Printed on: Mon Jul 25 2022, 08:48:49 PM(EST) Status: Currently Official on 26-Jul-2022 DocId: GUID-BFC6D11B-21C5-494E-A0C3-2EB88E2F297A_2_en-US

Printed by: Mahakam Beta Farma Official Date: Official as of 01-May-2013 Document Type: GENERAL CHAPTER @2022 USPC
DOI Ref: wzz7a DOI: https://doi.org/10.31003/USPNF_M99586_02_01
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á788ñ PARTICULATE MATTER IN INJECTIONS

This general chapter is harmonized with the corresponding texts of the European Pharmacopoeia and/or the Japanese
Pharmacopoeia. These pharmacopeias have undertaken not to make any unilateral change to this harmonized chapter. Portions
of the present general chapter text that are national USP text, and therefore not part of the harmonized text, are marked with
symbols (◆◆) to specify this fact.
Particulate matter in injections and parenteral infusions consists of extraneous mobile undissolved particles, other than gas
bubbles, unintentionally present in the solutions.
◆
As stated in Injections and Implanted Drug Products á1ñ, solutions for injection administered by the intramuscular or
subcutaneous route must meet the requirements of Particulate Matter in Injections á788ñ. This requirement has been indefinitely
postponed for products for veterinary use. Parenterals packaged and labeled exclusively for use as irrigating solutions are exempt
from the requirements of Particulate Matter in Injections á788ñ. Radiopharmaceutical preparations are exempt from the
requirements of Particulate Matter in Injections á788ñ. Parenteral products for which the labeling specifies use of a final filter prior
to administration are exempt from the requirements of Particulate Matter in Injections á788ñ, provided that scientific data are
available to justify this exemption.◆
For the determination of particulate matter, two procedures, Method 1 (Light Obscuration Particle Count Test) and Method 2
(Microscopic Particle Count Test), are specified hereinafter. When examining injections and parenteral infusions for subvisible
particles, Method 1 is preferably applied. However, it may be necessary to test some preparations by the Light Obscuration
Particle Count Test followed by the Microscopic Particle Count Test to reach a conclusion on conformance to the requirements.
Not all parenteral preparations can be examined for subvisible particles by one or both of these methods. When Method 1
is not applicable, e.g., in the case of preparations having reduced clarity or increased viscosity, the test should be carried out

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according to Method 2. Emulsions, colloids, and liposomal preparations are examples. Similarly, products that produce air or
gas bubbles when drawn into the sensor may also require microscopic particle count testing. If the viscosity of the preparation
to be tested is sufficiently high so as to preclude its examination by either test method, a quantitative dilution with an appropriate
diluent may be made to decrease viscosity, as necessary, to allow the analysis to be performed.
The results obtained in examining a discrete unit or group of units for particulate matter cannot be extrapolated with certainty
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to other units that remain untested. Thus, statistically sound sampling plans must be developed if valid inferences are to be
drawn from observed data to characterize the level of particulate matter in a large group of units.
◆
For the purpose of this chapter, small-volume parenteral is synonymous with small-volume injection, and large-volume
parenteral is synonymous with large-volume injection.◆
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METHOD 1 LIGHT OBSCURATION PARTICLE COUNT TEST

Use a suitable apparatus based on the principle of light blockage that allows for an automatic determination of the size of
particles and the number of particles according to size. The definition for particle-free water is provided in Reagents, Indicators,
and Solutions—Reagent Specifications.
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The apparatus is calibrated using dispersions of spherical particles of known sizes between 10 µm and 25 µm. These standard
particles are dispersed in particle-free water. Care must be taken to avoid aggregation of particles during dispersion. ◆System
suitability can be verified by using the USP Particle Count RS.◆

