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An Electronic Imaging System for Determining Droplet Size

and Dynamic Breakdown of Protein Stabilized Emulsions


ABSTRACT (Walstra 1965, 1968) and (Pearce and Kinsella, 1978), elec-
A computerized imaging system, consisting of a light microscope and trical conductivity measurements (Webb et al, 1970), centri-
a light sensitive diode array interfaced with a computer, was devel- fugation (Sherman, 1971), emulsion capacity tests (Swift et
oped and evaluated for monitoring emulsion droplet size distribution al., 1961), solubility studies (Voutsinas et al., 1983), the Coul-
and rate of emulsion droplet coalescence. This system compared fa- ter counter (Walstra and Oortwijn, 1969) and electron micros-
vorable with electron microscopic analyses and light scattering studies copy (Liboff et al., 1988). However, these methods lack the
for determining emulsion droplet size. Oil in water emulsions stabi- real time reliability of studying emulsion breakdown with light
lized with 0.05, 0.1, 0.2, 0.5, and 1.0% /3-lactoglobulinor casein microscopy (Halling, 1981). Many of these available methods
were studied. It was observed that P-lactoglobulin stabilized emul- are inadequate for protein stabilized emulsions. For example,
sionsformedsmallerdropletsthat coalescedat a slowerratethenthose the Coulter counter uses salts that can alter the conformation
stabilized by acid casein. Emulsions stabilized by less than 0.2% pro-
tein were extremely unstable. of the interfacial protein and may interfere with the state of
dispersion of the emulsion (Tadros and Vincent, 1986). Many
researchers now use light scattering methods to study emul-
INTRODUCTION sions because of their rapidity and reproducibility (Kinsella,
THE INCREASED PRODUCTION of formulated and fabri- Studies using light microscopes are limited by the lack of
cated foods requires the availability of reliable functional in- magnification that hinders accurate sizing of the smallest emul-
gredients, and the increased demand for functional ingredients sion droplets and is time consuming and tedious. The method
has dramatized a corresponding need for methods to charac- reported herein provides further magnification of the micro-
terize the functional properties of ingredients (Kinsella, 1984). scopic image and permits the sizing of smaller particles using
Emulsifying agents are of primary importance in imparting a light sensitive diode array. Because the emulsion is not ma-
stability to many food systems, such as milk, mayonnaise, nipulated, real time studies of emulsion breakdown can be
salad dressings, whipped toppings, and sauces. Emulsifiers accurately performed.
may function by one of several mechanisms; these include Protein based surface active agents are being increasingly
lowering the interfacial tension between the phases, surround- developed for use in food emulsions, and hence rapid and
ing the emulsion droplet with a charged layer and providing a reliable methods are needed to assestheir emulsifying activity
physical barrier to the approach of other droplets (Walstra, (EA) and emulsion stabilizing properties. The present research
1986). Proteins, which possessan amphiphilic nature by virtue describes the development of a method for determining EA
of their primary structurewith polar and apolardomains,are and monitoringthe breakdownof emulsionsstabilizedby j3-
useful emulsifyingagents.In additionto the electrostatxre- lactoglobulinand acid casein.
pulsion of charged groups, polypeptide loops and trains in an
interfacial film provide a barrier to the close approach of oil
droplets (Phillips, 1981). The breakdown of emulsions via co- MATERIALS & METHODS
alescenceof oil droplets,resultingin decreasedsurfacearea Materials
and flocculation is retarded by interfacial protein films (Dick-
inson and Stainsby, 1982). Creaming is slowed by homoge- The emulsions in this study were stabilized by either p-lactoglogulin
nizing the droplets to a small size to minimize the effects of or casein. Pure D-lactoglogulin (A & B from bovine milk, crystallized
the density differences between the phases (Walstra and Oot- an lyophilized) ‘and iGda\ole (crystalline) were both obtaiied from
the Simna Chemical Co. (St. Louis. MO\. Casein was nreoared bv
wijn, 1975). The wide range of hydrophobic and hydrophilic the hydrochloric acid precipitation of fresh raw skimmed’miik (Da<-
properties available in different proteins can be exploited in ies, 1936). Emulsions were made with pure peanut oil that did not
formulated food emulsion systems. Proteins also provide nu- contain added emulsifiers. The specific gravity of this oil was 0.91.
tritional benefits not available from lipid-based emulsifiers, The Staining solution of 0.1% Congo red, a hydrophilic dye, con-
and they are not subject to the concern and scrutiny provoked tained 5% gelatin (Bovine skin, type?II) which-was+buffere-d;o pH
by various classes of chemical emulsifiers. 7.0 with 0.05M imidazole buffer. The staining solution was main-
The use of proteins as emulsion stabilizers has been the tained as a liquid by keeping the solution at 90°C. The gelatin was
subject of many studies (Halling, 1981). The properties of obtained from Sigma Chemical Co., and pure Congo red was obtained
emulsions stabilized by proteins vary with the equipment used from Electron Microsconv Science (Fort Washington. PAL All other
.< _ I

