon emulsion separation has been limited to ultrasonic 13 vibration, as a means of purposely destabilizing a system. The effect of plastic packaging on mayonnaise has been restricted to the study of oxidation rates and plastic defamation resulting from adsorption of oil from a O/W emulsion. In general, the major factors that could affect emulsion failure are: - Improper ratio of oil and water phases - Incorrect choice or amount of emulsifier - Improper mixing - Impurities in oil, water, or emulsifier - Reaction between two or more components and or the container - Temperature extremes - Mechanical shock or vibration (Bennett,l977) 2.6 Testing Techniques various instruments have been devised for testing the consistency of the mayonnaise emulsion. One device used is a modified form of the Gardner mobilometer. The mobilometer is a special form of viscometer in which a plunger equipped with orifices is pressed through the sample under a constant force. Another device used is a special orifice-type viscometer which utilizes air pressure to discharge the sample through a standard orifice over a given time (Brown,1968). Probably the most widely used technique is measuring viscosity with 14 a Brookfield viscometer on a helipath stand. This measures the resistance of mayonnaise while continuously cutting downward into fresh sample. More complex methods developed for general use in emulsions have been developed. Turbidimetric methods, determining phase separation by reading light transmitted through a sample, have also been used to detect separation. Scanning electron microscopy has been used to monitor oil droplet size, configuration, and distribution. The incorporation of radioactive materials into the dispersed phase has been suggested by Banker (1962) as a method for determining rate separation and the general homogeneity of emulsions . Emulsion stability is coumonly measured in terms of the amounts of oil separating from an emulsion during centrifugation (Wang,l966) . Centrifugation is a direct and fundamental measure of demulsification. It was noted by Bennett (1977) , that the specific gravity of the two phases influences the stability of the emulsion. These non-visible changes in stability can be monitored by specific gravity measurements of the emulsion system. 3.0 EXPERIMENTAL DESIGN 3.1 Product The test product used for this study was regular mayonnaise. The ingredients in the test product (obtained from a major manufacturer) were: soybean oil, eggs, vinegar, water, egg yolks, salt, sugar, lemon juice, paprika, dried onion, dried garlic, calcium disodium EDTA, natural flavor. The oil content, specified. by the manufacturer, was 70 percent by weight. The resuts of the test may vary for a "lite" variety or "low cholesterol" type of mayonnaise due to different oil content. 3.2 Packages Container descriptions obtained from the manufacturer: 3.2.1) Glass jar The glass jar' was the 3202. wide mouth. with the following specifications. opening diameter- 2.0 in. bottom diameter- 3.0 in. inner diameter- 3.75 in. wall thickness .115‘I.01 in. 2.75 lb. weight full lid - metal 15 16 3.2.2) Plastic jar The plastic jar was injection blow molded PET. opening diameter - 3.0 in. bottom diameter - 3.0 in. inner diameter - 3.75 in. Mid-section wall thickness .022 in.t:.003 Shoulder wall thickness - .055 in. I .003 weight full 1.9 lb. lid - LDPE 3.2.3) Case pack The regular slotted container (RSC) was a C-flute, 175 lb. single wall. Its dimensions are 16x12x7 in.. Twelve jars are packed, four to a side and three on the ends of each case. 3.3 Equipment The testing equipment used in this study consisted of the following: i) The vibration system was a model 840 MTS electrohydraulic vibration table. ii) The pycnometer was a Fisher Scientific. iii) The balance was a Mettler model AE 160. iv) The centrifuge was a Sargent model CM, size 1. 17 3.4 Temperature Selection Three storage conditions were selected. Both plastic and glass jars were conditioned under these temperatures. 40 degrees Fahrenheit and 50% relative humidity to represent typical refrigerated truck conditions. 72 degrees Fahrenheit and 50% relative humidity to represent typical room conditions. 100 degrees Fahrenheit and 40% relative humidity to represent accelerated conditions. All conditioning temperatures were controlled within.t 2 degrees Fahrenheit. All relative humidities were controlled within t 4%. All vibration tests were done at 72 F and 50% relative humidity. A set of control samples not subjected to any vibration and described by 0.00 g, were maintained at each of the conditioning temperatures. All measurements were done at a mayonnaise temperature of 74 F I 1 F. 3.5 Vibration Selection For typical trucks, the vibration data shows that the input acceleration values most commonly occur near 0.59's (Ostrem,1985.) But, because of magnification factors, the g-levels typically experienced by stacked, filled shipping units is higher. Random vibration testing has shown the presence of transient truck 9- C levels as high as 10.0 g's (Marcondes 1988). The g-levels selected for this study were "typical levels", of 0.5 and 0.75 g's, and a 18 "worst case level" of 1.09. These levels represent continuous vibration forces experienced by a stack. Only plastic jars were vibrated at the 0.59. It was assumed that because 0.59 is a low level for stacked packages, there would be no qualitative change of significance in either container. For plastic it would be helpful to have this lower 9-level for establishing trends in specific gravity. It was not included in the statistical analysis. During the first ten days before vibration testing, a single sample case containing glass or plastic jars was vibrated at the assigned 9-levels to determine the resonant frequencies (state of maximum vibration magnification). Both plastic and glass resonant frequencies at 0.759 and 1.09 were determined by performing a frequency