Journal of Food Engineering 92 (2009) 324–330

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Journal of Food Engineering

journal homepage: www.elsevier.com/locate/jfoodeng

Laser diffraction particle sizing by wet dispersion method for spray-dried

infant formula
Byung-Man Kwak, Ji Eun Lee, Jang-Hyuk Ahn *, Tae-Hong Jeon
Research and Development Center, Namyang Dairy Co. Ltd., Gongju 314-914, Republic of Korea

a r t i c l e i n f o a b s t r a c t

Article history: A laser diffraction particle sizing method involving wet analysis could be adapted effectively to measure
Received 25 August 2008 the accurate particle size distribution of a spray-dried infant formula. Polar, polar aprotic and non-polar
Received in revised form 8 December 2008 solvents, such as ethanol, methanol, acetone, pentane, heptane and hexane, were tested as dispersants for
Accepted 10 December 2008
wet analysis. Non-polar solvents such as pentane, heptane and hexane found to be suitable dispersant
Available online 24 December 2008
because the shape of the infant formula particles in non-polar solvents was similar before and after
the measurement while the particles had dissolved in the other solvents. The particle size distributions
Keywords:
(PSD) of the infant formula determined by laser diffraction (Malvern Master Sizer, UK) using the dry anal-
Laser diffraction
Particle size
ysis method with air was unsuitable because some parts of the primary and aggregated infant formula
Infant formula particles had been destroyed. The PSD graph of the air dispersion was shifted toward a smaller particle
Agglomeration size from that of hexane dispersion. Overall, it is believed that laser diffraction particle sizing involving
Aggregation wet analysis with non-polar solvents may provide a suitable particle sizing method for infant formula
Wet analysis products that is better than an air dispersion method.
Dispersion Ó 2008 Elsevier Ltd. All rights reserved.

1. Introduction
Particle sizing of powder foods is widely used to control the vol-
ume of the products. By measuring the particle size of food prod-
ucts, it is possible to examine the relationship between the
particle size and bulk density (Eduardo and Lannes, 2007) or the
relationship between the particle size and solubility of the prod-
ucts (Lowe and Paterson, 1998). Recently, an evaluation of the par-
ticle size has been considered an important factor for developing
sub-micron particles (Jafari et al., 2007) or nano-particles (Jafari
et al., 2008). Traditionally, the tapping sieve method is the simplest
method used widely for particle sizing, which has been established
by international standard (ISO 5223, 1995). A tapping sieve meth-
od is convenient to handle but it does not provide certain particle
sizes due to the weighing of residual particles in each sieve.
An electrical sensing zone method was established to comple-
ment these limits for particle sizing using a sieve method (ISO
13319, 2000) and has been applied in many areas. Another popular
method, laser diffraction, was established as an international stan-
dard (IS0 13320-1, 1999) through a comparison study of sieving
and laser diffraction for raw materials in foodstuff (Dufour et al.,
1993). In addition, it has become the most utilized method for
measuring the particle size of various foods (Adhikari et al.,
2007; Moughal et al., 2000). Laser diffraction particle sizing is con-
* Corresponding author. Tel.: +82 41 856 0381; fax: +82 41 857 7933.
E-mail address: ppori5470@hotmail.com (J.-H. Ahn).
venient and provides various useful data, such as the particle size
distribution (PSD), volume mean diameter (VMD) and d(0.5) value,
using an internal statistical program. It also has many advantages,
such as versatility, short measurement time, simple to operate and
maintain, broad dynamic range and high reproducibility (Xu,
2000).
For laser diffraction, the test samples should be introduced into
the sample cell in which the laser light is transmitted through a
Fourier lens. In most cases, air is used as the dispersant to intro-
duce the samples into the cell through an inlet. However, it is
important to know if the structure of the particles is strong enough
to resist breakage caused by the air pressure. Two types of interna-
tional standard methods for laser diffraction have established in
dry and wet forms (ISO 13320-1, 1999). A manufacturer of laser
diffraction also recommends users select the appropriate disper-
sion method between dry and wet dispersion analysis (Kippax,
2005). One of the key points for laser diffraction particle sizing is
that it is important to present well dispersed particles, which
should be introduced to the sample cell at a suitable concentration
with minimal sample bias. Efficient sample dispersion is important
for achieving an optimum sample flow mode.
Generally, a dry analysis method with an air stream is used for
particle sizing for powders in the field of pharmaceuticals and
micronized powders (Xu and Guida, 2003). However, wet analysis
using solvents is more accurate in case of cohesive powders (Adi
et al., 2007). With other fragile powder samples, measurements
of the PSD of spray-dried infant formula by laser diffraction should

