Powder Technology 372 (2020) 394–403

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Powder Technology

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

Effect of storage conditions on physicochemical and microstructural

properties of skim and whole milk powders
Arissara Phosanam a,b, Jayani Chandrapala a, Thom Huppertz c,d, Benu Adhikari a,⁎, Bogdan Zisu e
a
School of Science, RMIT University, Bundoora, Melbourne, VIC 3083, Australia
b
Faculty of Natural Resources and Agro-Industry, Kasetsart University, Chalermphakiat Sakon Nakon Province Campus, 47000, Thailand
c
FrieslandCampina, Amersfoort 3818, the Netherlands
d
Food Quality and Design Group, Wageningen University, Wageningen 6708, the Netherlands
e
Spraying System Co., Fluid air, Melbourne, VIC 3029, Australia

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

Article history: This study examined the effects of storage conditions on physicochemical and microstructural characteristics of
Received 18 April 2020 whole milk powder (WMP) and low-, medium- and high-heat skim milk powder (SMP). Powders were stored at
Received in revised form 3 June 2020 three temperatures (25 °C, 35 °C and 45 °C) and three humidity levels (11%, 44% and 85%) for 21 days. WMP
Accepted 7 June 2020
exhibited lower equilibrium moisture content (EMC) than SMP at these storage conditions. SMP and WMP stored
Available online 10 June 2020
at 45 °C had lower EMC and monolayer moisture content than those stored at 25 °C and 35 °C. WMP had lower Tg,
Keywords:
crystallinity and caking strength than SMP. At 85% RH, the particles became irregularly shaped, caused by lactose
Milk powders crystallization. Crystallization of lactose and caking of powders became prominent when Tg decreased below the
Storage conditions storage temperature. Crystallization of lactose in these powders increased caking strength, Maillard reaction and
Glass transition surface free fat, particularly at high RH (85%) and high temperature (45 °C) conditions.
Crystallinity © 2020 Elsevier B.V. All rights reserved.
Surface-free fat
Caking

1. Introduction
Milk powder is convenient to use, saves transportation costs and can
be stored for a relatively long time (12–24 months) at ambient temper-
ature [1–3]. Spray drying is one of the most commonly applied tech-
niques for converting fluid milk into powder. Despite its many
advantages, spray drying process causes chemical and physical modifi-
cations in milk components that increase propensity of Maillard reac-
tion, milk fat oxidation and crystallization of lactose [4]. The nature
and extent of such modifications depends upon several parameters
such as the composition of milk and the storage and processing condi-
tions [5,6]. The rapid removal of water during spray drying results in
powders with an amorphous glassy matrix [7], which tends to convert
to a rubbery state when stored at high temperature and relative humid-
ity (RH) [8,9]. Powders in the glassy state can support their own weight
due to the high viscosity (1012 Pa.s) while those in the rubbery phase
display lower viscosity (106–108 Pa.s) [10] and greater molecular mobil-
ity, which can lead to undesirable phenomena such as lactose crystalli-
zation, Maillard reaction, and caking [11,12]. The existence of free fat on
surface of the powders and lactose crystallization play significant roles
in caking [13]. Rapid moisture removal during the spray-drying process
⁎ Corresponding author.
E-mail address: benu.adhikari@rmit.edu.au (B. Adhikari).
also results into excess fat on the powder surface and subsequent crys-
tallization of lactose hastens the migration of fat to surface and thus con-
tributes to caking [14].
Whole milk powder (WMP) and skim milk powder (SMP) are
widely used as ingredients in infant formula products, bakery products,
ice cream, yoghurt, dairy beverages, confectionary products and proc-
essed meats due to their desirable functional (e.g., solubility, emulsify-
ing, and foaming) and nutritional properties. The main compositional
difference between WMP and SMP is the fat content; WMP contains
~26–27% (w/w) fat whereas SMP contains <1.0% (w/w) fat [15]. Three
principal categories of SMP are distinguished: high-, medium- and
low-heat SMP, depending on the undenatured whey protein content
per grams of powder, known as whey protein‑nitrogen index (WPNI).
SMPs with a WPNI value ≥6.0 mg are referred to as low-heat (LH).
SMPs with WPNI values between 1.5 and 6.0 mg are identified as
medium-heat (MH), and SMPs with WPNI value ≤1.5 mg are defined
as high-heat (HH). This classification system of milk powders is used
commercially in various food applications [16]. Spray-dried SMPs can
be produced from milk heated at high (~120 °C for 240 s), medium
(~90 °C for 30 s) and low (~80 for <5 s) temperature conditions [17].
These processing conditions substantially affect the physicochemical
characteristics of milk powders. Thus, the storage condition for these
powders should be designed in such a way that the fresh-like physico-
chemical properties of the milk powders are maintained as long as
possible.