sweep from 3 Hertz to 50 Hertz. The resonant frequencies were easily pinpointed by visually locating the frequency of maximum movement of the RSC on the table. The point of maximum vertical container movement was choosen as the resonant frequency. The resonant frequency values obtained (see Table 3) for the cases at 0.75 and 1.0 9 are within the typical frequency range found in comercial truck shipments (ASTM D 4728- 87). During the test situations, the cases were vibrated at their resonant frequency (at a constant input of 0.5, 0.75 or 1.09) for a dwell time of 30 minutes each. For all vibration sessions, plastic and glass cases were taken from each temperature for each 9-level to be tested and evaluated. The cases were vibrated three 19 at a time, (eg. at 0.75 9, and 24 Hertz the 40F, 72F, 100F cases of plastic were on the table; next at 0.75 9, and 28 Hertz the 40F, 72F, lOOF cases of glass were on the table) all cases were laying flat on vibration table. Hence for any one test session 15 cases were vibrated: Plastic 3 temperatures x 3 g-level = 9 Glass 3 temperatures x 2 g-level = 6 total 15 cases 3.6 Time Periods In this research the three different groups of test conditions are referred to as Series A, B, and C. The set of containers under temperature control for 21 days are defined as Series A; for 26 days as Series B; and for 36 days as Series C. Series A and B had two vibration sessions to model a typical distribution situation: plant toldistribution center ride (first session) and distribution center to store ride (second session). The C-series was given twice as many vibration sessions to model a worst case situation. The maximum amount of time the cases spend in a typical distribution cycle is about five weeks. Hence, the maximum time under conditioning was designed.to be limited to about five weeks. 20 Table 1. Number of days of conditioning and number of vibration sessions given to the series. Series # of Days of vibration conditioning sessions - before vib. total one A 2 10 21 B 2 16 26 21 3.7 Methodology The methods of quantification of emulsion stability used for this study were percent weight by weight surface oiling and specific gravity. - Percent wt/wt separation. This is a fundamental and direct measure of demulsification (Wang,1966). Identically removed samples, from.the surface of the jar, are weighed and then centrifuged. The oil drawn out [of centrifuge tube is weighed in a disposable pipette. (The technique is described in section 3.8.) - Specific gravity. Subtle changes in the emulsion result from a change in the molecular matrix. As mayonnaise breaks down the relationship between the oil, water, and.air in the system changes. As the free energy of the system is lowered (in an attempt to become demulsified and more stable) bonding between these components changes. These non-visible changes in stability were monitored by specific gravity measurements (the technique is described in section 3.8). Hence, greater the amount of emulsion breakdown or "oiling", greater is the value:of specific gravity of the separated emulsion. Specific gravity is measured by using a pycnometer. A pycnometer is a precisely machine tooled stainless steel device used to weigh a fixed volume of any liquid (See Figure 1). 22 3.8 Sampling The critical portion of the product, from the consumer point of view, is the surface. Hence the tested portion of mayonnaise was taken from the first 1cm (depth) of surface for both specific gravity and percent oiling measurements. Eight jars per case were selected randomly for testing. Each jar was visually rated. Four of these jars were randomly assigned to be centrifuged; all eight were weighed by pycnometer. The procedure, identical for each series is described: i) Photographic documentation of open containers showing top surface (see Figure 2). i ii) Visual rating. Each jar was given either an acceptable or unacceptable rating depending on the amount of visible surface oil. The rating was done by the author. For the sample to be rejected it had to have oil overtly evident. iii) Centrifugation and specific gravity. Of‘the eight assigned containers four were chosen at random to be centrifuged. The procedure for centrifugation was as follows. From one side of the container, half of the top portion of mayonnaise was removed with a spatula down to about 1 cm. depth. With that half of the top portion removed, a specially cut plastic cupette was lowered into the jar (into the space where the 1 cm. of product had been removed) scooping out (by moving cup from center to edge of jar) the remaining first 1 cm. of untouched product. The scooped~ portion of mayonnaise was weighed. The weight of the sample varied from about 10 grams to about 15 grams. The sample was transferred 23 to a centrifuge tube and centrifuged at 3000 r.p.m. for 30 minutes. A disposable glass pipette was tared on the balance. Any visible oiling on top of centrifuge tube was drawn off in the disposable pipette and the pipette was weighed. The oil measurement is expressed as a percent. All eight containers had one sample taken from the first centimeter of surface depth for pycnometer weighing. . /‘\- Figure 1. Pycnometer 25 Plastic Glass Figure 2. Fresh and demulsified mayonnaise 26 The experimental design setup is described in Table 2. 27 Table 2. Total number of containers tested at each temperature, 9-leve1, and conditioning time. Days of conditioning 21 26 36 Plastic Glass Plastic Glass Plastic Glass Temp. 40 ' 72 100 g-level 0.00 4 4 4 4 4 4 0.50 12 - 12 - 12 - 0.75 ' 12 12 12 12 12 12 1.00 12 12 12 12 12 12 0.00 4 4 4 4 4 4 0.50 12 - 12 - 12 - 0.75 12 12 12 12 12 12 1.00 12 12 12 12 12 12 0.00 4 4 4 4 4 4 0.50 12 - 12 - 12 - 0.75 12 12 12 12 12 12 1.00 12 12 12 12 12 12 4.0 RESULTS AND DISCUSSION 4.1 Data The data for specific gravity may be found in Appendix B Tables B1-B9, and the raw data for percent oiling in Appendix C Tables C1-C6. 