EUROPEAN PHARMACOPOEIA 10.0 2.9.31.

Particle size analysis by laser light diffraction

A. Surface plate B. Die C. Neoprene gasket D. Punch E. Holder and shaft assembly F. Die underside

Figure 2.9.29.-1. – Typical apparatus used to obtain the compact for the determination of the intrinsic dissolution
Dimensions in millimetres
04/2019:20931 include laser light scattering in a wider angular range and
application of the Mie theory, in addition to the Fraunhofer
approximation and anomalous diffraction.
The technique cannot distinguish between scattering by single
particles and scattering by clusters of primary particles, i.e.
by agglomerates or aggregates. As most particulate samples
2.9.31. PARTICLE SIZE ANALYSIS BY contain agglomerates or aggregates and as the focus of interest
LASER LIGHT DIFFRACTION(12) is generally on the size distribution of primary particles, the
clusters are usually dispersed into primary particles before
The method is based on the ISO standards 13320-1(1999) and measurement.
9276-1(1998). For non-spherical particles, an equivalent sphere-size
distribution is obtained because the technique assumes
INTRODUCTION spherical particles in its optical model. The resulting
particle-size distribution may differ from those obtained
The laser light diffraction technique used for the determination by methods based on other physical principles (e.g.
of particle-size distribution is based on the analysis of the sedimentation, sieving).
diffraction pattern produced when particles are exposed
This chapter provides guidance for the measurement
to a beam of monochromatic light. Historically, the early
of size distributions of particles in different dispersed
laser diffraction instruments only used scattering at small
systems, for example, powders, sprays, aerosols, suspensions,
angles. However, the technique has since been broadened to
(12) This chapter has undergone pharmacopoeial harmonisation. See chapter 5.8. Pharmacopoeial harmonisation.

General Notices (1) apply to all monographs and other texts 373
2.9.31. Particle size analysis by laser light diffraction EUROPEAN PHARMACOPOEIA 10.0

