ORI GI NAL PAPER

Dairy Lecithin from Cheese Whey Fat Globule Membrane: Its

Extraction, Composition, Oxidative Stability, and Emulsifying
Properties
Dan Zhu

Srinivasan Damodaran
Received: 31 July 2012 / Revised: 14 September 2012 / Accepted: 18 September 2012 / Published online: 30 October 2012
AOCS 2012
Abstract An ethanol extraction method was studied for
the production of dairy lecithin from cheese whey-derived
milk fat globule membrane (MFGM). A two-step ethanol
extraction of MFGM involving rst extraction at pH 6.5,
followed by second extraction at pH 4.5 yielded 17.2 %
lipids. The extracted material contained about 90 % lipids,
4.5 % ash, and 1.2 % moisture. The phospholipid content
of the ethanol extract was 31 % and the remainder was
mostly neutral lipids. The phospholipid fraction contained
34 % sphingomyelin, 31 % phosphatidylcholine, 27 %
phosphatidylethanolamine, 4.6 % phosphatidylserine, and
3.1 % phosphatidylinositol. Since the ethanol extract con-
tained 31 % phospholipids, it can be technically termed as
dairy lecithin. The major fatty acid components were lin-
oleic acid (5.1 %), myristic acid (8.3 %), palmitic acid
(29 %), stearic acid (14 %), oleic acid (25 %), and the
remainder was minor fatty acids with chain length ranging
from C4:0 to C22:5. The dairy lecithin was semi-solid at
room temperature and exhibited a major phase transition at
about 35 C. Owing to its low polyunsaturated fatty acid
content, the dairy lecithin was reasonably stable to oxida-
tion as measured by the rate and extent of hexanal pro-
duction during 35 days of storage at 45 C. Oil-in-water
emulsions made with less than 2 % dairy lecithin (relative
to the total emulsion weight) were unstable; however,
emulsions made with greater than 4 % dairy lecithin were
very stable for more than 60 days at room temperature. The
results of this study indicated that a highly functional dairy
lecithin can be commercially produced using cheese whey-
derived MFGM as the starting material.
Keywords Dairy lecithin Cheese whey Milk fat
globule membrane Emulsion Oxidative stability
Phospholipid composition Phase transition
Introduction
Soy-derived lecithin is widely used as an emulsier in
food, pharmaceuticals, and cosmetic products. In the food
industry, lecithin is used as a multifunctional ingredient in
the manufacture of chocolate, bakery and instant products,
margarines, and mayonnaise [1, 2]. Commercial lecithin is
a concentrated mixture of phospholipids, mainly comprised
of phosphatidylcholine (PC), phosphatidylethanolamine
(PE) and phosphatidylinositol (PI), and other minor sub-
stances such as phosphatidic acid (PA), glycolipids, and
complex sugars dissolved in neutral oil.
Currently, lecithinfromsoybeanis the mainsource of food-
grade lecithin available to the food industry. However, an
alternative source of lecithin that can be regarded as a non-
GMO (non-Genetically Modied Organism) product would
be desirable to cater to the needs of a segment of the popula-
tion that is either allergic to soybean products or have an
aversion to GMO products. In this regard, the cheese whey-
derived milk fat globule membrane (MFGM) has the potential
to be a starting material for the production of dairy lecithin.
The MFGM-derived phospholipid has been receiving a
great deal of attention recently as it is considered to be a rich
source of bioactive lipids, especially phosphatidylserine (PS)
and sphingomyelins (SM) [3, 4]. Several studies have sug-
gested that consumption of PS and MFGM-derived SM
affects cell growth and development, improves memory,
reduces stress, suppresses Alzheimers disease, and enhances
brain development in infants [46], although the veracity of
such claims remains to be conrmed.
D. Zhu S. Damodaran (&)
Department of Food Science, University of Wisconsin-Madison,
1605 Linden Drive, Madison, WI 53706, USA
e-mail: sdamodar@wisc.edu
1 3
J Am Oil Chem Soc (2013) 90:217224
DOI 10.1007/s11746-012-2152-5
The MFGM is usually obtained from buttermilk. Isola-
tion of MFGM from buttermilk is a cumbersome process
[7, 8], which makes it uneconomical as a source of com-
mercial production of dairy lecithin. Recently, we have
developed a simple and straightforward process to separate
MFGM from cheese whey for the production of fat-free
whey protein isolate (WPI) and whey protein concentrate
[911]. This process is currently being used in commercial
production of fat-free WPI and the MFGM by-product
has not found a commercial value until now. Since the
MFGM is obtained in this process as a waste by-product,
commercial production of dairy lecithin from this
by-product should be economical depending on the isola-
tion method.
