USP 601 AEROSOLS, NASAL SPRAYS

Accessed from 108.250.52.
37 by aptuit on Fri Dec 07 08:51:21 EST 2012
232 〈591〉 Zinc Determination / Chemical Tests USP 35
〈591〉 ZINC DETERMINATION Physical Tests and

The need for a quantitative determination of zinc in the
Determinations
Pharmacopeial insulin preparations reflects the fact that the
element is an essential component of zinc-insulin crystals. In
common with lead, zinc may be determined either by the
dithizone method or by atomic absorption.
〈601〉 AEROSOLS, NASAL SPRAYS,
Dithizone Method METERED-DOSE INHALERS, AND
Select all reagents for this test to have as low a content of DRY POWDER INHALERS
heavy metals as practicable. If necessary, distill water and
other solvents into hard or borosilicate glass apparatus.
Rinse thoroughly all glassware with warm dilute nitric acid This general chapter contains test methods for propel-
(1 in 2) followed by water. Avoid using on the separator any lants, pressurized topical aerosols, nasal sprays, metered-
lubricants that dissolve in chloroform. dose inhalers, and propellant-free dry powder inhalers used
Special Solutions and Solvents— to aerosolize, or to aerosolize and meter, doses of powders
ALKALINE AMMONIUM CITRATE SOLUTION—Dissolve 50 g of dibasic for inhalation. Apply these methods, where indicated, in the
ammonium citrate in water to make 100 mL. Add 100 mL testing of the appropriate dosage forms.
of ammonium hydroxide. Remove any heavy metals that
may be present by extracting the solution with 20-mL por-
tions of Dithizone Extraction Solution (see Lead 〈251〉) until PROPELLANTS
the dithizone solution retains a clear green color, then ex-
tract any dithizone remaining in the citrate solution by shak- Caution—Hydrocarbon propellants are highly flammable and
ing with chloroform. explosive. Observe precautions and perform sampling and ana-
lytical operations in a well-ventilated fume hood.
CHLOROFORM—Distill chloroform in hard or borosilicate glass
apparatus, receiving the distillate in sufficient dehydrated al-
cohol to make the final concentration 1 mL of alcohol for General Sampling Procedure
each 100 mL of distillate.
DITHIZONE SOLUTION—Use Standard Dithizone Solution (see This procedure is used to obtain test specimens for those
Lead 〈251〉), prepared with the distilled Chloroform. propellants that occur as gases at about 25° and that are
STANDARD ZINC SOLUTION—Dissolve 625 mg of zinc oxide, ac- stored in pressurized cylinders. Use a stainless steel sample
curately weighed, and previously gently ignited to constant cylinder equipped with a stainless steel valve and having a
weight, in 10 mL of nitric acid, and add water to make capacity of not less than 200 mL and a pressure rating of
500.0 mL. This solution contains 1.0 mg of zinc per mL. 240 psi or more. Dry the cylinder with the valve open at
110° for 2 hours, and evacuate the hot cylinder to less than
DILUTED STANDARD ZINC SOLUTION—Dilute 1 mL of Standard
1 mm of mercury. Close the valve, cool, and weigh. Con-
Zinc Solution, accurately measured, with 2 drops of nitric nect one end of a charging line tightly to the propellant
acid and sufficient water to make 100.0 mL. This solution container and the other end loosely to the sample cylinder.
contains 10 µg of zinc per mL. Use this solution within 2 Carefully open the propellant container, and allow the pro-
weeks. pellant to flush out the charging line through the loose con-
TRICHLOROACETIC ACID SOLUTION—Dissolve 100 g of trichloroa- nection. Avoid excessive flushing, which causes moisture to
cetic acid in water to make 1000 mL. freeze in the charging line and connections. Tighten the fit-
Procedure—Transfer 1 to 5 mL of the preparation to be ting on the sample cylinder, and open the sample cylinder
tested, accurately measured, to a centrifuge tube graduated valve, allowing the propellant to flow into the evacuated
at 40 mL. If necessary, add 0.25 N hydrochloric acid, drop- cylinder. Continue sampling until the desired amount of
wise, to obtain a clear solution. Add 5 mL of Trichloroacetic specimen is obtained, then close the propellant container
Acid Solution and sufficient water to make 40.0 mL. Mix, valve, and finally close the sample cylinder valve. [Caution—
and centrifuge. Do not overload the sample cylinder; hydraulic expansion due
Transfer to a hard-glass separator an accurately measured to temperature change can cause overloaded cylinders to ex-
volume of the supernatant believed to contain from 5 to 20 plode.] Again weigh the charged sample cylinder, and calcu-
µg of zinc, and add water to make about 20 mL. Add 1.5 late the weight of the specimen.
mL of Alkaline Ammonium Citrate Solution and 35 mL of
Dithizone Solution. Shake vigorously 100 times. Allow the
chloroform phase to separate. Insert a cotton plug in the Approximate Boiling Temperature
stem of the separator to remove any water emulsified with
the chloroform. Collect the chloroform extract (discarding Transfer a 100-mL specimen to a tared, pear-shaped, 100-
the first portion that comes through) in a test tube, and mL centrifuge tube containing a few boiling stones, and
determine the absorbance at 530 nm, with a suitable weigh. Suspend a thermometer in the liquid, and place the
spectrophotometer. tube in a medium maintained at a temperature of 32°
Calculate the amount of zinc present by reference to a above the expected boiling temperature. When the ther-
standard absorbance-concentration curve obtained by using mometer reading becomes constant, record as the boiling
0.5 mL, 1.0 mL, 1.5 mL, and, if the zinc content of the temperature the thermometer reading after at least 5% of
sample extracted exceeds 15 µg, 2.0 mL of the Diluted Stan- the specimen has distilled. Retain the remainder of the spec-
dard Zinc Solution, corrected as indicated by a blank deter- imen for the determination of High-Boiling Residues.
mination run concomitantly, using all of the reagents but
no added zinc.
High-Boiling Residues, Method I
Allow 85 mL of the specimen to distill as directed in the
test for Approximate Boiling Temperature, and transfer the
Official from December 1, 2012

