Investigation of the aerodynamic characteristics of inhaler aerosols with an inhalation simulation machine

An experimental facility comprising of an inhalation simulation machine and a laser-Doppler anemometer has been developed to investigate the aerodynamic characteristics of inhaler aerosols. The machine has been used to simulate transient inhalation flows with different degrees of inspiratory effort. Measurements of the mean flow and of the turbulence levels and time scales have been obtained for three commercially-available dry powder inhalers at different inhalation pressures. The results provide an extensive characterisation of aerosol aerodynamics and can aid the assessment of inhalers under various inhalation conditions.     

Influence of flow rate on aerosol particle size distributions from pressurized and breath-actuated inhalers.

Particle size distribution of delivered aerosols and the total mass of drug delivered from the inhaler are important determinants of pulmonary deposition and response to inhalation therapy. Inhalation flow rate may vary between patients and from dose to dose. The Andersen Sampler (AS) cascade impactor operated at flow rates of 30 and 55 L/min and the Marple-Miller Impactor (MMI) operated at flow rates of 30, 55, and 80 L/min were used in this study to investigate the influence of airflow rate on the particle size distributions of inhalation products. Total mass of drug delivered from the inhaler, fine particle mass, fine particle fraction, percentage of nonrespirable particles, and amount of formulation retained within the inhaler were determined by ultraviolet spectrophotometry for several commercial bronchodilator products purchased in the marketplace, including a pressurized metered-dose inhaler (pMDI), breath-actuated pressurized inhaler (BAMDI), and three dry powder inhalers (DPIs), two containing salbutamol sulphate and the other containing terbutaline sulphate. Varying the flow rate through the cascade impactor produced no significant change in performance of the pressurized inhalers. Increasing the flow rate produced a greater mass of drug delivered and an increase in respirable particle mass and fraction from all DPIs tested.     

Next generation pharmaceutical impactor (a new impactor for pharmaceutical inhaler testing). Part I: Design.

A new cascade impactor has been designed specifically for pharmaceutical inhaler testing. This impactor, called the Next Generation Pharmaceutical Impactor (NGI), has seven stages and is intended to operate at any inlet flow rate between 30 and 100 L/min. It spans a cut size (D50) range from 0.54-microm to 11.7-microm aerodynamic diameter at 30 L/min and 0.24 microm to 6.12 microm at 100 L/min. The aerodynamics of the impactor follow established scientific principles, giving confident particle size fractionation behavior over the design flow range. The NGI has several features to enhance its utility for inhaler testing. One such feature is that particles are deposited on collection cups that are held in a tray. This tray is removed from the impactor as a single unit, facilitating quick sample turn-around times if multiple trays are used. For accomplishing drug recovery, the user can add up to approximately 40 mL of an appropriate solvent directly to the cups. Another unique feature is a micro-orifice collector (MOC) that captures in a collection cup extremely small particles normally collected on the final filter in other impactors. The particles captured in the MOC cup can be analyzed in the same manner as the particles collected in the other impactor stage cups. The user-friendly features and the aerodynamic design principles together provide an impactor well suited to the needs of the inhaler testing community.     

The effect of budesonide particle mass on drug particle detachment from carrier crystals in adhesive mixtures during inhalation.

The different fine particle fractions (FPFs) that are obtained, when different dry powder inhalers (DPIs) are used for the same powder formulation at the same flow rate, is the result of different powder de-agglomeration efficiencies for these DPIs. For adhesive mixtures, this is the efficiency with which the kinetic energy of the air flow through the DPI is converted into separation forces that detach drug particles from carrier crystals. We investigated the effect of drug particle diameter (mass) on drug-carrier separation during inhalation with three different inhalers (Sofotec Novolizer, Inhalator Ingelheim and a special test inhaler), at two different flow rates (30 and 60l/min). Two different size fractions were used as carrier material (45-63 and 100-150 microm). We measured decreasing amounts of residual drug on the carrier crystals after inhalation with increasing drug particle mass for all inhalers at both flow rates. The observed trends were the same for both carrier fractions. The decrease in residual drug on carrier is in agreement with increasing FPFs in an Erweka impactor. However, it has been calculated that the magnitude of the effect decreases with increasing de-agglomeration efficiency.     

Mass output and particle size distribution of glucocorticosteroids emitted from different inhalation devices depending on various inspiratory parameters.