There are several solvent-based methods available to
extract phospholipids from biological materials, including
extraction with near-critical dimethylether, ethanol, or
aqueous alcohol mixtures [1214]. Being generally regar-
ded as safe, ethanol is a preferred solvent for preparing
food-grade lecithin. Since PC is more soluble than other
phospholipids in ethanol, the resulting lecithin tends to be
enriched in PC, which is a good emulsier for oil-in-water
emulsions but not for water-in-oil emulsions [15, 16].
Furthermore, ethanol can be easily redistilled from the
extraction mixture and re-used in the process, which should
make the process economical.
The aim of this study was to investigate an ethanol-
based method for the extraction of phospholipids and other
lipids from whey-derived MFGM and to study the func-
tional properties, including the fatty acid composition,
thermal melting properties, oxidative stability, and emul-
sifying properties, of the extracted lipid.
Experimental Procedures
Materials
Claried and pasteurized mozzarella cheese whey was
obtained from a local cheese plant in Wisconsin. The pH of
the cheese whey was about 6.4. Absolute ethanol (200
proof) was from Decon laboratories, Inc. (King of Prussia,
PA). Ethyl pentanoate was from Sigma-Aldrich (St. Louis,
MO).
Isolation of MFGM from Cheese Whey
Isolation of MFGM from cheese whey was carried out as
described previously [10]. Briey, cheese whey (pH 6.4)
was rst ultraltered ve-fold using a spiral wound 10-kDa
molecular weight cut-off polysulfone membrane using a
benchtop Millipore ProFlux M12 Tangential Flow Filtration
System (Millipore Corp., Billerica, MA). Subsequently, the
retentate was dialtered three-fold in a continuous mode
using water. The nal conductivity of the dialtered
retentate was about 500 lS cm
-1
. The pH of the retentate
was adjusted to 4.2 and incubated for 30 min at 35 C
followed by centrifugation at 16,000g for 15 min at room
temperature. The sediment, which was the MFGM, was
re-suspended in water at pH 6.8 and lyophilized and stored
at -20 C until used.
One-Step Extraction of Dairy Lecithin from MFGM
at pH 6.5
In a typical lab-scale extraction, ethanol (720 mL) was
added to the dried MFGM (80 g) in a 1,000 mL beaker.
The mixture was rst subjected to sonication using a
Branson-450 sonier (Branson Ultrasonics, Danbury, CT)
equipped with a tip horn (18 mm diameter) at an amplitude
setting of 8, duty cycle of 90 %, and output energy 110 W.
The sonication was performed in an ice-water bath and the
temperature was maintained at *50 C for 25 min to
disrupt the membrane structure. The slurry was then
adjusted to pH 6.5 by adding 2 N Hcl and stirred with a
magnetic stirrer for 30 min at room temperature followed
by centrifugation at 16,000g for 10 min at room tempera-
ture. The clear, light yellow supernatant was collected. The
residue was re-suspended in 500 mL ethanol at pH 6.5 and
the above cycle was repeated once. The collected super-
natants were mixed together and evaporated in a rotary
evaporator at 60 C and 680 mm Hg pressure to remove
ethanol. The residue was then ushed with nitrogen at
60 C to remove the residual solvent.
Two-Step Extraction of Dairy Lecithin from MFGM
at pH 6.5 and 4.5
The negatively charged phospholipids, such as SM, PI, and
PS, are less soluble than the neutral phospholipids, such as
PC, in ethanol at pH 6.5 and therefore the efciency of
extraction of SM, PI, and PS might be limited at pH 6.5.
However, if the charge on these phospholipids can be
reduced by adjusting the pH to 4.5 (isoelectric pH of the
phosphate groups in PL), then they might become soluble
in ethanol. Thus, a two-step extraction was performed to
extract all phospholipids from MFGM: rst, the MFGM
was extracted at pH 6.5 as described above. The residue
obtained after the extraction was re-suspended in 300 mL
ethanol at pH 4.5 and stirred for 30 min at room temper-
ature and then centrifuged at 16,000 g for 10 min at room
temperature. This supernatant was combined with the
supernatants obtained at pH 6.5 and the combined extract
was evaporated in a rotary evaporator as described above.