Copyright (c) 2012 The United States Pharmacopeial Convention. All rights reserved.
Accessed from 108.250.52.37 by aptuit on Fri Dec 07 08:51:21 EST 2012
USP 35 Physical Tests / 〈601〉 Aerosols 233
centrifuge tube containing the remaining 15 mL of speci- pressure, and released upon activation of an appropriate
men to a medium maintained at a temperature 10° above valve system.
the boiling temperature. After 30 minutes, remove the tube
from the water bath, blot dry, and weigh. Calculate the
weight of the residue. Delivery Rate and Delivered Amount
Perform these tests only on containers fitted with continu-
High-Boiling Residues, Method II ous valves.
Delivery Rate—Select not fewer than four aerosol con-
Prepare a cooling coil from copper tubing (about 6 mm tainers, shake, if the label includes this directive, remove the
outside diameter × about 6.1 m long) to fit into a vacuum- caps and covers, and actuate each valve for 2 to 3 seconds.
jacketed flask. Immerse the cooling coil in a mixture of dry Weigh each container accurately, and immerse in a con-
ice and acetone in a vacuum-jacketed flask, and connect stant-temperature bath until the internal pressure is equili-
one end of the tubing to the propellant sample cylinder. brated at a temperature of 25° as determined by constancy
Carefully open the sample cylinder valve, flush the cooling of internal pressure, as directed under the Pressure Test be-
coil with about 50 mL of the propellant, and discard this low. Remove the containers from the bath, remove excess
portion of liquefied propellant. Continue delivering liquefied moisture by blotting with a paper towel, shake, if the label
propellant from the cooling coil, and collect it in a previ- includes this directive, actuate each valve for 5.0 seconds
ously chilled 1000-mL sedimentation cone until the cone is (accurately timed by use of a stopwatch), and weigh each
filled to the 1000-mL mark. Allow the propellant to evapo- container again. Return the containers to the constant-tem-
rate, using a warm water bath maintained at about 40° to perature bath, and repeat the foregoing procedure three
reduce evaporating time. When all of the liquid has evapo- times for each container. Calculate the average Delivery
rated, rinse the sedimentation cone with two 50-mL por- Rate, in g per second, for each container.
tions of pentane, and combine the rinsings in a tared 150- Delivered Amount—Return the containers to the con-
mL evaporating dish. Transfer 100 mL of the pentane sol- stant-temperature bath, continuing to deliver 5 second actu-
vent to a second tared 150-mL evaporating dish, place both ations to waste, until each container is exhausted. [NOTE—
evaporating dishes on a water bath, evaporate to dryness, Ensure that sufficient time is allowed between each actua-
and heat the dishes in an oven at 100° for 60 minutes. Cool tion to avoid significant canister cooling.] Calculate the total
the dishes in a desiccator, and weigh. Repeat the heating weight loss from each container. This is the Delivered
for 15-minute periods until successive weighings are within Amount.
0.1 mg, and calculate the weight of the residue obtained
from the propellant as the difference between the weights
of the residues in the two evaporating dishes. Pressure Test
Perform this test only on topical aerosols fitted with con-
Water Content tinuous valves.
Select not fewer than four aerosol containers, remove the
Proceed as directed under Water Determination 〈921〉, with caps and covers, and immerse in a constant-temperature
the following modifications: (a) Provide the closed-system bath until the internal pressure is constant at a temperature
titrating vessel with an opening through which passes a of 25°. Remove the containers from the bath, shake, and
coarse-porosity gas dispersion tube connected to a sampling remove the actuator and water, if any, from the valve stem.
cylinder. (b) Dilute the Reagent with anhydrous methanol to Place each container in an upright position, and determine
give a water equivalence factor of between 0.2 and 1.0 mg the pressure in each container by placing a calibrated pres-
per mL; age this diluted solution for not less than 16 hours sure gauge on the valve stem, holding firmly, and actuating
before standardization. (c) Obtain a 100-g specimen as di- the valve so that it is fully open. The gauge is of a calibra-
rected under General Sampling Procedure, and introduce the tion approximating the expected pressure and is fitted with
specimen into the titration vessel through the gas dispersion an adapter appropriate for the particular valve stem dimen-
tube at a rate of about 100 mL of gas per minute; if neces- sions. Read the pressure directly from the gauge.
sary, heat the sample cylinder gently to maintain this flow
rate.
Minimum Fill
Other Determinations Topical aerosols meet the requirements for aerosols under
Minimum Fill 〈755〉.
For those aerosols that use propellants, perform the tests
specified in the individual NF propellant monographs.
Leakage Test
AEROSOLS Perform this test only on topical aerosols fitted with con-
tinuous valves.
Because leaching of extractable substances into pres- Select 12 aerosol containers, and record the date and
surized formulations should be minimized, valve materials time to the nearest half hour. Weigh each container to the
and other components that are in contact with the product nearest mg, and record the weight, in mg, of each as W1.
meet the requirements under Elastomeric Closures for Injec- Allow the containers to stand in an upright position at a
tions 〈381〉 (Note that under Physicochemical Test Procedures temperature of 25.0 ± 2.0° for not less than 3 days, and
in 〈381〉 the directions for preparing a sample require pre- again weigh each container, recording the weight, in mg, of
extraction, which may cause an underestimate of the each as W2, and recording the date and time to the nearest
amount of extractables from a given component.) See also half hour. Determine the time, T, in hours, during which the
Aerosols under Pharmaceutical Dosage Forms 〈1151〉. containers were under test. Calculate the leakage rate, in
mg per year, of each container taken by the formula:
TOPICAL AEROSOLS (365)(24/T)(W1 − W2).
The following tests are applicable to topical aerosols con-
taining drug, in suspension or solution, packaged under Where plastic-coated glass aerosol containers are tested,
dry the containers in a desiccator for 12 to 18 hours, and

234 〈601〉 Aerosols / Physical Tests USP 35
allow them to stand in a constant-humidity environment for mum number of sprays per nostril as described on the label,
24 hours prior to determining the initial weight as indicated or instructions for use) collected at the beginning of unit life
above. Conduct the test under the same constant-humidity (after priming as described on the label, or instructions for
conditions. Empty the contents of each container tested by use) and at the label claim number of metered sprays, from
employing any safe technique (e.g., chill to reduce the in- each of 10 separate containers, must meet the following
ternal pressure, remove the valve, and pour). Remove any acceptance criteria: not more than 2 of the 20 doses are
residual contents by rinsing with suitable solvents, then rinse outside the range of 80% to 120% of label claim, and none
with a few portions of methanol. Retain as a unit the con- are outside the range of 75% to 125% of label claim, while
tainer, the valve, and all associated parts, and heat them at the mean for each of the beginning and end doses falls
100° for 5 minutes. Cool, weigh, record the weight as W3, within the range of 85% to 115% of label claim. If 3–6
and determine the net fill weight (W1 − W3) for each con- doses of the 20 doses collected are outside of 80% to 120%
tainer tested. [NOTE—If the average net fill weight has been of the label claim, but none are outside of 75% to 125% of
determined previously, that value may be used in place of label claim, and the means for each of the beginning and
the value (W1 − W3) above.] The requirements are met if the end doses fall within 85% to 115% of label claim, select 20
average leakage rate per year for the 12 containers is not additional containers for second-tier testing. For second-tier
more than 3.5% of the net fill weight, and none of the testing, the requirements are met if not more than 6 of the
containers leaks more than 5.0% of the net fill weight per 60 doses collected are outside the range of 80% to 120%
year. If 1 container leaks more than 5.0% per year, and if of label claim, none are outside the range of 75% to 125%
none of the containers leaks more than 7.0% per year, de- of label claim, and the means for each of the beginning and
termine the leakage rate of an additional 24 containers as end doses fall within the range of 85% to 115% of label
directed herein. Not more than 2 of the 36 containers leak claim.
more than 5.0% of the net fill weight per year, and none of
the 36 containers leaks more than 7.0% of the net fill
weight per year. Where the net fill weight is less than 15 g SAMPLING FOR DELIVERED-DOSE UNIFORMITY OF METERED-
and the label bears an expiration date, the requirements are DOSE NASAL SPRAYS
met if the average leakage rate of the 12 containers is not
more than 525 mg per year and none of the containers General Sampling Procedure—To ensure reproducible
leaks more than 750 mg per year. If 1 container leaks more in-vitro dose collection, it is recommended that a mechani-
than 750 mg per year, but not more than 1.1 g per year, cal means of actuating the pump assembly be employed to
determine the leakage rate of an additional 24 containers as deliver doses for collection. The mechanical actuation proce-
directed herein. Not more than 2 of the 36 containers leak dure should have adequate controls for the critical mechani-
more than 750 mg per year, and none of the 36 containers cal actuation parameters (e.g., actuation force, actuation
leaks more than 1.1 g per year. This test is in addition to speed, stroke length, rest periods, etc.). The test must be
the customary in-line leak testing of each container. performed on units that have been primed according to the
patient-use instructions. The test unit should be actuated in
a vertical or near vertical, valve-up, position. The two doses
Number of Discharges per Container collected at the beginning and end of the container life
should be the dose immediately following priming and the
Perform this test only on topical aerosols fitted with dose- dose corresponding to the last label claim number of doses
metering valves, at the same time as, and on the same con- from the container.
tainers used for, the test for Delivered-Dose Uniformity. Deter- For suspension products, the delivered dose should be de-
mine the number of discharges or deliveries by counting the livered into a suitable container (e.g., scintillation vial) in
number of priming discharges plus those used in determin- which quantitative transfer from the container under test
ing the spray contents, and continue to fire until the label can be accomplished. A validated analytical method is em-
claim number of discharges. The requirements are met if all ployed to determine the amount of drug in each delivered
the containers or inhalers tested contain not less than the dose, and data are reported as a percent of label claim. For
number of discharges stated on the label. solution products, the delivered dose can be determined
gravimetrically from the weight of the delivered dose, and
Delivered-Dose Uniformity the concentration and density of the fill solution of the
product under test.
The test for Delivered-Dose Uniformity is required for topi-
cal aerosols fitted with dose-metering valves. For collection METERED-DOSE INHALERS AND DRY
of the minimum dose, proceed as directed in the test for
Delivered-Dose Uniformity under Metered-Dose Inhalers and POWDER INHALERS
Dry Powder Inhalers, as described below, except to modify
the dose sampling apparatus so that it is capable of quan- The following tests are applicable to metered-dose inhal-
titatively capturing the delivered dose from the preparation ers that are formulated as suspensions or solutions of active
being tested. Unless otherwise stated in the individual mon- drug in propellants and dry powder inhalers presented as
ograph, apply the acceptance criteria for Metered-Dose In- single or multidose units. The following test methods are
halers and Dry Powder Inhalers as described below. specific to the aforementioned inhalers and may require
modification when testing alternative inhalation technolo-
gies (for example, breath-actuated metered-dose inhalers, or
NASAL SPRAYS dose-metering nebulizers). However, Pharmacopeial require-
ments for all dose-metering inhalation dosage forms require
The following test is applicable to nasal sprays, formulated determination of the delivered dose and Aerodynamic Size
as aqueous suspensions or solutions of drug, presented in Distribution. In all cases, and for all tests, prepare and test
multi-dose containers and fitted with dose-metering valves. the inhaler as directed on the label and the instructions for
In all cases, and for all tests, prepare and test the nasal use. When these directions are not provided by the product
spray as directed on the label and the instructions for use. manufacturer, follow the precise dose discharge directions
included in the tests below.
Delivered-Dose Uniformity
Unless otherwise directed in the individual monograph,
the drug content of the minimum delivered doses (mini-