emulsions, and gas bubbles in liquids, through analysis of relative positions. Note that only a limited angular range of
their angular light-scattering patterns. It does not address scattered light is collected by the lens(es) and, therefore, by
specific requirements of particle size measurement of specific the detector.
products.
DEVELOPMENT OF THE METHOD
PRINCIPLE The measurement of particle size by laser diffraction can give
A representative sample, dispersed at an adequate reproducible data, even in the sub-micron region, provided the
concentration in a suitable liquid or gas, is passed through instrument used and the sample tested are carefully controlled
a beam of monochromatic light, usually a laser. The light to limit variability of the test conditions (e.g. dispersion
scattered by the particles at various angles is measured by a medium, method of preparation of the sample dispersion).
multi-element detector. Numerical values representing the Traditionally, the measurement of particle size using laser
scattering pattern are then recorded for subsequent analysis. diffraction has been limited to particles in the range of
These scattering pattern values are then transformed, using an approximately 0.1 μm to 3 mm. Because of recent advances
appropriate optical model and mathematical procedure, to in lens and equipment design, newer instruments are capable
yield the proportion of total volume to a discrete number of of exceeding this range routinely. With the validation report
size classes, forming a volumetric particle-size distribution. the user demonstrates the applicability of the method for its
intended use.
INSTRUMENT Sampling. The sampling technique must be adequate to
The instrument is located in an environment where it is obtain a representative sample of a suitable volume for the
not affected by electrical noise, mechanical vibrations, particle-size measurement. Sample splitting techniques such
temperature fluctuations, humidity or direct bright light. as rotating riffler or the cone and quartering method may be
applied.
An example of a set-up of a laser light diffraction instrument
is given in Figure 2.9.31.-1. Other equipment may be used. Evaluation of the dispersion procedure. Inspect the
sample to be analysed, visually or with the aid of a
The instrument comprises a laser light source, beam microscope, to estimate its size range and particle shape. The
processing optics, a sample measurement region (or cell), a dispersion procedure must be adjusted to the purpose of the
Fourier lens, and a multi-element detector for measuring the measurement. The purpose may be such that it is preferable
scattered light pattern. A data system is also required for to deagglomerate clusters into primary particles as far as
deconvolution of the scattering data into a volumetric size possible, or it may be desirable to retain clusters as intact as
distribution and associated data analysis and reporting. possible. In this sense, the particles of interest may be either
The particles can enter the laser beam in 2 positions. In primary particles or clusters.
the conventional case the particles enter the parallel beam For the development of a method it is highly advisable to
before the collecting lens and within its working distance. In check that comminution of the particles does not occur,
so-called reversed Fourier optics the particles enter behind the and conversely, that dispersion of particles or clusters
collecting lens and thus, in a converging beam. The advantage is satisfactory. This can usually be done by changing
of the conventional set-up is that a reasonable path length for the dispersing energy and monitoring the change of the
the sample is allowed within the working distance of the lens. particle-size distribution. The measured size distribution must
The second set-up allows only small path lengths but enables not change significantly when the sample is well dispersed and
measurement of scattered light at larger angles, which is useful the particles are neither fragile nor soluble. Moreover, if the
when submicron particles are present. manufacturing process (e.g. crystallisation, milling) of the
The interaction of the incident light beam and the ensemble of material has changed, the applicability of the method must be
dispersed particles results in a scattering pattern with different verified (e.g. by microscopic comparison).
light intensities at various angles. The total angular intensity Sprays, aerosols and gas bubbles in a liquid should be
distribution, consisting of both direct and scattered light, measured directly, provided that their concentration is
is then focused onto a multi-element detector by a lens or a adequate, because sampling or dilution generally alters the
series of lenses. These lenses create a scattering pattern that, particle-size distribution.
within limits, does not depend on the location of the particles In other cases (such as emulsions, pastes and powders),
in the light beam. Hence, the continuous angular intensity representative samples may be dispersed in suitable liquids.
distribution is converted into a discrete spatial intensity Dispersing aids (wetting agents, stabilisers) and/or mechanical
distribution on a set of detector elements. forces (e.g. agitation, sonication) are often applied for
It is assumed that the measured scattering pattern of the deagglomeration or deaggregation of clusters and stabilisation
particle ensemble is identical to the sum of the patterns from of the dispersion. For these liquid dispersions, a recirculating
all individual single scattering particles presented in random system is most commonly used, consisting of an optical

1. Obscuration detector 5. Scattered light not collected by lens (4) 9. Working distance of lens (4)
2. Scattered beam 6. Particle ensemble 10. Multi-element detector
3. Direct beam 7. Light source laser 11. Focal distance of lens (4)
4. Fourier lens 8. Beam processing unit

Figure 2.9.31.-1. – Example of a set-up of a laser light diffraction instrument

374 See the information section on general monographs (cover pages)

EUROPEAN PHARMACOPOEIA 10.0 2.9.31. Particle size analysis by laser light diffraction

