PHARMACEUTICAL TECHNOLOGY

The Influence of Mechanical Processing of Dry Powder

Inhaler Carriers on Drug Aerosolization Performance
PAUL M. YOUNG, HAK-KIM CHAN, HERBERT CHIOU, STEPHEN EDGE, TERENCE H.S. TEE, DANIELA TRAINI
Advanced Drug Delivery Group, Faculty of Pharmacy, University of Sydney, Sydney, NSW 2006, Australia

Received 10 May 2006; revised 18 June 2006; accepted 10 July 2006

Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/jps.20933

ABSTRACT: The influence of processing on the performance of carrier material used in

dry powder inhalers was investigated. a-Lactose monohydrate crystals were processed by
ball milling for cumulative time durations and their properties evaluated. As expected,
milling reduced the median particle diameter while increasing fine particulate (<10 mm)
and amorphous levels. Recrystallization of these partially amorphous samples resulted
in a reduction in fines, elimination of amorphous material with little change in median
diameter. To study the effects of processing on aerosolization performance, blends of
lactose monohydrate with a model drug (nedocromil sodium trihydrate), were evaluated
using an in vitro multistage liquid impinger (MSLI) model. In general, milling and
storage of the carriers at high humidity (prior to blending) had a significant (ANOVA,
p < 0.05) effect on the fine particle fractions (FPF; <6.8 mm). These effects were attributed
predominantly to the fines content, showing a strong correlation between increased fines
and FPF (R2 0.974 and 0.982 for milled and recrystallized samples, respectively).
However, this relationship only existed up to 15% fines concentration, after which
agglomerate-carrier segregation was observed and FPF decreased significantly. These
results suggest that, after processing, high-dose drug formulation performance is
dominated by the presence of fines. 2007 Wiley-Liss, Inc. and the American Pharmacists
Association J Pharm Sci 96:13311341, 2007
Keywords: milling; surface activation; fines; aerosolization; dry powder; inhalation;
recrystallization; amorphous

INTRODUCTION surface area to mass ratio of the particulates.

Dry powder inhalers (DPI) are a novel route for
drug delivery, with the capability of targeting
disease states both locally (in the case of lung
diseases such as asthma), and systemically (e.g.
in the delivery of proteins and peptides). For
effective deposition in the lower airways and deep
lung, drug particles with aerodynamic particle
sizes of <5 mm are required. However, such
systems are highly cohesive due to the high
Correspondence to: Paul M. Young (Telephone: 61 2 9036
7035; Fax: 61 2 9351 4391; E-mail: py@pharm.usyd.edu.au)
Journal of Pharmaceutical Sciences, Vol. 96, 13311341 (2007)
2007 Wiley-Liss, Inc. and the American Pharmacists Association
Cohesive systems pose a problem for the desir-
ed deaggregation of particles as uncontrolled
agglomeration occurs naturally. Subsequently,
such agglomeration may lead to formulation
variations and a decrease in DPI efficacy. As a
consequence, large inert carrier systems are
employed as one method to overcome this pro-
blem, where the micron sized drug particles are
blended with larger inert material to reduce
agglomeration, improve flow and act as a diluent.
Ideally, during inhalation, the drug particles are
liberated from the carrier to penetrate the lower
airways while the carrier impacts on the ortho-
pharynx and is swallowed. As with all these

JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 96, NO. 5, MAY 2007 1331

1332 YOUNG ET AL.
systems, it is important to note, however, that the
energy supplied by the patient during inhalation
needs to be sufficient for liberation and aerosoli-
zation of the drug particulates.
Currently, lactose is the most popular material
approved for use in inhalation as carrier. A
naturally occurring disaccharide sugar, lactose is
found in milk, and is collected as a by-product
of whey produced during the manufacturing of
cheese. Lactose monohydrates for use in the
pharmaceutical industry, are produced through
precipitation from aqueous solution and is avail-
able in many particle size grades.1 Furthermore, to
obtain inhalation grade lactose monohydrate, the
crystals are processed through techniques such as
milling and sieving to produce the desired carrier
size characteristics.2 However, due to the indus-
trial scale of such processes, inter-batch, and inter-
supplier variations in the physicochemical proper-
ties of the carriers have been observed, leading to
variations in the aerosolization performance of the
final formulations.2 Such batch-to-batch variation
is most likely due to differences in fine particle
content, particle size distribution, surface mor-
phology, and amorphous content.
Intrinsic carrier particle size has been believed
for many years to play an important role in the
performance of DPI.36 Following a reduction in
physical particle diameter, a decrease in mean
mass aerodynamic diameter (MMAD) is observed.
In general, a decrease in MMAD has been asso-
ciated with an increase in drug dispersion. How-
ever, a recent study by Islam et al.,7 suggested
that the volume median diameter of carriers had
no significant effect on drug dispersion when
studied without the influence of fines content.
Earlier studies did not specify the fine particle
content, thus with a reduction in particle size, the
increasing concentration of fines may have been
masking the effect carrier size has on aerosol
performance. As fine particles are commonly
introduced inadvertently during the comminution
process, the influence of fines (e.g. carrier particles
with a diameter <15 mm) on DPI performance
has been studied extensively. Lucas et al.,8
Zeng et al.,6,9 Louey et al.,10,11 and Islam
et al.,7,12 have all demonstrated an improvement
in dispersion with the presence of fine particles.
More recently, Steckel et al.,13 indicated similar
findings, suggested that variation in small quan-
tities of fines (<5% below 15 mm diameter) in
different sized sieved fractioned lactose formula-
tions significantly influenced drug aerosolization
performance.
JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 96, NO. 5, MAY 2007
As discussed, processing methods such as
milling may induce variation in surface morphol-
ogy or result in increased amorphous content.
Surface morphology has been demonstrated to
directly influence the contact area between drug
particle and carrier, leading to variations in inter-
particulate adhesion. Several studies have
reported that variations in contact area, as a
result of differing surface structure, could poten-
tially compromise the aerosolization performance
of the drug particles.1416 In addition, the intro-
duction of amorphous material during high energy
mechanical processes is associated with a higher
surface adhesion energy compared to crystalline
surfaces.1719 As a consequence of raised adhesion
energy, poor deaggregation of drug particles is
observed.20 No studies have yet been reported in
relation to the direct impact of amorphous content
on drug dispersion from carriers. However, the
presence of amorphous material may cause pro-
blems, for example, due to the fusion of particles,
resulting in poor dispersion.1921
Milling is commonly used for processing of
powders in the pharmaceutical industry, and in
particular for inhalation drugs and carriers. When
carriers are milled, various changes to the physical
properties are induced. This is important since the
reliability of the DPI product mainly depends on
batch-to-batch consistency of the lactose monohy-
drate carrier. However, as mentioned previously,
variations between batches do occur, and this
study was initiated to investigate the effect of any
material changes induced by milling on DPI
performance. Further, as part of an ongoing study,
the influence of storage at high humidity prior to
blending was also investigated. The influence of
these processes on DPI aerosolization efficiency
was investigated using nedocromil sodium tri-
hydrate as a model drug system, and was corre-
lated with the physical properties of the carrier
particles.
MATERIALS AND METHODS