chemicals used were reagent grade and distilled deionized water was
for their preparation (Thorberg and Lundh, 1978). A standard Used throughout this study.
procedure has been improved by interfacing a single piston The imaging system consistedof a light sensitive diode array mounted
valve homogenizer with a computer to control energy input atop the right eyepiece of an American Optical AO-150 light micro-
(Haque and Kinsella, 1989). scope. The light microscope was equipped with a 15X eyepiece for
Several methods to characterize the stabilizing effects of examining fine emulsions and a 10X eyepiece for examining coarse
emulsifiers have been used. These include light scattering emulsions. The diode array is composed of a series of 32768 pixels,
3.56 per horizontal line and 128 per vertical line. The diode array was
interfaced to a Turbo-XT computer via an input/output board. The
computer was used to set the threshold light level for the diodes, set
Authors Klemaszewski and Kinsella are with the Inst. of Food the exposure time for the pixels and was used to gather the image
Science, Cornell Univ., Ithaca, NY 14853. Author Haque’s pres- data from the diode array. The computer set the exposure time foi the
ent address is Dairy Science Dept., Mississippi State Univ., Drawer pixels such that 25 to 30% of the pixels were transmitting 5 volts to
DD, Mississippi State, MS 39762. the computer. The exposure time was changed as needed by the com-

440-JOURNAL OF FOOD SCIENCE-Volume 54, No. 2, 1989

Emulsion preparation. A series of 8-lactoglogulin or acid casein
solutions. 0.05, 0.1. 0.2. 0.5. 0.75 and 1.0% (w/w). was made LID
in 50 mM imidazole’to pH 7.01 The emulsions were made by initially
dispersing 12 g of protein solution and 8 g of oil with a Janke-Kunkel
TP 18-10 turbo blender for 5 set prior to passage through the valve
homogenizer, as described by Haque and Kinsella (1989). The ho-
mogenizing system was kept at a constant 25°C using a circulating
water bath. Emulsions were obtained by passing the protein solution
and oil mixture through a valve homogenizer for 180 strokes, corre-
sponding to an energy input of 640 X 10” J.rne3. The emulsion was
transferred to a beaker and slowly stirred to prevent creaming. The
temperature of the emulsion was held constant at 25°C.
To monitor stability, i.e., changesin droplet size with time, aliquots
(0.1 mL) of the emulsion were removed at hourly intervals and mixed
in a testube with 0.2 mL of the staining solution at 90°C. The stained
emulsion was immediately placed on a microscope slide and spread
thinly to obtain a uniform layer of emulsion. A cover slip was placed
on the slide and the stained emulsion sample was then cooled on ice
to solidify the warm gelatin and prevent further coalescence of the
emulsion. Emulsions were then viewed using the imaging system, and
the image was printed out for measurement of the droplet diameters.
Coalescencewas observed by a decreasein the emulsion surface area
and an increase in the average size of the droplets with time.
Sample preparation for electron microscopy. To check the ac-
curacy of the imaging system, casein stabilized emulsions were viewed
using scanning electron microscopy @EM). Emulsions samples for
the electron microscope were prepared in the same manner as for the
imaging system. The methodology for sample preparation for SEM
has been decribed by Liboff et al. (1988).
Sample preparation for light scattering studies. Light scattering
methods, based on the Mie theory (Van De Hulst, 1957), have been
applied to the study of emulsions (Walstra, 1965) and Pearce and
Kinsella (1978). Emulsion samples for the light scattering studies were
prepared in the same manner as for the imaging system. For these
studies, the emulsion, from the homogenizer, was immediately diluted
from 50 to 5000 fold in a 0.1% SDS solution buffered to pH 7.0 by
0.05M imidazole-HCl to give an absorbance between 0.2 and 0.7.
The d, and emulsion surface area were determined by the spectro-
turbidimetric method of Walstra (1968) using a Gary 219 dual beam
spectrophotometer with automatic base-line correction to scan the
samples from 350 to 870 mm. The cuvettes used for this study allowed
a small acceptance angle of light to be passed through the sample.
Surface area of the emulsion was also determined from the turbidity
at 550 nm using the equations of Pearce and Kinsella (1978).
Calculations. By measuring the diameter of all the droplets on an
image, a histogram of the droplet size distribution for each view of
an emulsion was prepared. In order to make the experiment statisti-
cally valid, over 100 droplets were measured for each sample. The
average droplet size of the emulsions were determined from the his-
tograms prepared from the data obtained using the imaging system.
The average droplet size was determined using equation (l), where
d, is
d, = (1)
fig. 1-A typical micrograph of an oil (light) in water (dark) the average droplet diameter, nt is the number of droplets of size i
emulsion stabilized by 0.2% casein obtained using a comput- microns and N is the total number of droplets measured. The d, of
erized diode array imaging system to magnify light microscope the emulsion, described by Walstra (1965) as the average volume to
image (2940 x). surface ratio, was determined from the histograms obtained by the
imaging system using Eq. (2).
puter. The lower refractive index of the oil, compared to water, pro- d = X i3 ni l

