Powder Technology 179 (2007) 90 – 94

www.elsevier.com/locate/powtec

Laser diffraction particle sizing of cohesive lactose powders

Handoko Adi, Ian Larson, Peter Stewart ⁎
Department of Pharmaceutics, Victorian College of Pharmacy, Monash University, 381 Royal Pde, Parkville, Victoria 3052, Australia

Available online 27 January 2007

Abstract

The objective was to develop a suitable laser diffraction particle sizing method for cohesive lactose present either as agglomerates or adhered
onto coarse lactose carriers. Micronised lactose (ML) was prepared by fluid energy milling. Particle size distributions (PSD) were determined by
laser diffraction (Malvern Master Sizer S, U.K.). Ethanol, propan-2-ol, 1-butanol and isooctane were selected as dispersants. Sonication for 5 min
caused de-agglomeration of agglomerates initially formed in ethanol, propan-2-ol, and 1-butanol; the volume mean diameter (VMD) of ML in
ethanol, propan-2-ol, and 1-butanol was 2.9 μm, 2.8 μm and 2.5 μm, respectively. Coarse lactose (Foremost 95 (F95)) showed a mono-modal
distribution in all dispersants with a VMD of 117 μm in propan-2-ol. The robustness of particle sizing of ML in each dispersant was determined
over time. Propan-2-ol was the most suitable dispersant for ML, as the PSD (VMD of 2.8 ± 0.3 μm) did not change over a 3 h period. In
comparison, for ethanol, VMD of FL was stable only for 30 min, but then increased from 2.9 μm to 9.0 μm after 3 h. The particle size changes
over time occurring in ethanol were related to dissolution of fine lactose and possible re-crystallisation with subsequent increased VMD. ML
added to coarse lactose and existing as agglomerated particles or as particles adhered to the coarse lactose could be determined with the use of a
carefully selected sonication time to minimise coarse lactose comminution.
© 2007 Elsevier B.V. All rights reserved.

Keywords: Laser diffraction particle sizing; Cohesive lactose; Dispersants; Agglomeration; Particle adhesion

1. Introduction agglomeration and particle detachment in the medium is

In powder handling and processing, a detailed understanding
of particle size distributions is of crucial importance in
interpreting research, quality control, and product development
data. In powder industries, such as pharmaceutical, ceramics and
metallurgical industries, fine powders are difficult to handle due
to the cohesive properties of the particles. Cohesive fine powders
may contain agglomerates of “primary” particles that are held
together by particulate interactive forces, including electrostatic,
intermolecular and capillary forces and solid bridging [1]. These
same forces cause the adhesion of micronised fractions of fine
powders, i.e. particles less than about 15 μm, to the surface of
larger particles to form interactive units. De-agglomeration and
particle detachment from surfaces after powder wetting by a
dispersion medium depends in part on how much energy has
been provided to break inter-particulate bonding. Since
agglomerates or interactive units will be measured as large
particles by most particle sizing methods, complete de-
⁎ Corresponding author. Tel.: +61 3 99039517; fax: +61 3 99039583.
E-mail address: peter.stewart@vcp.monash.edu.au (P. Stewart).
0032-5910/$ - see front matter © 2007 Elsevier B.V. All rights reserved.
doi:10.1016/j.powtec.2007.01.018
necessary to ensure effective particle size measurement of all
particle distributions. A particle size analysis is useful only if the
sample is prepared so that the particles are in well-defined degree
of dispersion and do not re-agglomerate or adhere to the walls of
the sample container during the time required for analysis.
The preparation of a suspension of de-agglomerated particles
has been shown to be particularly difficult when working with
micronized, cohesive powders [2]. Dispersion processes are
intended to counteract the forces between particles by physico-
chemical and mechanical intervention. To achieve this, the
following three fundamental processes must take place [2]:
(1) wetting the solid; (2) de-agglomeration of particle agglom-
erates; and (3) stabilization of the dispersed suspension.
Mechanical means used to disperse particles include stirring,
shaking and ultrasonic treatment.
Lactose has been widely used in the pharmaceutical industry
as carrier and diluent, especially in the formulation of powder
mixtures for inhalation. It is a safe pharmaceutical excipient,
readily available, physico-chemically stable and compatible
with the majority of small molecular weight drugs [3]. Recent
studies have demonstrated that the presence of micronised
 H. Adi et al. / Powder Technology 179 (2007) 90–94 91