measuring cell, a dispersion bath usually equipped with represented by differently scaled and differently named
stirrer and ultrasonic elements, a pump, and tubing. numbers, e.g. obscuration, optical concentration, proportional
Non-recirculating, stirred cells are useful when only small number of total mass).
amounts of a sample are available or when special dispersion Determination of the measuring time. The time of
liquids are used. measurement, the reading time of the detector and the
Dry powders can also be converted into aerosols through the acquisition frequency are determined experimentally in
use of suitable dry powder dispersers, which apply mechanical accordance with the required precision. Generally, the time
force for deagglomeration or deaggregation. Generally, the for measurement permits a large number of detector scans or
dispersers use the energy of compressed gas or the differential sweeps at short time intervals.
pressure of a vacuum to disperse the particles to an aerosol,
Selection of an appropriate optical model. Most instruments
which is blown through the measuring zone, usually into the
use either the Fraunhofer or the Mie theory, though other
inlet of a vacuum unit that collects the particles. However, for
approximation theories are sometimes applied for calculation
free flowing, coarser particles or granules the effect of gravity
of the scattering matrix. The choice of the theoretical model
may be sufficient to disperse the particles adequately.
depends on the intended application and the different
If the maximum particle size of the sample exceeds the assumptions (size, absorbance, refractive index, roughness,
measuring range of the instrument, the material that is too crystal orientation, mixture, etc.) made for the test material.
coarse can be removed by sieving and the mass and percentage If the refractive index values (real and imaginary parts
of removed material are reported. However, after pre-sieving, for the used wavelength) are not exactly known, then the
note that the sample is no longer representative, unless Fraunhofer approximation or the Mie theory with a realistic
otherwise proven. estimate of the refractive index can be used. The former
Optimisation of the liquid dispersion. Liquids, surfactants, has the advantages that it is simple and it does not need
and dispersing aids used to disperse powders must : refractive index values ; the latter usually provides less-biased
particle-size distributions for small particles. For instance,
– be transparent at the laser wavelength and practically free if the Fraunhofer model is used for samples containing
from air bubbles or particles ; an appreciable amount of small, transparent particles,
– have a refractive index that differs from that of the test a significantly larger amount of small particles may be
material ; calculated. In order to obtain traceable results, it is essential
to document the refractive index values used, since small
– be non-solvent of the test material (pure liquid or
differences in the values assumed for the real and imaginary
pre-filtered, saturated solution) ; part of the complex refractive index may cause significant
– not alter the size of the test materials (e.g. by solubility, differences in the resulting particle-size distributions. Small
solubility enhancement, or recrystallisation effects); values of the imaginary part of the refractive index (about
– favour easy formation and stability of the dispersion ; 0.01-0.1 i) are often applied to allow the correction of the
absorbance for the surface roughness of the particles. It
– be compatible with the materials used in the instrument should be noted, in general, that the optical properties of the
(such as O-rings, gaskets, tubing, etc.) ; substance to be tested, as well as the structure (e.g. shape,
– possess a suitable viscosity to facilitate recirculation, surface roughness and porosity), bear upon the final result.
stirring and filtration. Validation. Typically, the validity of a procedure may be
Surfactants and/or dispersing aids are often used to wet the assessed by the evaluation of its specificity, linearity, range,
particles and to stabilise the dispersion. For weak acids and accuracy, precision and robustness. In particle-size analysis
weak bases, buffering of the dispersing medium at low or high by laser light diffraction, specificity as defined by ICH is
pH respectively can assist in identifying a suitable dispersant. not applicable as it is not possible to discriminate between
different components in a sample, nor is it possible to
A preliminary check of the dispersion quality can be
discriminate agglomerates from dispersed particles unless
performed by visual or microscopic inspection. It is also
properly complemented by microscopic techniques. Exploring
possible to take fractional samples out of a well-mixed stock
a linear relationship between concentration and response, or
dispersion. Such stock dispersions are formed by adding a
a mathematical model for interpolation, is not applicable to
liquid to the sample while mixing it with, for example, a glass
this procedure. Rather than evaluating linearity, this method
rod, a spatula or a vortex mixer. Care must be taken to ensure
requires the definition of a concentration range within which
the transfer of a representative sample and that settling of
the result of the measurements does not vary significantly.
larger particles does not occur. Therefore a sample paste is
Concentrations below that range produce an error due to a
prepared or sampling is carried out quickly from a suspension
poor signal-to-noise ratio, while concentrations above that
maintained under agitation.
range produce an error due to multiple scattering. The range
Optimisation of the gas dispersion. For sprays and dry depends mostly on the instrument hardware. Accuracy should
powder dispersions, a compressed gas free from oil, water be confirmed through an appropriate instrument qualification
and particles may be used. To remove such materials from and comparison with microscopy, while precision may be
the compressed gas, a dryer with a filter can be used. Any assessed by means of a repeatability determination.
vacuum unit should be located away from the measurement
zone, so that its output does not disturb the measurement. The attainable repeatability of the method mainly depends
on the characteristics of the material (milled/not milled,
Determination of the concentration range. In order to robust/fragile, width of its size distribution, etc.), whereas
produce an acceptable signal-to-noise ratio in the detector, the required repeatability depends on the purpose of the
the particle concentration in the dispersion must exceed a measurement. Mandatory limits cannot be specified in this
minimum level. Likewise, it must be below a maximum level chapter, as repeatabilities (different sample preparations) may
in order to avoid multiple scattering. The concentration range vary appreciably from one substance to another. However, it
is influenced by the width of the laser beam, the path length of is good practice to aim at acceptance criteria for repeatability
the measurement zone, the optical properties of the particles, such as srel ≤ 10 per cent [n = 6] for any central value of the
and the sensitivity of the detector elements. distribution (e.g. for x50). Values at the sides of the distribution
In view of the above, measurements must be performed at (e.g. x10 and x90) are oriented towards less stringent acceptance
different particle concentrations to determine the appropriate criteria such as srel ≤ 15 per cent [n = 6]. Below 10 μm, these
concentration range for any typical sample of material. (Note : values must be doubled. Robustness may be tested during the
in different instruments, particle concentrations are usually selection and optimisation of the dispersion media and forces.