https://doi.org/10.1016/j.powtec.2020.06.020
0032-5910/© 2020 Elsevier B.V. All rights reserved.
 A. Phosanam et al. / Powder Technology 372 (2020) 394–403
Despite continued research on milk powders, no reported study has
compared the physicochemical properties of SMP based on heat class
(high-, medium- and low-heat) and WMP differentiated by the milk
fat content when stored at very low and high RH conditions at mild tem-
perature. Thus, this study aimed to study effects of storage at three RHs
(11%, 44% and 85%) and three temperatures (25 °C, 35 °C and 45 °C) car-
ried out for 21 days on the moisture sorption isotherms, Tg, caking
strength, lactose crystallization, microstructure and the browning of
low-, medium- and high-heat SMP and WMP.
2. Materials and methods
2.1. Materials
LH-, MH- and HH-SMP and WMP were obtained from Saputo
Australia (Melbourne Victoria 3001, Australia). The declared protein
and the fat contents for SMPs were 33% (w/w) and 1% (w/w), respec-
tively, whereas WMP contained 26% (w/w) fat and 24% (w/w) protein.
Potassium carbonate (K2CO3), potassium acetate (CH3COOK), lith-
ium chloride (LiCl), sodium chloride (NaCl), potassium iodide (KI), po-
tassium chloride (KCl) and magnesium nitrate (Mg (NO3)2) were
provided by Sigma Aldrich Pty Ltd. (Castle Hill, NSW, Australia). All
chemicals are analytical-grade chemicals and were used without any
further modification/purification.
2.2. Determination of sorption isotherms
Approximately 2 g milk powders were placed in a pre-weighed plas-
tic cup and this cup was kept in desiccator containing saturated salt so-
lutions of LiCl, CH3COOK, K2CO3, Mg (NO3)2, KI, NaCl and KCl with
respective RHs of 11, 23, 44, 53, 69, 75 and 85%. The desiccator was
placed in an incubator pre-maintained at 25, 35 and 45 °C for 21 days
to ensure that samples were equilibrated to headspace RH. The powder
samples were then analyzed for the moisture content with an infrared
moisture analyzer (Ohaus Moisture Analyzer MB45). An equilibrium
moisture of powder samples for every temperature is plotted against
water activity (= %RH/100) for producing sorption isotherms. The
Guggenheim–Anderson–de Boer (GAB) model was applied to fit exper-
imental data through regression analysis to determine monolayer mois-
ture content (Mo; g water/100 g solid) [18]. The equilibrated samples
were used to carry out all the tests described below. The Tg of the sam-
ples was measured immediately (21st day).The remaining samples
were sealed in aluminum foil bags and stored at −18 °C before further
analysis.
2.3. Measurement of glass transition temperature
Five to ten milligrams of powder was weighed into DSC pans (Tzero-
T180823) and sealed using a DSC's sample press (Tzero-T080723). An
empty pan was used as a control/reference. The thermal analysis of
samples was conducted in four steps: (1) equilibration at −70 °C for 1
min; (2) heating from −70 to 150 °C at 5 °C /min; (3) cooling from
150 to −70 °C at 10 °C /min; and (4) heating from −70 to 150 °C at 5
°C /min. Tg was determined as the midpoint of transition from the sec-
ond heating thermogram using Universal Analysis 2000 software (TA
Instruments, USA).
2.4. Determining caking strength
Milk powder samples weighing 20 g were placed in plastic cups
(40.6 mm height × 33.0 mm diameter) then exposed to the tempera-
ture and RH settings described in section 2.2. The strength of cake was
analyzed by texture analyzer (TA-XT2 Plus, Stable Micro Systems Ltd.,
UK) equipped with a 5 mm diameter stainless steel cylindrical probe
and a 284.2 N load cell. The compression was conducted at 2 mm/s
395
probe speed with a 15 mm compression distance. The peak force was
used as the cake strength index.
2.5. Measurement of lactose crystallinity
Lactose crystallinity refers to the percentage of crystallized lactose in
the entire (bulk) milk powder. X-Ray diffraction (XRD) using a wide-
angle X-ray diffractometer (Bruker Axs D4 Endeavor Wide-angle XRD,
Germany) with Co-Kα radiation (40 kV, 35 mA) was used to determine
th lactose crystallinity (%) of the samples. About 1.5 g sample was placed
in sample holder and was scanned at 10–30° (2-theta) with increments
of 0.02°. The Diffrac.Eva.V4.3 software (Bruker) was used to analyze the
spectra and to identify the lactose peaks by matching the spectra of lac-
tose available in the chemical database.
2.6. Analysis of surface morphology
To analyze the morphology of powder particles, milk powders were
transferred to adhesive carbon tabs mounted on SEM stubs and gold
coated for 1 min under vacuum with a sputter coater (SPI-Module™
Sputter Coater, West Chester, USA). Gold coated samples were analyzed
with a scanning electron microscope (SEM, FEI Quanta 200,
Netherlands) functioning in high vacuum mode.
2.7. Measurement of colour parameters
The colour parameter of powders was measured in CIE L ∗ and b∗ col-
our space including lightness (L*, L* = 0, black; L* = 100, white) and
yellowness (b∗, positive values; blueness b∗, negative values). The CR-
400 chroma meter (Konica Minolta Sensing, Inc., Tokyo, Japan) cali-
brated with standard plate (No. 12133027) was used to determine
these parameters. Illuminant and observer conditions were D65 and
2°, respectively.
2.8. Measurement of surface free fat
A gram milk powder was placed in a sheet of filter paper (Whatman
no. 4) pre-folded in a glass funnel and then the sample was washed five
times with n-hexane at time interval of 10–15 s. The filtrate containing
extracted fat was evaporated on a heating block at 60 °C in a fume hood
until a constant weight of extracted fat residue was achieved. The
surface-free fat was calculated by Eq. (1)
B−A
Surface free fat ð%Þ ¼ x 100 ð1Þ
Weight of sample ðgÞ
where, B is the weight of sample and Petri dish (g) after drying, and A is
the weight of empty Petri dish (g).
2.9. Visualization of protein and fat distribution on surface of powders
Initially, about 1 mg of powder was placed on a glass-slide and ex-
cess was removed by tapping. SMP and WMP were stained with one
drop of equally mixed dye containing 0.1%(w/w) Fast Green and 0.1%
(w/w) Nile Red dissolved in polyethylene glycol 400. A coverslip was
used to protect the final mixture before viewing with a CLSM (Nikon
ECLIPSE 90i digital microscope, Japan). A 60× water immersion objec-
tive lens was used to determine the distribution of fat and protein on
the surface of powders dual-excitation at 488/633 nm following the
method of Tham et al. [19] with minor modifications.
2.10. Statistical analysis
IBM's statistical software (SPSS®, version 25, IBM Corp., Melbourne,
Australia) was used to statistically analyze results. Tests were conducted
in triplicate unless otherwise specified and results were reported as
396 A. Phosanam et al. / Powder Technology 372 (2020) 394–403