vided the minimum contrast required for droplets to be distinguished. x3

X i2 ni l
The use of Congo red accentuated the contrast. Data from the diode
array consisted of binary data reflecting the threshold light levels on The d, of the emulsion was also determined using the light scat-
each of the pixels. The computer converted these data to an image on tering method of Walstra (1965). This method involves gathering light
the screen. The image onthe screen was a magnification of the emul- scattering data over a range of wavelengths and fitting the absorbance
sion as seen by an experimenter through the eyepiece (Fig. 1). This obtained to equations for ideal theoretical turbidometric data for best
image was printed out for manual analyses. The magnification of the fit lines as described by Walstra (1965).
diode array and the light microscope was approximately 5500 fold Surface area of emulsions were calculated by three different meth-
with an approximate resolution of 0.18~ when the 15X eyepiece was ods. The method of Walstra (1968) uses the &s obtained from light
used. scattering data in equation (3), where 6 is the volume fraction of oil.
The imaging system was calibrated using an American Optical stage
micrometer marked in 0.1~ divisions. The calibration was confirmed 6.0
using latex microspheres of a known diameter. 2.04~. The micro- S. Area = -
spheres were obtained from the Epics division of Coulter Corp. (Hi-
aleah, FL). The surface area obtained from Eq. (3) is in units of meter2 per

Volume 54, No. 2, 1989-JOURNAL OF FOOD SCIENCE-441



f 20


Fig. 2-Scanning electron microscope view of emu(sion stabi- 1.6 2.2 2.7 3.3 3.8 4.3 4.9 5.4 6.0 6.5 7.0 7.6
lized by 0.3% casein obtained using the methods of Liboff et al.
(1988) (2704 x).
Dmplu Si (Micmns)

gram of emulsion. The method of Pearce and Kinsella (1978) gives b

the surface area of the emulsion in units of meter* per gram of rotein
directly from Eq. (4), where A is the absorbance at 550 nm, 8. 1s the

fraction of oil, C is the protein concentration of the solution used to

make the emulsion and 1 is the pathlength of the cuvette. The surface
area as calculated using Eq. (4) is also known as the emulsifying
activity (EA) for proteins.
The surface area of an emulsion can also be calculated from the
histogram obtained from the imaging system data. The total volume
and total surface area of all the droplets observed was determined
using Eq. (5) and (6), where r is droplet radius. The “dilution factor”
was determined using EQ. (7).