Materials
Micronized nedocromil sodium trihydrate (NST)
was obtained from Sonafi-Aventis (Cheshire,
England). Crystalline a-lactose monohydrate
(Lactochem1 crystals) was obtained from Borculo
Domo (Zwolle, The Netherlands). Water was
purified by reverse osmosis (MilliQ, Millipore,
Molsheim, France). Analytical grade chloroform
DOI 10.1002/jps

and n-octane were obtained from Biolab (Victoria,
Australia) and Fluka (Germany), respectively.
Preparation of Lactose Monohydrate Samples
Mechanical treatment of lactose monohydrate
samples was achieved by comminution in a
small volume ball mill (approximately 1 L)
containing 60 ceramic balls (mean diameter of
19.3
0.7 mm). Samples of lactose monohydrate
(approximately 100 g) were weighed into the ball
mill which was rotated at 42 rev min1 for
durations of 10, 20, 30, 40, 50, and 60 min. Each
sample was then collected and stored in tightly
sealed containers over phosphorus pentoxide
prior to sampling or blending.
In addition, to crystallize any amorphous
material, present in the milled samples, approxi-
mately 10 g of each milled lactose monohydrate
sample was transferred onto glass Petri-dishes
and stored (3 weeks) in tightly sealed containers
with a saturated solution of potassium chloride
(relative humidity, RH, of 85%). The samples were
regularly stirred to ensure moisture penetration
into the powder bed. After 3 weeks, each sample
was removed and transferred into containers with
phosphorous pentoxide (0% RH) for a minimum of
24 h before sampling or blending.
Physical Characterization
Particle Sizing of Processed Lactose
Monohydrate Samples and NST
Particle sizing was performed by laser diffraction
(Malvern Mastersizer S, Malvern Instruments
Ltd., Malvern, UK) using a 300RF lens and auto-
mated small volume dispersion sampling unit.
Approximately 200 mg of each lactose monohy-
drate sample or NST was dispersed in about
10 mL of chloroform, and added drop-wise into the
sampling unit containing chloroform until an
obscuration between 10% and 30% was obtained.
Size distributions were based on 2000 sweeps for
each sample, with refractive indices of 1.533,
1.358, and 1.444 for lactose monohydrate, NST
and chloroform, respectively. Each sample was
analysed in triplicate.
Scanning Electron Microscopy
Visualization of lactose monohydrate surface
morphology and the uniformity in blends were
investigated using scanning electron microscopy
(SEM) (XL30, Philips, Japan) at 10 keV. Each
DOI 10.1002/jps
DRUG AEROSOLIZATION PERFORMANCE 1333
sample was mounted on a carbon sticky tab and
platinum coated (10 nm thickness; Edwards
E306A Sputter Coater, UK), prior to analysis.
Images were obtained at magnifications of 1000
and 5000.
Amorphous Content Quantification of
Lactose Monohydrate
Organic Dynamic Vapor Sorption (Organic DVS)
was used to quantitatively determine the amor-
phous content in the milled lactose monohydrate
samples using a method described elsewhere.22
In simple terms, the technique measures the
adsorption of a dispersive molecule (n-octane) into
the surface of a sample as a function of partial
pressure. Since the relative adsorption of the
molecule into amorphous and onto crystalline
samples will vary, a calibration curve may be con-
structed by comparing the relative adsorption in
blends of 100% crystalline and 100% amorphous
samples. From this, the amorphous content of an
unknown sample may be determined. Measure-
ments were conducted using a DVS-1 (Surface
Measurement Systems, Alperton, UK), at 258C,
using n-octane as the organic probe. Approxi-
mately 100 mg of lactose monohydrate was
weighed into the sample pan and exposed to a
two-step octane partial pressure (p/po) cycle of
090%. Equilibrium at each step was deter-
mined by a dm/dt of 0.0002% min1. Each milled
sample (n 5), and the amorphous content calcu-
lated from the calibration data reported else-
where.22 In addition, a sample of the 60 min
recrystallized sample was analysed to ensure full
recrystallization.
Drug Content Determination
Quantification of NST content uniformity and
in vitro deposition was determined by UV spectro-
photometry (U-2000 spectrophotometer, Hitachi,
Japan) at a wavelength of 376.5 nm. Samples
were prepared and diluted appropriately in water.
The calibration plot for NST was linear over the
range 0.550.0 mg mL1 (R2 1.00). Lactose
monohydrate did not interfere with the analysis
at the wavelength used.
Dispersion Studies
Preparation of Blends
The influence of carrier milling on the drug
aerosolization efficiency was evaluated using 5%
w/w blends of NST. Blends of 1 g were prepared
JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 96, NO. 5, MAY 2007
1334 YOUNG ET AL.