0260-8774/$ - see front matter Ó 2008 Elsevier Ltd. All rights reserved.
doi:10.1016/j.jfoodeng.2008.12.005
 B.-M. Kwak et al. / Journal of Food Engineering 92 (2009) 324–330
be carried out with caution because they are comprised of specific
structures, such as primary particles, aggregates and agglomerates,
which are easily destroyed by pressure. In this study, therefore, we
investigated the effect of different air pressures on particle size
measurement of spray-dried infant formula using laser diffraction
involving dry analysis. In addition, laser diffraction particle sizing
using a wet dispersion method with several solvents was demon-
strated to measure the accurate particle sizes of spray-dried infant
formula. This is the first pioneering study on particle sizing for the
infant formula using laser diffraction.
2. Materials and methods
2.1. Materials
The spray-dried infant formula was provided by the Namyang
Dairy Co. Ltd., which is located in Gongju, Republic of Korea. The
tested infant formula is the most popular product in the Republic
of Korea and consists of 12%(w/w) total proteins, 27%(w/w) total
fats, 56%(w/w) total carbohydrates. Moreover, it contained 13
vitamins and nine minerals. The spray-dried infant formula con-
tains structures that allow milk proteins and lactose to encapsu-
late milk fats. The vitamins consisted of retinol, riboflavin,
thiamine, vitamin B6, ascorbic acid, cyanocobalamin, cholecalcif-
erol, phylloquinone, pantothenic acid, niacin, folic acid, biotin
and b-carotene. The minerals were composed of sodium, magne-
sium, potassium, calcium, iron, zinc, phosphorus, manganese and
copper. The values for the vitamins and minerals certified on the
product details per 100 g are as follows: retinol 510 lg RE, ribo-
flavin 0.6 mg, thiamine 0.45 mg, vitamin B6 0.3 mg, ascorbic acid
50 mg, cyanocobalamin 2 lg, cholecalciferol 8.8 lg, phylloqui-
none 30 lg, pantothenic acid 3 mg, niacin 5 mg NE, folic acid
100 lg, biotin 18 lg, b-carotene 100 lg, sodium 140 mg, magne-
sium 40 mg, potassium 440 mg, calcium 360 mg, iron 4 mg,
zinc 2.6 mg, phosphorus 200 mg, manganese 30 lg and copper
320 lg.
2.2. Chemicals
The solvents used to disperse the sample were methanol, etha-
nol, acetone, pentane, heptane and hexane. The ethanol, methanol,
pentane and heptanes were purchased from J.T. Backer Co.
(Philibsbug, NJ, USA). The hexane and acetone were obtained from
Thermo Fisher Scientific Inc. (Fair Lawn, NJ, USA). Hexane was add
to 1% lecithin for prevent adhesion to the equipment cell window.
2.3. Methods
2.3.1. Particle size analysis by dry method
The particle size distribution of the infant formula for dry anal-
ysis was estimated by laser diffraction (Malvern Mastersizer Sci-
rocco 2000, Malvern Instruments Ltd., UK). All procedures
including sample preparation, dispersion and analysis were carried
out according to the practice guide published by NIST (2001). In
the dry analysis method, air was used as the dispersion agent for
the infant formula particles from the inlet to sample cell. Approx-
imately 15 g of infant formula was loaded into a feeding tray. The
dispersion air pressure was adjusted to 0.1, 0.5, 1.0 and 2.0 bar,
respectively, in order to determine if particle attrition had oc-
curred. Between 1.0% and 3.0% obscuration was achieved through-
out the entire measurement duration. At least five repeat
measurements were obtained from each sample with the mean va-
lue. The residual value was always <1.0%. The analyzed infant for-
mula in the vacuum cleaner was collected for further studies when
dry analysis was completed.
325
2.3.2. Particle size analysis by wet method
The particle size distribution of the infant formula for wet anal-
ysis was measured by laser diffraction (Malvern Mastersizer Hydro
2000S, Malvern Instruments Ltd., UK). In the wet analysis method,
solvents were used to disperse the infant formula particles from
the inlet to sample cell. Approximately 1 g of infant formula was
dispersed in 10 ml of methanol, ethanol, acetone, pentane, hep-
tane, or hexane. The sample in each solvent was added drop-wise
to the sample area containing approximately 150 ml of dispersant
until there was between 10% and 20% obscuration. The refractive
index of each solvent was used as a reference index for statistical
calculation using the particle sizing program; ethanol (1.36), meth-
anol (1.33), hexane (1.38), heptane (1.39), pentane (1.36) and ace-
tone (1.36). The measurements were repeated at least five times to
ensure repeatability. All the residual value was always <1.0%, only