Efficient inhalation therapy depends on successful delivery of the drug to the lung. The efficacy of drug delivery is not only influenced by the characteristics of the inhalation device, but also by the patient's handling of the device and by the inspiratory maneuver achieved through the device. We analyzed the output characteristics of three different chlorofluorocarbon (CFC)-free breath-actuated inhalers for inhaled glucocorticosteroids (BUD Turbohaler, FP Diskus/Accuhaler and HFA-BDP Autohaler, respectively). Mass output and particle size distribution of drug aerosol delivered by the inhalers were determined depending on different inhalation parameters in vitro using an Andersen cascade impactor. We found that, beside the peak inspiratory flow (PIF), other factors such as flow acceleration and inhalation volume also have significant effects on aerosol generation with respect to mass output and particle size distribution. Thus, these parameters should be taken into account when a suitable device for an individual patient is to be selected. The dependency on inspiratory parameters was most pronounced for the dry powder inhalers. The Turbohaler showed by far the highest variance in particle output (fine particle fraction ranging from 3.4% to 22.1% of label claim), whereas the Diskus was less dependent on variations in inhalation (10.6% to 18.5% of label claim). The most constant aerosol output was found for the Autohaler, which also released the highest fine particle fraction (43.1% to 56.6% of label claim).     

Design and application of a new modular adapter for laser diffraction characterization of inhalation aerosols

An inhaler adapter has been designed for the characterization of the aerosol clouds from medical aerosol generators such as nebulizers, dry powder inhalers (dpis) and metered dose inhalers (mdis) with laser diffraction technology. The adapter has a pre-separator, for separation of large particles (i.e. carrier crystals) from the aerosol cloud before it is exposed to the laser beam. It also has a fine particle collector for measuring the emitted mass fraction of fines by chemical detection methods after laser diffraction sizing. The closed system enables flow control through the aerosol generators and all test conditions, including ambient temperature and relative humidity, are automatically recorded. Counter flows minimize particle deposition onto the two windows for the laser beam, which make successive measurements without cleaning of these windows possible. The adapter has successfully been tested for nebulizers, mdis and dpis. In a comparative study with ten nebulizers it was found that these devices differ considerably in droplet size (distribution) of the aerosol cloud for the same 10% aqueous tobramycin solution (volume median diameters ranging from 1.25 to 3.25 microm) when they are used under the conditions recommended by the manufacturers. The droplet size distribution generated by the Sidestream (with PortaNeb compressor) is very constant during nebulization until dry running of the device. Comparative testing of dpis containing spherical pellet type of formulations for the drug (e.g. the AstraZeneca Turbuhaler) with the adapter is fast and simple. But also formulations containing larger carrier material could successfully be measured. Disintegration efficiency of a test inhaler with carrier retainment (acting as a pre-separator) could be measured quite accurately both for a colistin sulfate formulation with 16.7% of a lactose fraction 106-150 microm and for a budesonide formulation with a carrier mixture of Pharmatose 325 and 150 M. Therefore, it is concluded that, with this special adapter, laser diffraction may be a valuable tool for comparative inhaler evaluation, device development, powder formulation and quality control. Compared to cascade impactor analysis, laser diffraction is much faster. In addition to that, more detailed and also different information about the aerosol cloud is obtained.     

Investigation of triboelectric charging in dry powder inhalers using electrical low pressure impactor (ELPI)

Electrostatics and triboelectrification phenomena in dry powder inhalers (DPI) are not well understood, but as shown in this study they may play an important role. Using model formulations of albuterol in lactose, the extent of triboelectrification in the operation of DPI was investigated using an electrical low pressure impactor (ELPI). An experimental apparatus was developed, the performance of the ELPI was evaluated for consistency and reproducibility, and compared to a conventional inertial impactor. Using a statistical experimental design the effects of lactose type, drug load, capsule fill, capsule material, and inhaler were assessed. DPI formulations appear to be subject to strong triboelectric effects. Charge separation can occur between different size fractions, i.e. different fractions can carry charges of different sign. In particular, lactose type, inhaler, and capsule material have a strong effect on the magnitude and polarity of the charge developed during DPI operation. The study suggests that the polarity of the aerosol can be controlled by choice of lactose type, capsule material, and inhaler, which could be exploited for targeting different lung physiologies.     