The extracted dairy lecithin was stored at -20 C until
used.
218 J Am Oil Chem Soc (2013) 90:217224
1 3
Compositional Analysis of the Dairy Lecithin
The total fat content of the ethanol extract was analyzed by
the Folch method [17]. The moisture content was deter-
mined by the Karl Fischer method using a 795 KFT Titrino
(Metrohm Ion analysis, Metrohm Ltd., CH9101, Herisau,
Switzerland). The ash content was determined according to
the standard method for the dairy products [18]. Triplicates
were performed for each analysis and the data were pre-
sented as means standard deviations.
Analysis of Nonpolar Lipids by Thin Layer
Chromatography (TLC)
The lecithin extract (25 mg) was dissolved in 1.0 mL of
CHCl
3
:MeOH (2:1, v/v) solvent. An aliquot of the sample
solution (10 lL) was spotted onto a 10 9 10 cm HPTLC
Silica 60 plate (EMD Chemicals Inc., Merck KGaA
Darmstadt, Germany). The lipids were separated by one-
dimensional TLC using a solvent mixture composed of
petroleum ether:ethyl ether:acetic acid (85:15:2, v/v) [14].
Lipids were visualized by exposure to iodine vapor. All
organic compounds were further detected with 10 % (v/v)
sulfuric acid in methanol followed by charring.
Fatty Acid Composition Analysis
The fatty acid composition of the extracted dairy lecithin
was performed by Rtech Laboratories (St. Paul, MN) based
on the Association of Ofcial Analytical Chemists Method
996.06 [19].
Phospholipid Analysis by
31
P-NMR Spectroscopy
Approximately 0.25 g of dairy lecithin was dissolved in
25 mL of a detergent solution, and the
31
P analysis was
performed at the Avanti Polar Lipids, Inc. (Alabaster, AL)
using a Bruker AVANCE III 400 MHz NMR spectrometer.
The
31
P response was calibrated with a dioleoyl phospha-
tidylcholine standard. The sample solution was assayed at
512 scans with chemical shift interpretation for identica-
tion against known dairy phospholipid standards.
Differential Scanning Calorimetry (DSC)
The thermal melting behavior of dairy lecithin was studied
using a differential scanning calorimeter (DSC) (Micro
DSC model VII, Setaram, Caluire, France) as follows: The
lecithin (50 mg) was accurately weighed into pre-weighed
DSC vessels (Hastelloy C276) and closed tightly with the
stopper. An empty DSC vessel was used as reference.
Samples were heated from 5 to 115 C at a constant
heating rate of 1 C/min. After the rst scan the sample
was cooled back to 5 C and re-scanned from 5 to 115 C.
The melting temperature (T
m
) was determined from the
thermogram using the Setsoft software (version 1.4) sup-
plied by the DSC manufacturer. The DSC was calibrated
using cyclohexane, phenyl ether and o-terphenyl standards
recommended by the DSC manufacturer. DSC analysis was
carried out in triplicates.
Emulsion Preparation
Oil-in-water emulsions were prepared as reported else-
where [20]. The dairy lecithin from MFGM was accurately
weighed (5320 mg) into a 7-mL plastic vial and mixed
with 0.4 g soybean oil. The mixture was incubated at 45 C
for 10 min to make ensure complete dispersion of the
sample in the soybean oil. To the mixture was added
1.6 mL of 10 mM phosphate buffer (pH 7.0, 0.02 %
NaN
3
).The mixture was pre-emulsied for 1 min in ice-
water bath using a Branson-450 sonier(Branson Ultra-
sonics, Danbury, CT) equipped with a microtip horn with
the setting 2 and output 8 %. The pre-emulsied mixture was
then passed through a high-pressure homogenizer (Emulsi-
Flex-B3, Avestin, Inc., Ottawa, Canada) at an operating
hydraulic pressure of 152 MPa with three passes. The volume
fraction of the oil phase (triglyceride) in the emulsion was
20 % w/w. Emulsions were made in triplicates.