the value of ethanol dispersion PSD was 1.2%. From the measure-
ment of the particle size distribution, various particle sizes such
as D[4, 3] (VMD, volume mean diameter), D[3, 2] (SMD, surface
mean diameter), d(0.1), d(0.9) and d(0.5) value were computed.
Each dispersed infant formula in the sample area was collected
and examined by optical microscopy to analyze the particle shape
and size.
2.3.3. Identification of dissolution in wet analysis
The main components, such as fat, protein and lactose, of the in-
fant formula were examined in order to evaluate whether the com-
ponents had dissolved in each solvent. Ten grams of the infant
formula were mixed with 100 ml of each solvent and the mixture
was shaken for 10 min at 280 rpm on an orbital shaker (LAB-LINE,
plateform 3524-49). The extracted solutions were passed through
Whatman No. 4 paper and collected in previously weighed flasks.
Each filtrate was evaporated using a rotary evaporator and the final
residue was used for analysis of protein and lactose.
The content of protein in the final residue was determined
according to Kjeldahl methods of AOAC 991.20 (2005) with slight
modification. The final residue was digested in H2SO4 with catalyst
(Kjeldahl tablets, Merck) and the protein content was determined
using Distillation unit (BÜCHI 339).
The content of lactose in the final residue was determined using
HPLC (High Performance Liquid Chromatography, Agilent 1100 ser-
ies) coupled with ELSD (Evaporative Light Scattering Detector
2000ES, Altech) according to the methods of Ferreira et al. (1998)
and AOAC 980.13 (2005). The final residue was dissolved in 5 ml
of distilled water. Subsequently, 5 ml of 95% ethanol and 0.5 ml
of 5% oxalic acid (w/v) were added and the volume was adjusted
to 25 ml with deionized water. After shaking of the mixture for
10 min, 10 ml of CHCl2 was added and centrifuged at 13,000 rpm
for 10 min. Ten microliters of the supernatant was injected to the
HPLC-ELS. The chromatographic separation was achieved with a
Prevail carbohydrate ES (Altech, 5 lm, 250 mm  4.6 mm i.d.).
The mobile phase used was acetonitrile–water (75–25, v/v) and
the flow rate was 1.0 ml/min.
Determination of the fat contents was carried out according to
the pentane method (Dieffenbacher and Lüthi, 1986) with six dif-
ferent solvents. Approximately 10 g of the infant formula was
added to 100 ml of each solvent. The mixture was then shaken
for 10 min at 280 rpm on an orbital shaker (LAB-LINE, plateform
3524-49). The extractions were passed through Whatman No. 4
paper and collected in previously weighted beaker. The filtrates
were evaporated on a hot plate until dryness. The total content
of fat was quantified by gravimetry.
2.3.4. Scanning electron microscope (SEM)
Field emission scanning electron microscopy (FE-SEM, FEI,
Sirion, USA) was used to examine the morphology and surface
appearance of the infant formula particles. The samples were
326 B.-M. Kwak et al. / Journal of Food Engineering 92 (2009) 324–330
attached to specimen stubs with two-sided adhesive tape and
Pt-coated with a sputter coater (BAL-TEC, SCD 005, Germany) at
30 mA for 150 s. The coated microcapsules were examined using
a Sirion SEM at 10 kV with a 1.5 nm resolution according to a
previously reported method (Rosenberg et al., 1985).
2.3.5. Microscopy analysis
Visualization of the infant formula particles was carried out
using an optical microscope with (OLYMPUS BX51, JAPAN) an O fil-
ter. Approximately 0.5 g of the infant formula was placed on a slide
glass and the infant formula was dispersed adequately. Images of
the infant formula particles were captured using a microscopy-
coupled digital camera (DIXI 3000). The observations were per-
formed at several magnifications.
For dry method, the infant formula particles such as single and
broken particles were counted under the microscope. The particle
counting was carried out before and after air dispersion at different
air pressures. Twenty views were obtained from each slide and
each view corresponded to the area of 2 mm  2 mm. More than
500 particles were observed on each slide and five slides were pre-
pared for each sample.
3. Results and discussion
3.1. Particle sizing of infant formula by dry analysis method
In the dry analysis method, the infant formula particles were
introduced to the sample cell using a compressed air stream. As
shown in Table 1, the particle size parameters such as d(0.1),
d(0.5), d(0.9), D[4, 3] and D[3, 2] values of the infant formula parti-