Relationship of stage mensuration data to the performance of new and used cascade impactors.

Cascade impaction is a standard test method for characterizing the quality of inhalable drug products. The sizes of the nozzles on each stage of the impactor are the critical dimensions for the performance of the impactor. Compendial reference methods call for periodic measurement of the size of the nozzles on each stage, a procedure known as stage mensuration. There is however currently no guidance on acceptable mensuration criteria. We aim to remedy this situation by providing a sound basis for understanding and using mensuration data, be it for acceptance criteria for new impactors or for the setting of mensuration tolerances for in-use impactors. We first show that multi-nozzle impactor stages behave as if all of the nozzles are equal in size to an effective diameter, , that is composed of the area-mean and areamedian diameters, W* and , calculated directly from the individual nozzle diameters for all nozzles on a given stage (equation 1): W= (W*)(2/3) x (W)(1/3) (1). Hence, the effective diameter provides an intuitive and technically sound basis for setting acceptance criteria for new and in-use impactors. We tabulate these criteria for the Mark II eight-stage Andersen cascade impactor and the Next Generation Pharmaceutical Impactor in a manner similar to the tables of critical impactor dimensions published in EP Supplement 5.1 and in USP 28. For two different impactors or for one impactor measured at two different times (e.g., at manufacture and in use), we find that the D50 values of a given stage are related to the effective diameters by D(50,2)/D(50,1)= (W(2)/W(1))(3/2) (2). Using the stage mensuration data for new, as-manufactured NGIs, we compare the D(50 )values of the first 125 as-manufactured NGIs with those of the archivally calibrated NGI. We further establish that the archivally calibrated NGI has D(50) values within 0.3% of an entirely perfect, hypothetical NGI with all nozzles equal to the nominal nozzle diameters. We also apply the equations to a specific mensurated impactor to show that a used impactor with some nozzles outside of the original manufacturing specifications can have the same aerodynamic performance as a new impactor.     

Cascade impactors for the size characterization of aerosols from medical inhalers: their uses and limitations.

Cascade impactors, including the multi-stage liquid impinger, are by far the most widely encountered means for the in vitro determination of the particle size distribution of aerosols from medical inhalers, both in product development, batch release and in applications with add-on devices. This is because they directly measure aerodynamic size, which is the most relevant parameter to describe particle transport within the respiratory tract. At the same time, it is possible to quantify the mass of active pharmaceutical ingredient in different size ranges independent of other non-physiologically active components of the formulation. We begin by providing an overview of the operating principles of impactors and then highlight the various configurations and adaptations that have been adopted to characterize the various classes of inhaler. We continue by examining the limitations of the cascade impaction method, in particular looking at potential sources of measurement bias and discussing both appropriate and inappropriate uses of impactor-generated data. We also present a synopsis of current developments, including the Next Generation Pharmaceutical Impactor, and automation of cascade impactors for routine inhaler performance measurements.     

Effect of inhalation flow rate on the dosing characteristics of dry powder inhaler (DPI) and metered dose inhaler (MDI) products.

Both the dose delivered from the device and the particle size of the medication are important parameters for inhalation products because they influence the amount of drug that is delivered to the patient's lung. The inspiratory flow rate may vary from dose to dose in a given patient and between patients. The Marple-Miller Cascade Impactor, a new multistage inertial impactor that operates at two flow rates (30 and 60 liters/min) with comparable particle size cut-offs, provides a means to study the effect of inhalation flow rate on the particle size distributions of inhalation products. The medication delivery, mass median aerodynamic diameter (MMAD), and fine particle mass were determined, in a randomized fashion, for albuterol, beclomethasone, budesonide, and terbutaline in both metered dose inhaler (MDI) and dry powder inhaler (DPI) products as a function of flow rate. In all cases, independent of drug or device used, the MDI products had a more reproducible respirable dose than the breath-actuated DPI products tested as a function of inhalation flow rate.     