General Notices (1) apply to all monographs and other texts 375
2.9.31. Particle size analysis by laser light diffraction EUROPEAN PHARMACOPOEIA 10.0

The change of the dispersing energy may be monitored by the particle size, which in turn is defined as the diameter of a
change in the particle-size distribution. volume-equivalent sphere. Q3(x) denotes the volume fraction
undersize at the particle size x. In a graphical representation,
MEASUREMENT x is plotted on the abscissa and the dependent variable Q3
on the ordinate. Most common characteristic values are
Precautions. The instructions given in the instrument manual calculated from the particle-size distribution by interpolation.
are followed : The particle sizes at the undersize values of 10 per cent, 50 per
– never look into the direct path of the laser beam or its cent, and 90 per cent (denoted as x10, x50, and x90 respectively)
reflections ; are frequently used. x50 is also known as the median particle
size. It is recognised that the symbol d is also widely used to
– earth all instrument components to prevent ignition of designate the particle size, thus the symbol x may be replaced
solvents or dust explosions ; by d.
– check the instrument set-up (e.g. warm-up, required Moreover, sufficient information must be documented about
measuring range and lens, appropriate working distance, the sample, the sample preparation, the dispersion conditions,
position of the detector, no direct bright daylight); and the cell type. As the results depend on the particular
– in the case of wet dispersions, avoid air bubbles, instrument, data analysis program, and optical model used,
evaporation of liquid, schlieren or other inhomogeneities these details must also be documented.
in the dispersion ; similarly, avoid improper mass-flow
from the disperser or turbulent air-flow in the case of dry CONTROL OF THE INSTRUMENT PERFORMANCE
dispersions ; such effects can cause erroneous particle-size
distributions. Use the instrument according to the manufacturer’s
instructions and carry out the prescribed qualifications at an
Measurement of the light scattering of dispersed sample(s). appropriate frequency, according to the use of the instrument
After proper alignment of the optical part of the instrument, and substances to be tested.
a blank measurement of the particle-free dispersion medium
must be performed using the same method as that used for the Calibration. Laser diffraction systems, although assuming
measurement of the sample. The background signal must be idealised properties of the particles, are based on first
below an appropriate threshold. The detector data are saved principles of laser light scattering. Thus, calibration in the
in order to substract them later from the data obtained with strict sense is not required. However, it is still necessary to
the sample. The sample dispersion is measured according to confirm that the instrument is operating correctly. This can
the developed method. be undertaken using any certified reference material that is
acceptable in industrial practice. The entire measurement
For each detector element, an average signal is calculated, procedure is examined, including sample collection, sample
sometimes together with its standard deviation. The dispersion, sample transport through the measuring zone,
magnitude of the signal from each detector element depends measurement, and the deconvolution procedure. It is essential
upon the detection area, the light intensity and the quantum that the total operational procedure is fully described.
efficiency. The co-ordinates (size and position) of the
detector elements together with the focal distance of the lens The preferred certified reference materials consist of spherical
determine the range of scattering angles for each element. particles of a known distribution. They must be certified
Most instruments also measure the intensity of the central as to the mass-percentage size distribution by an absolute
(unscattered) laser beam. The ratio of the intensity of a technique, if available, and used in conjunction with an agreed,
dispersed sample to that in its absence (a blank measurement) detailed operation procedure. It is essential that the real and
indicates the proportion of scattered light and hence the imaginary parts of the complex refractive index of the material
particle concentration. are indicated if the Mie theory is applied in data analysis. The
representation of the particle-size distribution by volume
Conversion of scattering pattern into particle-size will equal that of the distribution by mass, provided that the
distribution. This deconvolution step is the inverse of the density of the particles is the same for all size fractions.
calculation of a scattering pattern for a given particle-size
distribution. The assumption of spherical particle shape The response of a laser diffraction instrument is considered to
is particularly important as most algorithms use the meet the requirements if the mean value of x50 from at least
mathematical solution for scattering from spherical particles. 3 independent measurements does not deviate by more than
Furthermore, the measured data always contain some random 3 per cent from the certified range of values of the certified
and systematic errors, which may vitiate the size distributions. reference material. The mean values for x10 and x90 must not
Several mathematical procedures have been developed deviate by more than 5 per cent from the certified range of
for use in the available instruments. They contain some values. Below 10 μm, these values must be doubled.
weighting of deviations between measured and calculated Although the use of materials consisting of spherical particles
scattering patterns (e.g. least squares), some constraints is preferable, non-spherical particles may also be employed.
(e.g. non-negativity for amounts of particles), and/or some Preferably, these particles have certified or typical values
smoothing of the size distribution curve. from laser diffraction analyses performed according to an
The algorithms used are specific to each make and model agreed, detailed operating procedure. The use of reference
of equipment, and are proprietary. The differences in the values from methods other than laser diffraction may
algorithms between different instruments may give rise to cause a significant bias. The reason for this bias is that the
differences in the calculated particle-size distributions. different principles inherent in the various methods may
lead to different sphere-equivalent diameters for the same
Replicates. The number of replicate measurements (with non-spherical particle.
individual sample preparations) to be performed depends on
the required measurement precision. It is recommended to set Although the use of certified reference materials is preferred,
this number in a substance-specific method. other well-defined reference materials may also be employed.
They consist of substances of typical composition and
particle-size distribution for a specified class of substances.
REPORTING OF RESULTS
Their particle-size distribution has proven to be stable over
The particle-size distribution data are usually reported time. The results must comply with previously determined
as cumulative undersize distribution and/or as density data, with the same precision and bias as for the certified
distribution by volume. The symbol x is used to denote the reference material.