by geometric hand mixing 50.00 mg NST with particles with an MMAD < 6.8 mm) as a percentage
950.00 mg of lactose monohydrate sample in a of the total recovered dose. The total recovered
glass mortar using a spatula. Blend homogeneity dose (loaded dose) was calculated as the total
was performed by analysing 35.00
1.00 mg amount collected from the inhaler, throat, and all
samples of each blend (n 5, due to the relatively stages of the MSLI. Emitted dose was calculated as
small blend size) five times for each powder total recovered from all stages, postdevice.
mixture. In all cases, an acceptable degree of All samples were evaluated in triplicate and
homogeneity was achieved with the mean drug were randomized for formulation. Temperature
content across all blends being within 100.0
and humidity during the in vitro investigation was
3.0% of the theoretical value and each blend 258C and 45% RH, respectively.
exhibiting a coefficient of variance <5.0%. Ap-
proximately 35.00
1.00 mg samples of each
blend were manually filled into size 3 hard RESULTS AND DISCUSSION
gelatine capsules (Capsugel1, NSW, Australia)
and stored at 45% RH and 258C for 24 h prior to Particle Sizing of Processed Lactose Monohydrate
testing. Samples and NST
Particle size analysis of NST gave median
In Vitro Aerosolization Studies diameter of 1.10 mm with 90% particles less than
5.40 mm, suggesting the model drug was suitable
The influence of the carrier humidity conditioning
for inhalation and DPI studies.
on the aerosolization of NST from the freshly
The influence of milling time on the particle size
milled and milled-recrystallized lactose monohy-
distributions of both freshly milled and recrystal-
drate carrier was investigated using apparatus C,
lized lactose monohydrate samples was investi-
the MSLI (Copley Scientific, Nottingham, UK),
gated. As expected, the milling process resulted in
according to the method described in the British
a significant reduction (ANOVA, p < 0.05) in the
Pharmacopoeia 2005. Briefly, the apparatus con-
median particle diameter with respect to time
sisted of a USP throat, four stages, each stage
(Fig. 1). Such observations are in agreement
containing 20 mL of water, and a filter stage,
with previous investigations.2325 In general, a
which, at a flow rate of 60 L min1 produces
decrease in median diameter from 115
1 mm for
MMAD cut-off points of 13.0, 6.8, 3.1, and 1.7 mm
the untreated lactose monohydrate to 63
1 and
for stages 1, 2, 3, and 4, respectively. The flow
65
1 mm for freshly milled and recrystallized
rate through the MSLI was controlled by a GAST
lactose monohydrate samples was observed,
rotary vein pump and solenoid valve timer
respectively. It interesting to note however, that
(Copley Scientific). Prior to testing, a 60 L min1
incremental increases in mill time resulted in an
flow rate through the MSLI was set using a
calibrated flow meter.
The aerosolization performance of each blend
was investigated using a CyclohalerTM DPI
(Novartis, Surrey, UK). Briefly, a capsule of the
formulation was placed into the sample compart-
ment of the CyclohalerTM, inserted into a specially
constructed MSLI mouthpiece adapter, primed
and tested at 60 L min1 for 4 s. A 3 s delay prior to
testing was instigated to allow the pump to settle.
Deposited drug fractions were collected from the
DPI and MSLI stages using water. In addition,
the extracted solution for the filter stage was
filtered through a 0.22 mm PVDF filter (Millex GV,
Millipore, Billerica, MA) to remove traces of the
glass filter. The amount of active contained in each
aliquot was determined by UV spectrophotometry
using the method described previously. The FPF Figure 1. Influence of mill time on the median
was defined as the total amount of NST particles particle diameter for freshly milled (*) and recrystal-
deposited in stages 3, 4 and filter (corresponding to lized milled (*) lactose monohydrate samples.

JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 96, NO. 5, MAY 2007 DOI 10.1002/jps
 DRUG AEROSOLIZATION PERFORMANCE 1335

exponential decrease in median diameter (R2

0.996). Again, such observations are expected
since the probability of individual particles being
involved in fracturing processes diminishes as the
particles become smaller.25,26 The mean energy
required to cause fractures increases with deple-
tion in crystal cleavage planes while the magni-
tude of local stress from contact with the milling
material (in this case, ceramic balls) decreases. In
addition, increasing particle aggregation following
particle size reduction may occur. As a conse-
quence, energy may be expended in breaking up
the aggregates instead of the particles. In such
cases, particle size reduction will cease to occur at
some practical limit.25 Secondly, as the particles
become smaller and more numerous, friction
diminishes and the sample may behave as a
semi-solid. Larger particles may arch and protect
smaller particles from impact, whilst smaller
particles coat the grinding medium and cushion-
ing the larger particles from impact. This protec-
tion may prevent further particle size
reduction.27
Representative particle size distributions for
the freshly milled and recrystallized lactose
monohydrate samples are shown in Figure 2A
and B, respectively. From Figure 2A it can be seen
that large variations in the size distribution of both
freshly milled and recrystallized lactose monohy-
drate exist, particularly in relation to the small Figure 2. Particle size distributions of (A) freshly
particle fractions. To further investigate this, the milled lactose monohydrate and (B) recrystallized
variation in percentage particles less than 10 mm milled samples.
(classified as fines for the purpose of this paper)
with milling time was studied. Analysis of the fines
concentration with respect to mill time indicated a ities may allow water vapor to condense in the
significant (ANOVA, p < 0.05) increase from 4.4% capillaries that exist between individual, with
to 18.0% between 0 and 60 min mill time. Again, increased levels of fine particles resulting in an
such observations are expected, since it has been increased number of capillaries.28,29 Furthermore,
suggested that cleavage planes, commonly found highly soluble materials, such as lactose monohy-
in crystals, fracture into many fine particles. This drate, may undergo limited dissolution at inter-
results in few relatively large particles, a number particulate contact points with subsequent solidi-
of fine particles and relatively few particles of fication, thus resulting in solidliquid bridge
intermediate size.27 formation between particles, leading to particu-
In comparison, particle size distribution of the late fusion.30 Similarly, such particle fusion could
recrystallized milled samples suggested a reduced be facilitated by the presence of amorphous
rate of increase in the fines percentage with content on particulate surfaces recrystallising at
milling time (4.410.1%, between 0 and 60 min elevated humidity.19,21
mill time). Such observations are most likely due to
fusion between the mill-induced fines and larger
Scanning Electron Microscopy
lactose monohydrate particles. Fusion of particles
may be achieved by two possible mechanisms, Representative SEM images of the 0, 30, and
solidliquid bridge formation and/or particle 60 min mill time freshly milled lactose mono-
fusion through amorphous recrystallization. For hydrate samples are shown in Figure 3AC,
example, storage of samples at elevated humid- respectively. As expected, images were in good

DOI 10.1002/jps JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 96, NO. 5, MAY 2007
1336 YOUNG ET AL.
Figure 3. Representative SEM images of freshly
milled lactose monohydrate samples after (A) 0 min,
(B) 30 min, and (C) 60 min mill times.
correlation with the particle size analysis, con-
firming that increasing the mill time resulted in
both a decrease in median particle diameter with
a concurrent increase in fines material. Repre-
sentative high magnification images of the freshly
JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 96, NO. 5, MAY 2007
Figure 4. Representative SEM images of (A) 60 min
freshly milled lactose monohydrate blend and (B) 60 min
recrystallized lactose monohydrate blend.
milled and recrystallized mill samples are shown
in Figure 4A and B, respectively. From Figure 4A,
it can be seen that the samples from the 60 min
mill process exhibit discrete fine particulates
distributed across the surface of the larger lactose
monohydrate particles. In comparison, analysis of
the recrystallized 60 min mill time samples
(Fig. 4B), suggested many of the fine particulates
had become fused to the large lactose monohy-
drate particulate surfaces. Again such observa-
tions are in good agreement with particle size
analysis discussed previously.
Amorphous Content
As previously discussed, the degree of amorphous
content present in the milled lactose monohydrate
samples was determined using a novel organic-
DVS technique. As expected, an increase in mill
time resulted in an exponential increase in
DOI 10.1002/jps