Volume = I: F (5)

Surface Area = I: 47rr2 (6) 1.4 2.4 3.4 4.4 5.4 6.5 7.5 a.5 9.5 10.5
DmplU sii (haicmns)
Dilution Factor = Volume “Og Oil
Result of Eq. (5) Fig. J-Histograms of emulsion droplet size distribution in
emulsions stabilized by (a) 0.2% plactoglobulin after 0 hr; (b)
This dilution factor gave the factor with which the result of equation 0.2% casein after 0 hr.
(6) was multiplied to calculate the surface area of l.Og of oil. Values
of surface area obtained were all converted to meters2 per gram of
protein, as these were the units used for comparing the emulsifying
activity (EA) of proteins. emulsion stabilized by 0.2% P-lactoglobulin (Fig. 3a) to the
range of 1.4~ to 10.2~ for an 0.2% acid casein stabilized
emulsion (Fig. 3b). These two emulsions had comparable av-
erage diameters of approximately 5~.
THE EMULSIONS studied in these experiments were com- The change in emulsion stability with time was observed as
posed of a heterogenous population of dispersed spherical oil the tendency for the droplet size distribution to increase with
droplets, as shown by electron microscopy (Fig. 2). These time, as shown in Fig. 4, which are the combined histograms
emulsions, as observed using the imaging system, are shown obtained for 0.5% P-lactoglobulin stabilized emulsions 2 and
as printouts of what was observed through the light microscope 6 hr after the emulsion was prepared. The distribution mode
(Fig. 1). By measuring the diameter of the droplets on an for the sample taken at 2 hr was l.Op,, as compared to 1.4~
image printout, a histogram of droplet size distribution for each for the sample at 6 hr. The droplet size distribution of the
emulsion was prepared. The droplet sizes observed ranged from sample taken after 2 hr is skewed to the right slightly more
0.41~.for an emulsion stabilized by 1% P-lactoglobulin to drop- than the 6 hr sample. Since the droplet size distribution of this
lets larger than 40 p, in emulsions stabilized by dilute, 0.05% emulsion should be normal (Walstra, 1968), skewness indi-
P-lactoglobulin. cates that some droplets were smaller than the resolving power
The histograms revealed that the droplet size distributions of the imaging system. The smallest droplets observed using
were normally distributed with one mode (Fig. 3). This type the imaging system have diameters around 0.36~.
of distribution is usual for a simple emulsion system (Walstra, Data were gathered for similar emulsions stabilized with
1968). Emulsions stabilized by P-lactoglobulin were less het- specified concentrations of &lactoglobulin and acid casein as
erogeneous than those stabilized by acid casein, as shown by emulsifying agents, and the droplet size distribution of emul-
comparing the range of droplet sizes of 1.6~ to 7.6~ for an sions were studied by electron microscopic, light scattering

442-JOURNAL OF FOOD SCIENCE-Volume 54, No. 2, 1989


0.2 0.4 0.6 0.8 1.0

Protein Concentration(96)

0.3 0.6 1.0 1.4 1.7 2.1 2.4 2.8 3.2 3.5

‘Fig. 4-Histograms of emulsion droplet size distribution in

emulsions stabilized by 0.5% plactoglobulin after 2 hr and 6 hr
of coalescence.

