Windox 5

Particle size
and size distributions of UT-M and B-M with 8 h milling time were
determined by laser diffraction using a HELOS-CUVETTE 6 R1 (Sympatec).
The samples were dispersed in water (1 mL) in a 6 mL cuvette, diluted
with water (5 mL), and analyzed using Windox 5.8.0.0 software (Sympatec),
applying Fraunhofer theory. The particle size distribution of B-M,
40 and 100 min milling times, were determined by laser diffraction
using HELOS/KR-QUIXEL R3 (Sympatec). The pellet was resuspended in
1 mL of 96% (v/v) EtOH, diluted with 250 mL of distilled water, and
sonicated for 120 s. The analysis was performed in triplicate at ambient
temperature. The particle sizes are presented as the distribution
density and sum of distribution. The 10×, 50×, and 90×
were taken from the sum of distribution, which represent the upper
size limit of 10, 50, and 90% of the particles.

The PSD was assessed with a HELOS laser diffractometer (Sympatec GmbH, Clausthal-Zellerfeld, Germany) with the RODOS dispersing system. Pulmonary particles were dispersed at 3 bar and nasal particles at 0.2 bar to ensure particle integrity. Data analysis was performed by the Windox 5.8.0.0 software from Sympatec (Clausthal-Zellerfeld, Germany) utilising the Fraunhofer theory. The reported data is the mean value of three-fold determination. The span value was calculated with the following Equation (1):

Laser Diffraction (HELOS/KR, Sympatec GmbH, Germany) was used to evaluate the particle size distribution of the protein powders produced by spray drying (n = 3). The optical mode R2 (for a size range of 0.45–87.5 μm) was chosen. The dry dispersion system (RODOS, Sympatec GmbH, Germany) was coupled to a vibrating drain (Vibri, Sympatec GmbH, Germany) to disperse the powder samples for analysis. A dispersing pressure of 2 bar was suitable to disperse the particles during a sampling time of 120 s. Windox5 Software (Sympatec GmbH, Germany) was used to analyse the volumetric particle size distributions. The S/V ratio was calculated by dividing SMD (Sauter Mean Diameter) by the VMD (Volume Mean Diameter). The diameters Dv₁₀, Dv₅₀ and Dv₉₀ were used to calculate the span of the sample’s particle size distribution. Dv₁₀, for example, describes the 10% of particles that are of that specific size in the volume distribution, Dv₅₀ and Dv₉₀ describe 50% and 90%, respectively. The distribution width or span was calculated according to Eq. (6): $$span=\frac{{\mathit{Dv}}_{90}-{\mathit{Dv}}_{10}}{{\mathit{Dv}}_{50}}$$

The particle size distribution was measured by laser diffraction using a SymPatec HELOS equipped with a RODOS/ ASPIROS dry dispenser (Germany). All measurements were performed using the R5 (4.5 to 875 μm) and R3 (0.5 to 175 μm) lens for L₁₂ and L₁₈ designs respectively. The particles were dispersed under a 4-bar pressure. About 5 mg of each sample was placed in an ASPIROS glass vial. The particle size distribution (volume mean diameter, X10, X50, X90) were analyzed with WINDOX 5 software (SymPatec, Germany).

Laser diffraction was applied to determine the particle size distribution (PSD) of the initial and processed material. For this, a Helium Neon Laser Optical System (HELOS^®, Sympatec GmbH, Clausthal-Zellerfeld, Germany) was equipped with a RODOS^® dispersion module and a R4 lens with a measuring range of 0.5 to 350 µm. With compressed air (2 bar), the automatically fed (VIBRI^® dry dosing system, Sympatec GmbH) material was dispersed in an aerosol jet towards the measuring zone.
Evaluation of the size distribution was performed with the Windox 5 software (Sympatec GmbH) based on the Fraunhofer enhanced equation.

The particle size distribution of the raw materials and respective blends was evaluated using laser diffraction (HELOS/KR, Sympatec GmbH, Clausthal-Zellerfeld, Germany). The powders were placed on a vibrating chute (Vibri, Sympatec GmbH, Clausthal-Zellerfeld, Germany) and dispersed using a dry dispersing system (RODOS, Sympatec GmbH, Clausthal-Zellerfeld, Germany). The powders were sampled during 10 s and the measurements with an R2 (0.45–87.5 μm) and R5 lens (4.5–875 μm) triggered once an optical concentration (C_opt) of 0.5% was reached. To evaluate the pressure at which the particles de-agglomerate, pressure titration was applied. For this, the primary dispersion pressure (PDP) was manually adjusted in 0.2 bar steps in the range of 0.1–2.0 bar. Measurements were done in triplicate (n = 3) at 0.1, 1.5 and 2.0 bar. Before each measurement, the dispersing system was cleaned using sand and a reference measurement was taken. The resulting volumetric particle size distributions were calculated and analyzed using Windox 5 software (Sympatec GmbH, Clausthal-Zellerfeld, Germany).

The volumetric size distribution of the spray-freeze-dried powders was evaluated by laser diffraction. A laser diffractometer configured with a dosing unit for inhalers and nebulizers (HELOS/KR+INHALER, Sympatec GmbH, Clausthal-Zellerfeld, Germany) was used to determine the size distribution of the particles dispersed from a Breezhaler^® (Novartis AG, Basel, Switzerland) as previously described [20 (link)]. In brief, the Breezhaler^® was mounted onto the central unit of the INHALER module by inserting the mouthpiece into the adaptor horizontally. A suction airflow was provided by a vacuum pump at 60 L/min. Prior to measurement, a size 3 gelatin capsule (Capsugel^®, Morristown, NJ, USA) was loaded with 1.0 ± 0.1 mg powder and placed in a Breezhaler^®. The particle size distribution was calculated by the WINDOX 5 software (version 5.8.0.0, Sympatec GmbH, Clausthal-Zellerfeld, Germany) based on the enhanced Fraunhofer theory. Particle size data were expressed as D₁₀, D₅₀, and D₉₀, which represent the equivalent spherical volume diameters at 10%, 50%, and 90% cumulative volumes, respectively. Span was calculated as (D₉₀–D₁₀)/D₅₀. The experiment was performed in triplicate.