Powder Technology 262 (2014) 170176

Contents lists available at ScienceDirect

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

A study of jet-milling and spray-drying process for the physicochemical

and aerodynamic dispersion properties of amiloride HCl
Marija Djoki a, Kyriakos Kachrimanis b, Ljiljana Solomun a, Jelena Djuri a, Dragana Vasiljevi a, Svetlana Ibri a,
a
Faculty of Pharmacy, Department of Pharmaceutical Technology, University of Belgrade, Belgrade, Serbia
b
Department of Pharmaceutical Technology, Aristotle University of Thessaloniki, Thessaloniki, Greece

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

Article history: The aim of this investigation was to examine the effect of a jet-mill and spray-drying process on the physico-
Received 26 January 2014 chemical and aerodynamic dispersion properties of amiloride HCl particles. Micro-ne particles were prepared
Received in revised form 10 April 2014 by a spiral air jet-mill Hosokawa 50 AS and Mini Spray-Dryer B-191. A 23-1 fractional factorial experimental de-
Accepted 19 April 2014
sign was implemented to screen three jet-mill process parameters. Possible changes in the physicochemical
Available online 27 April 2014
properties of the material due to spiral jet-milling were examined by the determination of the particle size
Keywords:
distribution (PSD), powder true density, powder owability, diffuse reectance Fourier transform infrared
Jet-milling spectroscopy (DRIFTS) and scanning electron microscopy (SEM). The effect of different jet-milling parameters
Spray-drying and the spray-drying process on the aerosol dispersion characteristics was examined with a cascade impactor
Aerosol dispersion with a preseparator using the Aerolizer dry powder inhaler device (Novartis, Switzerland). The results show
Amiloride HCl that small changes in the particle size within the 1 to 5 m range had an impact on the physicochemical and
Particle size aerosol dispersion properties of jet-milled and spray-dried particles.
2014 Elsevier B.V. All rights reserved.

1. Introduction
Inhalation of aerosols as a means of pulmonary drug delivery has
been used for many years to deliver xenobiotics to the lungs for both
systemic and local effects. The ability to deliver active substances direct-
ly to the site of action enables lower doses compared to other routes of
administration with an equivalent therapeutic response and a lower
systemic exposure [1]. Inhalable aerosols for the treatment of lung
diseases are delivered largely using either pressurised metered dose in-
halers (MDIs) or dry powder inhalers (DPIs). DPIs are a widely accepted
inhaled dosage form, particularly in Europe where they are currently
used by an estimated 40% of patients to treat asthma and chronic ob-
structive pulmonary disease [2]. DPIs contain particles like those inves-
tigated in this study. The dispersion of drug particles as aerosols from
the powder formulations, containing either drug alone or excipient
Abbreviations: PSD, particle size distribution; DRIFTS, diffuse reectance Fourier trans-
form infrared spectroscopy; Cv, cumulative particle volume; ED, emitted dose; FPF, ne
particle fraction; MMAD, mass median aerodynamic diameter; GSD, geometric standard
deviation; MDIs, metered dose inhalers; DPIs, dry powder inhalers; EP, European
Pharmacopoeia; MSLI, multistage liquid impinger; ACI, Andersen cascade impactor; NGI,
Next generation impactor; FT-IR, Fourier transform infrared; CI, Carr's compressibility
index; HR, Hausner ratio; , angle of repose; tap, tapped density; bulk, bulk density;
SEM, scanning electron microscopy; SPSS, Statistical Products and Service Solutions;
DSCG, disodium cromoglycate.
Corresponding author at: Department of Pharmaceutical Technology, Faculty of
Pharmacy, Vojvode Stepe 450, 11221 Belgrade, Serbia. Tel.: +381 11 3951363;
fax: +381 11 3972840.
E-mail address: svetlana.ibric@pharmacy.bg.ac.rs (S. Ibri).
blends, is required for lung deposition. Various factors may affect parti-
cle dispersion from interactive mixtures, including the carrier size and
distribution [35], shape and surface roughness [6,7], drug-carrier
ratio [4,5,8], presence of ternary components [810] and mixing and
storage conditions [8]. Unfortunately, the choice of excipients used to
enhance a powder's aerosol behaviour is very limited for pulmonary ap-
plications. Alternative strategies involve the engineering of particles
with controlled and suitable properties, such as a narrow particle size
distribution and improved dispersibility [11]. In this study, particle
properties were controlled by three jet-mill process parameters and a
spray-drying process.
When using a DPI, a failure to inhale deeply and forcibly at the start
of the inhalation means that the drug particles generated are not suf-
ciently de-aggregated to enter the lungs and are simply deposited in
the mouth and oro-pharynx where they have no clinical efcacy [12].
A multistage impactor is usually the method of choice to characterise
the quality of the dose emitted from an inhaler. Multistage impactors
are described in the European Pharmacopoeia (EP), namely the multi-
stage liquid impinger (MSLI), the Andersen cascade impactor (ACI)
and the more recently developed next generation impactor (NGI) [13].
The ACI was used in this study.
Particles with aerodynamic diameters of 1 to 5 m are considered re-
spirable [14]. Respirable-sized particles are traditionally prepared by
jet-milling or spray-drying techniques, which are well-established and
validated techniques used to manufacture dry powders for inhalation.
Spray-drying is a technique involving atomisation of a liquid feed into
a hot gaseous medium to produce dried particulates [15]. Jet-milling is