0.0 0.2 0.4 0.6 0.8 1.0 1.2

8.0j T Pmein CImcentration(96)
3 6.0 -
Fig. 6-Relationship between methods for determination of sur-
face area for emulsions stabilized by different concentrations of
8 proteins. (a) casein (b) plactoglobulin. Surface area determined
using a--o imaging system; M-H method of Walstra (1965);
z 4.0 - A-A method of Pearce and Kinsella (1978).
2.0 -
From the data gathered in these experiments, it appeared that
the light scattering method was appropriate for emulsions with
average droplet diameters less than 7.0~~.The results obtained
0.0 0.2 0.4 0.6 0.8 1.0 1.2 from light scattering for finer emulsions were much more var-
Protein Concentration(%) iable than the data obtained from the imaging system, as the
Fig. 5-Effect of protein concentration and method of determi- standard error of the d,, was greater than OSp, for light scat-
nation on d,, of emulsions (as defined by Eq. (2). U-Q plac- tering data (Fig. 5), whereas it was less than 0.1~ for data
toglobulin stabilized emulsion, observed using imaging system; obtained from the imaging system. The correlation between
u plactoglobulin stabilized emulsion, monitored using the the imaging system and the light scattering method was quite
method of Walstra (1965); A-A casein stabilized emulsion, ob- high, 0.96, for emulsion droplets stabilized by greater than
served using imaging system; A-A casein stabilized emulsion, 0.3% of either protein. The a,, as determined by these two
monitored using the method of Walstra (1965). methods did not differ by more than 0.5l.~ for smaller droplets
(Fig. 5). This agreement was offset by the high variation of
the light scattering method.
methods and the imaging system. The imaging system was The surface area of the emulsions was determined using the
also used to study the breakdown of these emulsions. methods of Walstra (1965). Pearce and Kinsella (1978) and
Comparison with light scattering method. The average the imaging system. The results of these experiments showed
emulsion droplet volume to surface diameter (&), determined that the light scattering method of Walstra (1965) again showed
by light scattering (Walstra, 1968), was compared with the 4, greater disparity compared to the imaging system when study-
of the same emulsions obtained using the imaging system. The ing coarse emulsions (Fig. 6). At protein concentrations less
d,, of the emulsions was very comparable for emulsions sta- than 0.2%, this method indicated a finer emulsion than was
bilized by 0.2% protein or greater (Fig. 5). However, when actually present. Since the method of Walstra (1965) calculates
protein concentrations were reduced to less than 0.2%, this surface area directly from d.,,, using Eq. (3), it is expected that
light scattering method gave lower values for the &,. The 4% the same limitations apply to surface area calculations. At pro-
of an emulsion stabilized with 0.1% casein was determined to tein concentrations greater than 0.2%, the imaging system
be 25.8t~,according to the imaging system, but this light scat- compared favorably with the results obtained using the method
tering method gave a d,, of 4.91.L. of Walstra (1965). The results derived from the light scattering
It has been reported that the light scattering method used is method of Pearce and Kinsella (1978) were comparable to the
not accurate in studying coarse emulsions (Walstra, 1968). results obtained with the imaging system at protein concentra-

Volume 54, No. 2, 1989-JOURNAL OF FOOD SCIENCE-443


0.0 0.2 0.4 0.6 0.8 1.0 1.2

Protein Concentration (46) 0 2 4 6 a 10
Breakdown Tie (bows)

0.0 0.2 0.4 0.6 0.8 1.0 1.2

Protein Concentration (%) 0 2 4 6 8 10
Fig. 7-Effect of protein concentration on (A) average droplet Breakdown Tie (hours)
diameter and (B)‘emulsion surface area for emulsions stabilized
by M Plactoglobulin and A-A casein. Fig. 8-Effect of emulsion breakdown time on (a) average drop-
let diameter and (b) emulsion surface area for emulsions sta-
bilized by U-U 0.5% plactoglobulin; A-A 1.0% plactoglobulin;
W--W 0.5% casein; A-A 1.0% casein.
tions less than 0.2% and greater than 0.2%. The data obtained
from the imaging system and those by the method of Pearce
and Kinsella (1978) were comparable at all the casein concen- sequent interactions are also critically important (Graham and
trations studied (Fig. 6a), but emulsions stabilized with p- Phillips 1979; Kinsella, 1984).
lactoglobulin did not compare as favorably (Fig. 6b). The method The effects of protein concentration on unit surface area of
of Pearce and Kinsella (1978) had a higher variability than the emulsion were similar, with surface area increasing with in-
imaging system, but the variability was similar to that of the creased protein concentration. However, this was not the case
Walstra (1965) method. The method of Pearce and Kinsella for surface area per unit protein at different protein concentra-
(1978) is simple and is useful for estimating surface area of tions (Fig. 7b). As protein concentration increased, the EA
emulsions with mean droplet sizes in the range of 2t.1,to 15p, initially increased to a maximum at 0.5% protein and then
surface area. decreased. This effect may be caused by protein forming a
Initial emulsion characteristics. The average droplet size thicker film at the interface without an increase in surface area
of emulsions stabilized by S-lactoglobulin and acid casein, at directly proportional to the amount of available protein. At
640 x lo6 J-m-’ of energy input, decreased with increasing higher concentrations, the protein at the interface may not com-
protein concentration (Fig. 7a). Emulsions stabilized by less -pletely unfold. At lower protein concentrations, the protein
than 0.2% protein were initially unstable, with average droplet may unfold more extensively because it is thermodynamically
sizes much greater than 5t.~. At protein concentrations greater favorable to coat as much interfacial area as possible by un-
than 0.2%, B-lactoglobulin formed emulsions with smaller folding (Graham and Phillips, 1979; Halling, 1981).
emulsion droplets than acid casein. The average droplet size Emulsion stability. Data gathered using the imaging system
of the emulsions stabilized by 1.0% P-lactoglobulin was 1.14+, showed that the average droplet size of emulsions increased
as compared to 1.9111.for 1.0% acid casein stabilized emul- and the interfacial surface area decreased with time (Fig. 8).
sions. The effects of increasing protein concentration on drop- The average droplet diameter of an emulsion stabilized by 1.0%
let size were small at protein concentrations greater than 0.2%. p-lactoglobulin increased from 1.14~ to 1.3111 after 5 hr of
The average droplet size did not differ much between emul- storage at 25°C. The average droplet diameter of an emulsion
sions stabilized by 0.5% and 1.0% protein for the proteins stabilized by 1.0% acid casein increased from 1.91b to 2.39p,
studied. B-Lactoglobulin may form finer emulsions than acid in the same time period (Fig. 8a). The slopes of the bestfit
casein because it may adsorb at the interface more rapidly than lines for the breakdown of these two emulsions were 0.03t.r,/
acid casein and perform extensive molecular interactions. The hr for 1.0% P-lactoglobulin and O.O9p&r for 1.0% acid casein.
ability of a protein to adsorb at interfaces influences its ability By comparing these slopes, it was observed that acid casein
to form emulsions (Dickinson and Stainsby, 1982), but sub- stabilized emulsions had a faster breakdown rate than did p-