Optimized Inhalation Aerosols. II. Inertial Testing Methods for Particle Size Analysis of Pressurized Inhalers

Pressurized metered dose inhaler (MDI) output from three different albuterol formulations was characterized using three inertial separation devices. Results were compared for the Delron six-stage cascade impactor (DCI6), the Andersen Mark II eight-stage impactor (ACI8), and Copley's twin-stage liquid impinger (LI). None of the devices tested in this study was ideal in all respects. All devices could differentiate between formulations in terms of respirable doses (albuterol amount with aerodynamic diameters <5.5 through 6.4 µm). Only the high-flow rate LI could differentiate among all three formulations when data were presented in terms of respirable percentage (RP) of drug collected. Values for RP were in excellent agreement for the independently calibrated impactors when the same evaporation chamber was used atop the impactors. The LI appeared to overestimate values for RP in vivo. Results are discussed in light of the debate surrounding the revision of USP aerosol testing requirements. Rigorous specifications for evaporation chambers and methodologies are necessary for meaningful inter- and intra-laboratory comparison of results when any of these devices are used.     

Dry powder inhaler device influence on carrier particle performance

Dry powder inhalers (DPIs) are distinguished from one another by their unique device geometries, reflecting their distinct drug detachment mechanisms, which can be broadly classified into either aerodynamic or mechanical-based detachment forces. Accordingly, powder particles experience different aerodynamic and mechanical forces depending on the inhaler. However, the influence of carrier particle physical properties on the performance of DPIs with different dispersion mechanisms remains largely unexplored. Carrier particle trajectories through two commercial DPIs were modeled with computational fluid dynamics (CFD) and the results were compared with in vitro aerosol studies to assess the role of carrier particle size and shape on inhaler performance. Two percent (w/w) binary blends of budesonide with anhydrous and granulated lactose carriers ranging up to 300 μm were dispersed from both an Aerolizer? and Handihaler? through a cascade impactor at 60 L min(-1). For the simulations, carrier particles were modeled as spherical monodisperse populations with small (32 μm), medium (108 μm), and large (275 μm) particle diameters. CFD simulations revealed the average number of carrier particle-inhaler collisions increased with carrier particle size (2.3-4.0) in the Aerolizer?, reflecting the improved performance observed in vitro. Collisions within the Handihaler?, in contrast, were less frequent and generally independent of carrier particle size. The results demonstrate that the aerodynamic behavior of carrier particles varies markedly with both their physical properties and the inhalation device, significantly influencing the performance of a dry powder inhaler formulation.     

Cascade impactor data and the lognormal distribution: nonlinear regression for a better fit.

At present, cascade impactors are the instruments of choice for measuring the particle size distribution of aerosol present in the complex discharge from pharmaceutical inhalers. The distribution of drug captured in the cascade impactor may be most usefully represented by the lognormal distribution. Only two parameters must be extracted from the analysis of cascade impactor data in order to describe the distribution. These two parameters are the mass median aerodynamic diameter (MMAD) and the geometric standard deviation (GSD). A cumulative version of the lognormal curve or more frequently, a linearized version of the cumulative curve called a "log probability plot," is used as a surrogate for the lognormal curve. The probability plot has great appeal since a lognormal distribution yields a straight line on log probability paper. One may easily determine the apparent MMAD and GSD from this linear plot. However, when one plots a lognormal curve, using the MMAD and GSD derived from a log probability plot, over a histogram constructed from cascade impactor data, an obvious mismatch is frequently seen. In order to derive parameters that more truly reflect the impactor data, a computer program, which uses nonlinear regression to derive an MMAD and GSD for the lognormal curve, has been written. It is presented here.     

Aerodynamic particle size of metered-dose inhalers determined by the quartz crystal microbalance and the Andersen cascade impactor.

Given the rapid sizing capability and high sensitivity, the quartz crystal microbalance (QCM) cascade impactor has been evaluated for the size determination of metered-dose inhaler (MDI) aerosols. The effects of surfactants present in MDI formulations, crystal coating, particle bounce and crystal overloading on the QCM cascade impactor are investigated. To reduce particle bounce, it is necessary to coat the crystals and use new coated surfaces for each measurement. Mass median aerodynamic diameters (MMADs) obtained from the QCM cascade impactor are compared to those from the commonly used Andersen cascade impactor. For MDI formulations containing little or no surfactants, MMADs obtained from the QCM and Andersen cascade impactors are comparable. For MDI formulations containing a significant amount of surfactant (or any non-volatile excipients), the QCM cascade impactor measures the combined size distribution of the drug and non-volatile excipients. A technique is devised in this study to deduce the drug-only size distribution from the QCM impactor for surfactant-containing MDI formulations and show comparable results to the Andersen cascade impactor except for high drug load Intal. The QCM impactor has proved to be a useful tool for rapid size measurement of MDI formulations.     