mean ± standard deviation. To find significant difference in samples,

one-way analysis of variance (ANOVA) was conducted. Significant
groups were identified by post-hoc comparison tests (Duncan's Multi-
ple Range Test, DMRT) at 95% confidence level (P < .05).

3. Results and discussion

3.1. Moisture sorption isotherms

The moisture sorption isotherms of LH-, MH- and HH-SMP and WMP
at tested temperatures (25, 35, and 45 °C) are shown in Fig. 1. WMP
showed low equilibrium moisture content compared to SMP at all tem-
perature and relative humidity combinations. Since fat is not a water-
absorbing constituent, the samples containing higher percentage of fat
absorbed less moisture upon equilibration [20]. However, the equilib-
rium moisture content (EMC) on solids non-fat (SNF) basis of WMP
was higher than on total solid (TS) basis and higher in WMP than in
SMPs at the higher RH studied (>53%RH). Jouppila and Roos [21] stated
that water content after 24 h (SNF basis) was higher in milk powders
containing fat than in skim milk powder at the highest RH studied
(76.4%). This higher water content correlated with the delayed lactose
crystallization as the amount of adsorbed water released by the pow-
ders containing fat decreased at the beginning of storage. The sorption
isotherms showed that the EMC increases by increasing aw at a given
temperature. This is due to an increase in vapour pressure in the head-
space with increasing relative humidity when the temperature is con-
stant [22]. Also, the EMC increased linearly at aw values up to 0.44 and
then increased non-linearly above this aw for both SMPs and WMPs.
This could be due to the presence of characteristic multilayer sorption
and capillary condensation regions in SMP and WMP as they contain
amorphous lactose and protein. At low and intermediate aw values
(multilayer sorption region), equilibrium moisture content rises line-
arly with aw, whereas with higher aw values (capillary-condensation re-
gion) a rapid improvement in equilibrium moisture content was
experienced [23].
According to McMinn and Magee [24], a decrease of EMC at higher
temperatures is attributed to higher activation of water molecules
which results into freeing water molecules from binding sites and ulti-
mately decreasing the moisture content at equilibrium. Temperature af-
fects mobility of water molecules thereby affecting the dynamic
equilibrium between adsorbents phase and vapour phase. In food sys-
tems, at a given aw, higher temperature decreases equilibrium moisture
content [25]. The monolayer moisture content (Mo) in these milk pow-
ders varied from 3.0 to 4.0 g water/100 g solid. The results showed that
the Mo of SMPs and WMP decreased by increasing temperature(Table
1). Lower Mo at higher temperatures is ascribed by reducing number
of binding sites because of chemical and physical changes instigated
by temperatures [24].