444-JOURNAL OF FOOD SCIENCE-Volume 54, No. 2, 1989

lactoglobulin stabilized emulsions. This effect was observed at of emulsions stabilized by acid casein and p-lactoglobulin. The
both protein concentrations studied. The faster breakdown rate P-lactoglobulin gave emulsionsthat had a smaller averagedroplet
of the acid casein stabilized emulsions may reflect, in part, the size, larger surface area, and a slower rate of breakdown at
larger droplets of the emulsion initially. Large droplets provide protein concentrations greater than 0.2%. At protein concen-
a greater driving force for the creaming of the emulsion and trations less than 0.2%, the emulsions were extremely unsta-
possibly resulting in higher rate of droplet collisions. This could ble. The data gathered compared favorably with light scattering
result in an increase in the rate of coalescence (Walstra and studies and electron microscopy. The imaging system can also
Oortwign, 1975). In addition, caseins may form weaker films be used on systems other than oil in water emulsions.
around the fat globules which are desorbed or ruptured upon
contact of the droplets.
Increasing the protein concentration resulted in a retardation
of the rate of coalescence. Emulsions stabilized by 1.0% pro- Davies, W.L. 1936. The proteins of milk: Casein. In “The Chemistry of
Milk,” E.H. Tri (Ed.), D. Van Nostrand Co., Inc. New York.
tein had a slower rate of droplet size increase than those sta- Dickinson, E. an B Stamsby,
. G. 1982. “Colloids in Foods,” Applied Science
bilized by 0.5% protein (Fig. 8a). The slope of curve of Publishers, Barking, Essex, England.
Graham, D.E. and Phillips, MC. 1979. Proteins at li uid interfaces. !.
coalescenceof an emulsion stabilized by 0.5% acid casein was %;&;s of adsorptron and surface denaturatron. J. 2 011.Interface Ser.
O.llpJhr, as compared to O.O9tL/hrfor 1.0% acid casein. The . “. Z”“.