Particle Size Determination
The droplet size distributions of the emulsions were mea-
sured on the same day of preparation using a particle size
analyzer (90Plus, Brookhaven Instruments Corporation,
NY). The relative refractive index of the dispersed phase
and the continuous phase was 1.473 and 1.330, respec-
tively. The emulsion samples were stored at 23 C and the
particle size was determined periodically. The average
diameter based on volume-to-surface area method (d
32
)
was used to monitor changes in droplet size distribution
during storage. The mean diameter d
32
was dened as
d
32

P
n
i
d
3
i
P
n
i
d
2
i
1
where n
i
is the number of droplets of diameter d
i
. To prevent
multiple scatteringeffects, the emulsions were diluted, prior to
particle size measurements, with distilled water to keep the
concentration typically between 10
-5
and 10
-2
volume frac-
tion. Experiments were performed in triplicate and the results
are presented as means standard deviations.
Creaming
The emulsions were stored at 23 C in graduated glass
tubes. The total height of the emulsion (H
e
) and the height
J Am Oil Chem Soc (2013) 90:217224 219
1 3
of the serum layer (H
s
) were measured. The extent of
creaming was characterized as
Creaming index 100 x H
s
=H
e
: 2
Determination of Oxidative Stability
The dairy lecithin (40 mg) was weighed exactly into glass
vials (17 9 60 mm, Fisher Scientic, Pittsburgh, PA)
capped with Teon lined screw caps. The vials were stored
in dark incubation chambers maintained at 45 C. Vials
were withdrawn (in triplicates) at various time intervals for
analysis of aroma volatiles.
Analysis of Headspace Volatiles by Gas
Chromatography-Mass Spectrometry (GCMS)
Aroma volatiles in dairy lecithin samples were analyzed by
headspace GCMS analysis using the solid-phase mic-
roextraction (SPME) technique as described elsewhere
[11]. In a typical experiment, 2 mL of deionized water and
56 small glass beads were added to each vial containing
40 mg lecithin. The vials were re-capped and the disper-
sions (1.5 %) were vigorously stirred for 5 min at room
temperature using a magnetic stirrer. Then, to each vial was
added 25 lL of a stock solution (100 ppm) of ethyl pent-
anoate as an internal standard and the vial was then stop-
pered with a Teon septum and incubated in a Multi-Block
heater (Barnstead Lab-Line, Cardinal Health, Dublin, OH)
at 37 C. A SPME needle was inserted into the vial and the
SPME ber (50.3 lm DVB/Carbone/PDMS; Supelco,
Bellefonte, PA) was exposed to headspace for 40 min.
Volatiles were desorbed from the SPME ber at 220 C for
5 min in the injection inlet of Hewlett-Packard HP6890
series gas chromatograph (Hewlett-Packard, Palo Alto,
CA) and separated on a Restek RTX-5MS capillary column
(30 m 9 0.25 mm, 0.5 lm thickness) (Restek Inc., Belle-
fonte, PA) using helium as the carrier gas at a constant ow
rate of 0.9 mL/min. For GC analysis, GC oven temperature
was maintained at 35 C for 8 min, increased from 35 to
120 C at the rate of 5 C/min and held at 120 C for 1 min
and then ramped from 120 to 250 C at the rate of 10 C/
min and held at 250 C for 3 min. Volatiles eluting from
the column were routed to an Agilent 5973 mass spec-
trometer (Agilent Technologies, Inc., Santa Clara, CA) and
identied using NIST 98 mass spectral library, version 1.7
(National Institute of Standards and Technology, Gai-
thersburg, MD). Operating conditions for MS were: ion
source temperature of 230 C, electric multiplier tube
(EMT) voltage of 2752.9 V, and mass scan range of m/
z = 36250 atomic mass units at the rate of 2.76 scans/
min. Samples were analyzed in triplicates and volatile
compounds were quantied from peak area measurements
compared against the internal standard (ethyl pentanoate).
The amount of hexanal was calculated according to the
following equation:
Hexanal content lg/g A
X
=A
IS
Wt
IS
=Wt
S
: 3
where A
X
is the area of hexanal in the sample, A
IS
is the
area of internal standard in the sample, Wt
IS
is the amount
of internal standard (lg), and Wt
S
is the sample weight (g).
The content is expressed in lg/g [21].