Table 1
Monolayer moisture content (Mo, g water/100 g total solid) of of LH SMP, MH SMP, HH
SMP and WMP stored at 25 °C, 35 °C and 45 °C. Mo was determined by fitting GAB
equation.

Samples Temperature (°C)

25 35 45

LH SMP 3.5 3.2 3.0

Fig. 1. Equilibrium moisture content (EMC) of low heat (LH), medium heat (MH), and high MH SMP 3.9 4.0 3.0
heat (HH) skim milk powder (SMP) and whole milk powder (WMP) stored at 25 °C, 35 °C HH SMP 3.2 3.1 3.0
and 45 °C. The solid lines are best fit lines produced by fitting GAB equation to WMP 3.2 3.7 3.0
experimental data.
 A. Phosanam et al. / Powder Technology 372 (2020) 394–403 397

Table 2
Glass transition temperature (Tg) of LH SMP, MH SMP, HH SMP and WMP stored at 25 °C, 35 °C and 45 °C and equilibrated at 11% RH, 44% RH and 85% RH.

Samples Tg (midpoint), oC

LH SMP MH SMP HH SMP WMP

Control 55.0cB ± 0.0 58.8dB ± 1.6 54.5eB ± 2.3 49.0dA ± 0.6

25C 11RH 58.7cC ± 0.7 59.9cC ± 0.0 57.1eB ± 0.6 54.0eA ± 0.5
25C 44RH 13.3bB ± 1.26 13.4bB ± 0.0 15.7dB ± 1.4 9.3eA ± 0.3
25C 85RH −34.5aNS ± 4.12 −32.5aNS ± 0.0 −33.8aNS ± 2.1 −32.1bNS ± 2.0
35C 11RH 68.3dB ± 0.7 67.2dAB ± 0.0 65.9fAB ± 0.5 65.2fA ± 0.9
35C 44RH 13.3bB ± 0.2 11.1bA ± 0.0 10.6cA ± 0.6 10.8cA ± 0.4
35C 85RH −34.8aAB ± 0.4 −31.9aC ± 0.0 −33.8aB ± 0.2 −35.2aA ± 0.9
45C 11RH 70.2dC ± 0.4 68.8dB ± 0.1 68.7fB ± 0.3 66.3fA ± 0.2
45C 44RH 9.5bA ± 0.7 12.2bA ± 2.5 12.1cA ± 2.5 9.9cA ± 0.4
45C 85RH −32.1aB ± 3.6 −32.9aB ± 0.4 −28.4dA ± 0.0 −31.2bA ± 1.4

Values are presented as mean ± standard deviation (n = 2);

Values in a column with different lowercase superscript differ significantly (p < .05). Values in a row for a given parameter with different uppercase superscript
differ significantly (p < .05).