General Precautions
The test is carried out under conditions limiting particulate matter, preferably in a laminar flow cabinet.
Very carefully wash the glassware and filtration equipment used, except for the membrane filters, with a warm detergent
solution, and rinse with abundant amounts of water to remove all traces of detergent. Immediately before use, rinse the
equipment from top to bottom, outside and then inside, with particle-free water.
Take care not to introduce air bubbles into the preparation to be examined, especially when fractions of the preparation are
being transferred to the container in which the determination is to be carried out.
In order to check that the environment is suitable for the test, that the glassware is properly cleaned, and that the water to
be used is particle-free, the following test is carried out. Determine the particulate matter in five samples of particle-free water,
each of 5 mL, according to the method described below. If the number of particles of 10 µm or greater size exceeds 25 for the
combined 25 mL, the precautions taken for the test are not sufficient. The preparatory steps must be repeated until the
environment, glassware, and water are suitable for the test.

Method
Mix the contents of the sample by slowly inverting the container 20 times successively. If necessary, cautiously remove the
sealing closure. Clean the outer surfaces of the container opening using a jet of particle-free water, and remove the closure,
avoiding any contamination of the contents. Eliminate gas bubbles by appropriate measures such as allowing to stand for 2 min
or sonicating.
For large-volume parenterals, single units are tested. For small-volume parenterals less than 25 mL in volume, the contents
of 10 or more units are combined in a cleaned container to obtain a volume of NLT 25 mL; the test solution may be prepared
by mixing the contents of a suitable number of vials and diluting to 25 mL with particle-free water or with an appropriate
particle-free solvent when particle-free water is not suitable. Small-volume parenterals having a volume of 25 mL or more may
be tested individually.

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Printed on: Mon Jul 25 2022, 08:48:49 PM(EST) Status: Currently Official on 26-Jul-2022 DocId: GUID-BFC6D11B-21C5-494E-A0C3-2EB88E2F297A_2_en-US
Printed by: Mahakam Beta Farma Official Date: Official as of 01-May-2013 Document Type: GENERAL CHAPTER @2022 USPC
DOI Ref: wzz7a DOI: https://doi.org/10.31003/USPNF_M99586_02_01
2

Powders for parenteral use are reconstituted with particle-free water or with an appropriate particle-free solvent when
particle-free water is not suitable.
◆
For pharmacy bulk packages for parenteral use labeled “Not for Direct Infusion”, proceed as directed for small-volume
parenterals when the volume is 25 mL or more. Calculate the test result on a portion that is equivalent to the maximum dose
given in the labeling. For example, if the total bulk package volume is 100 mL and the maximum dose volume is 10 mL, then
the average particle count per mL would be multiplied by 10 to obtain the test result based on the 10-mL maximum dose.
[NOTE—For the calculation of test results, consider this maximum dose portion to be equivalent to the contents of one full
container.]
Products packaged with dual compartments meant to hold a drug product and a solvent should be prepared and tested as
directed for large-volume parenterals or small-volume parenterals, depending on container volume. Mix each unit as directed
in the labeling, activating and agitating to ensure thorough mixing of the separate components and drug dissolution.◆
The number of test specimens must be adequate to provide a statistically sound assessment. For large-volume parenterals
or for small-volume parenterals having a volume of 25 mL or more, fewer than 10 units may be tested, using an appropriate
sampling plan.
Remove four portions, NLT 5 mL each, and count the number of particles equal to or greater than 10 µm and 25 µm.
Disregard the result obtained for the first portion, and calculate the mean number of particles for the preparation to be
examined.

Evaluation
For preparations supplied in containers with a nominal volume of more than 100 mL, apply the criteria of Test 1.A.
For preparations supplied in containers with a nominal volume of less than 100 mL, apply the criteria of Test 1.B.

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For preparations supplied in containers with a nominal volume of 100 mL, apply the criteria of Test 1.B.
[NOTE—Test 1.A is used in the Japanese Pharmacopoeia.]
If the average number of particles exceeds the limits, test the preparation by the Microscopic Particle Count Test.
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Solutions for parenteral infusion or solutions for injection supplied in containers with a nominal content of more than
100 mL: The preparation complies with the test if the average number of particles present in the units tested does not exceed
25 per mL equal to or greater than 10 µm and does not exceed 3 per mL equal to or greater than 25 µm.