Fig. 1. Sampling apparatus for pressurized metered-dose inhalers.
Delivered-Dose Uniformity Unless otherwise specified in the individual monograph,

the requirements for dosage uniformity are met if not less
The test for Delivered-Dose Uniformity is required for inhal- than 9 of the 10 doses are between 75% and 125% of the
ers (e.g., metered-dose inhalers or dry powder inhalers) con- specified target-delivered dose and none is outside the
taining drug formulation (e.g., solution, suspension, or pow- range of 65% to 135% of the specified target-delivered
der) either in reservoirs or in premetered dosage units, and dose. If the contents of not more than 3 doses are outside
for drug formulations packaged in reservoirs or in preme- the range of 75% to 125% of the specified target-delivered
tered dosage units where these containers are labeled for dose, but within the range of 65% to 135% of the specified
use with a named inhalation device. (For inhalations pack- target-delivered dose, select 20 additional containers, and
aged in premetered dosage units, see also Uniformity of Dos- follow the prescribed procedure for analyzing 1 minimum
age Units 〈905〉.) Note that the target-delivered dose is the dose from each. The requirements are met if not more than
expected mean drug content for a large number of deliv- 3 results, out of the 30 values, lie outside the range of 75%
ered doses collected from many inhalers of the chosen to 125% of the specified target-delivered dose, and none is
product. In many cases, its value may depend upon the outside the range of 65% to 135% of the specified target-
manner in which the test for delivered dose is performed. delivered dose.
For metered-dose inhalers, the target-delivered dose is speci-
fied by the label claim, unless otherwise specified in the
individual monograph. For dry powder inhalers, where the SAMPLING THE DELIVERED DOSE FROM METERED-DOSE
label claim is usually the packaged or metered-dose of drug, INHALERS
the target-delivered dose is specified in the individual mono-
graph and is usually less than the label claim. Its value re- To determine the content of active ingredient in the dis-
flects the expected mean drug content for a large number charged spray from a metered-dose inhaler, use the sam-
of delivered doses collected from the product, using the pling apparatus described below, using a flow rate of 28.3 L
method specified in the monograph. of air per minute (±5%), unless otherwise stated in the indi-
Unless otherwise directed in the individual monograph, vidual monograph.
the drug content of the minimum delivered dose from each Apparatus A—The apparatus (see Figure 1) consists of a
of 10 separate containers is determined in accordance with
the procedure described below.

filter support base with an open-mesh filter support, such as ing the valve for a duration sufficient to ensure that the
a stainless steel screen, a collection tube that is clamped or dose has been completely discharged. Detach the inhaler
screwed to the filter support base, and a mouthpiece from Apparatus A, and disconnect the vacuum. Assay the
adapter to ensure an airtight seal between the collection contents of the apparatus for drug after rinsing the filter
tube and the mouthpiece. Use a mouthpiece adapter that and the interior of the apparatus with a suitable solvent.
ensures that the opening of the inhaler mouthpiece is flush
with the front face or 2.5-mm indented shoulder in the
sample collection tube, as appropriate. The vacuum connec- SAMPLING THE DELIVERED DOSE FROM DRY POWDER
tor is connected to a system comprising a vacuum source, INHALERS
flow regulator, and flowmeter. The source should be capa-
ble of pulling air through the complete assembly, including To determine the content of active ingredient emitted
the filter and the inhaler to be tested, at the desired flow from the mouthpiece of a dry powder inhaler, use Apparatus
rate. When testing metered-dose inhalers, air should be B (see Figure 2).
drawn continuously through the system to avoid loss of
drug into the atmosphere. The filter support base is de-
signed to accommodate 25-mm diameter filter disks. At the
airflow being used, the sample collection tube and the filter
disk must be capable of quantitatively collecting the Deliv-
ered Dose. The filter disk and other materials used in the
construction of the apparatus must be compatible with the
drug and the solvents that are used to extract the drug
from the filter. One end of the collection tube is designed
to hold the filter disk tightly against the filter support base.
When assembled, the joints between the components of the
apparatus are airtight so that when a vacuum is applied to
the base of the filter, all of the air drawn through the collec-
tion device passes through the inhaler.
Procedure—Prepare the inhaler for use according to the
label instructions. Unless otherwise specified in the individ- Fig. 2. Apparatus B: Sampling apparatus for dry powder
ual monograph, with the vacuum pump running, ensuring inhalers. (See Table 1 for component specifications.)
an airflow rate through the inhaler of 28.3 L of air per min-
ute (±5%), discharge the minimum recommended dose into
the apparatus through the mouthpiece adapter by depress-
Table 1. Component Specifications for Apparatus B (see Fig. 2)

Code Item Description Dimensions
A Sample collection tubea See Fig. 2 34.85-mm ID × 12-cm length
B Filterb See Fig. 2 47-mm glass fiber filter
C Connector (e.g., short metal coupling with ≥8-mm ID
low diameter branch to P3)
D Vacuum tubing (e.g., silicon tubing with an A length of suitable tubing ≥8 mm ID with an
outside diameter of 14 mm and internal volume of 25 ± 5 mL.
an internal diameter of 8 mm)
E Two-way solenoid valvec See Fig. 2 2-way, 2-port solenoid valve having an ID ≥8
mm and an opening response time of ≤100
milliseconds.
F Vacuum pumpd See Fig. 2 Pump must be capable of drawing the required
flow rate through the assembled apparatus
with the dry powder inhaler in the mouth-
piece adapter. Connect the pump to the sole-
noid valve using short and wide (≥10-mm ID)
vacuum tubing and connectors to minimize
pump capacity requirements.
G Timere See Fig. 2 The timer switches current directly to the sole-
noid valve for the required duration.
a An example being a Millipore product number XX40 047 00 (Millipore Corporation, 80, Ashby Road, Bedford, MA 01732), modified so that the
exit tube has an ID ≥ 8-mm, fitted with Gelman product number 61631.
b A/E (Gelman Sciences Inc., 600 South Wagner Road, Ann Arbor, MI 48106) or equivalent.
c ASCO product number 8030G13, Automatic Switch Company, 60 Hanover Road, Florham Park, NJ 07932.
d Gast product type 1023, 1423, or 2565 (Gast Manufacturing Inc., PO Box 97, Benton Harbor, MI 49022) or equivalent.
e Eaton Product number 45610-400 (Eaton Corporation, Automotive Products Division, 901, South 12th Street, Watertown, WI 53094) or equiva-
lent.
f An example being a PDM 210 pressure meter (Air-Neotronics Ltd., Neotronics Technology plc, Parsonage Road, Takeley, Bishop’s Stortford, CM22
6PU, UK), or equivalent.

g Parker Hannifin type 8FV12LNSS (Parker Hannifin plc., Riverside Road, Barnstable, Devon EX31 1NP, UK) or equivalent.
h Flow Coefficient, as defined by ISA S75.02 “Control valve capacity test procedure” in Standards and Recommended Practices for Instrumentation
and Control, 10th ed., Vol. 2, 1989. Published by Instrument Society of America, 67 Alexander Drive, P.O. Box 1227, Research Triangle Park, NC
27709, U.S.A.

Table 1. Component Specifications for Apparatus B (see Fig. 2) (Continued)

P1 Pressure tap See Fig. 2 2.2-mm ID, 3.1-mm OD flush with the internal
surface of the sample collection tube, centered
and burr free, 59 mm from its inlet. The pres-
sure taps P1, P2, and P3 must not be open to
the atmosphere during dose collection.
P1, P2, P3 Pressure measurementsf
H Flow-control valveg See Fig. 2 Adjustable regulating valve with maximum Cv ≥
1h.
a An example being a Millipore product number XX40 047 00 (Millipore Corporation, 80, Ashby Road, Bedford, MA 01732), modified so that the
exit tube has an ID ≥ 8-mm, fitted with Gelman product number 61631.
b A/E (Gelman Sciences Inc., 600 South Wagner Road, Ann Arbor, MI 48106) or equivalent.
c ASCO product number 8030G13, Automatic Switch Company, 60 Hanover Road, Florham Park, NJ 07932.
d Gast product type 1023, 1423, or 2565 (Gast Manufacturing Inc., PO Box 97, Benton Harbor, MI 49022) or equivalent.
e Eaton Product number 45610-400 (Eaton Corporation, Automotive Products Division, 901, South 12th Street, Watertown, WI 53094) or equiva-
lent.
f An example being a PDM 210 pressure meter (Air-Neotronics Ltd., Neotronics Technology plc, Parsonage Road, Takeley, Bishop’s Stortford, CM22
6PU, UK), or equivalent.