http://dx.doi.org/10.1016/j.powtec.2014.04.066
0032-5910/ 2014 Elsevier B.V. All rights reserved.
 M. Djoki et al. / Powder Technology 262 (2014) 170176 171

nozzle for injectable air (0.80.9 mm), the diameter of the ring
Nomenclature
nozzle for grinding air (0.70.9 mm) and the air pressure for milling
(46 bar), while the feed rate (10 g/min) was kept constant.
angle of repose ()
Spray-dried particles were prepared from aqueous feed solutions
d50 50th percentile geometric particle size (m)
using a Mini Spray-dryer B-191 (Buichi, Flawil, Switzerland). The
tap particle tap density (g/cm3)
atomising gas rate at which the spray-dryer was operated was set to
bulk particle bulk density (g/cm3)
800 l/h, and the aspirator setting was 100%. Nitrogen was used as the
d84 ratio of the aerodynamic particle size at the 84th
atomisation gas. The particle size distribution was obtained using the
percentile (m)
following parameters: feed concentration (0.35% w/w), feed rate
d16 ratio of the aerodynamic particle size at the 16th
(12 ml/min), atomisation pressure (2 bar), inlet temperature (150 C)
percentile (m)
and outlet temperature (95 C), to obtain respirable particles compara-
ble with jet-milled particles. The mentioned spray-drying parameters
resulted in spherical hollow spray-dried particles of amiloride HCl that
a size reduction technique whereby high-energy comminution of
are useful for inhalation therapy or systemic absorption via the respira-
particles occurs by particleparticle and particlewall collisions under
tory tract, as disclosed previously in the literature [28].
the inuence of high-velocity jets of compressed gases. Limited control
Particle size distributions were determined by a laser diffraction-
over the size, shape and morphology can be exercised [16]. Therefore, it
based Malvern Mastersizer 2000 particle size analyser (Malvern
is very important to improve the milling process efciency and quality
Instruments, Worcestershire, UK). Particles were dispersed using a
by determining the optimal milling parameters. The effects of the
Scirocco feeder instrument (Scirocco 2000 dry powder feeder, Malvern
main mechanical milling parameters, namely the milling time, process
Instruments) with 1 bar pressure and a feed rate of 25%. The results
control agent, ball to powder ratio and milling speed, in the planetary
shown are the average values of two measurements.
ball milling of nanocrystalline Al 2024 powder are optimised by the
The d10, d50 and d90 values were determined. However, the laser
Taguchi method. The mean particle size (d50) is used to evaluate the
diffraction provides a measurement of the geometrical rather than aero-
effect of the process parameters on the mechanical milling process
dynamic particle size, and the apparent particle density and dynamic
[17]. The Taguchi method was used in this study with the d50 value
shape factors of drug agglomerates are not considered.
and emitted dose as responses. The particle size and microhardness of
The true density of all jet-milled samples was measured using
the composite powders are strongly affected by milling time in a plane-
helium pycnometry (Quantachrome ultrapycnometer 1000, USA). The
tary ball mill, as well as the reinforcement size and reinforcement
reported values are the average of three measurements.
content [18,19]. The mechanical milling process is inuenced by a
Fourier transform infrared (FT-IR) spectra were used to investigate
new process control agent technique, and an articial neural network
any possible changes in the jet-milled amiloride HCl samples that may
model for the prediction of the effect of a gradual process control
have occurred at the molecular level during the jet-milling processes.
agent is developed [20]. In this study, the diameter of an injector nozzle
These spectra were recorded using a Shimadzu IR Prestige-21 Fourier
for injectable air, the diameter of a ring nozzle for grinding air and the
transform infrared spectrophotometer (Shimadzu Corporation, Kyoto,
air pressure for the milling effects on the physicochemical and aerody-
Japan). The spectrophotometer was coupled with a horizontal Golden-
namic dispersion properties were examined.
Gate MKII single reection ATR system (Specac, Kent, UK). The samples
The advantages of jet-milling include the absence of contamination
were analysed directly in the bulk form. The scanning range was
due to an autogenous grinding mechanism, low wear rate, and small
6504000 cm1, the resolution was 4 cm1 and the number of scans
footprint, which leads to a reduction in the space requirements, low
per spectrum was 32.
noise and the ability of grinding heat-sensitive materials [2124]. How-
The static powder ow was characterised using the angle of repose,
ever, the inuence of the jet-milling process parameters on aerosol
apparent density, Carr's compressibility index (CI) and the Hausner
dispersion powder formulations containing drug alone has yet to be
ratio (HR), determined from the tapped (tap) and bulk densities (bulk):
examined [25].
The present study uses amiloride HCl as a model compound for  
CI tap bulk =tap  100% 1
drug-only powder formulations. Aerosolised amiloride has been
shown to improve mucociliary clearance in patients with cystic brosis
[26]. Either polymorphic dihydrate A or B may be received when HR tap =bulk : 2
purchasing raw amiloride HCl due to an insignicant difference in the
solubility and dissolution rate and the high similarity of their FTIR spec-
The angle of repose was determined by measuring the height of the
tra, as well as their melting points and hydroscopicity [27]. The objective
cone of powder and calculating the angle of repose () from the follow-
of this study was to examine the inuence of the jet-mill and spray-
ing equation:
drying process on the physicochemical and aerosol dispersion charac-
teristics of amiloride HCl particles with an appropriate size and tan height=0:5  base: 3
structure for DPI use.
The apparent density was measured in a 100-ml glass graduated cyl-
2. Experimental procedure inder (ISOLAB Laborgerte GmbH, Wertheim, Germany) using an auto-
matic tapper (Erweka SVM, Erweka GmbH, Germany). The apparent
2.1. Materials and methods volume was determined after 10, 500 and 1250 taps. The apparent den-
sity was reported after 1250 taps. The tapped and bulk densities were
Amiloride hydrochloride (batches no. 3100/02/10), used as the measured in a 100-ml glass graduated cylinder (ISOLAB Laborgerte
model drug, was supplied by Sifavitor srl (Milano, Italy). Amiloride GmbH, Wertheim, Germany) using an automatic tapper (Erweka SVM,
HCl is physically stable under normal storage conditions only in the Erweka GmbH, Germany). The tapped density was determined after
dihydrate phase, and this is the commercially available form. 400 taps. Low CI and HR values are an indication of better ow behav-
To investigate pulmonary deposition of the jet-milled samples, iour. Carr's index values of less than 25% and Hausner ratio values less
amiloride HCl was jet-milled with a spiral air jet-mill Hosokawa 50 AS than 1.25 indicate acceptable ow properties.
(Hosokawa Micron Ltd., Cheshire, England). The resulting particle size Amiloride HCl was quantied by ultraviolet (UV) absorbance mea-
distributions were adjusted by varying the diameter of the injector sured at a wavelength of 362 nm in a UVvisible CamSpec M330
172 M. Djoki et al. / Powder Technology 262 (2014) 170176