4.2 Calculations The data collected was used to calculate as specific gravity and percent oiling following the procedure outlined in Appendix A. Specific gravity is the ratio of the mass of mayonnaise to the mass of an equal volume of water at the same temperature. The percent surface oil is the weight of oil measured after centrifugation divided by the weight of sample centrifuged. To derive the most accurate comparison between the containers, the first two series (A and B) were compared independently from the third (C) because of the different number of vibration sessions. Because of its subjective nature, the results of visual data are presented only as simple percentage comparisons. The 0.59 level is not included in any statistical evaluation. The 0.5 g averaged numbers for specific gravity and percent oiling have been given with Tables 4 and 5. Where they are presented it is strictly to help in establishing trends. The analysis of variance was done on a statistical program called Statistical Package for the Social Sciences (S.P.S.S.). From the" 28 29 calculations two important values are given the F-value and the mean squared error (MSE). The MSE allows development of a minimum significant difference (MSD) value. (See Appendix E for derivation of MSD.) The MSD value allows a comparison of mean values for plastic containers (of specific gravity and percent oiling) to mean values for glass containers and see if container types differ significantly in their response to the vibration levels. In addition, a Pearson correlation was done for both Series A-B data and Series C data to find out how well the specific'gravity measure relates to the percent oiling. 4.3 Quantitative Results Table 3 shows the resonant frequencies obtained for the glass and plastic cases. Tables 4 shows the means calculated from the eight measurements of specific gravity for both plastic and glass jars. Table 5 shows the means calculated from the four measurements of the percent surface oil for both containers. Following Table 5, is an explanation of the data analysis. A summary of the statistical analysis is provided in Tables 6 and 7. 30 Table 3. Resonant frequencies of the test samples. Series Container g-level Vibration Sessions Resonant Frequency (Hz) 1 2 3 4 . A Plastic 0.50 44 44 - - 0.75 24 23 - - 1.00 24 24 - - A Class 0.75 28 28 - - 1.00 24 22 - - B PlaStic 0.50 44 44 - - 0.75 27 27 - - 1.00 24 23 - - B Class 0.75 26 27 - 4 1.00 24 22 - - c Plastic 0.50 44 44 44 44 0.75 27 27 26 26 1.00 23 23 26 26 C Class 0.75 26 26 26 23 1.00 24 22 22 22 *The primary resonance frequency will vary for the various test conditions. This is due to the effect of storage temperature on the corrugated fibers and crushing of flutes during the first dwell. Table 4 . 31 and glass test containers evaluated. Averaged specific gravity of mayonnaise for all plastic Plastic Glass Series Acc. Temperature (F) 40 72 100 40 72 100 A 0.00 .8910 .8910 .8940 .8900 .8900 .8971 0.50 .8913 .8946 .9050 - - - 0.75 .8912 .8887 .9064 .8932 .8947 .8989 1.00 .8968 .8989 .9089 .8959 .9055 .9127 B 0.00 .8977 .8961 .9020 .8970 .8966 .9012 0.50 .8996 .8968 .9178 - - - 0.75 .9080 .8989 .9188 .8960 .8945 .9047 1.00 .9139 .9010 .9178 .9026 .9073 .9167 C 0.00 .8976 .9005 .9120 .9010 .8960 .9028 0050 09019 08982 09147 - - - 0.75 .9011 .8976 .9162 .8991 .8954 .9049 1.00 .9122 .9136 .9211 .9030 .9106 .9213 32 Table 5. Averaged percent surface oil of mayonnaise for all plastic and glass test containers evaluated. Plastic Glass Series Acc. Temperature (F) 40 72 100 40 72 100 A ' 0.00 0 0 0 0 0 0 0.50 - 0 0 0 - - - 0.75 0 0 1.00 0 0 0 1.00 .68 .50 3.97 .66 3.32 5.32 B 0.00 0 0 0 0 0 0 0.50 0 0 0 - - - 0.75 0 0 .76 0 0 0 1.00 .65 1.33 5.04 1.19 3.19 3.62 C 0.00 0 0 0 0 0 0 0.75 .50 .75 .70 0 1.07 .98 1.00 .94 1.71 5.60 1.56 4.94 6.37 33 4.3.1 Series A88 A 4-way analysis of variance was done on specific gravity and percent oiling to discover if there was significant interaction between the independent variables (temperature, time of conditioning, g-1evel, and container type) and the (dependant variables (specific gravity and oiling) used as the stability indicators. The statistical evaluation is provided in Appendix D Tables D1 and D2. The F-value and the significance of F value show a high level of interaction between the dependant variables and all independent variables. Hence, no general conclusion about differences between plastic and glass should be made from these observations. Any comparison of plastic to glass should be specific to a particular temperature, 9-level and time of conditioning if one wants to draw conclusions about differences with statistical confidence. The difference between the plastic and glass mean values (for specific gravity or percent oiling at any time-temperature-vibration combination) is compared with the MSD value calculated at a 95% confidence level (See appendix E). If the difference between the plastic and glass mean value is greater than or equal to the MSD value then one can say, with 95% confidence, that for the stability measure (specific gravity or percent oiling) at a particular 9- level, temperature, and length of storage, one type of container of mayonnaise demulsified more than the other. The comparisons 34 between containers in the A and 8 series are summarized in Table 6. 35 Table 6. Comparison of Series A-B containers for significant differences. Series Temp. g-level Significant Container Difference extreme* 5.6. 40 0.75 n 1.00 n 72 0.75 y glass 1.00 n 100 0.75 y plastic 1.00 n P.O. 40 0.75 n 1.00 n 72 0.75 n 1.00 y glass 100 0.75 y plastic 1.00 y glass 5.6. 40 0.75 y plastic 1.00 y plastic 72 0.75 n 1.00 y glass 100 0.75 y plastic 1.00 n P.O. 40 0.75 n 1.00 y glass 72 0.75 n 1.00 y glass 100 0.75 n 1.00 y plastic S.G. specific Gravity Comparison P.O. Percent Oiling Comparison *The container showing higher demulsification. 