Figure 5. Influence of mill time on the degree of
amorphous content in freshly milled lactose monohy-
drate samples. Milled-recrystallized lactose monohy-
drate samples were completely crystalline (0.00%
amorphous content; not shown).
amorphous content (Fig. 5) (R2 0.999), correlat-
ing with the concurrent decrease in median
diameter observed by size analysis. Again, such
observations may be expected, since amorphous
content was introduced into the sample by surface
molecular modification during the milling pro-
cess.17 As the degree of comminution decreases, it
is logical to conclude that the introduction of
surface molecular damage would follow suit. In
comparison, organic-DVS analysis of the 60 min
mill time recrystallized sample suggested a
completely crystalline material (0.0% amorphous
content). Since the milling process, would most
likely induce amorphous domains on the surface
of the lactose, differences in interfacial forces
between NST and the freshly milled or recrystal-
lized system should exist. The freshly milled
sample, under the experimental conditions used
here, would be thermodynamically unstable and
would have surface amorphous domains with a
degree of molecular mobility relative to the
environmental conditions (45% RH). Clearly,
under such conditions, the force of interaction
would be higher and result in a reduced FPF
when compared to the re-crystallized lactose
system.
In Vitro Aerosolization Performance
The aerosolization performance of NST from
blends of milled and recrystallized lactose mono-
hydrate was studied using a MSLI. Analysis of the
deposition data suggested milling resulted in no
DOI 10.1002/jps
DRUG AEROSOLIZATION PERFORMANCE 1337
Figure 6. Relationship between mill time and fine
particle fraction for freshly milled (*) and recrystallized
milled (*) lactose monohydrate samples. *R2 relation-
ship for freshly milled samples between 0 and 40 min
mill time.
significant difference in either loaded or emitted
dose across all mill times and for either freshly
milled or recrystallized lactose monohydrate
samples (ANOVA, p > 0.05). Mean loaded doses
of 1614
79 and 1602
81 mg and emitted doses of
1264
58 and 1274
41 mg were observed for
freshly milled and recrystallized samples, respec-
tively. Since no difference in loaded or emitted
dose was observed between all samples, the
influence of milling and recrystallization of
lactose monohydrate carriers on the aerosoliza-
tion performance could be confidently evaluated.
The FPF (MMAD < 6.8 mm) of the loaded dose
was used as a measure of DPI performance. The
influence of mill time on the FPF of both freshly
milled and recrystallized lactose monohydrate
samples is shown in Figure 6. In general, the
comminution process had a significant in-
fluence on aerosolization performance of NST in
both freshly milled and recrystallized samples
(ANOVA, p < 0.05). Furthermore, with the
changes in the physical properties of the carrier
induced by ball milling, a linear relationship1
was observed between FPF and milling time for
both freshly milled and recrystallized samples
(R2 0.954 and 0.938 for freshly milled and
recrystallized samples, respectively). It is inter-
esting to note however, a deviation from linearity
for FPF occurred for the freshly milled samples,
after 40 min mill time. In addition, analysis of FPF
1
Linear analysis for freshly milled samples is only applied
between 0 and 40 min mill times.
JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 96, NO. 5, MAY 2007
1338 YOUNG ET AL.
between freshly milled and recrystallized samples
suggested after 10 min mill time, significant
differences in FPF over the linear region between
20, 30, and 40 min mill times was observed
(ANOVA, p < 0.05).
Clearly, the process of milling induces signifi-
cant variation in aerosolization performance with
an apparent linear relationship between mill time
and FPF over the range 040 and 060 min for
freshly milled and recrystallized lactose mono-
hydrate samples, respectively. As previously dis-
cussed, such observations may be attributed to
many physical characteristics including amor-
phous content, variation in median diameter and
an increase in fines. The relationships between
such factors are discussed in more detail below.
Influence of Median Particle Diameter on
In Vitro Performance
As discussed earlier, multiple changes in the
physical properties of the carrier system were
introduced into the system while milling. The
most obvious change was the significant reduction
in median particle diameter. Since the median
particle diameter for both freshly milled and
recrystallized samples was similar, analysis of
the FPF, with respect to lactose monohydrate
median diameter, suggested a poor relationship
for both samples. In general, R2 values of 0.894
and 0.823 were observed for freshly milled and
recrystallized samples, respectively (Fig. 7).
Thus, a relationship between NST aerosolization
Figure 7. Influence of median particle diameter on
fine particle fraction of freshly milled (*, R2 0.894)
and recrystallized milled (*, R2 0.823) lactose mono-
hydrate samples.
JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 96, NO. 5, MAY 2007
efficiency and median particle size was not as
evident.
A review of the literature revealed conflicting
reports concerning the relationship between med-
ian particle size and FPF. As previously discussed,
many early studies have suggested that reducing
the median particle diameter of carriers signifi-
cantly improves FPF,4,5,31 however, it should be
remembered that with any energy induced particle
size reduction processes, fine particles are intro-
duced. Furthermore, no detailed information
concerning the fines content was given in these
studies. Interestingly, two recent study by Islam
et al.,7,12 and Steckel et al.,13 investigating the
influence of particle size on DPI performance,
reported no significant relationship between FPF
and decreasing median particle diameter, which
correlates with the data presented here.
Influence of Amorphous Content on
In Vitro Performance
The presence of meta-stable, that is amorphous,
material in the surface of DPI carriers may have
implications for product performance. An appar-
ent relationship between amorphous content and
FPF for freshly milled samples was observed
(R2 0.921). However, since recrystallized sam-
ples contained no detectable amorphous content
there would be no relationship between FPF and
amorphous content (e.g. a plot of % amorphous
content against FPF in the recrystallized sam-
ples, would result in all data sitting at 0%).
Furthermore, previous reports have suggested
increased amorphous content in DPI systems
resulted in decreased aerosolization perfor-
mance.17,19,21 Clearly such relationships need to
be studied further, in isolation of other physical
factors.
Influence of Fines Content on In Vitro Performance
As previously discussed, a significant increase in
fines with increased mill time was observed for
both freshly milled and recrystallized lactose
monohydrate samples. The relationship between
fine lactose monohydrate content and FPF is
shown in Figure 8. As with the variation of FPF
and mill time FPF (Fig. 6), a linear relationship
between FPF and the percentage of fines (<10 mm)
present in each sample was observed. However, it
is interesting to note that the relationship was
more significant with R2 values of 0.974 and 0.982
DOI 10.1002/jps