spectrophotometer (CamSpec Limited, Sawston, Cambridge, UK). The Table 2

amiloride HCl deposited on the eight stages and lter was washed The quantitative factor effects for d50 value and emitted dose.

with 100 ml 0.13% NaCl. A volume of 4.9 ml of 0.1 M HCl was added to
40 ml of each of the eight stages and lter washout, and the solution
was brought to 50 ml with 0.13% NaCl. The results are reported as
percentages.
The aerodynamic particle size distribution was measured by a
Spraytec in line with an ACI (Malvern Instruments, Worcestershire,
UK) non-viable cascade impactor consisting of eight stages together
with a nal lter operating at an airow rate of 28.3 l/min. The stages
were clamped together. The Aerolizer (Pzer) dry powder inhalation
device was lled with 20 mg powder for each test and mounted to the
Spraytec through a USP throat device. The data output from the Spraytec
included a cumulative particle volume (Cv) as a measure of the cloud
particle mass, as well as the particle sizing information as a function of
time. Drug deposition in the ACI on stages 07 and the lter was deter-
mined by UV spectrophotometric analysis. A suitable quantication
method was previously described [29]. The amount of amiloride HCl de-
posited in the eight stages and nal lter was quantied. The emitted
dose (ED) was dened as the mass of particles delivered from the inhal-
er, expressed as a percentage of the total amount of amiloride HCl col-
lected. The ne particle fraction (FPF) was dened as the mass of
particles deposited in stage 2 and lower stages, expressed as a percent-
age of the total amount of amiloride HCl collected. The mass median
aerodynamic diameter (MMAD) was calculated as the 50th percentile
of the aerodynamic particle size distribution by mass. The geometric
standard deviation (GSD) was calculated as the ratio of the particle
size at the 84th percentile to the 16th percentile, using the following
equation [30]:
1=2
GSD d84 =d16
where GSD is the geometric standard deviation, d84 is the ratio of the
particle size at the 84th percentile, and d16 is the ratio of the particle
size at the 16th percentile.
For the three measurements of each of the ve samples, the statisti-
cal signicance of the differences between the emitted dose, FPF, MMAD
and GSD of amiloride HCl at the various stages of the ACI was deter-
mined by analysis of variance, using SPSS for Windows (Statistical Prod-
ucts and Service Solutions Inc., Chicago, Illinois, USA). The absence of
differences was chosen as the H0-hypothesis.
The surface morphology was examined by a scanning electron mi-
croscope (Jeol JSM-6390LV, Jeol, Peabody, USA). The particles were cov-
ered with gold to perform these measurements.
2.2. Experimental design
A 2(3-1) screening experimental design was conducted to assess the
inuence of altering three jet-milling process variables on the particle
mean diameter, as well as the emitted dose of the aerosol dispersion.
The factors and levels investigated for each factor are given in Table 1.
The values for the parameters were selected according to the
manufacturer's recommendation, keeping in mind the basic principles
Factor Factor effects
d50 Emitted dose
1
A 0.17 0.91
B2 5.000E003 0.28
C3 0.45 7.13
A1 diameter of injector nozzles for injectable air.
B2 diameter of ring nozzles for grinding air.
C3 air pressure.
of jet-milling. The factor effects were calculated using Design Expert
software (Stat-Ease, Minneapolis, USA).
3. Results and discussion
3.1. Screening study
After the application of the 2(3-1) fractional factorial design, the
factor effects on the d50 value and the emitted dose as responses are
shown in Table 2.
The results of the examined responses from the samples obtained
according to the 2(3-1) fractional factorial design (FFD) were tted into
the linear model, and the following equations (in terms of coded
factors) were calculated:
Y 1 d50 2:340:17  X 1 5:000 E003  X 2 0:45  X 3 5
Y 2 ED 76:470:91  X 1 0:28  X 2 7:13  X 3 : 6
4
Lower diameters of the injector nozzle for injectable air, lower
diameters of the ring nozzle for grinding air, and lower air pressure
for grinding all resulted in higher d50 values.
Regarding the factor effects on the emitted dose of the aerosol dis-
persion as the response, which are shown in Table 2, it is seen that a
lower diameter of injector nozzle for injectable air, a lower diameter
of ring nozzle for grinding air, and a lower air pressure for grinding
resulted in higher results for the emitted dose of aerosol dispersion.
3.2. Physicochemical characterisation
3.2.1. Particle size distribution
Jet-milled and spray-dried particles were produced with the d50 and
GSD values shown in Table 3. The results indicate that the geometric
particle diameter is inuenced by the air pressure of grinding because
the geometric particle diameters were higher for lower air pressure of
grinding (4 bar). Thus, by using lower air pressure for grinding, a higher
geometric particle diameter is obtained because lower pressurised
grinding air leads to poor acceleration and larger powder particles.
The lower geometric particle diameter (d50 = 1.71 m, 2.07 m) was
achieved by higher air pressure for grinding (6 bar). For the same air
pressure for grinding, a larger diameter injector nozzle for injectable
air (0.9 mm) generated lower geometric particle diameters. Thus, a