36 4.3.2 Series C This analysis follows the same procedure of comparing MSD to mean difference as jpreviously' explained. Recall C-series had four vibration sessions. A 3-way analysis of variance is applied to this data set, because time ceased to be an independent variable. The statistical evaluation is given in Appendix D (Tables D3 and D4). Table 7 summarizes the comparisons between containers in the C series. 37 Table 7. Comparison of Series C containers for significant differences. Series Temp. g-level Significant Container Difference extreme* S G. C 40 0.75 n 1.00 y plastic 72 0.75 n 1.00 n 100 0.75 y plastic 1.00 n P O. C 40 0.75 n 1.00 n 72 0.75 y glass 1.00 y glass 100 0.75 n 1.00 n 5.6. Specific Gravity Comparison P.O. Percent Oiling Comparison *The container showing higher demulsification. 38 4.3.3 Correlations The Pearson correlation shows the degree of linear relationship between the two measurements of emulsion stability. The correlation was done using the S.P.S.S. program. A +1 value would be a perfect correlation whereas a -1 value would describe a perfect inverse relationship. To say with any significance that the relationship between increases in specific gravity correlate with increases in percent surface oil, the Pearson number should be 2: +.5 (Gill,1978). For the comparison of Series A&B the Pearson correlation is +.67. For the comparison Series C the Pearson correlation is +.70. Hence, there is a decent correlation between surface oiling and the specific gravity, both describe emulsion stability . 4.3.4 Comparisons Recall, that because of a high amount of interaction between container type and the various independent factors, only specific comparisons should be made. Only one general conclusion can be made: the results suggest that at 1.0 g-level, glass response is consistently worse. Of the 8 comparisons where glass proved to be more extreme, 6 were at 1.09. Specific comparisons show that one container does not fail more. consistently, at a particular g-level/temperature combination, than the other. Nor. do the results show that one container fails more regularly than the another. 39 A hypothesis as to why glass containers exhibit a consistently worse response at 1.0 9: glass tending to fail at a higher rate than plastic at 1.0 g is due to a "water hamer" like effect. These g-forces are completely transferred to the product because of the rigidity of glass, unlike in plastic where the container flexing absorbs some of the shock. 4.3.5 Stability Trends Figures 3,4 and 5 compare the trends in specific gravity for the three series using those calculated means from Table 4. Figures 6,7 and.8 compare the trends in surface oiling for the three series using those calculated means from Table 5. 40 < sequom sou zuu>euu uuuuoeae cu access .m eosm.m E mooted“: Enacted—co... cop Nb ov _ emu-6.-..-- 38.... 32o 6:8. «use 2.8... . . AN: w I - I — III I II Iflilliil. .33. -4 -.- l -l . .. . ...u4» a. / m / j! ==3H A w . - .i . omen-o if 3. .- I. - . -. Sand .. -. I!!! I... -. -3... .. -- . 886 -. 3...!!- _ . 886 .- . . 886 T -- -- -- : .. 85.6 .i- -_ . 85.6 ..- -3. -- _ . . .!. .-I--.-!. .l- .l.m....-..3..w....H!3..H-M.H.H.....H-. 1-.--..33--. . . _ °°N¢.C moo; @ who m cod 2: Ema: - o _ .. c... 3.2.... - -......-..!w- -. i3 886 Ame-:9 oggoods 41 GOP _ mmma . _ flnfl . - AA //r ”L m season sou zun>euu oququeda EA smooch .c «Lam‘s C moocmoE 9.222.th ozwm. -.."" o _ one.” - ozmmE .um: slunnue.. me An -I. 3C. CV mum-O .- .4 l- .-_ .I.I Dal-ll I i'l. -. --..2.-...-E. 0.: I .‘I’tl‘u 4’1 .III .oll. I‘ll'l No flood z: Juan - o a 03mm?— / N I l. -Il.....u . 693.6 @236 Ommcd OOOGO Omcmd OO—md cm-md chmd 9356 Annals) oggoads 42 u mouuom uou Auu>oum uuuuooaa cu meson? .n scam.m E woocmoov oesumcooth cop N5 ov _ 3.5. ems-a-..- mmo- ...w........_._ .35. 6:8... 1 /...l L / . 1..” , .3“ A - E. /.-: ._ /:. _ ommmo -- .. 8on -- .-- - ones-o I- 3 3.. ....I.| I . .- ooooo - I... 3. ll... .......l.. . once-o - 3.....l 3.. ll..- .............. i .- 856 L. .- . t. ........-.....-....l...: . -. .. om—md .. !. .- -. .........s. g .. . . 838 go; VA who m 86 z: Ema... - o . ml!!! . -, 3......1JJJ... SNQAV Annals oggoads 43 < magnum sou dog—«o assumed cu moses? .o unsaum C woocmoo. confided—ooh. «A Ov "XQMmgn. uwmizmw "SQmQSA— \ .Wmixmw-.L nxammzn. ¥ .. ..- H! 4 - .3... - .. . .. . -- . / 4.9.. 9.. 9.. .- -- I 5....- o.~ .. - oo l I............... o... .. - ow 3.- .. 3.. -. .-.-. 3.... .. oo 59.. Z who W. Ema: - o_ -. a--- - -3.-- . . . os sumo wowed 44 oop- wwaé .292“. I- 0.7.,0040 4..- m meuoem sou meduuo oeoooee cu moons? E woocooo. obs—Eooth we woo-O --.-A1rll..n_..- / .IIII 'III. A cows-E--- maso- cows-a -II'I 0IIIII 4"-'I"I'lll. II ..ll.I .A 4.32.1 Gd 0.— ON nfimu nfias O6 c6 . OH sumo IUOOJSd 45 cow. o aouuom yaw mcuuuo accouum :« assume .m ou:w_m C 82963 929.352. Nu ow .56 .. . 9%....- . -mmm_a..---.w --mmmi . .438: 2.3... .. .. ../ L / mu. - ink..- / V... , -_ % ’0'..-- LI' I III: . I. 3.5 a... man: - o _ 2.-...-.. .r .-..N I-.!I|.||.I..Io 6.6 O. — ON 6.0 O...‘ O...” 66 ON fiuugo mamad 46 Comparing plastic to glass. When comparing the mean value (see Table 4) for specific gravity between identically treated containers: in all categories plastic shows a higher mean in 13 of 18 observations and glass in S of 18 observations made. In the critical categories, (0.75 g at 40 and 72 F) of six comparisons plastic mean value is higher four of six times. These trends are outlined in Figures 3,4 and 5. -Comparing the controls. The comparison between the specific gravity of the controls (0.00 9, same temperature) and the vibrated containers, using the same MSD number, shows that: in the plastic category (Series A-B) 9 of 16 observations *were significantly’ different. In the glass category (Series 11-8) 6 of 16 observations were significantly different. In the plastic category (Series C) 3 of 6 observations were significantly different. In the glass category (Series C) 2 of 6 observations were significantly different. Although the objective of the research. was to compare plastic to glass, comparing the controls shows that mayonnaise in plastic has a higher trend towards demulsification. These comparisons can also be made in Figures 3,4 and 5. These comparisons imply that low levels of vibration influence mayonnaise stability in plastic more so than in glass. 