g Parker Hannifin type 8FV12LNSS (Parker Hannifin plc., Riverside Road, Barnstable, Devon EX31 1NP, UK) or equivalent.
h Flow Coefficient, as defined by ISA S75.02 “Control valve capacity test procedure” in Standards and Recommended Practices for Instrumentation
and Control, 10th ed., Vol. 2, 1989. Published by Instrument Society of America, 67 Alexander Drive, P.O. Box 1227, Research Triangle Park, NC
27709, U.S.A.
This apparatus is capable of sampling the emitted doses at a uum pump is worn or of insufficient capacity. Critical (sonic)
variety of airflow rates. flow conditions in the flow-control valve are required in or-
Apparatus B—The apparatus is similar to that described der to ensure that the volumetric airflow drawn from the
in Figure 1 for testing metered-dose inhalers. In this case, mouthpiece is unaffected by pump fluctuations and changes
however, the filter and collection tube have a larger internal in airflow resistance of the inhaler. Remove the inhaler from
diameter to accommodate 47-mm diameter filter disks. This the mouthpiece adapter and without disturbing the flow-
feature enables dosage collection at higher airflow rates—up control valve, measure the airflow rate drawn from the
to 100 L of air per minute—when necessary. A mouthpiece mouthpiece, Qout, by connecting a flowmeter to the mouth-
adapter ensures an airtight seal between the collection tube piece adaptor in an airtight fashion. Use a flowmeter cali-
and the mouthpiece of the dry powder inhaler being tested. brated for the volumetric flow leaving the meter in an air-
The mouthpiece adapter must ensure that the tip of the tight fashion to directly determine Qout or, if such a meter is
inhaler mouthpiece is flush with the open end of the sample unobtainable, calculate the volumetric flow leaving the
collection tube. Tubing connectors, if they are used, should meter (Qout) using the ideal gas law. For example, for a
have an internal diameter greater than or equal to 8 mm to meter calibrated for the entering volumetric flow (Qin), use
preclude their own internal diameters from creating signifi- the formula:
cant airflow resistance. A vacuum pump with excess capac-
ity must be selected in order to draw air, at the designated Qout = QinP0 / (P0 – ∆P)
volumetric flow rate, through both the sampling apparatus
and the inhaler simultaneously. A timer-controlled, low resis- where P0 is the atmospheric pressure and ∆P is the pressure
tance, solenoid-operated, two-way valve is interposed be- drop over the meter. If the flow rate is greater than 100 L
tween the vacuum pump and the flow-control valve to con- of air per minute, adjust the flow-control valve until Qout
trol the duration of flow. This type of valve enables 4.0 L of equals 100 L per minute; otherwise, record the value of
air (±5%) to be withdrawn from the mouthpiece of the in- Qout, and leave the flow-control valve undisturbed. Define
haler at the designated flow rate. Flow control is achieved the test flow duration, T = 240/Qout, in seconds, so that a
by ensuring that critical (sonic) flow occurs in the flow-con- volume of 4.0 L of air (±5%) is withdrawn from the inhaler
trol valve (absolute pressure ratio P3/P2 ≤ 0.5 under condi- at the test flow rate Qout, and adjust the timer controlling
tions of steady-state flow). the operation of the two-way solenoid valve accordingly.
Prime or load the inhaler with powder for inhalation accord-
Procedure—Operate the apparatus at an airflow rate ing to the labeled instructions. With the vacuum pump run-
that produces a pressure drop of 4 kPa (40.8 cm H2O) over ning and the solenoid valve closed, insert the inhaler
the inhaler to be tested and at a duration consistent with mouthpiece horizontally into the mouthpiece adapter. Dis-
the withdrawal of 4 L of air from the mouthpiece of the charge the powder into the sampling apparatus by activat-
inhaler. [NOTE—If the flow rate and duration are defined ing the timer controlling the solenoid valve and withdraw-
otherwise in the monograph, adjust the system to within ing 4.0 L of air from the inhaler at the previously defined
5% of those values.] Determine the test flow rate using Ap- airflow rate. If the labeled instructions so direct, repeat the
paratus B as follows. Insert an inhaler into the mouthpiece operation so as to simulate the use of the inhaler by the
adapter to ensure an airtight seal. In cases where the drug patient (e.g., inhale two or three times, if necessary, to
packaging modifies the inhaler’s resistance to airflow, use a empty the capsule). Repeat the whole operation n −1 times
loaded, drug-free inhaler (with previously emptied packag- beginning with the text, “Prime or load the inhaler with
ing). In other cases, use an unloaded (drug free) inhaler. powder,” where n is the number of times defined in the
Connect one port of a differential pressure transducer to the labeling as the minimum recommended dose. Detach the
pressure tap, P1, and leave the other open to the atmos- dry powder inhaler from the sampling apparatus, and dis-
phere. Switch on the pump, and open the two-way sole- connect the vacuum tubing, D. Assay the contents of the
noid valve. Adjust the flow-control valve until the pressure apparatus for drug after rinsing the filter and the interior of
drop across the inhaler is 4.0 kPa (40.8 cm H2O). Ensure the apparatus with a suitable solvent. Where specified in
that critical (sonic) flow occurs in the flow-control valve by individual monographs, perform this test under conditions
determining the individual values for absolute pressure, P2 of controlled temperature and humidity.
and P3, so that their ratio P3/P2 is less than or equal to 0.5.
If this criterion cannot be achieved, it is likely that the vac-

Delivered-Dose Uniformity over the Entire DRY POWDER INHALERS

Contents
Apparatus—Use Apparatus B as directed in Sampling the
The test for Delivered-Dose Uniformity over the Entire Con- Delivered Dose from Dry Powder Inhalers under Delivered-Dose
tents is required for inhalers (e.g., metered-dose inhalers or Uniformity at the appropriate airflow rate for testing.
dry powder inhalers) containing multiple doses of drug for- Procedure—Proceed as directed for Procedure in Sam-
mulation (e.g., solution, suspension, or dry powder) either pling the Delivered Dose from Dry Powder Inhalers under De-
in reservoirs or in premetered dosage units (e.g., blisters), livered-Dose Uniformity. A single dose is defined as the num-
and for drug formulations packaged in reservoirs or in multi- ber of actuations stated in the product labeling as the
ple-dose assemblies of premetered dosage units that have a minimum recommended dose. Select a single inhaler and
predetermined dose sequence, where these multiple-dose follow the labeled instructions for loading with powder, dis-
assemblies are labeled for use with a named inhalation de- charging and cleaning throughout. Collect a total of 10
vice. The test for Delivered-Dose Uniformity over the Entire doses—three doses at the beginning, four in the middle [(n/
Contents also ensures that multidose products supply the to- 2) − 1 to (n/2) + 2, where n is the number of minimum
tal number of discharges stated on the label. Unless other- recommended doses on the label], and three at the end—of
wise directed in the individual monograph, the drug con- the labeled contents following the labeled instructions. Prior
tent of at least 9 of the 10 doses collected from one inhaler, to collecting each of the doses to be analyzed, clean the
in accordance with the procedure below, are between 75% inhaler as directed in the labeling.
and 125% of the target-delivered dose, and none is outside
the range of 65% to 135% of the target-delivered dose. If
the contents of not more than 3 doses are outside the Particle Size
range of 75% to 125%, but within the range of 65% to
135% of the target-delivered dose, select 2 additional inhal- The particle or droplet size distribution in the spray dis-
ers, and follow the prescribed procedure for analyzing 10 charged from metered-dose inhalers, and the particle size
doses from each. The requirements are met if not more distribution in the cloud discharged from dry powder inhal-
than 3 results, out of the 30 values, lie outside the range of ers, are important characteristics used in judging inhaler
75% to 125% of the target-delivered dose, and none is performance. While particle size measurement by micros-
outside the range of 65% to 135% of the target-delivered copy can be used to evaluate the number of large particles,
dose. agglomerates, and foreign particulates in the emissions of
metered-dose inhalers (e.g., Epinephrine Bitartrate Inhalation
Aerosol), whenever possible this test should be replaced with
METERED-DOSE INHALERS a method to determine the aerodynamic size distribution of
the drug aerosol leaving the inhaler. The aerodynamic size
Apparatus—Use Apparatus A as directed in Sampling the distribution defines the manner in which an aerosol deposits
Delivered-Dose from Metered-Dose Inhalers under Delivered- during inhalation. When there is a log-normal distribution,
Dose Uniformity at a flow rate of 28.3 L of air per minute the aerodynamic size distribution may be characterized by
(±5%). the mass median aerodynamic diameter (MMAD) and geo-
Procedure—A single dose is defined as the number of metric standard deviation (GSD). The aerodynamic size dis-
sprays specified in the product labeling as the minimum rec- tribution of the drug leaving metered-dose and dry powder
ommended dose. Select a single metered-dose inhaler, and inhalers is determined using Apparatus 1, 2, 3, 4, 5, or 6 as
follow the labeled instructions for priming, shaking, clean- specified in this chapter. A fine particle dose or fine particle
ing, and firing the inhaler throughout. Unless otherwise pre- fraction can also be determined as that portion of the in-
scribed in the patient instructions, shake the inhaler for 5 haler output having an aerodynamic diameter less than the
seconds, and fire one minimum recommended dose to size defined in the individual monograph. This may be ex-
waste. Wait for 5 seconds, and collect the next dose. Detach pected to correlate with the drug dose or that fraction of
the inhaler from Apparatus A, and disconnect the vacuum. the drug dose that penetrates the lung during inhalation.
Assay the contents of the apparatus for drug after rinsing Individual monographs may also define the emitted frac-
the filter and the interior of the apparatus with a suitable tions of the delivered dose in more than one aerodynamic
solvent. Collect two more doses, allowing at least 5 seconds size range.
between doses. Discharge the device to waste, waiting for
not less than 5 seconds between actuations (unless other-
wise specified in the individual monograph), until (n/2) + 1 AERODYNAMIC SIZE DISTRIBUTION
minimum recommended doses remain, in which n is the
number of minimum recommended doses on the label. Col- Cascade impaction devices classify aerosol particles and
lect four more doses, allowing at least 5 seconds between droplets on the basis of those particles’ aerodynamic diame-
doses, unless otherwise specified in the individual mono- ters. The principle of their operation, whereby they separate
graph. Discharge the device to waste, as before, until three aerosol particles and droplets from a moving airstream on
doses remain. Collect the final three doses, allowing at least the basis of particle or droplet inertia, is shown in Figure 3.
5 seconds between doses. Note that the rate of discharges
to waste should not be such to cause excessive canister
cooling.