TEST 1.B
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Solutions for parenteral infusion or solutions for injection supplied in containers with a nominal content of less than
100 mL: The preparation complies with the test if the average number of particles present in the units tested does not exceed
6000 per container equal to or greater than 10 µm and does not exceed 600 per container equal to or greater than 25 µm.
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METHOD 2 MICROSCOPIC PARTICLE COUNT TEST

Use a suitable binocular microscope, a filter assembly for retaining particulate matter, and a membrane filter for examination.
The microscope is adjusted to 100 ± 10 magnifications and is equipped with an ocular micrometer calibrated with an
objective micrometer, a mechanical stage capable of holding and traversing the entire filtration area of the membrane filter,
and two suitable illuminators to provide episcopic illumination in addition to oblique illumination.
The ocular micrometer is a circular diameter graticule (see Figure 1) and consists of a large circle divided by crosshairs into
quadrants, transparent and black reference circles 10 µm and 25 µm in diameter at 100 magnifications, and a linear scale
graduated in 10-µm increments. It is calibrated using a stage micrometer that is certified by either a domestic or international
standard institution. A relative error of the linear scale of the graticule within ±2% is acceptable. The large circle is designated
the graticule field of view (GFOV).

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Printed on: Mon Jul 25 2022, 08:48:49 PM(EST) Status: Currently Official on 26-Jul-2022 DocId: GUID-BFC6D11B-21C5-494E-A0C3-2EB88E2F297A_2_en-US
Printed by: Mahakam Beta Farma Official Date: Official as of 01-May-2013 Document Type: GENERAL CHAPTER @2022 USPC
DOI Ref: wzz7a DOI: https://doi.org/10.31003/USPNF_M99586_02_01
3

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Figure 1. Circular diameter graticule. The large circle divided by crosshairs into quadrants is designated the graticule field of
view (GFOV). Transparent and black circles having 10-µm and 25-µm diameters at 100× are provided as comparison scales for
particle sizing.

Two illuminators are required. One is an episcopic brightfield illuminator internal to the microscope, the other is an external,
focusable auxiliary illuminator that can be adjusted to give reflected oblique illumination at an angle of 10°–20°.
The filter assembly for retaining particulate matter consists of a filter holder made of glass or other suitable material, and is
equipped with a vacuum source and a suitable membrane filter.
The membrane filter is of suitable size, black or dark gray in color, nongridded or gridded, and 1.0 µm or finer in nominal
pore size.

General Precautions
The test is carried out under conditions limiting particulate matter, preferably in a laminar flow cabinet.
Very carefully wash the glassware and filter assembly used, except for the membrane filter, with a warm detergent solution,
and rinse with abundant amounts of water to remove all traces of detergent. Immediately before use, rinse both sides of the
membrane filter and the equipment from top to bottom, outside and then inside, with particle-free water.
In order to check that the environment is suitable for the test, that the glassware and the membrane filter are properly
cleaned, and that the water to be used is particle-free, the following test is carried out. Determine the particulate matter of a
50-mL volume of particle-free water according to the method described below. If more than 20 particles 10 µm or larger in size
or if more than five particles 25 µm or larger in size are present within the filtration area, the precautions taken for the test are
not sufficient. The preparatory steps must be repeated until the environment, glassware, membrane filter, and water are suitable
for the test.