47 -An explanation. Emulsions in which water is the continuous medium may be expected to show a high conductivity; emulsions in which the oil phase is the continuous medium may be expected to show little, or no,conductance (Becher,1977). The possibility exists that there is a electrostatic discharge being transferred to the product as a result of vibrational friction between the PET and the fluid medium causing droplet breakdown (Hart,l985). This charge could affect the electrical double layer forces which act as a major stabilizing force between phases. Similarly, if a charge was being transferred to the product it may be affecting the electrical forces of attraction between oil droplets (van der Waals forces), promoting coalescence. This charging would be negligible in glass containers because of the inert nature of glass. Another factor present in plastic and not in glass is the sidewall flexing of plastic containers, observed even at very low g-levels. This results in wave propagation from the side wall into the emulsion resulting in increased turbulent motion. This additional energy of motion could promote increased Brownian motion. Increases in Brownian motion, as pointed out, will have a tendency to increase the rate of coalescence. Lastly, it may promote settling of the dispersed onion and garlic particles which help in the stability of the emulsion by reducing surface tension forces. 48 4.4 Qualitative Results The data in Table 8 describes the number of containers out of a total of eight examined, that.passed the visual test. Figures 9,10 and 11 compare the trends in visual acceptability for the three series. 49 Table 8. Visual pass-fail rating for all plastic and glass test containers. Plastic Glass Series Acc. Temperature (F) 40 72 100 40 72 100 0.75 7 8 6 8 8 8 1.00 6 8 0 6 5 O B 0.50 8 8 8 - - - 0.75 8 8 3 8 8 8 1.00 8 5 0 4 4 0 C 0.50 8 8 8 - - - 0.75 8 8 3 8 6 2 1.00 4 S 0 6 l 0 50 < aouuom you uaou Aeson> acuooaa auscucueoo accuse: .o ausaqm E mooamocv Santos—co... cop Nh ow mam—G O=mfl_n_ ”mm—mu "v—uwfl—l ”mm-G "Hana—Q .... .22“. . ...M: «...--....- - -M..h...-....-/.22---2 ....Wm.“ ...-M222 L - -A2....: . .... I C ..m J 2.2.2! 2.2222 ...2 .22 e - .- o P w :2. .- - 2 . on u 1.0. -- -- 2 .. Om mud n 22.. ...... r.- .2, . .. av s S. 1-...-- -- - .. on u 1 5 .. . 2.2. 2. 1 O0 m s 2 2|- - _ 22222 H . .22 I .- ch m l; ...2 , .. m H... 8 l ...... .... - ..--2.- . -- m 6m m... .... -- -2.-...H...-..|:I|.._:_2-.II:I:2|-- 2-2-22.2-- -.h “...-n... . . ._ OQP ace; 2 who U 86 E. Qua: - o _ . w .- 22 . - -222... -..... hf... I’ll: 52. fl mfldhflm hOu HUGH Hflaflflsr Qfidfllfln— IBISunp-OU uflflouam .O— 9333‘..— E 82on 2293th 8' «h 3 .85 23-2--.-..---mmamd- ems... _ .35 ._ 2.2.... .2 Op Om cm 2 .. am Oh CG CG co — 2222 2222 222222222222 A8... 229... M 8... ..-: an? - o . _ a . ..I'!" l0... - III'IIII.ItlIII 1391 lensgA Bugssed warned 51 a magnum now have mooou> «nuance auoc«eucoo ucsouam .c. agenda E $9on 9393th OS Nu ov mama oemmfii-..-.-mmflw-.-- amen. mama. . 2.3... _ ‘ -_ . .... /» a f; V. x ._ / o d .- .... o. m . m - ow u 1&- 2 on d - e- w 8. .- 2. on u U 5 .---....- H.22 .2 . 2 cm A .2. .....- mm on m .222 .HHHM n6 2..-- , H m. on I .2, .2 2:.“ N. _22222- 2:222 nHHm .Umw 03 2 1... - 22.4..-- . . .....u..2u......... - -.h. .. . . .. - .. N. Go— noo; 2 £6 E 8.8 :.= .35.. - o _ l . 52 o nouuom you away unsnu> mcuoooo ouocumusoo accused ... ouswuu E «@233 9398th cow N5 0? «use _ Sign .. --.mmfiwsm pew-«E- _ mama _ 2.35 .. z--- .»i. .. a. .. / o “W H . ? - e w 2.. 2 cm . u , 1. ..... l...||2l2|1 O” w H, as S S. 222 2. HHHH fix“ nu H...- ..H, 5 - H22. ... ”.22 CG A 2, 2 ...-2.. m... 2- 22 HI: 22H Oh % ...22 m on .I. 221 .mm- -222- .--.- 2, H22...- cm 2... 2. S 22 1. .2 222.222-- 2.-.... . 669 58.22. MoooEjman o: 2.Hfi2Wa-2 ~ -22.2HU.22.uw.284 53 4.4.1 Comparing qualitative results The visual rating implies there is very little difference between the two containers. In most observations, no difference was found using visual rating of the containers. The only consistent observation was that mayonnaise in PET jars had a higher acceptance at the 1.0 g-level than did glass jars. 5.0 CONCLUSION Specific gravity and surface oil were used as indicators of emulsion stability. The following are the conclusions of this study : i) Vibration forces encountered in typical shipping environments have a significant effect on emulsion breakdown of mayonnaise. Temperature and storage time accelerate this phenomenon. ii) There was no consistent significant difference (for a 95% confidence level) in the amount of surface oil or specific gravity of mayonnaise between identically tested plastic and glass containers. iii) Specific gravity was an acceptable means to monitor emulsion stability. It showed that there were no consistent differences between glass and plastic in the critical categories,(the closest to typical transit conditions: 0.75 g at 40 or 72?). iv) The percent surface oiling was the most conclusive way in which to judge emulsion breakdown. It was concluded that the percent surface oil had to be greater than 1.0% to be obvious (detectable). 