Figure 8. Relationship between percentage lactose
monohydrate fines content (<10 mm), fine particle
fraction for freshly milled (*) and recrystallized milled
(*) lactose monohydrate samples. *R2 relationship for
freshly milled samples where fines are less than 15%.
being observed for freshly milled and recrystal-
lized lactose monohydrate samples, respectively.
Again, as with the variation of FPF with mill time
data, linearity for the freshly milled samples only
existed up to a certain extent (equivalent to
40 min or 13.9% fines), after which, the FPF
decreased.
More importantly, when compared to the influ-
ence of mill time, the relationship between the
fines content of either freshly milled or recrystal-
lized samples and FPF indicated no significant
difference with respect to fines content (ANOVA,
p < 0.05). In simple terms, samples with less than
15% fines resulted in no significant difference in
FPF between samples of milled or recrystallized
lactose monohydrate carriers containing similar
fines percentage. Such observations correlate with
previous investigations. Zeng et al.,6,9 and Islam
et al.,7,12 reported a significant reduction in FPF
with a reduction in fines content. The FPF was
returned to the original level when the fines
removed were restored, and further improved with
increasing fines content. Various studies reported
a trend of improved FPF when fine particles
(lactose monohydrate, glucose or PEG 6000) were
introduced to coarse carrier mixtures, irrespective
of coarse carrier particle size.8,10,11,32
In general, two mechanisms have been pro-
posed to explain such observations: the active site
theory and multiplet/agglomeration theory.
In simple terms, the active site theory suggests
that the increasing carrier fine concentration
DOI 10.1002/jps
DRUG AEROSOLIZATION PERFORMANCE 1339
occupies high energy-active sites, thus promot-
ing drug adhesion to occur at relatively lower
energy-passive sites.6,811,33 Clearly, the result of
such an effect would be the easier detachment
of drug particles and thus increased FPF.
However, although the existences of such sites
have been experimentally verified in recent
publications,1619,33 it is suggested that at such
high drug loadings (5% w/w) active site effects
would be minimal. Alternatively, adhesion and
redistribution of ingredients, when fines are
present, may produce a mixed agglomerate of
drug and fine material (forming multiplets or
agglomerates). Such agglomeration may result in
improved drug aerosolization, since, due to simple
physics, dispersion of larger agglomerates from
larger carrier particles will be achieved at lower
forces, when compared to typical individual drug
carrier systems.10 In the case of such high drug
loadings, the authors propose that this mechanism
would most likely dominate FPF.
Although such general theories are attractive, a
deviation from linearity at high fine concentra-
tions (>15% fines) still exists. One of the most
likely explanations for the observed decrease in
FPF in samples with fines >15% is a mixed
agglomerate theory, where at high fines concen-
trations drug-lactose monohydrate agglomerates
fail to adhere to the larger lactose monohydrate
carrier particles and become segregated. Such
segregation results in a biphasic system in that the
typical ordered mix of micronized drug/lactose
monohydrate becomes a two-component blend:
the carrier/drug agglomerate particles and the
free agglomerates. Such a blend may result in
deviation from an expected agglomeratecarrier
relationship. To further investigate the potential
for such segregation, the particulate structure of
formulations containing different levels of fine
lactose monohydrate were investigated. Represen-
tative SEM images of formulations containing
18.0% (60 min freshly milled lactose monohydrate)
and 10.1% (60 min recrystallized lactose mono-
hydrate) fines are shown in Figure 9A and B,
respectively.
When comparing samples containing >15%
fines (Fig. 9A), large drug-fine agglomerates, that
were clearly separated from the coarse carrier,
were present in the system. In contrast, with
moderate fines content (Fig. 9B), a homogenous
blend of mixed drug-fine agglomerates adhering to
the coarse carrier surface was depicted. From such
observations, it is reasonable to assume, that a
critical agglomerate size may exist in drug-fine-
JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 96, NO. 5, MAY 2007
1340 YOUNG ET AL.
Figure 9. Representative SEM images of (A) freshly
milled lactose monohydrate blends containing >15%
fines and (B) recrystallized lactose monohydrate blends
containing <15% fines.
carrier blends where segregation may result in a
reduction in aerosolization performance. Indeed,
in recent studies similar relationships were
observed.10
Finally, although a direct relationship between
fine concentration and aerosolization performance
was observed (<15% fines), a discrepancy still
exists when comparing the regression slopes of
freshly milled and recrystallized lactose mono-
hydrate samples (Fig. 8) (slope 1.56 and 1.08 for
freshly milled and recrystallized samples, respec-
tively). It is envisaged that this variation in slope
may be due to increased adhesion between the
drug particles and amorphous regions. It is also
important to note, that in this study, drug particles
were blended with the lactose postrecrystalliza-
tion. It is envisaged that, if blended prior to
recrystallization, the drug particles would possibly
fuse to the lactose surface as with the fines,
JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 96, NO. 5, MAY 2007
markedly reducing the FPF. However, these
parameters needed to be studied in isolation (i.e.
without the presence of fines) and are suggested
for future investigation.
CONCLUSION
Industrial processing of carrier materials used in
DPI systems induces changes in the physical
properties of the particles. Here, a simple ball
milling process was used to produce particles
which exhibited a reduced particle size, increased
levels of fines and amorphous material. In addi-
tion, variation in storage conditions of the pro-
cessed excipient was also shown to induce
changes in fines and amorphous content. When
used in a high dose DPI system, significant
changes in the FPF were observed with increasing
milling times. The relationships between milling
time, physical property of the carrier and FPF
were investigated. In general, the strongest
relationship between carrier physical property
and FPF was observed when considering the fines
content. Such a relationship was independent of
storage conditions, with increasing fines (<15%)
resulting in a linear increase in FPF. Subse-
quently, the presence of fines was shown to play
the predominating role in influencing DPI perfor-
mance in this system. Furthermore, increasing
fine content above 15% resulted in a deviation
from linearity and may be related to changes in
overall formulation characteristics. Finally, there
appears to be some evidence that the presence of
amorphous content may contribute to a decreased
FPF. However, like the effect of particle size,
further investigation is required to study the
effect of these two parameters on dry powder
dispersion in isolation, without the masking effect
of fines content.
REFERENCES
1. Edge S, Kibbe A, Kussendrager K. 2006. Lactose,
Monohydrate. In: Rowe RC, Sheskey PJ, Owen
SC, editors. Handbook of pharmaceutical excipi-
ents, 5th Edition. London: Pharmaceutical Press.
pp 389395.
2. Steckel H, Markefka P, teWierik H, Kammelar R.
2004. Functionality testing of inhalation grade
lactose. Eur J Pharm Biopharm 57:495505.
3. Bell JH, Hartley PS, Cox JS. 1971. Dry powder
aerosols. I. A new powder inhalation device.
J Pharm Sci 60:15591564.
DOI 10.1002/jps