Fig. 1. Scanning electron micrographs of lactose particle. (A) micronised lactose (Magnification of ×3.0K), (B) non-micronised lactose (Magnification of ×1.0K) and
(C) non-micronised lactose (Magnification of ×200K).

lactose was the key factor in controlling drug dispersion from 2.3. Particle size analysis method
drug-lactose mixtures of inhalation [4,5].
The selection of a reliable particle size analysis methodology The particle size of lactose was measured by laser diffraction
to measure the particle size distribution of micronised lactose (Malvern Mastersizer S, Malvern Instruments Ltd., U.K.) using
and coarse lactose containing adhered cohesive lactose is the 300 RF lens and the small volume sample presentation unit
essential to characterize the influence of fine lactose on the drug (capacity 150 ml). Approximately 500 mg of lactose powder
dispersion in respiratory drug delivery. Various strategies to was dispersed in 5 ml of propan-2-ol with the aid of a sonication
disperse lactose for laser diffraction particle size analysis have
been employed including: dispersion of lactose powder in
chloroform containing a few drops of span 85 and sonication for
1 min [3]; dispersion in ethanol with sonication for 3 min [5];
dispersion of lactose in butan-1-ol with the aid of sonication for
3 min [4]. However, method validation, such as the effect of
dispersants and sonication time on the size measurement of fine
lactose, has not been disclosed. The objective of this study,
therefore, was to investigate the influence of the interactive
behaviour of micronised lactose on its particle size analysis and
to develop a suitable methodology for particle size measure-
ment of cohesive lactose samples using laser diffraction.

2. Experimental

2.1. Materials

Aeroflo95 (F95) (Foremost Farm, USA); Inhalac 120

(In120) (Meggle GmbH, Germany) and Lactose Edible 90
(Lactose New Zealand) were used. Propan-2-ol (Merck Pty.
Ltd., Australia), Absolute Ethanol (CSR, Australia), n-butyl
alcohol and 2,2,4-trimethylpentane (Isooctane) (Chem-supply
Pty. Ltd., Australia) were the dispersants and used as supplied.

2.2. Milling of lactose

Lactose Edible 90 was milled using a fluid energy mill (K-tron

Soder, USA). Injector pressure of 862 kPa and grinding pressure Fig. 2. Particle size distribution of micronised (A) and non-micronised (Aeroflo
of 690 kPa were employed at feed rate of 400 rpm to produce 95) (B) lactose in different dispersants without sonification using laser
micronised lactose. diffraction.
92 H. Adi et al. / Powder Technology 179 (2007) 90–94

Fig. 4. The influence of storage time in different dispersants on the volume mean
diameter of micronised lactose determined using laser diffraction.

2.5. Statistical analysis

Volume mean diameter were compared using a One Way

Analysis of Variance (ANOVA) (SPISS, USA), with probability
values of less than 0.05 considered as statistically significant.

3. Results and discussion

3.1. Particle sizing of micronised and non-micronised lactose

Micronised lactose (VMD 2.5 μm), used in the development

of a suitable particle sizing method of cohesive lactose by laser
diffraction, was prepared using fluid energy milling and.