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Printed on: Mon Jul 25 2022, 08:48:49 PM(EST) Status: Currently Official on 26-Jul-2022 DocId: GUID-BFC6D11B-21C5-494E-A0C3-2EB88E2F297A_2_en-US
Printed by: Mahakam Beta Farma Official Date: Official as of 01-May-2013 Document Type: GENERAL CHAPTER @2022 USPC
DOI Ref: wzz7a DOI: https://doi.org/10.31003/USPNF_M99586_02_01
4

Method
Mix the contents of the samples by slowly inverting the container 20 times successively. If necessary, cautiously remove the
sealing closure. Clean the outer surfaces of the container opening using a jet of particle-free water, and remove the closure,
avoiding any contamination of the contents.
For large-volume parenterals, single units are tested. For small-volume parenterals less than 25 mL in volume, the contents
of 10 or more units are combined in a cleaned container; the test solution may be prepared by mixing the contents of a suitable
number of vials and diluting to 25 mL with particle-free water or with an appropriate particle-free solvent when particle-free water
is not suitable. Small-volume parenterals having a volume of 25 mL or more may be tested individually.
Powders for parenteral use are constituted with particle-free water or with an appropriate particle-free solvent when
particle-free water is not suitable.
The number of test specimens must be adequate to provide a statistically sound assessment. For large-volume parenterals
or for small-volume parenterals having a volume of 25 mL or more, fewer than 10 units may be tested, using an appropriate
sampling plan.
Wet the inside of the filter holder fitted with the membrane filter with several mL of particle-free water. Transfer to the filtration
funnel the total volume of a solution pool or of a single unit, and apply a vacuum. If needed, add stepwise a portion of the
solution until the entire volume is filtered. After the last addition of solution, begin rinsing the inner walls of the filter holder by
using a jet of particle-free water. Maintain the vacuum until the surface of the membrane filter is free from liquid. Place the
membrane filter in a Petri dish, and allow the membrane filter to air-dry with the cover slightly ajar. After the membrane filter
has been dried, place the Petri dish on the stage of the microscope, scan the entire membrane filter under the reflected light
from the illuminating device, and count the number of particles that are equal to or greater than 10 µm and the number of
particles that are equal to or greater than 25 µm. Alternatively, partial membrane filter count and determination of the total

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filter count by calculation is allowed. Calculate the mean number of particles for the preparation to be examined.
The particle sizing process with the use of the circular diameter graticule is carried out by estimating the equivalent diameter
of the particle in comparison with the 10 µm and 25 µm reference circles on the graticule. Thereby the particles are not moved
from their initial locations within the graticule field of view and are not superimposed on the reference circles for comparison.
The inner diameter of the transparent graticule reference circles is used to size white and transparent particles, while dark
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particles are sized by using the outer diameter of the black opaque graticule reference circles.
In performing the Microscopic Particle Count Test, do not attempt to size or enumerate amorphous, semiliquid, or otherwise
morphologically indistinct materials that have the appearance of a stain or discoloration on the membrane filter. These materials
show little or no surface relief and present a gelatinous or film-like appearance. In such cases, the interpretation of enumeration
may be aided by testing a sample of the solution by the Light Obscuration Particle Count Test.
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Evaluation
For preparations supplied in containers with a nominal volume of more than 100 mL, apply the criteria of Test 2.A.
For preparations supplied in containers with a nominal volume of less than 100 mL, apply the criteria of Test 2.B.
For preparations supplied in containers with a nominal volume of 100 mL, apply the criteria of Test 2.B.
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[NOTE—Test 2.A is used in the Japanese Pharmacopoeia.]

TEST 2.A
Solutions for parenteral infusion or solutions for injection supplied in containers with a nominal content of more than
100 mL: The preparation complies with the test if the average number of particles present in the units tested does not exceed
12 per mL equal to or greater than 10 µm and does not exceed 2 per mL equal to or greater than 25 µm.

TEST 2.B
Solutions for parenteral infusion or solutions for injection supplied in containers with a nominal content of less than
100 mL: The preparation complies with the test if the average number of particles present in the units tested does not exceed
3000 per container equal to or greater than 10 µm and does not exceed 300 per container equal to or greater than 25 µm.

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