Statistical Analysis
A completely randomized design was used in the study. For
the emulsion functionality, the treatments were the con-
centration of dairy lecithin and storage time; the responses
were the emulsied particle sizes and creaming index. For
the oxidative stability, the treatment was storage time; and
the response was the concentration of hexanal. All treat-
ments were performed in triplicate. Data were analyzed
using JMP

Pro 10.0.0 software (Copyright 2012 SAS

Institute Inc., Cary, NC). One-way analysis of variance
(ANOVA) was performed to determine the treatment
effects. When signicant treatment effects (P\0.05) were
found, their means were separated by the Tukey test.
Results and Discussion
Composition of Dairy Lecithin
Previously, we reported that the total lipid content of the
MFGM isolate as determined by the Mojonnier method
(AOAC, 1996) was about 1719 % (w/w) and the protein
content was 6673 % and the remainder was ash, moisture
and lactose [11]. In the present study, the yield of total
lipids resulting from one-step ethanol extraction of MFGM
at pH 6.5 was 14.7 0.6 %. However, in the two-pH step
ethanol extraction of MFGM, rst at pH 6.5 and then at pH
4.5, the yield of total lipid was about 17.2 1.1 % on dry
weight basis, which indicated that even after the two-step
ethanol extraction a small amount of some lipids remained
in the residue.
Compositional analysis of the dairy lecithin obtained by
the two-step ethanol extraction showed that it contained
89.9 2.3 % total lipids, 1.2 0.02 % moisture, and
4.6 0.1 % ash.
Figure 1 shows the TLC prole of dairy lecithin
obtained from two-step ethanol extraction of MFGM. In
addition to phospholipids, the extract contained signicant
amounts of neutral lipids (TAGs, DAGs, MAGs, choles-
terol, and cholesterol esters) and free fatty acids. A com-
parison of the intensity of the PL band against the rest of
the neutral lipids in the sample suggests that the PL content
of the lecithin sample was less than 50 % of the total lipids.
220 J Am Oil Chem Soc (2013) 90:217224
1 3
Figure 2 shows a typical
31
P-NMR spectrum of dairy lec-
ithin extracted from MFGM. The phospholipid content
determined from the
31
P NMR of the lecithin sample
extracted by the two-step method is presented in Table 1.
The total phospholipid content of the sample was about
31 % (w/w) and the remainder was mostly neutral lipids.
These results are similar to the value reported by Boyd
et al. [22] for lipids extracted from freeze-dried whey
powder using the Mojonnier method. The major phospho-
lipids in the present lecithin sample were PC (*31 %), PE
(*27 %), and SM (*34 %) and the minor components
were PS and PI at about 4.6 and 3.1 %, respectively. These
values were very different from the phospholipid compo-
sition of lipids extracted from whey protein concentrate [22]
in which PC and PE were reported to be the major phos-
pholipids at 33 and 60 % level, respectively, with a very
small amount of SM at about 1.7 %. The reason for the dif-
ference is that in the latter study the neutral lipids were rst
removed by extraction with hexane, followed by extraction of
the phospholipids using an ethanolwater mixture [22].
The phospholipid content of the lecithin sample
extracted from MFGM using the one-step ethanol extrac-
tion method is also shown in Table 1. The total phospho-
lipid content of this lecithin sample was also about 31 %,
which was same as the lecithin sample obtained by the two-
step extraction method (Table 1). Thus, although the yield
of total lipids in the two-step ethanol extraction method
was higher than the one-step extraction method (i.e., 17.2
vs. 14.7 %), neither the neutral lipid to phospholipid ratio
nor the composition of the phospholipid was altered by
these extraction methods.
Fatty Acid Composition of Dairy Lecithin
The fatty acid composition of the dairy lecithin obtained
by the two-step extraction method is shown in Table 2.
The major fatty acids were linoleic (5.1 %), myristic
(*8.3 %), palmitic (*29 %), stearic (*14 %) and oleic
(*25 %) acids. The content of other fatty acids, such as
C4:0, C14:1, C16:1 and C18:3, was very low. Of the total
fatty acids, the mono-unsaturated fatty acids content was
about 20 % and the saturated fatty acid content was about
46 %. The total fat content of the lecithin, based on the
fatty acid content expressed as triglyceride, was about
79 % (Table 2). This implies that the reminder of the fat
in the dairy lecithin was cholesterol and cholesterol esters.
Thus, the distribution of phospholipids, neutral lipids, and
cholesterol in the dairy lecithin sample was about 32, 47,
and 21 %, respectively.