Fig. 4a. Apparatus 1: Expanded view of induction port for use with metered-dose and dry powder inhalers.
Because the dimensions of the induction port used to con-

nect inhalers to the cascade impactors and impingers
(shown in Apparatus 1, 2, 3, 4, 5, and 6) also define the
mass of drug that enters the aerodynamic sizing device,
these are carefully defined and, where possible, are held
constant between each apparatus (see Figures 4, 6, 7, 8, and
9).
Fig. 4. Apparatus 1: Assembly of induction port and en-

trance cone mounted on cascade impactor.
Because the size distributions produced by different im-

pactors are often a function of impactor design and the
airflow rate through them, there is a need to standardize
the instruments that are used to test inhalers (i.e., Apparatus
1 or 6 for metered-dose inhalers) or to provide guidelines
on system suitability where different apparatuses may be
used (i.e., Apparatus 2, 3, 4, or 5 for dry powder inhalers).
Because of the varied nature of the formulations and de-
Fig. 3. Schematic representation of the principle of opera- vices being tested, the cascade impaction system and tech-
tion of cascade impactors. (A single jet per impactor stage is nique selected for testing an inhaler should fulfill a number
shown. Impactors with multiple jets in each stage function of criteria.
in the same manner.)
Stage Mensuration—Manufacturers of cascade impaction
devices provide a definitive calibration for the separation

Fig. 4b. Apparatus 1: Expanded view of the entrance cone for mounting induction port on the Andersen cascade impactor
without preseparator. Material may be aluminum, stainless steel, or other suitable material. Surface roughness (Ra) should be
approximately 0.4 µm.
Fig. 5. Apparatus 2, 3, 4, or 5: General control equipment. (See Table 3 for component specifications.)
characteristics of each impaction stage in terms of the rela- monograph, the selected technique should ensure that not
tionship between the stage collection efficiency and the aer- more than 5% of the inhaler’s total delivered drug mass
odynamic diameter of particles and droplets passing (into the impactor) is subject to loss between the impaction
through it as an aerosol. Calibration is a property of the jet device’s sample collection surfaces. In the event that inter-
dimensions, the spatial arrangement of the jet and its collec- stage drug losses are known to be greater than 5%, either
tion surface, and the airflow rate passing through it. Be- the procedure should be performed in such a way that wall
cause jets can corrode and wear over time, the critical losses are included along with the associated collection
dimensions of each stage, which define that impaction plate, or an alternative apparatus should be used. As an ex-
stage’s calibration, must be measured on a regular basis. ample, the following procedures described for Apparatus 1
This process, known as stage mensuration, replaces the and 3 have been written to include wall losses along with
need for repetitive calibration (using standard aerosols) and the associated collection plate. Provided, however, that such
ensures that only devices that conform to specifications are losses are known to be less than or equal to 5% of the total
used for testing inhaler output. The process involves the delivered drug mass into the impactor and that there are no
measurement and adjustment of the critical dimensions of instructions to the contrary in an individual monograph, the
the instrument. technique may be simplified by only assaying drug on the
Interstage Drug Loss (wall losses)—Where method varia- collection plates.
tions are possible and there is no apparatus specified in the

more than 125% of the average minimum recommended

dose determined during testing for Delivered-Dose Uniform-
ity. This is not a test of the inhaler but serves to ensure that
the test results are valid.
Use one of the multistage impaction devices shown be-
low, or an equivalent, to determine aerodynamic particle
size distributions of drugs leaving the mouthpieces of me-
tered-dose or dry powder inhalers. Apparatus 1 and 6
[Figures 4 and 9 (without preseparator), respectively] are in-
tended for use with metered-dose inhalers at a single airflow
rate. Apparatus 2, 3, 4, and 5 (Figures 6, 7, 8, and 9, respec-
tively) are intended for use with dry powder inhalers at the
appropriate airflow rate, Qout, determined earlier, provided
that the value of Qout falls in the range 30–100 L per
minute.
NOTE—If Qout is greater than 100 L per minute, testing
should be performed with Qout set at 100 L per minute; if
Qout is less than 30 L per minute, testing is performed with
Qout at 30 L per minute.
Apparatus 1 for Metered-Dose Inhalers—Use this appa-
ratus, or an equivalent, at a flow rate of 28.3 L per minute
(±5%), as specified by the manufacturer of the cascade
impactor.
Design—The design and assembly of this apparatus and
the induction port to connect the device to an inhaler are
shown in Figures 4, 4a, and 4b1.
Critical engineering dimensions applied by manufacturers
to the stages of Apparatus 1 are provided in Table 2. During
use, some occlusion and blockage of jet nozzles may occur
and therefore, “in use” mensuration tolerances need to be
justified.
Fig. 6. Apparatus 2: Assembly of induction port, stage col- Table 2. Critical Dimensions for the Jet Nozzles of
lector, and filter holder. Apparatus 1
(Marple-Miller impactor, Model 160 with USP induction
port.) Stage # Number of Jets Nozzle Diameter (mm)
0 96 2.55 ± 0.025
1 96 1.89 ± 0.025
Re-Entrainment—Where method variations are possible, 2 400 0.914 ± 0.0127
the selected technique should seek to minimize particle re-
entrainment (from an upper to a lower impaction stage) on 3 400 0.711 ± 0.0127
stages that contribute to size fractions defined in the indi- 4 400 0.533 ± 0.0127
vidual monograph, especially where this may affect the 5 400 0.343 ± 0.0127
amounts of drug collected. Minimizing the number of sam- 6 400 0.254 ± 0.0127
pled doses, the use of coated particle collection surfaces, 7 201 0.254 ± 0.0127
and proving that multiple-dose techniques produce statisti-
cally similar results to those from smaller numbers of doses, Procedure—Set up the multistage cascade impactor as de-
are all methods that can be used for this purpose. In the scribed in the manufacturer’s literature with an after filter
event that re-entrainment cannot be avoided, the number below the final stage to capture any fine particles that oth-
of doses collected, the time interval between doses, and the erwise would escape from the device. To ensure efficient
total duration of airflow through the cascade impaction de- particle capture, coat the particle collection surface of each
vice should be standardized. Under these circumstances, the stage with glycerol, silicone oil, or other suitable liquid typi-
presentation of impaction data should not presume the va- cally deposited from a volatile solvent, unless it has been
lidity of the impactor’s calibration (i.e., aerodynamic diame- demonstrated to be unnecessary. Attach the induction port
ter ranges should not be assigned to drug masses collected and mouthpiece adapter to produce an airtight seal be-
on specific stages). tween the inhaler mouthpiece and the induction port as
By using appropriate assay methods and a suitable men- shown in Figure 4. Use a mouthpiece adapter that ensures
surated impaction device, aerodynamic particle size distribu- that the tip of the inhaler mouthpiece is flush with the open
tions can be determined for drugs leaving the mouthpieces end of the induction port. Ensure that the various stages of
of metered-dose or dry powder inhalers. If temperature or
humidity limits for use of the inhaler are stated on the label, 1 A suitable cascade impactor is available as Model Mk II from Thermo-Elec-
it may be necessary to control the temperature and humid- tron, 27 Forge Parkway, Franklin, MA 02038. The impactor is used without
the preseparator. The inhaler is connected to the impactor via the induction
ity of the air surrounding and passing through the device to port, atop the entrance cone shown in Figure 4. If an equivalent impactor is
conform to those limits. Ambient conditions are presumed, employed, the induction port in Figure 4a should be used, although the
unless otherwise specified in individual monographs. entrance cone (Fig. 4b) should be replaced with one to fit the impactor in
question. Note that the internal surfaces of the induction port (Fig. 4a) are
Mass Balance—In addition to the size distribution, good designed to fit flush with their counterparts in the entrance cone (Fig. 4b).
analytical practice dictates that a mass-balance be per- This design avoids aerosol capture at the junction of the two pipes.
formed in order to confirm that the amount of the drug
discharged from the inhaler, which is captured and meas-
ured in the induction port-cascade impactor apparatus, is
within an acceptable range around the expected value. The
total mass of drug collected in all of the components (mate-
rial balance) divided by the total number of minimum rec-
ommended doses discharged is not less than 75% and not