4. French DL, Edwards DA, Niven RW. 1996. The
influence of formulation on emission, deaggrega-
tion and deposition of dry powders for inhalation.
J Aerosol Sci 27:769783.
5. Steckel H, Muller BW. 1997. In vitro evaluation of
dry powder inhalers II: influence of carrier particle
size and concentration on in vitro deposition. Int J
Pharm 154:3137.
6. Zeng XM, Martin GP, Tee SK, Ghoush AA, Marriott
C. 1999. Effects of particle size and adding
sequence of fine lactose on the deposition of
salbutamol sulphate from a dry powder formula-
tion. Int J Pharm 182:133144.
7. Islam N, Stewart P, Larson I, Hartley P. 2004.
Effect of carrier size on the dispersion of salmeterol
xinaofoate from interactive mixtures. J Pharm Sci
93:10301038.
8. Lucas P, Anderson K, Stanniforth JN. 1998.
Protein deposition from dry powder inhalers: Fine
particle multiplets as performance modifiers.
Pharm Res 15:562.
9. Zeng XM, Martin GP, Tee SK, Marriott C. 1998.
The role of fine particle lactose on the dispersion
and deaggregation of salbutamol sulphate in an air
stream in vitro. Int J Pharm 176:99110.
10. Louey MD, Stewart PJ. 2002. Particle interactions
involved in aerosol dispersion of ternary interactive
mixtures. Pharm Res 19:15241531.
11. Louey MD, Razia S, Stewart PJ. 2003. Influence of
physico-chemical carrier properties on the in vitro
aerosol deposition from interactive mixtures. Int J
Pharm 252:8798.
12. Islam N, Stewart P, Larson I, Hartley P. 2004.
Lactose surface modification by decantation: Are
drug-fine lactose ratios the key to better dispersion
of salmeterol xinafoate from lactose-interactive
mixtures? Pharm Res 21:492499.
13. Steckel H, Markefka P, teWierik H, Kammelar R.
2006. Effect of milling and sieving on functionality
of dry powder inhalation products. Int J Pharm
309:5159.
14. Zeng XM, Martin GP, Marriott C, Pritchard J.
2000. The influence of carrier morphology on drug
delivery by dry powder inhalers. Int J Pharm 200:
93106.
15. Flament MP, Leterme P, Gayot A. 2004. The
influence of carrier roughness on adhesion, content
uniformity and the in vitro deposition of terbutaline
sulphate from dry powder inhalers. Int J Pharm
275:201209.
16. Young PM, Cocconi D, Colombo P, Bettini R, Price
R, Steele DF, Tobyn MJ. 2002. Characterization of
a surface modified dry powder inhalation carrier
prepared by particle smoothing. J Pharm Phar-
macol 54:13391344.
17. Price R, Young PM. 2005. On the physical trans-
formations of processed pharmaceutical solids.
Micron 36:519524.
DOI 10.1002/jps
DRUG AEROSOLIZATION PERFORMANCE 1341
18. Buckton G, Darcy P. 1999. Assessment of disorder
in crystalline powdersa review of analytical
techniques and their application. Int J Pharm
179:141158.
19. Young PM, Price R. 2004. The influence of humidity
on the aerolisation of micronised and SEDS
produced salbutamol sulphate. Eur J Pharm Sci
22:235240.
20. Podczeck F, Newton JM, James MB. 1997. Varia-
tions in the adhesion force between a drug and
carrier particles as a result of changes in the
relative humidity of the air. Int J Pharm 149:151
160.
21. Ward GH, Schultz RK. 1995. Processed-induced
crystallinity changes in albuterol sulphate and its
effect on powder physical stability. Pharm Res
12:773779.
22. Young PM, Chiou H, Tee T, Traini D, Chan HK,
Thielmann F, Burnett D. 2007. The use of organic
vapor sorption to determine low levels of amor-
phous content in processed pharmaceutical pow-
ders. Drug Dev Ind Pharm 33:9197.
23. Chen Y, Ding Y, Papadopoulos DG, Ghadiri M.
2004. Energy-based analysis of milling alpha-
lactose monohydrate. J Pharm Sci 93:886895.
24. Kwan CC, Chen YQ, Ding YL, Papadopoulos DG,
Bentham AC, Ghadiri M. 2004. Development of a
novel approach towards predicting the milling
behaviour of pharmaceutical powders. Eur J
Pharm Sci 23:327336.
25. Berg TGO, Avis LE. 1970. Exploratory experiments
on kinetics of comminution. Powder Technol 4:27
31.
26. Piret EL. 1953. Fundamental aspects of grinding.
Chem Eng Prog 49:5662.
27. Parrott EL. 1974. Milling of pharmaceutical solids.
[Review] [95 refs]. J Pharm Sci 63:813829.
28. Coelho MC, Harnby N. 1978. Moisture bonding in
powders. Powder Technol 2:201205.
29. Schubert H. 1984. Capillary forces: Modelling and
application in particulate technology. Powder
Technol 37:105116.
30. Young PM, Price R, Tobyn MJ, Buttrum M, Dey
F. 2003. Effect of humidity on aerosolization of
micronized drugs. Drug Dev Ind Pharm 29:959
966.
31. Bell JH, Hartley PS, Cox JS. 1971. Dry powder
aerosols. I. A new powder inhalation device.
J Pharm Sci 60:15591564.
32. Zeng XM, Pandhal KH, Martin GP. 2000. The
influence of lactose carrier on the content homo-
geneity and dispersibility of beclomethasone dipro-
pionate from dry powder aerosols. Int J Pharm
197:4152.
33. Young PM, Edge S, Traini D, Jones MD, Price R,
El-Sabawi D, Urry C, Smith C. 2005. The influence
of dose on the performance of dry powder inhalation
systems. Int J Pharm 296:2633.
JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 96, NO. 5, MAY 2007