Table 1
Composition of the 2(3-1) factorial design plan (4 batches to prepare, runs respectively).
Table 3
Run Diameter of Diameter of ring Air pressure for d10, d50 and d90 values of jet-milled and spray-dried particles.
injector nozzle nozzle for grinding milling (bar)
Sample d10 (m) d50 (m) d90 (m)
for injectable air (mm)
air (mm) JM1 1 0.84 1.71 4.01
JM1 2 0.80 2.07 4.92
Coded Real Coded Real Coded Real
JM1 3 0.87 2.62 6.58
1 1 0.8 1 0.7 +1 6 JM1 4 0.86 2.96 7.31
2 1 0.8 +1 0.9 1 4 SD2 0.85 2.89 7.16
3 +1 0.9 +1 0.9 +1 6
JM1 jet-milled.
4 +1 0.9 1 0.7 1 4
SD2 spray-dried.
 M. Djoki et al. / Powder Technology 262 (2014) 170176
Table 4
True density of unmilled and jet-milled amiloride HCl.
Sample True density (g/cm3)
UM1 1.71
JM2 1 1.67
JM2 2 1.68
JM2 3 1.72
JM2 4 1.73
UM1 unmilled.
JM2 jet-milled.
larger diameter injector nozzle allows a better supply of pressurised in-
jectable air through a venture, higher acceleration and production of
ner particles. The diameter of the ring nozzles for grinding had the
lowest inuence on the characteristics of the particle size distribution,
when the mentioned values for the other process parameters were
used.
3.2.2. True density measurement
The true densities of unmilled and jet-milled amiloride HCl are
presented in Table 4. True density values higher than 1.71 g/cm3 for
jet-milled samples indicate a high level of crystallinity compared with
the raw amiloride HCl sample.
3.2.3. Fourier transition infrared (FT-IR)
The main regions for identifying the amiloride HCl infrared spectrum
were at 3250 and 3150 cm1 assigned to N\H stretching, at 1680 cm1
due to C\O stretching and at 1240 cm1 due to N\(C6H6) stretching.
FTIR scans on the jet-milled and unmilled samples were indistinguish-
able with respect to the polymorphism. Both polymorphs A and B had
identical infrared spectra, corresponding to a previously published
spectrum (Fig. 1) [31].
3.2.4. Powder ow
The angle of repose, apparent density, bulk and tapped densities,
Carr's compressibility index and Hausner ratio values of the jet-milled
amiloride HCl samples are listed in Table 5. For jet-milled particles,
greater owability (lower angle of repose, CI and HR) was observed
for larger particles (d50 = 2.62 and 2.96 m) compared to smaller par-
ticles (d50 = 1.71 and 2.07 m). Higher jet-milled particles (d50 =
2.62 and 2.96 m) exhibited good powder ow behaviour, and particle
sizes closer to 1 m owed poorly. Similar observations were previously
reported [32,33]. For more poorly owing materials (d50 = 1.71 and
2.07 m), there are frequently greater interparticulate interactions,
Fig. 1. The IR spectra of unmilled and milled samples of amiloride HCl.
173
and a greater difference between the bulk and apparent densities was
observed. The loose aggregates, made of large particles (d50 of 2.62 m
SD (g/cm3) and 2.96 m) exhibited good ow and entrainment properties and
were easily disaggregated within the airstream. (See Table 6.)
0.001
0.002
0.001 3.3. Aerosol dispersion characterisation
0.001
0.002
3.3.1. Cumulative particle volume (Cv)
The cumulative particle volume (Cv) proles as a measure of the
cloud particle mass, as well as particle sizing information as a function
of time, are shown in Fig. 2. The powder release proles of the jet-
milled samples (d50 of 2.96 m and 2.62 m) and spray-dried sample
(d50 of 2.89 m) show a higher concentration compared to the jet-
milled samples (d50 of 1.71 m, 2.07 m). The peak concentration oc-
curred at the same time, illustrating better powder emptying.