cles decreased with increasing air pressure. The D[4, 3] decreased
from 177.8 lm to 147.6 lm with increasing air pressure from
0.1 bar to 2.0 bar, and the d(0.5) value also decreased from
160.7 lm to 135.5 lm. This suggests that the sticky agglomerates
had been separated effectively by the high air pressure. Destruc-
tion of the primary particles or aggregates may be another reason
for the decreasing particle size with increasing air pressure.
According to the laser diffraction manufacturer’s application notes
(Malvern Instruments, 2005. Application Note MRK524-02), the
particle size of a fragile powder could change gradually with
increasing air pressure from 0 bar to 2 bar.
Spray-dried infant formula consists of specific structures, such
as primary particles, aggregates and agglomerates (Fig. 1). Aggre-
gates and agglomerates are well known in that they are formed
by inter-particle bond energies ranging from weak van der Waals
forces to stronger solid-state necks in the case of fumed silica
(SiO2) produced by industrial aerosol processes involving flame,
hot-wall, and spray pyrolysis reactors (Tsantiris and Pratsinis,
2004). The importance of distinguishing between aggregates and
agglomerates was well explained in case of titania (TiO2) pigment
particles (Teleki et al., 2008).
Optical microscopy and SEM were used to inspect the morpho-
logical changes in the infant formula particles before and after air
dispersion. Since optical microscopy had limited magnification and
resolution, BS3406 recommends sizing by optical microscopy for
particles that are a minimum of 10 times larger than the resolution
Table 1
Summary of particle size parameters of the infant formula at different air pressures.
Air pressure (bar) d(0.1) d(0.5)
0.1 74.9 ± 0.6 160.7 ± 1.5
0.5 72.4 ± 0.1 156.5 ± 0.3
1.0 68.3 ± 0.1 149.4 ± 0.4
2.0 54.7 ± 0.6 135.5 ± 1.0
Values are expressed as mean ± standard deviation of five determinations.
Fig. 1. Scanning electron micrograph of the infant formula particles (magnification
of 200).
limit of the objective lens in the microscope (British Standard Insti-
tute, 1993). In a previous study (Zingerman et al., 1992), although
an optical microscopy/image analysis technique was used and par-
ticles below a certain size, 0.5 lm, were disregarded, the relevance
of the experimental results was not adversely affected by scanning
electron microscopy/image analysis system. The optical micros-
copy did not provide full details of the fine infant formula particu-
lates adhering to the surface of the larger infant formula particles
while scanning electron microscopy provided the full details. How-
ever, the limited magnification and resolution showed whether the
infant formula particles had broken up. In addition, the most
important reason for using optical microscopy was to capture seri-
al micrographs if the agglomerates and aggregates had separated
in a natural way. The separation of loosely bound agglomerates
and the detachment of some aggregates from larger ones in a dis-
persant, such as hexane, were observed by optical microscopy. This
is explained more clearly in the next section using a wet analysis
method with several solvents.
A relevant determination of the particle size distribution re-
quires the analysis of a sufficient amount of particles. Vigneau
et al. (2000) noted that a number of 500 particles, five slides each
containing about 100 particles, were determined as the minimal
number of particles required for assessing a reliable particle size
distribution using microscope. In this study, five slides each con-
taining about 500 particles were chosen to determine particle
shape and size distribution for each sample. As the result of obser-
vation, the number of single particles significantly increased after
air dispersion, especially at 1.0 and 2.0 bar. The dispersed single
particles by air at 1.0 bar were pointed with arrows in Fig. 2. The
single and broken particle numbers had risen with increasing air
pressure (Table 2). The increase in the number of single particles
is believed to be caused by the dispersion of small particles ad-
hered to the agglomerates. In case of broken particles, more parti-
cle destruction occurred with higher air pressures. These changes
in number of single and broken particles were probably related
d(0.9) D[4, 3] D[3, 2]
308.2 ± 2.3 177.8 ± 1.3 124.0 ± 1.0
300.8 ± 1.4 173.4 ± 0.6 118.9 ± 0.2
283.0 ± 0.8 163.5 ± 0.4 111.5 ± 0.3
262.8 ± 3.9 147.6 ± 1.6 90.3 ± 1.4
 B.-M. Kwak et al. / Journal of Food Engineering 92 (2009) 324–330 327