54 55 v) Visual rating is still the best means of indicating emulsion instability since this represents a direct correlation between consumer acceptance/rejection of product. Appendix A Specific gravity is the ratio of the mass of mayonnaise to the mass of an equal volume of water at the same temperature. The raw data collected was changed into specific gravity as follows: wt. of pycnometer with sample product = Wp wt. of pycnometer empty = We wt. of pycnometer with water = Ww Specific Gravity of sample = (Wp-We)/(Ww-We) (A-l) (We and Ww are shown in Appendix G) Percent Oiling The calculation was done as follows: net wt. of surface oil drawn = We net wt. of centrifuged sample = Ws Percent surface oil = (Wo/Ws) * 100. (A-Z) 56 Weight of mayonnaise in pycnometer at 74 57 Appendix B P (in grams). Table 31. Series A (40 F). g-level Plastic Glass 0.00 44.0110 43.9997 44.0070 43.9948 44.0121 44.0212 44.0551 43.9732 0.50 44.0618 44.0178 44.0285 44.0495 44.0120 43.9015 43.9904 44.0348 0.75 43.9194 43.9918 43.9310 44.0618 43.9868 44.0389 44.0334 44.0483 43.9614 44.0322 44.0988 44.0384 43.9287 44.0478 43.9574 44.0220 1.00 44.1638 44.0648 43.9878 44.0384 43.9863 44.0777 44.1488 44.0906 44.0668 44.0588 44.0820 44.0748 44.0687 44.0620 44.0595 44.0599 58 Table 82. Series A (72 F). g-level Plastic Glass 0.00 44.0071 43.9675 44.0325 44.0315 44.0141 44.0028 43.9825 43.9962 0.50 43.9698 44.0708 43.9888 44.2130 44.0118 44.0263 44.0520 44.0753 0.75 43.9635 44.0620 43.9465 44.0301 43.9570 44.0554 44.0963 44.0438 43.9028 44.0801 44.0086 44.0534 43.9647 44.0348 44.0234 44.0624 1.00 44.0518 44.0495 44.0518 44.0398 44.1849 44.1508 44.0743 44.0452 44.1439 44.0482 44.0998 44.0930 44.0888 44.1168 44.1138 44.1399 59 Table 83. Series A (100 F). g-level Plastic Glass 0.00 44.0824 44.1112 44.0404 44.0825 44.0173 44.0782 44.0345 44.0473 0.50 44.1692 44.1729 44.1466 44.2082 44.1791 44.1578 44.1871 44.1658 0.75 44.2222 44.1003 44.1916 44.0698 44.2078 44.1074 44.1488 44.1262 44.1730 44.1108 44.2088 44.0906 44.1559 44.0980 44.1808 44.1043 1.00 44.2398 44.2518 44.2468 44.3018 44.1405 44.2108 44.2440 44.3078 44.2295 44.2268 44.1928 44.2878 44.2188 44.2460 44.2338 44.2698 60 Table 34. Series B (40 F). g-level Plastic Glass 0.00 44.1201 44.1129 44.0935 44.0815 44.0586 44.0445 44.0751 44.0759 0.50 44.1223 44.1379 44.0020 44.1630 44.1292 44.1088 44.1182 44.1200 0.75 44.2650 43.9394 44.1000 44.1010 44.3075 44.0351 44.2444 44.1455 44.2410 44.0967 44.2512 44.1090 44.2220 44.0673 44.2318 44.0752 1.00 44.2602 44.0170 44.2640 44.1810 44.4132 44.2300 44.3606 44.1283 44.2500 44.0350 44.2730 44.2054 44.2472 44.1602 44.3888 44.1986 61 Table 85. Series B (72 F). g-level Plastic Glass 0.00 44.1029 44.1062 44.0335 44.0818 44.0798 44.0451 44.0566 44.0629 0.50 44.0630 43.9969 44.1238 44.0263 44.1600 44.0920 44.0532 44.1012 0.75 44.1287 44.1500 44.0872 43.9020 44.0920 43.8954 44.1670 44.0050 44.0465 44.1065 44.0732 44.0764 44.1008 44.1657 44.0820 44.1046 1.00 44.1077 44.1524 44.0545 44.2488 44.1830 44.2180 44.0650 44.1130 44.2170 44.1999 44.1572 44.2030 44.1111 44.2762 44.1138 44.1782 62 Table 86. Series B (100 F). g-level Plastic Glass 0.00 44.1619 44.0949 44.1286 44.1288 44.1329 44.1651 44.0872 44.1252 0.50 44.3390 44.3326 44.3350 44.3390 44.3266 44.3130 44.3300 44.3501 0.75 44.3132 44.0933 44.2950 44.1797 44.3330 44.1636 44.3460 44.2313 44.3100 44.1727 44.2909 44.1616 44.3590 44.1746 44.3181 44.1686 1.00 44.4610 44.3220 44.5000 44.3197 44.5044 44.3230 44.4604 44.2680 44.4973 44.3250 44.5056 44.2880 44.4670 44.3021 44.4686 44.3195 63 Table 37. Series C (40 F). g-level Plastic Glass 0.00 44.1080 44.1624 44.0873 44.1362 44.0831 44.0883 44.0644 44.1138 0.50 44.0670 44.0984 44.1078 44.1484 44.1720 44.1222 44.2030 44.1720 0.75 44.0654 44.0691 44.1600 44.0937 44.1382 44.1252 44.1454 44.0930 44.1654 44.0630 44.1237 44.0993 44.1008 44.1697 44.1290 44.1085 1.00 44.3660 44.0610 44.1113 44.1385 44.2770 44.0988 44.3030 44.1310 44.2645 44.1167 44.2421 44.1035 44.2342 44.1214 44.2468 44.1108 64 Table B8. Series C (72 F). g-level Plastic Glass 0.00 44.0888 44.0808 44.1233 44.0682 44.1510 44.0730 44.1173 44.0260 0.50 44.0555 43.8897 43.9874 43.9500 43.9431 44.0120 43.9650 44.0096 0.75 44.0598 44.0630 44.1331 44.1700 44.1380 44.0810 44.1196 44.0566 43.9583 43.9810 44.1881 44.0060 43.9900 44.0638 44.1021 44.0606 1.00 44.2995 44.1470 44.2653 44.2312 44.2891 44.2790 44.2624 44.2428 44.2430 44.2430 44.2879 44.2350 44.2743 44.2790 44.2622 44.2370 65 Table 89. Series C (100 F). g-level Plastic Glass 0.00 44.2190 44.1873 44.2866 44.1096 44.2557 44.1348 44.2519 44.1528 0.50 44.1980 44.3481 44.3826 44.3329 44.3237 44.5247 44.3190 44.2380 0.75 44.2750 44.1164 44.2833 44.1771 44.3230 43.9944 44.3003 44.2121 44.2270 44.1641 44.4110 44.2400 44.3179 44.2901 44.2830 44.1706 1.00 44.3770 44.3244 44.3787 44.4004 44.3410 44.3282 44.3455 44.3511 .44.3884 44.3580 44.3522 44.3575 44.3588 44.4167 44.3511 44.3623 Weight of surface oil versus weight of centrifuged sample Under no circumstances did controls (ie. measurable surface oil , grams). tables. Table C1. Series A (plastic only). Appendix C (in 0.0 g ) have a g-level temp. sample oil percent weight weight oiling 0.50 40 10.4253 0.00 0.00 11.4530 0.00 0.00 12.7855 0.00 0.00 12.1445 0.00 0.00 72 10.5670 0.00 0.00 9.8781 0.00 0.00 11.2456 0.00 0.00 13.6556 0.00 0.00 100 14.3133 0.00 0.00 11.2313 0.00 0.00 12.9710 0.00 0.00 11.9873 0.00 0.00 0.75 40 10.2635 ,0.00 0.00 14.3324 0.00 0.00 11.4535 0.00 0.00 11.5675 0.00 0.00 72 9.2425 0.00 0.00 11.1600 0.00 0.00 13.4653 0.00 0.00 12.8966 0.00 0.00 100 13.0600 0.10 0.77 10.3511 0.08 0.77 16.3160 0.35 1.70 11.6456 0.11 0.90 66 hence the category is omitted in the Table C1 (Cont'd) 67 g-level temp. sample oil percent weight weight oiling 1.00 40 .14.2580 0.10 0.68 12.3690 0.14 1.14 10.6750 0.05 0.50 16.2000 0.08 0.49 72 10.6756 0.00 0.00 15.2211 0.04 0.69 9.5540 0.07 0.73 12.2340 0.06 0.50 100 13.1866 0.56 4.25 11.3530 0.37 3.28 9.2971 0.41 4.37 15.5822 0.61 3.90 Table C2. Series B (plastic only). 68 g-level temp. sample oil percent weight weight oiling 0.50 40 12.5363 0.00 0.00 12.5433 0.00 0.00 11.9650 0.00 0.00 13.7557 0.00 0.00 72 12.8764 0.00 0.00 13.5635 0.00 '0.00 11.5689 0.00 0.00 9.1231 0.00 0.00 100 13.0987 0.00 0.00 9.6441 0.00 0.00 10.0992 0.00 0.00 12.3805 0.00 0.00 0.75 40 11.2450 0.00 0.00 9.7867 0.00 0.00 10.3243 0.00 0.00 12.1234 0.00 0.00 72 11.3453 0.00 0.00 13.4566 0.00 0.00 13.2432 0.00 0.00 9.8746 0.00 0.00 100 15.2170 0.09 0.63 15.7500 0.16 1.00 16.0734 0.10 0.63 14.5671 0.11 0.77 1.00 40 15.1625 0.09 0.60 11.3800 0.06 0.53 17.1341 0.10 0.66 16.6880 0.13 0.75 72 10.2210 0.20 1.96 ‘ 12.0700 0.10 0.83 11.9331 0.20 1.66 14.1152 0.11 0.79 100 15.1001 0.66 4.34 15.9900 0.94 5.90 16.3211 0.89 5.44 15.3251 0.62 4.02 Table C3. Series C (plastic only). 