3.3.2. Emitted dose (ED)
The obtained emitted doses express the drug mass exiting the device
after inhalation. Signicant differences were observed due to the d50
using the Aerolizer (p b 0.001) in the ED of the jet-milled samples
(d50 of 2.62 m and 2.96 m) and the spray-dried sample (d50 of 2.89
m) compared to the jet-milled particles (d50 of 1.71 m, 2.07 m),
where a lower ED was obtained with the smaller jet-milled particles
(Fig. 3).
The results indicate that the emitted dose, as well as the geometric
particle diameter, is inuenced by the air pressure of grinding. The emit-
ted dose was higher for a lower air pressure of grinding (4 bar). Thus, by
using a lower air pressure for grinding, a higher emitted dose of larger
powder particles is obtained. A lower emitted dose was achieved by a
higher air pressure for grinding (6 bar).
Jet-milled particles (d50 of 1.71 m, 2.07 m), where a lower ED was
obtained, align with one another and have a large contact area, giving
rise to high cohesive forces and producing large agglomerates that are
not delivered completely from the inhaler.
The EDs of jet-milled samples (d50 of 2.62 m and 2.96 m) and the
spray-dried sample (d50 of 2.89 m) were higher due to particles that
did aerosolise effectively and aligned with the airow, allowing them
to pass through the impactor [34,35]. Therefore, the ability of the pow-
der to be uidised by the airow through an inhaler is indicated by the
emitted dose.
3.3.3. Fine-particle fraction (FPF)
The obtained ne-particle fraction corresponds to the percentage of
the mass of drug particles that have an aerodynamic diameter of less
than 5 m. Such particles can be deposited in the deep lung after inha-
lation. Signicantly higher FPF (p b 0.001) was obtained with large-
sized jet-milled particles (d50 of 2.62 and 2.96 m) and spray-dried par-
ticles (d50 of 2.89 m) compared to jet-milled particles (d50 of 1.71 m,
2.07 m). Signicantly lower results were observed due to the d50 using
the Aerolizer (p b 0.001) in the FPF of the jet-milled sample (d50 of
2.62 m) compared to the jet-milled sample (d50 of 2.96 m) and
spray-dried sample (d50 of 2.89 m). The FPF results are shown in
Fig. 4. The larger particles of the jet-milled sample (d50 of 2.96 m)
and spray-dried sample (d50 of 2.89 m) result in higher FPF [32] and
are less cohesive with lower specic surface areas and van der Waals
forces per unit mass of particles. These particles are expected to separate
easier and to provide more ne particles. The maximum FPF is thus the
result of a balance of the particle cohesion and inhaler efciency against
the available ne particles at a specic air ow. These results support a
previous study where lower FPFs were observed with smaller particle
sizes of spray-dried mannitol and disodium cromoglycate (DSCG)
particles [36,37].
3.3.4. Mass median aerodynamic diameter (MMAD)
The obtained mass median aerodynamic diameter of an aerosol pre-
sents the particle diameter that has 50% of the aerosol mass residing
174 M. Djoki et al. / Powder Technology 262 (2014) 170176

Table 5
Angle of repose, apparent density, bulk and tapped densities, Carr's compressibility index and Hausner ratio of jet-milled amiloride HCl samples.

Sample d50 (m) Angle of repose () Apparent density (g/ml) Bulk density (g/ml) Tapped density (g/ml) Compressibility Index Hausner ratio

JM1 1 1.71 45.3 0.381 0.281 0.379 25.86 1.35

JM1 2 2.07 42.5 0.298 0.227 0.295 23.05 1.30
JM1 3 2.62 40.0 0.314 0.251 0.312 19.55 1.24
JM1 4 2.96 39.1 0.317 0.256 0.315 18.73 1.23

JM1 jet-milled.