376 See the information section on general monographs (cover pages)

EUROPEAN PHARMACOPOEIA 10.0 2.9.32. Porosity and pore-size distribution by mercury porosimetry

Qualification of the system. In addition to the calibration, inter-connectivity, the internal and external surface area,
the performance of the instrument must be qualified at regular powder granulometry, bulk and tapped density could also be
time intervals or as frequently as appropriate. This can be inferred from volume-pressure curves ; however, these aspects
undertaken using any suitable reference material as mentioned of the technique do not fall under the scope of this chapter.
in the previous paragraph. Practical considerations presently limit the maximum applied
The qualification of the system is based on the concept that absolute pressure reached by some equipment to about
the equipment, electronics, software and analytical operations400 MPa, corresponding to a minimum equivalent pore
constitute an integral system, which can be evaluated as an diameter of approximately 0.003 μm. The maximum diameter
entity. Thus the entire measurement procedure is examined, will be limited for samples having a significant depth due to
including sample collection, sample dispersion, sample the difference in hydrostatic head of mercury from the top to
transport through the measuring zone, and the measurement the bottom of the sample. For most purposes this limit may
and deconvolution procedure. It is essential that the total be regarded as 400 μm.
operational procedure is fully described. Inter-particle and intra-particle porosity can be determined,
In general, unless otherwise specified in the individual but the method does not distinguish between these porosities
monograph, the response of a laser diffraction instrument where they co-exist.
is considered to meet the requirements if the x50 value does The method is suitable for the study of most porous materials.
not deviate by more than 10 per cent from the range of values Samples that amalgamate with mercury, such as certain
of the reference material. If optionally the values at the sides
metals, may be unsuitable for this technique or may require
of the distribution are evaluated (e.g. x10 and x90), then these
a preliminary passivation. Other materials may deform or
values must not deviate by more than 15 per cent from the compact under the applied pressure. In some cases it may
certified range of values. Below 10 μm, these values must be be possible to apply sample-compressibility corrections and
doubled. useful comparative data may still be obtained.
NOTE : for calibration of the instrument, stricter requirements
Mercury porosimetry is considered to be comparative, as
are laid down in the paragraph Calibration. for most porous media a theory is not available to allow
an absolute calculation of results of pore-size distribution.
Therefore this technique is mainly recommended for
development studies.
07/2008:20932
Mercury is toxic. Appropriate precautions must be observed
to safeguard the health of the operator and others working in
the area. Waste material must also be disposed of in a suitable
manner, according to local regulations.