Thermal Melting Prole of Dairy Lecithin
Due to the high saturated to unsaturated fatty acid ratio
of dairy lecithin (1.5:1.0), the dairy lecithin is semi solid
at room temperature. In contrast, because of its high
unsaturated fatty acid content (about 10 % C18:1, and
59 % C18:2 [22]) soy lecithin is typically a liquid at
room temperature. The thermal melting prole of the
dairy lecithin (Fig. 3) showed a major endothermic peak
- Cholesterol
esters
- TAGs
- FFAs
- Cholesterol
- DAGs
- MAGs
- PLs
A B
Fig. 1 Thin layer chromatography of dairy lecithin. a Sample stained
with iodine; b Charring method
1.0 0.0 -0.5 -1.0
0
.
2
1

S
M
0
.
2
6

P
E
0
.
7
0


P
I
0
.
8
1

P
C
0
.
4
7

P
S
Chemical shift (ppm)
Fig. 2
31
P NMR spectrum of dairy lecithin. Peak identication: SM
sphingomyelin, PE phosphatidylethanolamine, PS phosphatidylser-
ine, PI phosphatidylinositol, PC phosphatidylcholine
J Am Oil Chem Soc (2013) 90:217224 221
1 3
at 35 C and two minor endothermic peaks at about
12 C and 45 C. When the sample was re-scanned after
cooling back to 5 C (broken line, Fig. 3), the sample
exhibited a major broad transition at 2535 C, a minor
transition at 9 C, and a broad and shallow transition at
4050 C. In general, the thermogram qualitatively
shifted to a lower temperature during re-scanning com-
pared to the rst scan, indicating that the packing order
of lipids in the solid state had been altered after the
initial melting. Nevertheless, the thermogram suggested
that the dairy lecithin essentially remained as a solid fat
at temperatures below 25 C, compared to the liquid
state of soy lecithin. This property of dairy lecithin may
provide some unique advantages over soy lecithin in
certain food applications where thin solid lm coating of
lecithin might be desirable.
Oxidative Stability of Dairy Lecithin
Since the polyunsaturated fatty acid content of dairy leci-
thin is very low (Table 2), it is reasonable to predict that
the oxidative stability of dairy lecithin would be much
better than lecithin from other sources. Lipid oxidation of
MFGM generates several volatile compounds [11]; how-
ever, hexanal production was used in this study as an
indicator of oxidative stability of dairy lecithin. The time
evolution of hexanal production in dairy lecithin during
storage at 45 C is shown in Fig. 4. A small spike in
hexanal production was observed during the rst two days
of storage, followed by a lag time for about 10 days,
beyond which a second phase of hexanal production
occurred until it reached a saturation value after 20 days.
The maximum amount of hexanal produced after 35 days
of storage of the sample at 45 C was about 3 lg/g. Since
the lecithin sample was in the liquid state at the 45 C
storage temperature, the bimodal production of hexanal
Fig. 3 Differential scanning calorimeter thermogram of dairy leci-
thin. The solid line represents the rst scan and the dashed line is the
rescan of the same sample
Table 1 Phospholipid content and composition of ethanol soluble extract from MFGM under two different pH extraction conditions
Phospholipid Ethanol extraction at pH 6.5 only Combined ethanol extraction at pH 6.5 and 4.5
g/100 g % of total PL g/100 g % of total PL
SM 9.94 32.33 10.57 34.03
PE 8.85 28.79 8.49 27.36
PS 1.21 3.93 1.42 4.58
PI 0.85 2.76 0.96 3.09
PC 9.90 32.21 9.61 30.95
Total 30.75 31.05
Phospholipid contents were determined by
31
P NMR. The values shown are from single NMR analysis for each sample
Table 2 Fatty acid composition of dairy lecithin
Fatty acid % Fatty acid %
C04:0 Butyric 2.1 C17:1 Margaroleic 0.2
C06:0 Caproic 1.2 C18:0 Stearic 13.7
C08:0 Caprylic 0.6 C18:1 Oleic 24.9
C10:0 Capric 1.4 C18:1T Elaidic 4.3
C12:0 Lauric 1.9 C18:2 Linoleic 5.1
C13:0 Tridecanoic 0.1 C18:2T Linoelaidic 0.7
C14:0 Myristic 8.3 C18:3 Linolenic 0.4
C14:1 Myristoleic 0.5 C20:0 Arachidic 0.2
C15:0 Pentadecanoic 1.1 C20:3 Eicosatrienoic 0.4
C16:0 Palmitic 29.1 C20:4 Arachidonic 0.5
C16:1 Palmitoleic 1.1 C21:0 Heneicosanoic 0.6
C16:1T Palmitelaidic 0.4 C22:5 Docosapentaenoic 0.2
C17:0 Margaric 0.7
cis-Monounsaturated fat 20.32 Saturated fat 45.77
cis-Polyunsaturated fat 5.19 Total fat (as triglycerides) 79.15
Omega 3 fat 0.78 trans fat 4.04
Omega 6 fat 4.42
Based on single sample analysis
222 J Am Oil Chem Soc (2013) 90:217224
1 3
cannot be attributed to phase distribution in the sample. It
is likely that the free fatty acids, which are in considerable
amounts in the sample (Fig. 1), may initially undergo rapid
oxidation as a result of a greater access of the double bonds
to oxygen, followed by oxidation of phospholipid- and
TAG-bound fatty acids. We have observed a similar
bi-modal hexanal production during oxidation of intact
MFGM isolated from cheese whey [11].