69 g-level temp. sample oil percent weight weight oiling 0.50 40 11.8540 0.00 0.00 12.0896 0.00 0.00 10.4567 0.00 0.00 9.0566 0.00 0.00 72 13.7532 0.00 0.00 11.4797 0.00 0.00 12.4560 0.00 0.00 11.1112 0.00 0.00 100 12.7340 0.00 0.00 13.6757 0.00 0.00 14.1231 0.00 0.00 9.7839 0.00 0.00 0.75 40 11.6450 0.05 0.55 17.3240 0.07 0.41 13.4000 0.05 0.45 12.3400 0.05 0.65 72 13.4350 0.10 0.75 14.2424 0.11 0.70 15.7886 0.14 0.90 16.3450 0.10 0.60 100 16.2000 0.11 0.65 12.2502 0.07 0.60 13.4330 0.07 0.49 11.3456 0.12 1.10 1.00 40 10.5140 0.13 1.24 15.4030 0.20 1.28 16.7300 0.07 0.45 16.2540 0.15 0.90 72 12.3701 0.15 1.23 13.7981 0.24 1.77 11.3542 0.36 3.07 14.3801 0.13 0.91 100 16.4390 1.02 6.22 15.4343 0.97 6.25 12.9401 0.69 5.32 13.6544 0.58 4.21 70 Table C4. Series A (glass only). g-level temp. sample oil percent weight weight oiling 0.75 40 12.2340 0.00 0.00 13.5644 0.00 0.00 12.6755 0.00 0.00 14.2342 0.00 0.00 72 13.3450 0.00 0.00 9.4324 0.00 0.00 12.8940 0.00 0.00 13.3443 0.00 0.00 100 11.2342 0.00 0.00 14.2311 0.00 0.00 12.9780 0.00 0.00 16.1238 0.00 0.00 1.00 40 10.2630 0.07 0.68 14.8003 0.05 0.34 14.3030 0.18 1.26 14.3691 0.06 0.40 72 10.5770 0.47 4.41 15.7831 0.57 3.59 12.5643 0.55 4.38 11.4565. 0.10 0.90 100 10.7600 0.51 4.77 13.6251 0.79 5.77 12.3520 0.73 5.88 14.7445 0.71 4.85 71 Table CS. Series 8 (glass only). g-level temp. sample oil percent weight weight oiling 0.75 40 12.2130 0.00 0.00 12.7981 0.00 0.00 11.2200 0.00 0.00 16.0202 0.00 0.00 72 11.0243 0.00 0.00 9.1200 0.00 0.00 12.8988 0.00 0.00 16.2323 0.00 0.00 100 12.3000 0.00 0.00 14.4325 0.00 0.00 11.4552 0.00 0.00 12.3333 0.00 0.00 1.00 40 16.6040 0.32 1.62 9.3180 0.06 0.69 14.4560 0.17 1.20 12.9412 0.08 0.91 72 13.8002 0.46 3.33 15.3325 0.57 3.72 17.8350 0.50 2.83 14.1560 0.42 2.96 100 13.4706 0.51 3.76 9.0900 0.33 3.60 15.1551 0.61 4.00 13.8347 0.43 3.11 72 Table C6. Series C (glass only). g—level temp. sample oil percent weight weight oiling 0.75 40 14.1000 0.00 0.00 14.1411 0.00 0.00 11.7981 0.00 0.00 17.1667 0.00 0.00 72 15.0000 0.15 1.01 10.2801 0.12 1.16 12.3841 0.12 1.00 18.5622 0.21 1.13 100 9.6003 0.08 0.83 11.3650 0.11 0.97 12.0830 0.20 1.65 13.5051 0.07 0.50 1.00 40 14.3852 0.09 0.61 16.3400 0.15 0.89 10.3921 0.13 1.29 15.8450 0.36 2.28 72 14.9003 0.77 5.14 10.8470 0.52 4.80 14.7802 0.71 4.81 12.5432 0.49 3.94 100 13.9821 1.09 7.82 12.5580 0.60 4.79 14.3772 1.00 6.95 12.2623 0.67 5.48 Appendix D Table D1. Series A and B 4-way ANOVA test results. Specific gravity by container type (CT).9-1evel (GL),temperature (TEMP.),and time of exposure (TE). Sun of Mean Sig. Squares DP Square F of E Main Effects 1466158.3 5 .002932316 144.42 .000 CT 75366.7 1 .000753668 37.12 .000 CL 252590.1 1 .002525901 124.4 .000 TEMP. 727686.4 2 .003638432 179.2 .000 TE 410516.0 1 .004105150 202.2 .000 2-way inter. 240485.9 9 .000267207 13.16 .000 or or. 5525.5 1 .000055255 2.72 .101 CT TEMP. 77200.9 2 .000386004 19.01 .000 CT TE 94518.7 1 .000945187 46.55 .000 CL TEMP. 17711.1 2 .000088555 4.36 .014 CL TE 6533.3 1 .000065333 3.22 .075 TEMP. TE 38996.3 2 .000194982 9.60 .000 3-Way inter. 55161.9 7 .000078803 3.88 .001 CT GL TEMP. 11918.6 2 .000059593 2.93 .056 CT GL TE 3485.0 1 .000034850 1.72 .192 CT TEMP. TE 38704.8 2 .000193524 9.53 .000 CL TEMP. TE 1053.5 2 .000005267 .26 .772 4-Way inter. 37988.3 2 .000189941 9.35 .000 CT-TE-GL-TEMP 37988.3 2 .000189941 9.35 .000 Explained 1799794.S 23 .000782519 38.54 .000 Residual .00341114 168 .000020304 Total 1799794.S 191 .000112089 73 Table D2. Percent oiling by container type (TEMP.),and time of exposure (TE). Appendix D Series A and B 4-way ANOVA test results. (CT) .g-level (GL) , temperature Sum of Mean Sig. Squares DP Square F of F REES—Effécts 181.31 5* 36.26 143.156 .000 CT .81 1 .81 3.18 .079 CL 107.94 1 107.94 426.12 .000 TEMP. 72.82 2 36.41 143.73 .000 TE .09 1 .09 .36 .550 2-way inter. 61.12 9 6.79 26.81 .000 CT GL 5.05 1 5.05 19.94 .000 CT TEMP. 5.96 2 2.98 11.77 .000 CT TE .02 1 .02 .07 .788 CL TEMP. 46.24 2 23.12 91.27 .000 CL TE .55 1 .55 2.16 .146 TEMP. TE 3.23 2 1.61 6.37 .003 3-Way inter. 8.08 7 1.15 4.55 .000 CT GL TEMP. 2.41 2 1.21 4.77 .011 CT GL TE .45 l .45 1.76 .189 CT TEMP. TE 3.07 2 1.53 4.16 .004 CL TEMP. TE 2.11 2 1.05 4.16 .020 4-Way inter. 5.01 2 2.51 9.89 .000 CT-TE-GL-TEMP 5.01 2 2.51 9.89 .000 Explained 255.53 23 11.11 43.86 .000 Residual 17.98 72 .253 Total 273.51 95 2.91 Table D3. (TEMP.).‘ Appendix D Series C 3-way ANOVA test results. Specific gravity by container type (CT),g-level (GL),temperature Main Effects CT GL ' TEMP. 2-way inter. CT GL CT TEMP. GL TEMP. 3-Way'inter. CT GL TEMP. Explained Residual Total Sum of Mean Sig. Squares DP Square F of F 746532.04 4 .001866330 56.34 .000 41126.76 1 .000411268 12.42 .001 331702.59 1 .003317026 100.18 .000 373702.68 2 .001868513 56.43 .000 24953.18 5 .000049906 1.51 .196 2430.09 1 .000024301 .73 .394 3409.15 2 .000017046 .51 .599 19113.94 2 .000095569 2.89 .061 79560.06 2 .000397800 12.01 .000 79560.06 2 .000397800 12.01 .000 851045.28 11_ .000773677 23.37 .000 .00278124 84 .000033110 851045.28 95 .000118860 76 Appendix D Table D4. Series C 3-way ANOVA test results. Percent oiling by container type (CT),g-level (GL),temperature (TEMP.). Sum of Mean Sig. Squares DP Square F of E _—Main Effects 17"_T1. 3 4 42—.84 T1. 31 . 000 CT 10.54 1 10.54 22.47 .000 CL 102.42 1 102.42 218.30 .000 TEMP. 61.22 2 30.61 65.24 .000 2-way inter. 44.04 5 8.81 18.77 .000 CT GL 1.91 1 1.91 4.08 .051 CT TEMP. 6.40 2 3.20 6.82 .003 CL TEMP. 34.20 2 17.10 36.45 .000 3-Way inter. 