Fig. 3. Effect of sonication time on the particle size distribution of micronised

(A) and non-micronised (Aeroflo95) (B) lactose in propan-2-ol using laser
diffraction.

in a water bath for 3 min. The sonicated sample was added drop-
wise into sample cell containing 150 ml of propan-2-ol until an
obscuration between 10–30% was obtained. Size measurement
of each sample was performed using 2000 sweeps and analysed
with the reference refractive index of lactose (1.533) and
propan-2-ol (1.378) and estimated imaginary refractive index of
lactose of 0.1. The average particle size distribution was
measured from 5 replicates of each sample. The percentage of
particles below 5 μm and 10 μm were determined using
cumulative frequency distribution (undersized) curve. The
residual value was always below 1%. Others dispersants
were: Absolute ethanol (refractive index of 1.361), 1-butanol
(1.399) and isooctane (1.389). Sonication times (0, 0.5, 1, 3, 5,
8, and 15 min) and the robustness of the particle size analysis
were determined over a period of 0 to 3 h. The average particle
size was described by the volume mean diameter (VMD).

2.4. Scanning electron microscopy

Powder samples were mounted on metal sample plates. The

samples were gold coated with a sputter coater (BAL-TEC SCD
005, Japan) using an electrical potential of 2.0 kV and 25 mA for
3 min. The particles were examined at several magnifications
under a Hitachi S-570 scanning electron microscope (Tokyo,
Japan) operating at 10 kV.
Fig. 5. The influence of storage time in the propan-2-ol (A) and absolute alcohol
(B) on the particle size distribution of micronised lactose determined using laser
diffraction.
 H. Adi et al. / Powder Technology 179 (2007) 90–94 93

The scanning electron micrograph of the micronised lactose Table 2

powder showed that the powder contained agglomerates of Summary of particle size parameters of mixture of Inhalac 120 and 5%ML
(standard deviations are within brackets)
particles of varying sizes (Fig. 1A), while the SEM of non-
micronised lactose (Aeroflo 95) showed coarse, dispersed Sonication time (min) d10% (μm) VMD (μm) D90% (μm) b10 μm (%)
particles with some very fine lactose either adhered on the 1 72.6 (2.4) 124.8 (0.4) 180.4 (2.2) 1.8 (0.1)
surface of the coarse lactose particles (Fig. 1B) or as 2 50.8 (4.1) 115.2 (1.3) 177.8 (2.8) 4.8 (0.1)
3 21.8 (1.6) 101.5 (1.2) 161.7 (3.6) 6.2 (0.5)
agglomerates with the coarse lactose (Fig. 1C). PSD of
4 7.7 (0.5) 100.5 (2.3) 169.6 (2.7) 12.7 (0.7)
micronised lactose (ML) and non-micronised lactose (Aeroflo 5 7.2 (0.2) 102.8 (0.4) 172.4 (0.6) 13.4 (0.2)
95) was determined using different dispersants. 6 2.9 (0.1) 68.4 (1.0) 144.0 (1.5) 30.4 (0.4)
As expected, when the samples were not sonicated, the PSD 7 2.6 (0.1) 67.4 (1.2) 146.7 (2.8) 30.5 (0.4)
of ML appeared as multi-modal distributions in different 8 1.6 (0.1) 49.3 (1.7) 130.3 (2.4) 42.7 (1.1)
9 1.3 (0.1 42.6 (0.6) 122.2 (1.1) 47.6 (0.4)