Inertial sizing of aerosol inhaled from two dry powder inhalers with realistic breath patterns versus constant flow rates

A procedure is developed that allows particles inhaled with realistic breath patterns to be sized by cascade impaction at a constant flow rate. This procedure is then used to examine the difference between particle sizes obtained with constant flow rate (step profile) versus actual-subject breath patterns for two dry powder inhalers DPIs; the Ventodisk® and Spiros® inhalers (delivering salbutamol sulphate). Aerosol inhaled from the DPIs by a breath simulator was combined with make-up air to provide 300 l/min. to a pair of virtual impactors. These impactors separate out particles in the nominal diameter range of 1–10 μm for sizing at 30 l/min. by a MOUDI cascade impactor, with filter collection of particles outside this range. Breathing patterns of ten subjects ranging in age from 6 to 17 years of age were measured and recorded using whole-body plethysmography while these volunteers inhaled through Ventodisk® and Spiros® inhalers. Particle sizes with four of these breath patterns, as well as several constant flow rate step profiles, were then obtained using the sizing apparatus with a realistic mouth–throat intake. Our results show that as long as the constant flow rates were near typical values occurring in the actual-subject breaths, particle sizes obtained with constant flow rates were not significantly different (P>0.01) from those occurring with actual-subject breath patterns. Significant differences are present if constant flow rates unrepresentative of those expected during particle uptake with the actual-subject patterns are used with the Ventodisk®. These results show that judiciously chosen constant flow rates give rise to inertial particle size measurements that are equivalent to those obtained during actual-subject inhalation for the two types of DPIs tested.     

In vitro/in vivo comparisons in pulmonary drug delivery

Establishing clear relationships between in vitro and in vivo data for inhaled drug products is an important goal. In vitro aerodynamic particle size distributions (APSDs) are expected to have some predictive power not only for drug deposition, but also for clinical effects. APSD data obtained by cascade impaction have been compared with lung deposition data measured in gamma scintigraphy studies. Whole-lung deposition correlated significantly with fine particle fraction (FPF) across a range of inhaler devices. FPF, defined in terms of aerosol <5.8 microm or <6.8 microm diameter, systematically overestimated lung deposition for virtually all inhalers. Lung deposition showed closer numerical equivalence to the percentage of the aerosol dose smaller than 3 microm diameter. Correlations exist between APSD data and whole-lung deposition, which may allow the greater use of APSD data for comparing inhaler devices. Agreement between in vitro and in vivo data may be improved by measuring APSD in ways that more closely mimic clinical use, including the use of impactor inlets that simulate the human upper airway anatomy. At the present time there are few published data that relate APSD to the clinical response of inhaled drugs in an unambiguous way.     

Novolizer: how does it fit into inhalation therapy?

Inhalation therapy is the preferred route of administration of anti-asthmatic drugs to the lungs. However, the vast majority of patients cannot use their inhalers correctly, particularly pressurised metered dose inhalers (pMDIs). The actual proportion of patients who do not use their inhalers correctly may even be under-estimated as GPs tend to over-estimate correct inhalation technique. Dry powder inhalers (DPIs) have many advantages over pMDIs. Unlike pMDIs, they are environmentally-friendly, contain no propellant gases and, more importantly, they are breath-activated, so that the patient does not need to coordinate actuation of the inhaler with inspiration. Three key parameters for correct inhaler use should be considered when evaluating existing or future DPI devices and especially when choosing the appropriate device for the patient: (1) usability, (2) particle size distribution of the emitted drug and (3) intrinsic airflow resistance of the device. The Novolizer is a breath-activated, multidose, refillable DPI. It is easy to use correctly, has multiple feedback and control mechanisms which guide the patient through the correct inhalation manoeuvre. In addition, the Novolizer has an intelligent dose counter, which resets only after a correct inhalation and may help to monitor patient compliance. The Novolizer has a comparable or better lung deposition than the Turbuhaler at similar or higher peak inspiratory flow (PIF) rates. A flow trigger valve system ensures a clinically effective fine particle fraction (FPF) and sufficient drug delivery, which is important for a good lung deposition. The FPF produced through the Novolizer is also relatively independent of flow rate and the device shows better reproducibility of metering and delivery performance compared to the Turbuhaler. The low-to-medium airflow resistance means that the Novolizer is easy for patients to use correctly. Even children, patients with severe asthma and patients with moderate-to-severe chronic obstructive pulmonary disease (COPD) have no problems to generate the trigger inspiratory flow rate required to activate the Novolizer. The Novolizer uses an advanced DPI technology and may improve patient compliance.     