3.2. Effect of temperature and storage RH on Tg
Tg is a critical parameter in assessing stability of stored dairy pow-
ders as Tg process is closely related to their stability [26–29]. It is re-
ported that the lactose content dominates the glass transition
behavior in milk powders [30]. Thus, we presumed that the Tg measured
in milk powder samples indicated Tg of lactose. Tg is a significant caking
indicator in milk powder with higher Tg indicating better stability and
delayed lactose crystallization [19]. Tg values for low-, medium-, high-
heat SMPs and WMP equilibrated at different temperatures RH values
are provided in Table 2. The Tg for all powder samples decreased with
increasing RH irrespective of temperature indicating increased moisture
uptake and plasticizing effect of absorbed water. Tg values of WMPs
were lower than that of SMPs at any given storage temperature and RH.
Tg is considered an essential characteristic of milk powders and is
highly dependent upon RH and storage temperature. Results show
that Tg of the samples decreased when the headspace RH increased,
which is consistent with the reported effect of RH on the Tg of dairy
ingredients [31]. Tg of carbohydrates and proteins declines with an in-
creasing water content which acts as plasticizer and consequently in-
creasing molecular mobility, which initiates the glass transition
process at a lower temperature [11,32]. The Tg of various milk powders
was expressed as a function of SNF, a close similarity was observed to
values for amorphous lactose [33].
3.3. Effect of the temperature and storage RH on crystallinity
Lactose in spray-dried SMP and WMP is converted to the amorphous
glassy state because of rapid drying. The glassy form of lactose is ther-
modynamically unstable and highly hygroscopic [34]. A higher mobility
of lactose molecules at high moisture content and temperature supports
its crystallization. The effect of temperature and RH on crystallinity of
LH-, MH-, and HH-SMP and WMP are shown in Fig. 2. The X-ray
diffractograms of control powder had no characteristic peaks (Fig. 2 A
and B) indicating an amorphous glassy state. Crystallinity was negligible
in all powders stored at 11% RH, irrespective of the storage temperature.

Fig. 2. Diffraction patterns of control medium-heat (MH) SMP (A), control full cream milk powder (WMP) (B), MH SMP stored at 45 °C and 85% RH (C), and WMP stored at 45 °C and 85%
RH (D).
398 A. Phosanam et al. / Powder Technology 372 (2020) 394–403
Fig. 3. Powder crystallinity of low-heat (LH) SMP, medium-heat (MH) SMP, high-heat
(HH) SMP and full cream milk powder (WMP) stored at 25 °C, 35 °C and 45 °C and 11%
RH, 44% RH and 85% RH. Columns within the same temperature with different
superscript differ significantly (P˂0.05).
Crystallinity was not detected in any of the powders stored at 44% RH
and 25 °C. However, the crystallinity of all the powders increased signif-
icantly at 85% RH when stored at 25 °C (Fig. 3). All powders stored at 35
°C and 45 °C at 44% and 85% RH showed significant (p < .05) crystallin-
ity. The crystallinity in powders increased further when stored at 85%
RH than 44% RH irrespective of temperature (Fig. 2 C, D and Fig. 3).
The rate of crystallization of amorphous sugars depends on moisture
content and storage temperatures above Tg (i.e., T-Tg) [20]. Interest-
ingly, WMP showed lower crystallinity compared to SMPs at 44% and
85% RH at 35 °C and 45 °C, which was likely due to lower lactose content
in WMP. These observations agree with that of Jouppila and Roos [36],
who defined that the presence of milk fat reduced the rate of lactose
crystallization. It is also consistently shown that milk fat (or plant oil)
overrepresents the surface composition of spray dried powders [37].
Thus, lipid-rich surface layer of WMP is expected to dissuade the ad-
sorption of water molecules. Lactose crystallization and caking are in-
terrelated [12].
3.4. Effect of temperaure and storage RH on cake strength
Spray dried milk powders are unstable because of their amorphous
structure and glassy state and have a tendency of sorbing water and cre-
ating caked powder which occurs when they are exposed to high RH
and temprature [35]. The effects of RH and temperature on caking
strength of powders is represented in Fig. 4. Caking was not observed
in any of powders stored at 11% RH at all storage temperatures; how-