Fig. 2. Scanning electron micrographs of the infant formula particles (A) before air dispersion (magnification of 95) and (B) after air dispersion at 1.0 bar (magnification of
100).

Table 2
Changes in number of single and broken particles before and after the air dispersion.

Slide No. Before After

Single Broken Single Broken
0.1 bar 1 41 4 52 5
2 43 5 49 6
3 42 3 41 4
4 49 4 51 6
5 45 4 48 5
Total 220 (8.4 ± 0.7) 20 (0.8 ± 0.1) 241 (9.2 ± 0.8) 26 (1.0 ± 0.2)
0.5 bar 1 41 3 50 4
2 43 4 53 7
3 46 3 49 5
4 42 4 51 6
5 38 4 43 5
Total 210 (8.1 ± 0.5) 18 (0.7 ± 0.1) 246 (9.5 ± 0.7) 27 (1.0 ± 0.2)
1.0 bar 1 39 4 64 14
2 50 4 64 13
3 40 3 65 12
4 45 5 74 14
5 47 3 69 11
Total 221 (8.4 ± 0.9) 19 (0.7 ± 0.2) 336 (12.8 ± 0.9) 64 (2.4 ± 0.3)
2.0 bar 1 41 4 74 28
2 45 3 80 29
3 38 4 80 29
4 37 3 65 26
5 44 4 77 28
Total 205 (7.9 ± 0.7) 18 (0.7 ± 0.1) 376 (14.4 ± 1.2) 140 (5.4 ± 0.2)

Values are expressed in the sum of the counted particle numbers from five slides.
The expressed values within brackets are in terms of percent (%) mean ± standard deviations to the total counted particle numbers of five slides.

to the result of particle size distribution which decreased with
increasing air pressure (Table 1). Increasing the air pressure
seemed to lead aid dispersion of the infant formula but increased
breakage of the particles was also observed. In the case of using
2.0 bar, a better dispersion of the infant formula had occurred
but 2.0 bar was not suitable air pressure for determination of the
PSD of the infant formula in that the number of broken particles
had increased by 4.7%. The broken particle after air dispersion at
2.0 bar is shown in Fig. 3. The exposed cross-section, pointed as
B, is believed to be caused by breakage of particles at a certain area
such as A, where two particles are loosely bound. According to the
laser diffraction manufacturer’s application notes (Malvern Instru-
ments, 2005. Application Note MRK524-02), dry characterization
of fragile particles such as infant formula was not recommended
because dispersion and break-up of the particles occurred simulta-
neously with increasing air pressure. Therefore, a laser diffraction
particle sizing method involving dry analysis was not suitable for
the measurement of the particle size distribution for spray-dried
infant formula.
3.2. Particle sizing of infant formula by wet analysis method
An appropriate dispersant should be selected by considering
several criteria for successful wet analysis. The dispersant should
not dissolve the samples during the measurement but should
wet the sample particles well to effectively separate sticky agglom-
erates. In addition, the dispersant should stabilize the sample par-
ticles to prevent re-agglomeration. Since infant formula is
dissolves easily in water, it was anticipated that water would not
be a suitable dispersant. Hence, several solvents were examined.
The dispersants were chosen according to their polarity. Polar, po-
lar aprotic and non-polar solvents, such as methanol, ethanol, ace-
tone, pentane, heptane and hexane, were examined to determine if
they are suitable dispersants. After laser diffraction particle sizing
using the various dispersion agents, the particles were examined
by optical microscopy to determine if there were changes in shape.
As shown in Table 3, all d(0.5) values and D[4, 3] by the polar
dispersants, such as methanol and ethanol were smaller than those
dispersed by the non-polar dispersants. Despite the initial belief
328 B.-M. Kwak et al. / Journal of Food Engineering 92 (2009) 324–330