Fig. 7. Apparatus 3: Expanded views of top for the Andersen preseparator adapted to the USP induction port. Material may
be aluminum, stainless steel, or other suitable material; interior bore should be polished to surface roughness (Ra) approxi-
mately 0.4 µm.
the cascade impactor are connected with airtight seals to seconds and discharge one delivery to waste. With the vac-
prevent leaks. Turn on the vacuum pump to draw air uum pump running, insert the mouthpiece into the mouth-
through the cascade impactor, and calibrate the airflow piece adapter and immediately fire the minimum recom-
through the system with an appropriate flowmeter attached mended dose into the cascade impactor. Keep the valve
to the open end of the induction port. Adjust the flow- depressed for a duration sufficient to ensure that the dose
control valve on the vacuum pump to achieve steady flow has been completely discharged. If additional sprays are re-
through the system at the required rate, and ensure that quired for the sample, wait for 5 seconds before removing
the airflow through the system is within ±5% of the flow the inhaler from the mouthpiece adapter, shake the inhaler,
rate specified by the manufacturer. Unless otherwise pre- reinsert it into the mouthpiece adapter, and immediately fire
scribed in the patient instructions, shake the inhaler for 5 the next minimum recommended dose. Repeat until the re-

quired number of doses have been discharged. The number achieve a steady flow through the system at the required
of minimum recommended doses discharged must be suffi- rate, Qout, so that Qout is within ±5% of the value deter-
cient to ensure an accurate and precise determination of mined during testing for Delivered-Dose Uniformity. Ensure
Aerodynamic Size Distribution. [NOTE—The number of mini- that critical flow occurs in the flow-control valve, at the air-
mum recommended doses is typically not greater than 10.] flow rate to be used during testing, by using the following
After the last dose has been discharged, remove the inhaler procedure. With the inhaler in place, and the intended flow
from the mouthpiece adapter. Rinse the mouthpiece running, measure the absolute pressure on both sides of the
adapter and induction port with a suitable solvent, and di- flow-control valve (P2 and P3 in Figure 5). A ratio of P3/
lute quantitatively to an appropriate volume. Disassemble P2 ≤ 0.5 indicates critical flow. Switch to a more powerful
the cascade impactor, place each stage and its associated pump, and remeasure the test flow rate if P3/P2 > 0.5. Ad-
collection plate or filter in a separate container, and rinse just the timer controlling the operation of the two-way sole-
the drug from each of them. [NOTE—If it has been deter- noid valve so that it opens this valve for a duration of T
mined that wall losses in the impactor are less than or equal seconds as determined during testing for Delivered-Dose Uni-
to 5%, then the collection plates only may be used.] formity. Prime or load the dry powder inhaler with powder
Dilute each quantitatively to an appropriate volume. Us- for inhalation according to the labeled instructions. With the
ing the method of analysis specified in the individual mono- vacuum pump running and the two-way solenoid valve
graph, determine the mass of drug collected in each of the closed, insert the inhaler mouthpiece, held horizontally, into
components. To analyze the data, proceed as directed under the induction port mouthpiece adapter. Discharge the pow-
Data Analysis. der into the apparatus by opening the two-way solenoid
Apparatus 2 for Dry Powder Inhalers— valve for a duration of T seconds. After the two-way sole-
Design—The design and assembly of Apparatus 2, and noid valve has closed, remove the inhaler from the mouth-
the induction port to connect the device to an inhaler, are piece adapter. If additional doses are required for the sam-
shown in Figure 6.2 [NOTE—The induction port is shown in ple, reload the inhaler according to the labeled instructions,
detail in Figure 4a.] The impactor has five impaction stages reinsert the mouthpiece into the mouthpiece adapter, and
and an after filter. At a volumetric airflow rate of 60 L per repeat the operation until the required number of doses
minute (the nominal flow rate, Qn), the cutoff aerodynamic have been discharged. After discharge of the last dose,
diameters D50,Qn of Stages 1 to 5 are 10, 5, 2.5, 1.25, and switch off the vacuum pump.
0.625 µm, respectively. The after filter effectively retains Rinse the mouthpiece adapter and induction port with a
aerosolized drug in the particle size range up to 0.625 µm. suitable solvent, and quantitatively dilute to an appropriate
Set up the multistage cascade impactor with the control volume. Disassemble the cascade impactor, and place the
system as specified in Figure 5. To ensure efficient particle after filter in a separate container. Rinse the drug from each
capture, coat the particle collection surface of each stage of the stages and the filter, and quantitatively dilute each to
with glycerol, silicone oil, or other suitable liquid typically an appropriate volume. Using the method of analysis speci-
deposited from a volatile solvent, unless it has been demon- fied in the individual monograph, determine the mass of
strated to be unnecessary. Assemble the impactor as de- drug collected in each of the components. Determine the
scribed in the manufacturer’s literature with an after filter cutoff diameters of each of the individual stages of the im-
below the final stage to capture any fine particles that oth- pactor, at the value of Q = Qout employed in the test by the
erwise would escape from the device. Attach the induction formula:
port and mouthpiece adapter to produce an airtight seal D50,Q = D50,Qn(Qn/Q) / , (Eq. 1)
1
2
between the inhaler mouthpiece and the induction port.

Use a mouthpiece adapter that ensures that the tip of the where D50,Q is the cutoff diameter at the flow rate, Q, em-
inhaler mouthpiece is flush with the open end of the induc- ployed in the test, and the subscript, n, refers to the nomi-
tion port. Ensure that the various stages of the cascade im- nal values determined when Qn equals 60 L per minute.
pactor are connected with airtight seals to prevent leaks. Thus, when Q equals 40 L per minute, the cutoff diameter
Turn on the vacuum pump, open the solenoid valve, and of Stage 2 is given by the formula:
calibrate the airflow through the system as follows. Connect
a flowmeter to the induction port. Use a flowmeter cali- D50,40LPM = 5 µm × [60/40]1/2 = 6.1 µm.
brated for the volumetric flow leaving the meter to directly
determine Qout, or, if such a meter is unobtainable, calculate
the volumetric flow leaving the meter (Qout) using the ideal General Procedure—Perform the test using Apparatus 2 at
gas law. For example, for a meter calibrated for the entering the airflow rate, Qout determined earlier, during testing for
volumetric flow (Qin), use the formula: Delivered-Dose Uniformity, provided Qout is less than or equal
to 100 L per minute. [NOTE—If Qout is greater than 100 L
Qout = QinP0/(P0 – ∆P) per minute, use an airflow rate of 100 L per minute.] Con-
nect the apparatus to a flow control system that is based
where P0 is the atmospheric pressure and ∆P is the pressure upon critical (sonic) flow as specified in Figure 5 (see also
drop over the meter. Adjust the flow-control valve to Table 3).
2 The cascade impactor is available as the Model 160 Marple-Miller Impactor
from MSP Corporation, Minneapolis, MN. The inhaler should be connected
to the impactor via the induction port, shown in Figure 4a.