ever, strong caking was observed in SMPs stored at 85% RH and 25 °C,
44% RH and 85% RH at 35 °C and 45 °C. WMP had a lower caking
strength compared to SMPs at 44% RH and 85% RH at all three temper-
atures, which can be attributed to their high fat content and a low lac-
tose crystallinity [20,36]. Caking occurs when amorphous glassy
powders transition to rubbery state at above their Tg, which greatly in-
creases the mobility of water and facilitates the formation of inter-
particle liquid and solid bridges. Humidity-induced caking is a common
mechanism for caking which occurs when the increased moisture con-
tent depresses the Tg below the storage temperature [37,38]. Powder
particles containing low molecular weight carbohydrates such as lac-
tose become sticky when their temperature is 10–20 °C their Tg,
known as sticky-point temperature [39,40]. Increased molecular mobil-
ity on the surface of rubbery solid particles favors cohesive interactions
among each other making powders cohesive. The cohesive interaction/
force of sticky particles increases initially as a response to increase in
temperature and/or moisture content. Thus, the strength of cohesive
stickiness, measured in this work as caking strength, initially rises to a
maximum level and then decreases due to continuous drop in viscosity
caused by the increase in sorbed moisture and/or temperature. Thus,
caking of amorphous powder particles depends strongly on the sur-
rounding temperature and RH [32]. The behavior of amorphous and
crystalline particles is different when they are exposed to moist air.
When exposed to higher RH, the amorphous particles readily bridge/
stick/sinter with each other due to their hygroscopic nature while the
crystalline particles are less sensitive to variation in RH [41,42]. Caking
of amorphous solid particles can be explained by sintering mechanism
in which molecular mass from neighboring particles diffuses/transfers
towards each other to fill the void/gap between them. Sintering in
amorphous solids can only occur when the surrounding temperature
becomes higher than their Tg because molecules acquire higher ability
to flow. The sintering mechanism also explains that the free specific sur-
face energy of powder systems decreases when the gap/void between
two neighboring particle decreases; thus, the sintering or caking pro-
cess is thermodynamically favored. The rate formation of sinter bridges
(tendency to cake) depends on the molecular mobility. The molecular
mobility of rubbery solids depends on their viscosity, which in itself de-
pends on the temperature difference between the powder and their Tg,
i.e. (T-Tg) [43,44]. Thus, the crystallization of lactose in milk powders oc-
curs when (T-Tg) is positive. The crystallization of lactose within sinter
bridges has potential to alter the caking dynamic which occurs when
amorphous lactose becomes rubbery. It is expected that when the lac-
tose crystallizes in the sinter bridges, it will affect the caking strength
[43,44].
The other factor that affects the moisture adsorption and subsequent
crystallization and caking of milk powders is their surface composition.
It has been reported that up to 91.8% of the surface of whole milk pow-
der can be covered by milk fat [20]. As the surface of WMP is expected to
be mostly covered by fat, these powders are expected to have lower lac-
tose crystallinity and caking strength compared to SMPs due to the for-
mation of fewer lactose (solid) bridges due to higher surface fat.
3.5. Effect of temperature and storage RH on powder microstructure
SEM is a technique which is used to assess in the microstructure of
powder samples. The microstructure of medium-heat SMPs stored at
25 °C, 35 °C and 45 °C at 11%, 44% and 85% RH are shown in Fig. 5. The
particle of medium-heat SMP stored at 25 °C and 11% RH (Fig. 5, A-1)
appeared to have an irregular surface with both shallow and deep
folds and an absence of sharp crystals. MH-SMPs stored at 25 °C and
44% RH (Fig. 5, A-2) showed agglomeration of rough-surfaced particles
Fig. 4. Caking strength of low-heat (LH), medium-heat (MH), high-heat (HH) SMPs and
whole milk powder (WMP) stored at 25 °C, 35 °C and 45 °C and 11% RH, 44% RH and
85% RH. Bars in the same temperature with different superscript letters differ
significantly (P˂0.05).
 A. Phosanam et al. / Powder Technology 372 (2020) 394–403 399