dispersants (absolute ethanol, propan-2-ol, 1-butanol and iso-
10 1.0 (0.1) 36.6 (0.4) 114.4 (0.8) 49.5 (0.3)
octane), due to the particle agglomeration (Fig. 2A). The result
obtained was consistent with the SEM image, where agglom-
erates of micronised particle were seen. (Fig. 1A). All 3 min of sonication. From 3 to 15 min there was no significant
dispersants except iso-octane showed PSD below about difference in the VMD of ML (2.5 μm) (P N 0.05). It could be
10 μm and agglomerate distributions above about 100 μm. concluded that ultrasonic treatment was effective in dispersing
Iso-octane showed a small distribution below 10 μm and two the agglomerates after 3 min. Similar results were also found
agglomerate distributions. [2,7,8], although the mechanisms involved has not yet been
On the other hand, PSD of Aeroflo 95 were consistent with completely clarified. Sonication for 5 min was considered
different dispersants (ethanol, propan-2-ol and 1-butanol), and optimum for both propan-2-ol and 1-butanol. Isooctane was
showed monomodal distribution with no agglomeration ob- found to be an unsatisfactory dispersant for both ML and F95,
served (Fig. 2B). Unusual results were observed with iso-octane, where spurious VMD of 55.5 μm for ML and 152.04 μm for
with a PSD significantly different from the other dispersants. F95, and tri-modal distributions were obtained.
Particulate interactions, including agglomeration observed in The PSD of non-micronised lactose, Aeroflo 95, became
ML after milling, depend on the balance between the interactive broader, with a greater proportion of fine particles observed
and detachment forces present in the powder system. Micronisa- during sonification. This occurred in all dispersants and is
tion decreased particle mass and thus adhesion or cohesion demonstrated by its behaviour in propan-2-ol (Fig. 3B);
becomes dominant. The inter-particulate forces between particles however, little change in the PSD occurred after 5 min
become more dominant as the particle size reduced to less than suggesting that sonication energy detached the fine particles
20 μm [1]. In addition, post-milling changes can occur because from the carrier surface, resulting in a broader distributions. The
the milled particles have highly energetic surfaces that can readily VMD of Aeroflo 95 after sonication for 15 min resulted in a
adsorb small amounts of water [6], inducing partial dissolution significant decrease in particle size for both propan-2-ol and
that results in re-crystallization at the particle surface as it attempts 1-butanol with the VMD reduced from 117 μm to 70.2 μm and
to lower its energy state. Alternatively, the plasticizing effect of 117 μm to 82.2 μm, respectively (P b 0.05). This observation
the water may lower the glass transition temperature of the was consistent with the SEM data where fine lactose was
amorphous material at the surface and result in re-crystallization. observed on the surface of the coarse lactose (Fig. 1B).