Table 6
Summary of cascade impaction data (mean SD, n = 3).

Sample d50 (m) Emitted dose (%) Fine particle fraction (%) MMAD (m) GSD (m)

JM1 1 1.71 68.15 1.58 39.78 1.34 7.30 0.02 2.74 0.01
JM1 2 2.07 70.54 1.12 41.25 1.03 7.27 0.01 2.37 0.01
JM1 3 2.62 82.97 2.37 45.67 1.64 5.53 0.01 1.68 0.00
JM1 4 2.96 84.23 1.35 50.62 1.17 5.71 0.00 1.51 0.00
SD2 2.89 83.74 2.08 50.19 1.24 5.61 0.01 1.58 0.01

JM1 jet-milled.
SD1 spray-dried.

above and 50% of its mass below it. Signicant differences were ob- because the obtained aerodynamic diameter relates the particle to the
served using the Aerolizer (p b 0.001) in the MMAD of each jet- diameter of a sphere of unit density that has the same settling velocity
milled sample (d50 of 1.71 m, 2.07 m, 2.62 m and 2.96 m) and as the particle of interest, regardless of its shape or density.
spray-dried sample (d50 of 2.89 m). The results are shown in Fig. 5.
The MMAD increased with increases in the d50 of both the jet-milled
(d50 of 2.62 and 2.96 m) and spray-dried particles (d50 of 2.89 m). 3.3.5. Geometric standard deviation (GSD)
However, signicantly higher MMAD was obtained for the jet-milled Signicant differences (p b 0.001) were observed in the GSD for
particles (d50 of 1.71 m and 2.07 m). In general, a decrease in each jet-milled sample (d50 of 1.71 m, 2.07 m, 2.62 m and 2.96 m)
MMAD has been associated with an increase in drug dispersion. Higher and spray-dried sample (d50 of 2.89 m). For jet-milled particles,
MMADs were obtained with increasing d50 for the jet-milled (d50 =
2.62 m and 2.96 m) and spray-dried particles (d50 = 2.89 m),
which was attributed to the proportionality between the aerodynamic
and geometric diameters [32].
This was not true for the larger MMADs, which ranged between 5
and 7 m, of the jet-milled particles (d50 = 1.71 m, 2.07 m), probably
due to the characteristics of the powders that exhibited excellent forma-
tion of particle aggregates. Consequently, the laser diffraction technique
has been proved to be an important tool for initial formulation and pro-
cess screening for a DPI formulation. Moreover, the MMAD assessment
could be useful, especially in the process control and quality control of
nished products, as it allows a rapid screening of many products

Fig. 3. Emitted dose of jet-milled and spray-dried particles obtained by dispersing various
samples containing amiloride HCl (JM jet-milled, SD spray-dried, n = 3 error bars indicate
SD).

Fig. 4. Fine particle fractions of jet-milled and spray-dried particles obtained by dispersing
Fig. 2. Cv proles of jet-milled samples and spray-dried sample (JM jet-milled, SD various samples containing amiloride HCl (JM jet-milled, SD spray-dried, n = 3 error bars
spray-dried). indicate SD).
 M. Djoki et al. / Powder Technology 262 (2014) 170176 175

Fig. 5. MMADs of jet-milled and spray-dried particles obtained by dispersing various samples
containing amiloride HCl (JM jet-milled, SD spray-dried, n = 3 error bars indicate SD).

lower GSDs were obtained with larger jet-milled particles (d50 of 2.62
and 2.96 m) and spray-dried particles (d50 of 2.89 m). Lower GSDs
were observed in particles with d50 between 2.5 and 3 m, which was
indicative of less variable aerosol dispersion, and higher GSDs were ob-
served with d50 values closer to 1 m (Fig. 6). These results support a
previous study where lower GSDs were observed in particles with d50
between 2 and 5 m, and higher GSDs were observed with d50 values
closer to 1 m [32]. A larger GSD for the jet-milled samples (d50 of
1.71 m and 2.07 m) implies a longer large particle size tail in the dis-
tribution [38]. The GSD for a well-functioning stage should ideally be
less than 1.2 (the GSD for an ideal size fractionator would be 1.0,
which indicates a monodisperse aerosol) [39].

3.4. Visualisation of particles

To investigate the effect of the jet-mill and spray-drying process on

the particle surface morphology, scanning electron microscopy was per-
formed. Fig. 7 shows the micrographs of the amiloride HCl samples
taken before and after jet-milling and spray-drying. The bulk sample ap-
pears to consist of many large and irregularly shaped plates. In contrast,
the jet-milled sample shows small particles of more uniform angular
shape. The spray-dried particles were small, round, and spherical with
smooth surfaces. The size of the primary particles observed by SEM
was consistent with the size data obtained by laser diffraction.