Table 3. Component Specifications for Figure 5

A Connector (e.g., short metal coupling with ≥ 8-mm ID
low diameter branch to P3)
B Vacuum tubing (e.g., silicon tubing with an A length of suitable tubing ≥8 mm ID with an internal vol-
outside diameter of 14 mm and ume of 25 ± 5 mL.
an internal diameter of 8 mm)
C Two-way solenoid valvea See Fig. 5 2-way, 2-port solenoid valve having an ID ≥8 mm and an
opening response time of ≤100 milliseconds.
D Vacuum pumpb See Fig. 5 Pump must be capable of drawing the required flow rate
through the assembled apparatus with the dry powder in-
haler in the mouthpiece adapter. Connect the pump to the
solenoid valve using short and wide (≥ 10-mm ID) vacuum
tubing and connectors to minimize pump capacity require-
ments.
E Timerc See Fig. 5 The timer switches current directly to the solenoid valve for
the required duration.
P2, P3 Pressure measurements Determine under steady-state flow conditions with an abso-
lute pressure transducer.
F Flow control valved See Fig. 5 Adjustable regulating valve with maximum Cv ≥ 1.
aAn example being ASCO product number 8030G13 (Automatic Switch Company, 60 Hanover Road, Florham Park, NJ 07932) or equivalent. See
also Footnote h in Table 1.

bGast product type 1023, 1423, or 2565 (Gast Manufacturing Inc., PO Box 97, Benton Harbor, MI 49022) or equivalent.
cAn example being Eaton Product number 45610-400 (Eaton Corporation, Automotive Products Division, 901 South 12th Street, Watertown, WI
53094) or equivalent.
dParker Hannifin type 8FV12LNSS, or equivalent (Parker Hannifin plc, Riverside Road, Barnstable, Devon EX31 1NP, UK). See also Footnote h in
Table 1.
Table 4. Component Units of Multistage Liquid Impinger (see Fig. 8)

Code1 Item Description Dimensions2
A,H Jet tube Metal tube screwed onto partition wall sealed by gas- see Figure 8a
ket (C), polished inner surface
B,G Partition wall Circular metal plate, diameter 120
Thickness see Figure 8a
C Gasket e.g., PTFE to fit jet tube
D Impaction plate Porosity O sintered-glass disk,
Diameter see Figure 8a
E Glass cylinder Plane polished cut glass tube
Height, including gaskets 46
Outer diameter 100
Wall thickness 3.5
Sampling port (F) diameter 18
Stopper in sampling port ISO 24/25
J Metal frame L-profiled circular frame with slit
Inner diameter to fit impaction plate
Height 4
Thickness of horizontal section 0.5
Thickness of vertical section 2
K Wire Steel wire interconnecting metal frame and sleeve
(two for each frame)
Diameter 1
L Sleeve Metal sleeve secured on jet tube by screw
Inner diameter to fit jet tube
Height 6
Thickness 5
M Gasket e.g., silicone to fit glass cylinder
N Bolt Metal bolt with nut (six pairs), length 205
Diameter 4
P O-ring Rubber O-ring, diameter × thickness 66.34 × 2.62
Q O-ring Rubber O-ring, diameter × thickness 29.1 × 1.6
R Filter holder Metal housing with stand and outlet see Figure 8b
S Filter support Perforated sheet metal, diameter 65
1See Fig. 8.
2Measurements in mm unless otherwise stated.

Table 4. Component Units of Multistage Liquid Impinger (see Fig. 8) (Continued)

Code1 Item Description Dimensions2
Hole diameter 3
Distance between holes (center-points) 4
T Snap-locks
U Multi-jet tube Jet tube (H) ending in multijet arrangement see inserts Figure 8a
V Outlet Outlet and nozzle for connection to vacuum Internal diameter ≥ 10 (Figure 8b)
1See Fig. 8.
2Measurements in mm unless otherwise stated.
Fig. 8a. Apparatus 4: Details of jet tube and impaction plate.

Inserts show end of multi-jet tube U leading to Stage 4.
(See Table 5 for dimension specifications.)
Fig. 8. Apparatus 4: Schematic of multistage liquid im-

pinger. (See Table 4 for component specifications.)
Fig. 8b. Apparatus 4: Expanded view of Stage 5. (See Table

4 for component specifications.)
Under steady flow conditions, at the appropriate volumet-

ric airflow rate through the entire apparatus, ensure that
critical (sonic) flow occurs in the flow control valve by deter-
mining the individual values for absolute pressure, P2 and
P3, so that their ratio P3/P2 is less than or equal to 0.5.
Coat the particle collection surface of each of the stages of
the cascade impactor to ensure that particles that have im-
pacted on a given stage are not re-entrained in the flowing

Table 5. Apparatus 4: Dimensions1 of Jet Tube with Impaction Plate (see Fig. 8a).
Filter
Type Code2 Stage 1 Stage 2 Stage 3 Stage 4 (Stage 5)
Distance 1 9.5 (−.0, +.5) 5.5 (−.0, +.5) 4.0 (−.0, +.5) 6.0 (−.0, +.5) n.a.
Distance 2 26 31 33 30.5 0
Distance 3 8 5 5 5 5
Distance 4 3 3 3 3 n.a.
Distance 5 0 3 3 3 3
Distance 63 20 25 25 25 25
Distance 7 n.a. n.a. n.a. 8.5 n.a.
Diameter c 25 14 8.0(±0.1) 21 14
Diameter d 50 30 20 30 n.a.
Diameter e 27.9 16.5 10.5 23.9 n.a.
Diameter f 31.75 (−.05, +.00) 22 14 31 22
Diameter g 25.4 21 13 30 21
Diameter h n.a. n.a. n.a. 2.70 (±.05) n.a.
Diameter j n.a. n.a. n.a. 6.3 n.a.
Diameter k n.a. n.a. n.a. 12.6 n.a.
Radius4 r 16 22 27 28.5 0
Radius4 s 46 46 46 46 n.a.
Radius4 t n.a. 50 50 50 50
Angle w 10° 53° 53° 53° 53°
Angle u n.a. n.a. n.a. 45° n.a.
Angle v n.a. n.a. n.a. 60° n.a.
1Measurements in mm with tolerances according to ISO 2768-m, unless otherwise stated.
2See Fig. 8a.
3Including gasket.
4Relative centerline of stage compartment.
n.a.: not applicable.
airstream, unless this has been shown to be unnecessary. priate precautions (alternative solvents, use of vapor traps,
Analyze the data as directed under Data Analysis. minimal pump operating times, etc.) to ensure operator safety
Apparatus 3 for Dry Powder Inhalers— during testing.] Attach a molded mouthpiece adapter to the
Design—Apparatus 3 is identical to Apparatus 1 (Figure 4), end of the induction port to produce an airtight seal be-
except that the manufacturer’s preseparator is added atop tween the inhaler mouthpiece and the induction port. Use a
Stage 0 to collect large masses of noninhalable powder prior mouthpiece adapter that ensures that the tip of the inhaler
to their entry into the impactor, and the outlet nipple, used mouthpiece is flush with the open end of the induction
to connect to vacuum tubing B (Figure 5), is replaced with port. Ensure that the various stages of the cascade impactor
one having an internal diameter ≥ 8 mm. To connect the are connected with airtight seals to prevent leaks.
preseparator of the impactor to the induction port (Figure Turn on the vacuum pump, open the two-way solenoid
4a), a specially designed top for the preseparator must be valve, and calibrate the airflow through the system as fol-
used. This is shown in Figure 7.3 The impactor, therefore, lows. Prime or load the dry powder inhaler with powder for
has eight stages, a preseparator (to collect large particu- inhalation according to the labeled instructions. With the
lates), and an after filter. At a volumetric airflow rate of 28.3 vacuum pump running and the two-way solenoid valve
L per minute (the nominal flow rate, Qn), the cutoff aerody- closed, insert the inhaler mouthpiece, held horizontally, into
namic diameters D50,Qn of Stages 0 to 7 are 9.0, 5.8, 4.7, the induction port mouthpiece adapter. Once the inhaler is
3.3, 2.1, 1.1, 0.7, and 0.4 µm, respectively. The after filter positioned, discharge the powder into the apparatus by acti-
effectively retains aerosolized drug in the particle size range vating the timer and opening the two-way solenoid valve
up to 0.4 µm. Connect the cascade impactor into the con- for the required duration, T ± 5%, as determined during
trol system specified in Figure 5. Omit Stage 6 and Stage 7 testing for Delivered-Dose Uniformity. After the two-way sole-
from the impactor if the test flow rate, Qout, used during noid valve has closed, remove the inhaler from the mouth-
testing for Delivered-Dose Uniformity was greater than or piece adapter. If additional doses are required for the sam-
equal to 60 L per minute. To ensure efficient particle cap- ple, reload the inhaler according to the labeled instructions,
ture, coat the particle collection surface of each stage with reinsert the mouthpiece into the mouthpiece adapter, and
glycerol, silicone oil, or other suitable liquid typically depos- repeat the operation until the required number of doses
ited from a volatile solvent, unless it has been demonstrated have been discharged. After discharge of the last dose, re-
to be unnecessary. Assemble the impactor as described in move the inhaler from the mouthpiece adapter, and switch
the manufacturer’s literature with an after filter below the off the vacuum pump.
final stage to capture any fine particles that otherwise would Carefully disassemble the apparatus. Using a suitable sol-
escape from the device. Place an appropriate volume (up to vent, rinse the drug from the mouthpiece adapter, induc-
10 mL) of an appropriate solvent into the preseparator, or tion port, and preseparator, and quantitatively dilute to an
coat the particle collection surfaces of the preseparator to appropriate volume. Rinse the drug from each stage, and
prevent re-entrainment of impacted particles. [Caution— the impaction plate immediately below, into appropriately
Some solvents form flammable vapor-air mixtures that may be sized flasks. Quantitatively dilute each flask to an appropri-
ignited during passage through a vacuum pump. Take appro- ate volume. Using the method of analysis specified in the
individual monograph, determine the mass of drug col-
3The cascade impactor is available as the Andersen 1ACFM Non-Viable Cas- lected in each of the samples. The aerodynamic cutoff diam-
cade Impactor (Mark II) from Thermo-Electron, 27 Forge Parkway, Franklin, eters of the individual stages of this device, in the airflow
MA 02038. The impactor is used with the preseparator.
range between 30 and 100 L per minute, are currently not