1.45 2 .73 1.55 .228 CT GL TEMP. 1.45 2 .73 1.55 .228 Explained 216.85 11 19.71 42.02 .000 Residual 15.95 36 .47 Total 232.80 47 5.17 Appendix E Series A and 3 Specific Gravity. The MSD for a confidence of 95%: Alpha = .05/2 Degrees of freedom = 168 8 repetitions From the T-table T= 1.974 (two containers*(MSE)/two containers‘ 4 of repetitions)E.5 SED (2(MSE)/8)E.S MSE is given by the print out as .0000203044 SED = (2*(.0000203044)/8)E.5 = .002255 MSD = T’SED = 1.974 *.00225 =.0044 Therefore the difference between the plastic and glass containers,given identical treatment, must be greater than or equal to .0044. Percent Oiling. Here the formulas are the same as above; T and MSE are different. T at alpha = .05/2 72 degrees of freedom 4 repetitions From the T-table T: 1.993 MSE is given in the printout as .253 SED = (2(.253)/4)E.5 = .35567 MSD = 1.993 *.3SS67 = .709 Therefore the difference between the plastic and glass _ containers,given identical treatment, must be greater than or equal to .709%. 77 78 Appendix E (cont'd) Series C Specific Gravity. The MSD for a confidence of 95%: Alpha = .05/2 Degrees of freedom = 84 8 repetitions From the T-table T=1.99 SED = (two containers*(MSE)/two containers* 4 of repetitions)E.5 = (2(MSE)/8)E.5 MSE is given by the print out as .00003311 SED = (2*(.00003311)/8)E.5 = .00288 MSD = T*SED = 1.99 * .00288 =.0057 Therefore the difference between the plastic and glass containers,given identical treatment, must be greater than or equal to .0057. Percent Oiling. Here the formulas are the same as above; T and MSE are different. T at alpha = .05/2 36 degrees of freedom 4 repetitions From the T-table T= 2.028 MSE is given in the printout as .469 SED = (2(.469)/4)E.S = .4842 MSD .4842 * 2.028 .982 Therefore the difference between the plastic and glass. containers,given identical treatment, must be greater than or equal to .982%. Appendix F The standard deviations of specific gravity measurement. 79 Plastic Glass Series G's temperature 40 72 100 40 72 100 0.00 .0019 .0018 .0024 .0017 .0023 .0022 0.50 .0041 .0064 .0027 - - - 0.75 .0041 .0050 .0031 .0026 .0014 .0013 1.00 .0055 .0039 .0052 .0044 .0039 .0037 0.00 .0022 .0026 .0027 .0024 .0022 .0025 0.50 .0039 .0045 .0019 - - - 0.75 .0051 .0031 .0020 .0053 .0066 .0032 1.00 .0058 .0048 .0042 .0068 .0044 .0018 0.00 .0016 .0022 .0024 .0027 .0021 .0028 0.50 .0032 .0054 .0044 - - - 0.75 .0030 .0067 .0045 .0028 .0047 .0059 1.00 .0061 .0015 .0059 .0020 .0035 .0027 80 Appendix F (cont'd) The standard deviations of the surface oil measurements. Plastic Glass Series G's temperature 40 72 100 40 72 100 A 0.00 0 0 0 0 0 0 0.50 0 0 0 - - - 0.75 0 0 .44 0 0 0 1.00 .30 .33 .48 .40 1.60 .58 B 0.00 0 0 0 0 0 0 0.50 0 0 0 - - - 0.75 0 0 .17 0 0 0 1.00 .10 .59 .89 .40 .40 .37 C 0.00 0 0 0 0 0 0 0.50 0 0 0 - - - 0.75 .11 .16 .27 0 .10 .48 1.00 .38 .98 .96 .70 .51 1.30 Appendix 6 Table A1. Pycnometer weight empty (in grams). 33.6374 33.6376 33.6377 33.6378 33.6375 33.6376 33.6376 Average was used in calculation of specific gravity. Table A2. Weight of pycnometer with distilled water at 24 C. 45.2763 45.2789 45.2787 45.2771 45.2781 45.2761 45.2769 45.2779 45.2775 Average was used in calculation of specific gravity. 81 REFERENCES "ASTM D4728-87 Standard test method for random vibration testing of shipping containers",Annual Book of A5174 Standards,1987. Banker,G. "Monitoring settling in emulsions by incorporation of radioactive materials", J. Pharm. Sci.,51,1962. Becher,P. "Encyclopedia of Emulsion Technology: Basic Theory", New York: Marcel Dekker Inc.,1983 Becher,P. "Encyclopedia of Emulsion Technology: Applications", New York: Marcel Dekker Inc.,1985. Becher,P. "Emulsions : Theory and Practice",Huntington: Krieger Publication Inc. 1977. Bennett,H. "Practical Emulsions", Third Ed. New York: Chemical Publishing Inc.,1968. Brown,A. "Utilization of a pressurized viscometer for measuring emulsion consistency", Chem. Ind.,30, 1968. Carroll,B. "The Stability of Emulsions and Mechanisms of Emulsion Breakdown", Sur. Col. Sci.,9,1976. Chang, C.,Fennema, 0. "Electron Microscopy of Mayonnaise.", Inst. Can. Sci. Technol. Aliment.,5,1972. Corran,J. "Some observations on a typical food emulsion", Emulsion Technology. Chemical Publishing Co. ,Erooklyn,MY, 1946 p176- 192. Fennema,o. "Food chemistry",Marcel Dekker, Inc.,NY,1985. Friberg,s. "Emulsion stability in food emulsions",Marcel Dekker Inc.,NY,1976. Gill,J. "Design and Analysis of Experiments",Iowa State Univ. Press.1978. Vol.1 Gill,J. "Design and Analysis of Experiments",1owa State Univ. Press.1978. Vol.3 p. Gillespie, T.,Rideal,E. "The effect of temperature on the interfacial films of the dispersed phase in a emulsion", Trans. Faraday Soc.,52,1956. Hart, D. "Triboelectric Characterization of Packaging Materials", Proceedings of the Electrical Overstress, Electrostatic Discharge Symposium. 1985. p 61. 82 83 King,D. "Emulsion stability", Trans. Faraday Soc.,37,1941. Liem,S.,Woods,D. "Review of Coalescence Phenomena", AIChE.,70, 1974. Marcondes,J. , "Dynamic Analysis of a Less Than Truckload Shipment", A Thesis. Michigan State Univ. 1988. Ostrem,F. "An Assessment of the Common Carrier Shipping Environment", AS'IM Committee on Packaging, First Ed.,1985. Reddy,S. "Experimental Study of Simultaneous Flocculation and Creaming", J. of Colloid and Interface Sci.,82,1981. Richmond,M.,Harte,B.,Gray,I.,Stine,C. "Physical Damage of Yogurt. The Role of Secondary Packaging on Stability of Yogurt", J. of Food Protection, Vol.48, No. 6 Rochow,I. "Breaking Emulsions By Freezing", Ind. Eng. Chem. ,28,1936. Tomoyoshi. "Dispersion State of Protein Stabilized Emulsions", SEM Inc. AMF O'Hare,Vol.3, 1983. Tung,M.,Jones,I.. "Microstructure of mayonnaise and salad dressing",SEM Inc. AMF O'Hare,Vol.3,1983 p523-530. U.S. Food and Drug Administration, Fed. Reg. Sec. 25,1964. Wang,J.,Kinsella,J. "Functional Properties of Novel Proteins", J. of Food Sci. ,4l,l966. HICHIG 9 3129300 4 9 WWW!) I)))fifl)’;)1§@))@@li))“