4. Conclusion

The particle size distributions of the obtained samples were suitable

for respiratory drug delivery. The d50 values differed between the jet-
milled samples, whereas minimal differences were observed in other
physicochemical properties, except in the powder ow. The results of
the aerosol dispersion characterisation indicate that the preparation of
particles with the best characteristics for lung deposition, by jet-

Fig. 7. SEM micrographs of bulk sample (a), jet-milled sample 4 (b) and spray-dried sample
of amiloride HCl.

milling, is favoured by lower air pressure for grinding (4 bar), a diame-

Fig. 6. GSDs of jet-milled and spray-dried particles obtained by dispersing various samples
containing amiloride HCl (JM jet-milled, SD spray-dried, n = 3 error bars indicate SD).
ter of the injector nozzle for injectable air in the range of 0.8 to 0.9 mm
and a diameter of the ring nozzle for grinding air in the range 0.7 to
0.9 mm, which resulted in higher EDs and FPFs and better ow charac-
teristics. The main limitation of the aerosol dispersion of respirable-
sized particles was the aggregation of jet-milled particles with d50
values close to 1 m, which were obtained with higher air pressure for
176 M. Djoki et al. / Powder Technology 262 (2014) 170176
grinding (6 bar). The optimisation of this parameter will enable a re-
duction of the agglomeration of particles and a reduction of adhesive
properties, consequently allowing for better dispersion of the formula-
tion and attainment of higher emitted dose values. In conclusion, these
results suggest that the jet-milling method of micronisation and spray-
drying method are appropriate techniques to control particle physico-
chemical and aerosol dispersion characteristics. Greater aerosol disper-
sion with less variation was achieved using particles with d50 values in
the 2.5 to 3 m range. Small changes in the particle size within the respi-
rable range (1 to 5 m) seemed to produce a major impact on the aero-
sol dispersion of powders. The aggregate size and behaviour played an
important role in the powder ow and aerosol dispersion.
Acknowledgments
This work was supported by Project TR 34007, Ministry of Science,
Republic of Serbia.
References
[1] A.L. Adjeji, P.K. Gupta, Inhalation Delivery of Therapeutics Peptides and Proteins,
rst edition Marcel Dekker, New York, 1997.
[2] P.J. Atkins, Dry powder inhalers: an overview, Respir. Care 50 (2005) 13041312.
[3] N.M. Kassem, K.K.L. Ho, D. Ganderton, The effect of air ow and carrier size on the
characteristics of an inspirable cloud, J. Pharm. Pharmacol. 41 (1989) (14 pp.).
[4] M.A. Braun, R. Oschmann, P.C. Schmidt, Inuence of excipients and storage humidity
on the deposition of disodium cromoglycate (DSCG) in the twin impinger, Int. J.
Pharm. 135 (1996) 5362.
[5] H. Steckel, B.W. Mller, In vitro evaluation of dry powder inhalers II: inuence of
carrier particle size and concentration on in vitro deposition, Int. J. Pharm. 154
(1997) 3137.
[6] N.M. Kassem, D. Ganderton, The inuence of carrier surface on the characteristics of
inspirable powder aerosols, J. Pharm. Pharmacol. 42 (1990) (11 pp.).
[7] Y. Kawashima, T. Sarigano, T. Hino, H. Yamoto, H. Takeuchi, Effect of surface mor-
phology of carrier lactose on dry powder inhalation property of pranlukast hydrate,
Int. J. Pharm. 172 (1998) 179188.
[8] D. Ganderton, The generation of respirable clouds from coarse powder aggregates, J.
Biopharm. Sci. 3 (1992) 101105.
[9] P. Lucas, K. Anderson, J.N. Staniforth, Protein deposition from dry powder inhalers:
ne particle multiplets as performance modiers, Pharm. Res. 15 (1998) 562569.
[10] X.M. Zeng, G.P. Martin, S.K. Tee, C. Marriot, The role of ne particle lactose on the
dispersion and deaggregation of salbutamol sulphate in an air stream in vitro, Int.
J. Pharm. 176 (1998) 99110.
[11] G. Pilcer, K. Amighi, Formulation strategy and use of excipients in pulmonary drug
delivery, Int. J. Pharm. 392 (2010) 119.
[12] M.L. Everard, S.G. Devadason, P.N.L. Souf, Flow early in the inspiratory manoeuvre
affects the aerosol particle size distribution from a Turbuhaler, Respir. Med. 91
(1997) 624628.
[13] European Pharmacopoeia, Section 2.9.18 Preparations for Inhalation: Aerodynam-
ic Assessment of Fine Particles, fth edition Council of Europe, Strasbourg, 2005.