that the infant formula particles would partially dissolve in polar

solvents, the polar dispersants were used in the wet analysis meth-
od to compare the changes in shape with the non-polar dispersed
infant formula.
The infant formula particles partially changed in shape in polar
and aprotic solvents (Fig. 4) whereas the particle shape did not
change in non-polar solvents (Fig. 5). Although the shape of the in-
fant formula maintained in the non-polar dispersants, we mea-
sured the main components, such as fat, protein and lactose,
whether the components had dissolved in each dispersant. The
ingredients dissolved in each dispersant were presented in Table
4. The determined contents of fat, protein and lactose in polar sol-
vents were much higher than those in aprotic and non-polar sol-
vents. In non-polar solvents, only fat was extracted among the
analyzed ingredients. The main components should be close to
zero in the ideal dispersant for the determination of particle size.
Polar and aprotic solvents are not suitable dispersant for wet anal-
ysis, since the infant formula had dissolved in those solvents.
Fig. 3. Scanning electron micrographs of the broken particle after air dispersion at In case of non-polar solvents such as pentane, heptane and hex-
2.0 bar (magnification of 2000). ane, only fat was extracted with low level, 0.60–0.65% (g/100 g, w/
w). The extracted fat content by pentane method, generally used
for extraction of surface free fat, was 0.64% and it was similar to
that by heptane and hexane; therefore, the detected fat by heptane
Table 3 and hexane was regarded as free fat on the surface of the infant for-
Summary of particle size parameters of the infant formula in each dispersant. mula. This small amount of the extracted fat may not affect the
Dispersant d(0.1) d(0.5) d(0.9) D[4, 3] D[3, 2]
particle size distribution when using wet analysis. Furthermore,
the infant formula in non-polar solvents was dispersed adequately
Methanol 66.0 ± 2.4 173.2 ± 3.0 361.5 ± 3.8 195.3 ± 2.8 100.0 ± 3.5
Ethanol 72.5 ± 2.1 174.3 ± 2.8 361.2 ± 4.5 197.6 ± 2.4 120.1 ± 3.3
without any formation of re-agglomeration throughout the entire
Acetone 79.7 ± 0.2 180.8 ± 0.4 338.8 ± 1.3 196.0 ± 0.5 125.9 ± 0.7 measurement duration. In case of ethanol, methanol and acetone,
Pentane 72.3 ± 0.7 185.9 ± 0.6 353.5 ± 1.1 200.7 ± 0.7 109.1 ± 1.8 the detected fat contents were higher than those in pentane
Heptane 86.2 ± 0.3 188.8 ± 0.6 355.7 ± 2.1 206.0 ± 0.7 136.9 ± 0.4 extraction. This showed that the fat encapsulated in the infant for-
Hexane 82.2 ± 0.2 187.9 ± 0.3 355.4 ± 1.8 204.6 ± 0.7 132.5 ± 0.3
mula was eluted by polar and aprotic solvents. Therefore, the non-
Values are expressed as mean ± standard deviation of five determinations. polar dispersants including pentane, heptane and hexane would be

Fig. 4. Optical micrographs of the infant formula particles dissolved in different dispersion agents (A) methanol, (B) ethanol and (C) acetone (magnification of 200).