effects of protein concentration on emulsion stability may also Halling, P.J. 1981. Protein stabilized foams and emulsions. CRC Crit. Rev.
Food Sci. Nutr. 13: 166.
reflect, in part, the slightly larger droplet size present at lower Haqu; 2 and Kinsella J E 1969 Emulsip ~~m&,i;;;~~eina De-
stabilizing protein concentrations(Walstra and Oortwign, 1975). velo ‘mint of a stand&&d method J 00”
Kinse la, J.E. 1984. Milk proteins: Ph sicochemical and unctional prop-
Stability associatedwith concentrationmay also reflect a thicker erties. CRC Critical Rev. Food Sci. r?utr. 21(3): 197.
interfacial protein layer providing a stronger barrier to the ap- Klemarzewski, J.L. and Kinsella, J.E. 1987. Unpublished data. Institute
of Food Science, Cornell University, Ithaca, NY.
proach of other emulsion droplets and minimizing coalescence Liboff, M., Goff, H.D., Haque, Z., Jordan, W.K., and Kinsella, J.E. 1988.
(Dickinson and Stainsby, 1982). Changes in the ultrastructure of emulsions as a result of electron mi-
The surface area per gram of emulsion decreased with in- preparation procedures. Food Microstructure. In press.
P::X. N. and Kinsella, J.E. 1978. Emulsifying properties of proteins:
creasing breakdown time (Fig. 8b). Surface area was calcu- evaluation of a turbidimetric technique. J. Agric. Food Chem. 26: 716.
lated in meter* per gram of emulsion to avoid confusion when Phillips, MC. 1981. Protein conformation at liquid interfaces and its role
in stabilizing emulsions. Food Technol. 3550.
comparing the coalescence rates of emulsions stabilized by Sherman, P. 1971. Accelerated tests of emulsion stability. Soap Perfumery
differing levels of protein concentration. The EA of an emul- Cosmetics 44: 693.
Swift, C.E., Lockett, C., and Fryar, A.J. 1961. Comminuted meat emul-
sion stabilized by 0.5% protein would be greater than the EA sions: The capacity of meats for emulsifying fat. Food Technol. 15: 468.
of an emulsion with 1.0% protein if the emulsions were iden- Tadros, T.F. and Vincent, B. 1986. Emulsion Stability. In “En clopedia
tical in droplet size distribution. Data for the average diameter of Emulsion Technology,” P. Becher (Ed.), Marcel Dekker, r nc., New
of emulsions was more precise than the surface area data ob- Tornberg, E. and Lundh, G. 1978. Functional characterization of protein
tained, even though both were calculated from the same data stabilized emulsions: standardized emulsifying procedure. J. Food Sci.
43: 1653.
obtained from the imaging system. This resulted from the in- Van De Hulst, H.C. 1957. “Light Scattering by Small Particles,” John
ordinate contributions of small droplets to surface area as op- Wiley and Sons, New York.
Voutsinas, L., Cheung, E., andNakai, S. 1983. Relationships of hydropho-
posed to volume in equations (5) and (6). To obtain the same biit2v6 to emulslfylng properties of heat denatured protems. J. Food Sa.
degreeof precision for surfaceareacalculations, a greaternumber
of observations must be taken as compared to the average We.&& P.J. 1965. Light scattering by milk fat globules. J. Coil. Interface
Sc-i 97, AOR
I-_. -. . __“.

diameter. Walstra! P.J. 1968. Estimating globule-size distribution of oil-in-water

Other applications. The imaging system was used to study emulsions by spectroturbidimetry. Br. J. Appl. Phys. 6: 1187.
Walstra, P. J. 1986. Formation of emulsions. In “Encyclopedia of Emulsion
emulsions that had the oil phase dyed with Sudan black, a Technology.” P. Becher, (Ed.), Marcel Dekker, Inc., New York.
hydrophobic dye. This was-done p;ior to emulsification,-so Walstraj P.J. and Oort\?ngn, H. 1969. Estimating globule-size distribution
;&&-m-water emulsions by Coulter Counter. Neth. Mdk Dany J. 29:
the lipid phase gave greater contrast. The imaging system gave -__.
similar data when either Sudanblack or congo red was used to Walstra, P.J. and Oortwign, H. 1975. Effect of globule size and concentra-
increase contrast. This indicated that the imaging system could tion on creaming in pasteurized milk. Neth. Milk Dairy J. 29: 263.
Webb, N.B., Ivey, F.J., Crai H.B., and Jones, V.A. 1970. The measure-
be used for the study of emulsions where the oil phase was ment of emulsion capacity 6 y electrical resistance. J. Food Sci. 35: 601.
continuous. With the use of high and low magnification eye- MS received 2/19/88; revised 7/11/88; accepted g/30/88.
pieces, the imaging system could be used to study a wide range
of fine and coarseemulsions (Klemaszewski and Kinsella, 1987).

USING THE DIODE assistedimaging system described in this Suuuortfrom the National Dairy ResearchBoard is aclmowledaed.
paper, it was possible to study the droplet sizes and breakdown

Volume 54, No. 2, 1989-JOURNAL OF FOOD SCIENCE-445