Emulsifying Properties of Dairy Lecithin
The effect of lecithin concentration on the average size
(d
32
) of droplets formed in oil-in-water emulsions is shown
in Fig. 5. The droplet size gradually decreased from about
1 lm to about 0.28 lm as the dairy lecithin concentration
was increased from 0.25 to 12 % (w/w). The emulsions
made with \2 % dairy lecithin concentration were very
unstable and they phase separated within 24 h (Fig. 6). The
rate of creaming of the emulsion made with 2 % dairy
lecithin is shown in Fig. 7 as an example. The creaming
index of this emulsion reached about 50 % within 14 days
of storage at room temperature and remained at that level
thereafter. On the other hand, emulsions made with lecithin
concentration C4 % were very stable for more than
60 days at room temperature (Fig. 6); there was no
observable creaming and the average droplet size remained
stable at 0.4 lm. Thus, the minimum concentration of dairy
lecithin required to produce a kinetically stable 20 % oil-
in-water emulsion appears to be about 4 % (w/w) of
emulsion. This value corresponds to about 20 % lecithin
relative to the weight fraction of the dispersed phase. Since
the phospholipid content of the dairy lecithin sample was
about 32 %, this translates to about 6 % of dairy phos-
pholipids relative to the weight of the dispersed oil phase.
The results presented here provide a simple commer-
cially feasible ethanol extraction method for the extraction
of phospholipid-rich lipids from whey-derived MFGM.
The MFGM is a by-product waste in the manufacture of
fat-free WPI [10], and therefore dairy lecithin extraction
Fig. 4 Time evolution of hexanal production in dairy lecithin stored
at 45 C
Fig. 5 Effect of dairy lecithin concentration on the droplet size (d
32
)
of freshly made 20 % oil-in-water emulsion. The lecithin concentra-
tion represents weight percentage relative to the total emulsion weight
Fig. 6 Photographs of 20 % oil-in-water emulsion stabilized by
0.2512 % dairy lecithin at 4 h, 24 h, and 8 weeks storage at room
temperature
Fig. 7 The effect of storage time at room temperature on the
creaming index of the 20 % oil-in-water emulsion stabilized by 2 %
dairy lecithin
J Am Oil Chem Soc (2013) 90:217224 223
1 3
from this waste as the starting material should be eco-
nomical. In addition, recycling of ethanol used in the
process will also make the process very cost effective.
Since the phospholipids in dairy lecithin mainly contains
more saturated and mono-unsaturated fatty acids (C16:0
and C18:0, and C18:1) and negligible amount of poly-
unsaturated fatty acids than it is in soy lecithin, the oxi-
dative stability of dairy lecithin is expected to be better
than soy lecithin. Furthermore, in addition to its use as an
emulsier in oil-in-water emulsions, the high melting
temperature of dairy lecithin may present an opportunity to
use it in other food and pharmaceutical applications where
thin solid emulsier lm coating is desirable.
Acknowledgments This research was funded by a grant from the
Graduate School at the University of Wisconsin-Madison under the
Innovation and Economic Development Research program.
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