Multijet and multistage aerosol concentrator: design and performance analysis.

A multijet and multistage aerosol concentrator was designed and fabricated with two virtual impactors in a series. Collection efficiency, internal loss, and concentration factors were calculated at ambient conditions for each stage. The total inlet flow rate of the aerosol concentrator was set at 1000 L/min(-1), while the minor flow rate for the first stage was at 6.0% of the total inlet flow and the minor flow rate of the second stage was at 6.7% of the first stage minor flow. The aerosol concentrator was calibrated using polystyrene latex particles in aerodynamic sizes ranging from 0.5 to 10 microm. Several configurations of the multijet acceleration nozzles and multitube receptors were designed in this study. The effects of the different designs were subsequently evaluated through experimentation. It was found that a properly designed multijet and multistage aerosol concentrator can significantly improve aerosol concentration performance. Results showed that the concentration factor increases from 1 to 240 over the particle size range studied. Applications of the multijet and multistage aerosol concentrator with high-volume flow rate can vary widely, from detection of biological aerosols at low concentration, laboratory aerosol sampling, clean room monitoring, and ambient aerosol measurements.     

Time-of-flight aerodynamic particle size analyzers: their use and limitations for the evaluation of medical aerosols.

Time-of-flight (TOF) aerosol analyzers are a class of instruments that measure the aerodynamic diameter of individual particles following a controlled acceleration in a well-defined flow field. Two instruments have been used to analyze the size of medical aerosols: Aerosizer particle size analyzer (TSI Particle Instruments/Amherst, Amherst, MA), Aerodynamic Particle Sizer (APS) aerosol spectrometer (TSI) Both instruments are capable of sizing several thousand particles a second, making it possible to obtain aerodynamic particle size distributions in a few seconds compared with up to 1 hour per measurement using compendial methods that are based on either the multistage liquid impinger or cascade impactor. This rapidity makes TOF analysis attractive for product development, as many different variables can potentially be investigated during a short period of time. The data thus obtained should be used with caution, however. Several issues, most notably the lack of a direct relationship with the mass of drug substance present and the vulnerability of the measurements to coincidence effects when sampling concentrated aerosols, may severely limit the value of data from many aerosol delivery systems, especially pressurized metered dose inhalers (pMDIs). A review of the literature illustrating the issues that are involved and providing guidance on the most appropriate uses of these analyzers is presented.     

<Emphasis Type="BoldItalic">In Vitro</Emphasis> Performance Testing of the Novel Medspray® Wet Aerosol Inhaler Based on the Principle of Rayleigh Break-up

Purpose

A new inhaler (Medspray®) for pulmonary drug delivery based on the principle of Rayleigh break-up has been tested with three different spray nozzles (1.5; 2.0 and 2.5 μm) using aqueous 0.1% (w/w) salbutamol and 0.9% (w/w) sodium chloride solutions.

Materials and methods

Particle size distributions in the aerosol were measured with the principles of time of flight (APS) and laser diffraction (LDA).

Results

The Medspray® inhaler exhibits a highly constant droplet size distribution in the aerosol during dose emission. Droplets on the basis of Rayleigh break-up theory are monodisperse, but due to some coalescence the aerosols from the Medspray® inhaler are slightly polydisperse. Mass median aerodynamic diameters at 60 l.min^?1 from APS are 1.42; 1.32 and 1.27 times the theoretical droplet diameters (TD’s) and median laser diffraction diameters are 1.29; 1.14 and 1.05 times TD for 1.5; 2.0 and 2.5 μm nozzles (TD: 2.84; 3.78 and 4.73 μm respectively).

Conclusions

The narrow particle size distribution in the aerosol from the Medspray® is highly reproducible for the range of flow rates from 30 to 60 l.min^?1. The mass median aerodynamic droplet diameter can be well controlled within the size range from 4 to 6 μm at 60 l.min^?1.