2.9.32. POROSITY AND PORE-SIZE PRINCIPLE

DISTRIBUTION OF SOLIDS BY The technique is based on the measurement of the mercury
volume intruded into a porous solid as a function of the
MERCURY POROSIMETRY applied pressure. The measurement includes only those pores
into which mercury can penetrate at the pressure applied.
INTRODUCTION
A non-wetting liquid penetrates into a porous system only
In general, different types of pores may be pictured as
under pressure. The pressure to be applied is in inverse
apertures, channels or cavities within a solid body, or as space
proportion to the inner diameter of the pore aperture. In
(i.e. interstices or voids) between solid particles in a bed,
the case of cylindrical pores, the correlation between pore
compact or aggregate. Porosity is a term that is often used
diameter and pressure is given by the Washburn equation :
to indicate the porous nature of solid material, and is more
precisely defined as the ratio of the volume of accessible pores
dp = -
4·σ cos θ
and voids to the total volume occupied by a given amount p
of the solid. In addition to the accessible pores, a solid may
contain closed pores, which are isolated from the external dp = pore diameter, in metres ;
surface and into which fluids are not able to penetrate. The
characterisation of closed pores, i.e. cavities with no access to σ = surface tension, in newtons per metre ;
an external surface, is not covered in this chapter. θ = contact angle of mercury on the sample, in degrees ;
Porous materials may take the form of fine or coarse p = applied pressure, in pascals.
powders, compacts, extrudates, sheets or monoliths. Their
characterisation usually involves the determination of the total APPARATUS
pore volume or porosity as well as the pore-size distribution.
The sample holder, referred to as penetrometer or dilatometer,
It is well established that the performance of a porous solid has a calibrated capillary tube, through which the sample
(e.g. its strength, reactivity, permeability or adsorbent power) can be evacuated and through which mercury can enter.
is dependent upon its pore structure. Many different methods The capillary tube is attached to a wider tube in which the
have been developed for the characterisation of pore structure. test sample is placed. The change in the volume of mercury
In view of the complexity of most porous solids, it is not intruded is usually measured by the change in capacitance
surprising to find that the results obtained are not always in between the mercury column in the capillary tube and a metal
agreement and that no single technique can be relied upon to sleeve around the outside of the capillary tube. If precise
provide a complete picture of the pore structure. The choice measurements are required the expected total void and pore
of the most appropriate method depends on the application volume of the sample should be between 20 per cent and
of the porous solid, its chemical and physical nature and the 90 per cent of the internal volume of the capillary tube. Since
range of pore-size. different materials exhibit a wide range of open porosities,
This chapter provides guidance for measurement of porosity a number of penetrometers with different capillary tube
and pore-size distribution by mercury porosimetry. It is a diameters and sample volumes may be required. A typical
comparative test, usually destructive, in which the volume of set-up for a mercury porosimeter instrument is given in
mercury penetrating a pore or void is determined as a function Figure 2.9.32.-1. The porosimeter may have separate ports
of an applied hydrostatic pressure, which can be related to for high- and low-pressure operation, or the low-pressure
a pore diameter. Other information such as pore shape and measurement may be carried out on a separate unit.

General Notices (1) apply to all monographs and other texts 377