Fig. 5. Optical micrographs of the infant formula particles dissolved in different dispersion agents (A) pentane, (B) heptane and (C) hexane (magnification of 100).
 B.-M. Kwak et al. / Journal of Food Engineering 92 (2009) 324–330
Table 4
Fat, protein and lactose content of solvent extractions of infant.
Dispersant Fat Protein
Methanol 12.19 ± 0.08 0.04 ± 0.00
Ethanol 14.94 ± 0.10 0.04 ± 0.00
Acetone 1.48 ± 0.02 0.01 ± 0.00
Pentane 0.64 ± 0.01 ND
Heptane 0.65 ± 0.01 ND
Hexane 0.60 ± 0.01 ND
Values are expressed as mean ± standard deviation of five determinations.
Values are expressed in terms of percent (%, w/w).
ND, not detected.
Fig. 6. Comparison of the particle size distribution for the infant formula by air
dispersion at 1.0 bar and hexane dispersion.
appropriate dispersant in wet analysis for particle sizing of the in-
fant formula particles.
A comparison of the particle size distribution by dry analysis
with that by wet analysis showed that the particle size distribution
decreased from hexane dispersion to air dispersion at 1.0 bar
(Fig. 6). This shows that the volume mean diameter (VMD) of the
infant formula by air dispersion at the 1.0 bar was determined to
be 163.5 lm, which is 41.1 lm smaller than the VMD (204.6 lm)
determined by hexane dispersion. The d(0.5) value, 149.4 lm, by
air dispersion at 1.0 bar was estimated to be 38.5 lm smaller than
that from the hexane dispersion, 187.9 lm. These results are prob-
ably related to the destruction of the infant formula particles by air
pressure, which was observed and counted under microscope.
According to ISO (1999) and NIST (2001), any deviations from
non-spherical powders can cause bias and errors to be introduced
in the particle size and size distribution results due to the theoret-
ical assumptions of particle sphericity. Since non-spherical parti-
cles such as infant formula have different cross-sections in
different orientations and particles are measured in all possible
orientations, some broadening of the particle size distribution
can be obtained compared to the equivalent distribution. More-
over, the median and mean diameter may be shifted, often to a lar-
ger size. Dufour et al. (1993) also reported the discrepancies of the
particle sizing of irregular shape particles. In case of our study, it
could be also differences from the true value. Particle sizing, how-
ever, using laser diffraction is very useful since it gives a quick and
easy assess to measurement of particle size. It also provides an
easy way to standardization for the quality control in the particle
producing industrial field. Furthermore, as our knowledge, there
was no investigation related to particle sizing method using laser
diffraction for agglomerated and fragile particles such as infant
formula.
329
4. Conclusion
Lactose The measurement of the PSD of commercial spray-dried infant
6.68 ± 0.04 formula should be carried out with caution since it consists of frag-
2.13 ± 0.01 ile particles formed in aggregated and agglomerated structure. In
0.02 ± 0.00 dry method using air stream as a dispersant, the dispersion of
ND
loosely bound agglomerates and particle destructions caused by
ND
ND detachment of a single particle from main aggregates simulta-
neously occurred with increasing air pressure. In laser diffraction
particle sizing, there was a limit to distinguish which one between
particle dispersion and destruction caused the decreasing particle
size with increasing air pressure. Since the laser diffraction method
involving dry analysis could not provide optimal instrumental con-
dition, the particles should be observed by microscopy whether
they were destructed or not. As we demonstrated, a laser diffrac-
tion particle sizing method involving dry analysis was not suitable
method for spray-dried infant formula because particle dispersion
and break-up occurred simultaneously.
On the other hand, particle destruction was not observed in wet
analysis. However, there was a dissolution problem in wet analysis
with some dispersants. Choosing a suitable dispersant which
should not dissolve the infant formula was the most important fac-
tor to carry out a successful PSD measurement using wet analysis.
While the main components of the infant formula dissolved in the
polar or polar aprotic dispersants such as ethanol, methanol and
acetone, they did not dissolve in non-polar solvents. Non-polar sol-
vents such as pentane, heptane and hexane were found to be suit-
able dispersants since the main components did not dissolve, only
small amount of the surface free fat dissolved. All of the PSD by wet
analysis using non-polar solvents was larger than that by dry
method even at lowest air pressure where neither dispersion nor
destruction occurred.
Overall, a laser diffraction particle sizing method using non-po-
lar dispersant provides a suitable particle sizing technique for in-
fant formula products that is superior to the air dispersion
method. We expect the proposed condition for wet analysis would
provide basic information on particle sizing for agglomerated par-
ticles such as infant formula using laser diffraction. This is the first
pioneering study on particle sizing for the infant formula using la-
ser diffraction.
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