[14] J. Heyder, J. Gebhart, G. Rudolf, C.F. Schiller, W. Stahlhofen, Deposition of particles in
the human respiratory tract in the size range 0.00515 m, J. Aerosol Sci. 17 (1986)
811825.
[15] K. Masters, Spray Drying Handbook, fth edition Longman Scientic and Technical,
New York, 1991.
[16] R.J. Malcolmson, J.K. Embleton, Dry powder formulations for pulmonary delivery, J.
Pharm. Sci. Technol. Today 1 (1998) 394398.
[17] A. Canakci, F. Erdemir, T. Varol, A. Patir, Determining the effect of process parame-
ters on particle size in mechanical milling using the Taguchi method: measurement
and analysis, Measurement 46 (2013) 35323540.
[18] T. Varol, A. Canakci, Effect of particle size and ratio of B4C reinforcement on proper-
ties and morphology of nanocrystalline Al2024B4C composite powders, Powder
Technol. 246 (2013) 462472.
[19] T. Varol, A. Canakci, Synthesis and characterization of nanocrystalline Al 2024B4C
composite powders by mechanical alloying, Philos. Mag. Lett. 93 (2013) 339345.
[20] A. Canakci, T. Varol, S. Ozsahin, Analysis of the effect of a new process control agent
technique on the mechanical milling process using a neural network model: mea-
surement and modeling, Measurement 46 (2013) 18181827.
[21] H. Berthiaux, C. Chiron, J. Dodds, Modelling ne grinding in a uidized bed opposed
jet mill: part II: continuous grinding, Powder Technol. 106 (1999) 8897.
[22] N. Midoux, P. Hoek, L. Pailleres, J.R. Authelin, Micronization of pharmaceutical sub-
stances in a spiral jet mill, Powder Technol. 104 (1999) 113120.
[23] J. Kolacz, Process efciency aspects for jet mill plant operations, Miner. Eng. 17
(2004) 12931296.
[24] F. Mller, R. Polke, G. Schdel, Spiral jet mills: hold up and scale up, Int. J. Miner. Pro-
cess. 4445 (1996) 315326.
[25] P.R. Byron, Prediction of drug residence times in regions of the human respiratory
tract following aerosol inhalation, J. Pharm. Sci. 75 (1986) 433438.
[26] P.J. Noone, J.A. Regnis, X. Liu, K.L. Brouwer, M. Robinson, L. Edwards, M.R. Knowles,
Airway deposition and clearance and systemic pharmacokinetics of amiloride fol-
lowing aerosolization with an ultrasonic nebulizer to normal airways, Chest 112
(1997) 12831290.
[27] M.J. Jozwiakowski, S.O. Williams, R.D. Hathaway, Relative physical stability of the
solid forms of amiloride HCl, Int. J. Pharm. 91 (1993) 195207.
[28] M. M. Van Oort, M. J. Sacchetti. Inventors: Glaxosmithkline Corporation, assignee.
Process and device for inhalation of particulate medicaments, US Patent Application
2003000592, 2003.
[29] M.L. Everard, S.G. Devadason, V.B. Sunderland, P.N.L. Souef, An alternative aerosol
delivery system for amiloride, Thorax 50 (1995) 517519.
[30] Anaesthetic and respiratory equipment nebulizing systems and components, In-
ternational Standard ISO 27427, 2nd edition, 2010.
[31] D.J. Mazzo, Amiloride hydrochloride, in: K. Florey (Ed.), Analytical Prole of Drug
Substances, 15, Academic Press, Orlando, 1986, pp. 134.
[32] M.D. Louey, M. Van Oort, A.J. Hickey, Aerosol dispersion of respirable particles in
narrow size distributions produced by jet-milling and spray-drying techniques,
Pharm. Res. 21 (2004) 12001206.
[33] Z. Wang, M. Kwauk, H. Li, Fluidization of ne particles, Chem. Eng. Sci. 53 (1998)
377395.
[34] H. Harris, Rapid Preformulation Screening of Drug Candidates for Dry Powder Inhaler
Preparation, (dissertation) University of Bath, Bath, 2008.
[35] M.D. Louey, M. Van Oort, A.J. Hickey, Aerosol dispersion of respirable particles in
narrow size distributions using drug-alone and lactose-blend formulations, Pharm.
Res. 21 (2004) 12071213.
[36] N. Chew, H.-K. Chan, Inuence of particle size, air ow and inhaler device on the
dispersion of mannitol powders as aerosols, Pharm. Res. 16 (1999) 10981103.
[37] N.Y.K. Chew, D.F. Bagster, H.-K. Chan, Effect of particle size, air ow and inhaler de-
vice on the aerosolisation of disodium cromoglycate powders, Int. J. Pharm. 206
(2000) 7583.
[38] C.J. Musante, J.D. Schroeter, J.A. Rosati, T.M. Crowder, A.J. Hickey, T.B. Martonen, Fac-
tors affecting the deposition of inhaled porous drug particles, J. Pharm. Sci. 91
(2002) 15901600.
[39] V.A. Marple, B.A. Olson, K. Santhanakrishnan, J.P. Mitchell, S.C. Murray, B.L. Hudson-
Curtis, Next generation pharmaceutical impactor (a new impactor for pharmaceuti-
cal inhaler testing). Part II: archival calibration, J. Aerosol Med. 16 (2003) 301324.