Fig. 5. SEM micrographs of MH SMP: (1), (2), (3) powders stored at 11%, 44%, 85% RH, respectively at 25 °C (A), 35 °C (B) and 45 °C (C) compared with control (N). Scale bar = 50 μm.

Fig. 6. SEM micrographs of WMP: (1), (2), (3) powders stored at 11%, 44%, 85% RH, respectively at 25 °C (A), 35 °C (B) and 45 °C (C) compared with control (N). Scale bar = 50 μm.
400 A. Phosanam et al. / Powder Technology 372 (2020) 394–403
and an absence of lactose crystals. In contrast, large numbers of needle-
like lactose crystals were found on surface of these SMPs when stored at
25 °C and 85% RH (Fig. 5, A-3), and 45 °C at all tested RHs (Fig. 5, C-1, C-2
and C-3).
SEM micrographs of WMP stored at 25 °C and 45 °C are shown in Fig.
6. WMP stored at 25 °C and 45 °C and 11% and 44% RH (Figs. 6, 25-A, B
and 45-A, B) showed a regular and smooth surface. The smooth surface
and absence of sharp crystals indicated that lactose remained in the
amorphous state. The powder particles were also clustered and agglom-
erated into noticeably larger structures (Fig. 6) compared to SMPs (Fig.
5). Storage of WMP at 25 °C and 85% RH (Figs. 6, 25-C) formed promi-
nent clusters of agglomerated particles with increasingly irregular and
rougher surface indicating that crystallization of lactose has occurred.
The agglomerated clusters were more pronounced at 45 °C and 85%
RH (Figs. 6, 45-C).
WMP had relatively smooth and uniformly-sized, and agglomerated
powder particles compared to SMPs due to the high fat coverage on the
surface [45–47]. A high proportion of fat on the surface makes WMP
particles weakly sticky and facilitated their clustering through fat bind-
ing. The storage of high fat containing powder at high temperature and/
or in a humid atmosphere resulted in release of oil from the inner par-
ticle to the surface, which increased surface fat coverage. It has been
shown that the particle structure is altered when amorphous lactose
undertakes crystallization [20]. When this occurs, fat is excluded from
the ordered structure of lactose crystals and expelled to the powder sur-
face [12,19]. Therefore, an increase in free fat on the powder surface oc-
curs as a result of lactose crystallization.
3.6. Effect of temperature and storage RH on the colour of powders
Colour is an essential parameter that influences consumer accept-
ability. The lightness and yellowness of SMP and WMP powders stored
under the tested conditions are presented in Figs. 7 and 8, respectively.
The control milk powders were lighter and less yellow than the stored
Fig. 7. Lightness (L*) of low-heat (LH), medium-heat (MH), high-heat (HH) SMPs and whole milk powder (WMP) stored at 25 °C, 35 °C and 45 °C and 11% RH, 44% RH and 85% RH. Columns
within the same sample having different superscript differ significantly (P˂0.05).
powders. As expected, the lightness values of control SMPs were slightly
higher than that of control WMP which were yellowish due to the
higher fat content. Milk fat contains a fat-soluble beta carotene which
imparts the yellow colour. After 21 days of storage, both the SMP and
WMP powders developed brown colour, and this was especially pro-
nounced at 85% RH at all storage temperatures. The formation of
brown colour is associated with Maillard reaction, which occurs be-
tween a protein containing lysine and reducing sugar such as lactose
[48]. As shown in Fig. 8, the lightness values of SMP and WMP powders
were lower than that of control powders. On the other hand, lightness
value for both SMP and WMP were lowest in samples stored at 85%
RH. The yellowness value is useful for monitoring the browning reac-
tions in milk powders [49]. Control and powders stored at 25 °C and
11% RH had the lowest yellowness. On the other hand, SMPs and
WMP stored at 45 °C and 85% RH had the highest yellowness.
Yellowness is the indicator of non-enzymatic browning and it further
increased at higher RHs (Fig. 8) due to the late-stage Maillard reaction
during storage. The Maillard reaction between lactose and milk proteins
produces lactulosyl-lysine conjugates. The extent of Maillard reaction
induced browning, as indicated by the change in yellowness values,
confirmed that it increases with temperature and RH of the storage
[50]. It has been reported that at a given RH, the browning rate increases
with the increase in storage temperature [51]. Our results showed that
powders stored at 85% RH containing a higher proportion of crystallized
lactose (Fig. 3) resulted into higher degree of Maillard reaction induced
browning as compared to those containing lower proportion of crystal-
lized lactose (e.g., control powders, SMP and WMP stored at 11%RH). In-
creased crystallization of lactose increased the rate of Maillard reaction
induced browning due to release of water expelled during the crystalli-
zation process [52]. The higher free moisture content made the milk
proteins and lactose more mobile and available for Maillard reaction.
Higher temperatures and RHs are essential preconditions that induce
Maillard reactions and browning in WMP and SMP. Fat in WMC can
slow down the Maillard reaction by lowering the propensity to absorb
 A. Phosanam et al. / Powder Technology 372 (2020) 394–403 401

Fig. 8. The yellowness (b*) of low-heat (LH), medium-heat (MH), high-heat (HH) SMPs and full cream milk powder (WMP) stored at 25 °C, 35 °C and 45 °C and 11% RH, 44% RH and 85%
RH. Columns within the same sample having different superscript differ significantly (P˂0.05).