3.2. Effect of sonication time 3.3. PSD changes of micronised lactose in different dispersant

The effect of the sonication time was investigated using The VMD changes of micronized lactose dispersed after
different dispersants (ethanol, propan-2-ol, 1-butanol and iso- sonication for 5 min were compared during storage over
octane) for the period of 0 to 15 min. PSD of ML was 180 min in ethanol, propan-2-ol and 1-butanol (Fig. 4).
significantly changed from bimodal distribution to the mono- Propan-2-ol was the most suitable dispersants for the ML,
modal distribution after 3 min, demonstrated by its behaviour in where the VMD of 2.8 ± 0.3 μm did not change over 3 h. The
propan-2-ol (Fig. 3A). Sonication gave a significant reduction PSD confirmed that the distribution was also consistent over the
in agglomeration for all dispersants. The VMD of ML was 3 h period (Fig. 5A). Both d10% and d90% were stable after 3 h
reduced in propan-2-ol, ethanol and 1-butanol from 53.9 to in the propan-2-ol, the value ranging from 1.2 to 1.3 μm for and
2.4 μm; 17.6 to 2.7 μm and 57.2 to 2.5 μm respectively after 4.8 to 5.4 μm respectively. There was no evidence of
agglomeration. VMD of the ML was not significantly changed
Table 1 even after 20 h of storage in propan-2-ol (VMD of 3.1 μm).
Summary of particle size parameters of Inhalac 120 and micronised lactose The VMD of the ML in1-butanol was stable for the 60 min
(Standard deviations are within brackets)
(VMD of 2.9 ± 0.03 μm); however, the VMD had increased
Sample d10% (μm) VMD (μm) D90% (μm) b10 μm (%) from 2.9 to 4.8 μm after 180 min (P b 0.05) (Fig. 4). The d10% of
Inhalac 120 115.8 (0.3) 162.1 (0.3) 215.3 (0.7) 1.4 (0.2) the ML was consistent during storage (1.2 ± 0.04 μm); however,
5.5 μm ML 1.2 (0.1) 5.5 (0.1) 10.4 (0.1) 89.6 (0.3) the d90% increased slightly from 4.8 to 5.6 μm and some
Inhalac and 5% of ML 103.1 (2.9) 152.7 (1.2) 209.3 (3.6) 5.8 (0.6)
agglomeration was evident after 180 min at about 50 μm.
94 H. Adi et al. / Powder Technology 179 (2007) 90–94
The re-agglomeration of particles is initiated through
particulate collisions because of their Brownian motion. If the
attractive forces prevail during the interaction, agglomerates can
develop. The attractive forces are mostly due to London-Van der
Waals forces; repulsive forces originate from several sources [9]
including electrostatic repulsion, steric hindrance or repulsive
hydration forces in the interfacial region. The key requirement
for stabilisation is that the net repulsive term exceeds the net
attractive term.
In absolute ethanol, the VMD of ML was stable for 30 min.
The VMD significantly increased from 2.9 to 9.0 μm after
180 min (P b 0.05). The rapid increase in the VMD was due to
particle dissolution and agglomeration. In Fig. 5B, the change in
PSD was clearly seen, with d90% increasing from 4.7 to 9.0 μm
while the d10% was constant (1.2 ± 0.1 μm). Particle agglom-
eration occurred with the development of a second distribution
around 50 μm after 60 min.
3.4. Recovery of added fine lactose to a coarse lactose carrier
Inhalac 120 was used as model carrier due to its relatively
low percentage of inherent fine lactose (FL) b10 μm (Table 1).
FL (5.5 μm; 5%) was mixed with Inhalac 120 and was expected
to form cohesive fine lactose agglomerates or adhere on the
carrier surface. In order to completely detach and de-
agglomerate the fine lactose in the presence of the Inhalac
120 (i.e. recover 5.8% of FL b 10 μm), an appropriate sonication
time should be used to disperse the FL, but not to comminute
the lactose carrier.
The effect of sonication to the inhalac 120 was investigated.
No significant change in percentage of FL b 10 μm (2.2%)
occurred during the sonication times from 1-5 min; however
further sonication to 20 min increased the percentage of FL
b 10 μm indicating possible Inhalac 120 break up.
During sonication of lactose mixture containing 5% added
ML, the percentage of FL b 10 μm significantly increased from
1.8% at 1 min to 6.2% (P b 0.05) after 3 min. Sonication of
lactose mixture for 3 min was considered to be optimum since
added fine lactose was recovered and previous studies on
Inhalac 120 alone showed no comminution in this time-frame.
The percentage of FL b10 μm showed no significant different
(P N 0.05) between the recovered FL from the Inhalac 120 and
ML mixture and expected theoretical concentration of FL
b 10 μm in the mixture (6.2% compared to 5.8%, respectively).
Further sonication resulted in the comminution of the coarse
lactose (Inhalac 120) particles (Table 2).
4. Conclusion
The study has developed an easily standardized particle size
analysis method using propan-2-ol and 3 min of sonication to
achieve dispersion. Sonication was essential for fine lactose de-
agglomeration and detachment from carrier particles. Sonica-
tion over 3 min was necessary to recover all ML from the coarse
lactose; however longer sonication time may result in the coarse
particle break up. The particle size changes over time occurring
in ethanol and 1-butanol were probably related to dissolution
and possible re-crystallisation with subsequent increased VMD.
Acknowledgments
Handoko Adi was supported by Departmental Scholarship,
Department of Pharmaceutics, Monash University. All lactose
samples were donated by Foremost Farms, USA; Meggle
GmbH, Germany; and Lactose New Zealand, NZ. We would
like to extend our thanks to Micro-powders Ltd. for the
micronised lactose.
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