moisture. Also, high fat content on particles' surface increases surface
hydrophobicity, thus decreasing wettability of particle and limiting
crystallization of lactose [21].
3.7. Effect of temperature and storage RH on surface free fat
Free fat in powders is the fat that is not entirely covered by protec-
tive matrix. It is an indicator of damage to membrane of the milk fat
globules [53]. The free fat can alter the desirable properties of dairy
powder such as oxidative stability and flowability [54]. The free fat ex-
tracted from MH-SMP and WMP at differences storage treatments is
presented in Table 3. The free fat of control and stored MH-SMP and
WMP at 25 °C and 11% RH (the most stable powders), and 45 °C and
85% RH (the most unstable powders) was similar except in WMP stored
at 45 °C and 85%RH which increased significantly (P < .05). In powders
containing amorphous lactose, milk fat is encapsulated in a matrix com-
prised of amorphous lactose and protein. Crystallization of lactose re-
leases encapsulated fat by damaging the membrane of the milk fat
globule and also rupturing the protective matrix [55]. The free fat of
control and stored MH-SMP did not increase significantly (P < .05) as
shown in Table 3, possibly due to the very low-fat content in MH-
SMP (1%).
3.8. Effect of temperature and storage RH on fat and protein distribution on
surface
CLSM is used to visualize the microstructure of food including
powders. The effect of storage parameters on surface composition
of SMP and WMP in terms of fat and protein distribution is investi-
gated using CLSM (Fig. 9). The control MH SMP powders exhibited a
continuous protein phase containing a small number of very fine fat
droplets (Fig. 9A). Similar observation was found for MH SMP pow-
ders stored at 11% RH of 25 °C and 45 °C (Fig. 4B and D). The control
and stored WMP at 25 °C and 11%RH showed a continuous protein
phase containing a large number of fat droplets (Fig. 9, F and G).
The lipid components initially embedded inside particles tended
to migrate towards surface when stored at high temperature and
high RH as observed in MH-SMP stored at 85% RH with 25 °C and
45 °C (Fig. 9C and E) and WMP stored at 85%RH of 25 °C (Fig. 9H
and J) and 45 °C and 11%RH and 45 °C (Fig. 9I). A higher fat content
observed on surface of particle after storage represents the aging-
induced fat migration. Higher intensity of red colour on particle sur-
face shows an increased fat coverage. This coverage of particle sur-
face by fat resulted in particle bridging (agglomeration) which is
responsible for the fat-induced caking [57]. Such increased fat local-
ization on surface of SMP and WMP is also aided by lactose
crystallization.

Table 3 4. Conclusion
Surface free fat (%) of medium-heat SMP and WMP stored at 25 °C and 45 °C
and equilibrated at 11% RH 85% relative humidity.
The fat content in milk powders affects their physicochemical is con-
Treatments Samples sidered during storage. This study quantified and explained the effect of
MH SMP WMP fat content on the properties of whole milk powder (WMP, 26% fat) and
Control 0.2 ±0.0 a
1.2 ±0.1a skim milk powders (SMPs, 1% fat) stored at three temperatures (25 °C,
25C 11RH 0.4 ±0.1a 1.6 ±0.2a 35 °C and 45 °C) and RHs (11%, 44% and 85%) for 21 days. Moisture sorp-
45C 85 RH 0.7 ±0.1a 16.5 ±1.0b tion isotherms, changes in Tg, crystallinity, surface fat content and cak-
Values are mean ± standard deviation; n = 3. Values of the surface free fat ing strength were measured. SMPs and WMP stored at 45 °C
with different superscript are significantly different (P < .05). equilibrated at the lowest moisture content (EMC) compared to those
402 A. Phosanam et al. / Powder Technology 372 (2020) 394–403
Fig. 9. Confocal laser microscopic images showing fat and protein distribution on the surface of MH SMP (A-E) and WMP (F-J) stored at 25 °C and 45 °C and 11% RH and 85% RH compared
with control powder at magnification 60×. (scale bar = 10 μm). Labelled by Nile Red (red) = fat and labelled by Fast Green (green) = protein.
stored at 25 °C and 35 °C. WMP had lower EMC than SMPs at all tested
temperatures and RHs. Tg of both WMP and SMP decreased with in-
creasing storage RH. WMP had lower Tg and lower crystallinity than
SMPs in all cases due to lower lactose content in the former. The caking
strength of SMPs were higher than that of WMP and SMP stored at low
RH (11%) showed particles with a regular-shaped surface, while those
stored at high temperature or/and RHs (85% RH at 25 °C and 44% RH
and 85% RH at 35 °C and 45 °C) had particles with higher surface rough-
ness and irregularity due to lactose crystallization. Maillard reaction was
more prominent in both SMP and WMP at higher storage RH and tem-
perature. Ambient temperature (25 °C) and lower RH (11% and 44%)
were found to be suitable for storage of both WMP and SMP. The heat
class (LH-, MH- and HH-heat) did not have significant influence on
the physicochemical and microstructural properties of SMPs as lactose
and fat contents determined these properties.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influ-
ence the work reported in this paper.
Acknowledgement
The first author acknowledges the scholarship support from The
Royal Thai Government (Ministry of Science and Technology of Thai-
land). The authors wish to thank Saputo Dairy Australia for providing
powder samples.
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