well established. Do not use the formula in Equation 1 to through the system as follows. Connect a flowmeter, cali-
calculate cutoff diameters. brated for the volumetric flow rate leaving the meter, to the
Procedure—Proceed as directed in the General Procedure induction port. Adjust the flow-control valve to achieve a
under Apparatus 2, except to use Apparatus 3. steady flow through the system at the required rate, Qout, so
Apparatus 4 for Dry Powder Inhalers— that Qout is within ±5% of the value determined during test-
NOTE—Apparatus 4, the multistage liquid impinger, has a ing for Delivered-Dose Uniformity. Ensure that critical flow oc-
small number of stages and is used extensively outside the curs in the flow-control valve, at the value of Qout to be
USA. It is provided here for the benefit of users in countries used during testing, using the following procedure. With
other than the USA. the inhaler in place, and the intended flow running, meas-
ure the absolute pressure on both sides of the flow-control
Design—The design and assembly of Apparatus 4 are valve (P2 and P3 in Figure 5). A ratio of P3/P2 ≤ 0.5 indi-
shown in Figs. 8, 8a, and 8b.4 The induction port, used to cates critical flow. Switch to a more powerful pump, and
connect the device to an inhaler, is shown in Fig. 4a. The remeasure the test flow rate if P3/P2 > 0.5. Adjust the timer
device is a multi-stage liquid impinger consisting of impac- controlling the operation of the two-way solenoid valve so
tion Stages 1, 2, 3, and 4 and an integral after filter (Stage that it opens that valve for the same duration, T, as used
5). The collection stages of the liquid impinger (see Fig. 8 during testing for Delivered-Dose Uniformity. Dispense 20 mL
and Table 4) are kept moist, unlike those of traditional im- of a solvent, capable of dissolving the drug, into each of the
pactors, such as Apparatus 1, 2, 3, 5, and 6; wetting may four upper stages of Apparatus 4, and replace the stoppers.
produce an effect similar to coating the stages of Apparatus [Caution—Some solvents form flammable vapor-air mixtures
2, 3, 5, and 6 at certain flow rates, although this should be that may be ignited during passage through a vacuum pump.
confirmed by demonstrating control over re-entrainment as Take appropriate precautions (alternative solvents, use of vapor
described earlier. An impaction stage comprises an upper traps, minimal pump operating times, etc.) to ensure operator
horizontal metal partition wall (B) through which a metal safety during testing.] Tilt the apparatus to wet the stoppers,
inlet jet tube (A) with its impaction plate (D) is protruding; thereby neutralizing their electrostatic charge. Adjust the
a glass cylinder (E) with sampling port (F), forming the ver- timer controlling the operation of the two-way solenoid
tical wall of the stage; and a lower horizontal metal partition valve so that it opens the valve for the same duration, T, as
wall (G) through which a jet tube (H) connects to the lower used during testing for Delivered-Dose Uniformity. Prime or
stage. The tube into Stage 4 (U) ends in a multi-jet arrange- load the dry powder inhaler with powder for inhalation ac-
ment. The impaction plate (D) is secured in a metal frame cording to the labeled instructions. With the vacuum pump
(J), which is fastened by two wires (K) to a sleeve (L) se- running and the two-way solenoid valve closed, insert the
cured on the jet tube (C). For more detail of the jet tube inhaler mouthpiece, held horizontally, into the induction
and impaction plate, see Fig. 8a. The horizontal plane of the port mouthpiece adapter. Discharge the powder into the
collection plate is perpendicular to the axis of the jet tube apparatus by activating the timer and opening the two-way
and centrally aligned. The upper surface of the impaction solenoid valve for the required duration, T ± 5%. After the
plate is slightly raised above the edge of the metal frame. A two-way solenoid valve has closed, remove the inhaler from
recess around the perimeter of the horizontal partition wall the mouthpiece adapter. If additional doses are required for
guides the position of the glass cylinder. The glass cylinders the sample, reload the inhaler according to the labeled in-
are sealed against the horizontal partition walls with gaskets structions, reinsert the mouthpiece into the mouthpiece
(M) and clamped together by six bolts (N). The sampling adapter, and repeat the operation until the required number
ports are sealed by stoppers. The bottom side of the lower of doses have been discharged. After discharge of the last
partition wall of Stage 4 has a concentric protrusion fitted dose, switch off the vacuum pump.
with a rubber O-ring (P) that seals against the edge of a Dismantle the filter stage of Apparatus 4. Carefully remove
filter placed in the filter holder. The filter holder (R) is a the filter, and extract the drug with solvent. Rinse the
basin with a concentric recess in which a perforated filter mouthpiece adapter and induction port with a suitable sol-
support (S) is flush-fitted. The filter holder is designed for vent, and quantitatively dilute to an appropriate volume.
76-mm diameter filters. The whole impaction stage assem- Rinse the inside of the inlet jet tube to Stage 1 (Figure 8),
bly is clamped onto the filter holder by two snap locks (T). allowing the solvent to flow into the stage. Rinse the drug
The impinger is equipped with an induction port (Fig. 4a) from the inner walls and the collection plate of each of the
that fits onto the Stage 1 inlet jet tube. A rubber O-ring on four upper stages of the apparatus, into the solution in the
the jet tube provides an airtight connection to the induction respective stage, by tilting and rotating the apparatus, while
port. An elastomeric mouthpiece adapter to fit the inhaler ensuring that no liquid transfer occurs between the stages.
being tested provides an airtight seal between the inhaler Using the method of analysis specified in the individual
and the induction port. monograph, determine the mass of drug collected in each
At a volumetric airflow rate of 60 L per minute (the nomi- of the six volumes of solvent. Ensure that the method cor-
nal flow rate, Qn), the cutoff aerodynamic diameters D50,Qn rects for possible evaporation of the solvent during the test.
of Stages 1 to 4 are 13.0, 6.8, 3.1, and 1.7 µm, respec- This may involve the use of an internal standard (of known
tively. The after filter effectively retains aerosolized drug in original concentration in the solvent and assayed at the
the particle size range up to 1.7 µm. Ensure that Apparatus same time as the drug) or the quantitative transfer of the
4 is clean and free of drug solution from any previous tests. liquid contents from each of the stages, followed by dilution
Place a 76-mm diameter filter in the filter stage, and assem- to a known volume. Determine the cutoff diameters of each
ble the apparatus. Use a low pressure filter capable of quan- of the individual stages of the impactor, at the value of Q =
titatively collecting the passing drug aerosol, which also al- Qout employed in the test by the formula:
lows a quantitative recovery of the collected drug. Set up
Apparatus 4 using the control system as specified in Figure 5. D50,Q = D50,Qn (Qn/Q)1/2
Attach the induction port (Figure 4a) and mouthpiece
adapter to produce an airtight seal between the inhaler where D50,Q is the cutoff diameter at the flow rate, Q, em-
mouthpiece and the induction port. Use a mouthpiece ployed in the test, and the subscript, n, refers to the nomi-
adapter that ensures that the tip of the inhaler mouthpiece nal values determined when Qn equals 60 L of air per min-
is flush with the open end of the induction port. Ensure that ute. Thus, when Q equals 40 L of air per minute, the cutoff
the various stages of the apparatus are connected with air- diameter of Stage 2 is given by the formula:
tight seals to prevent leaks. Turn on the vacuum pump,
open the two-way solenoid valve, and calibrate the airflow D50,40LPM = 6.8 µm × (60/40)1/2 = 8.3 µm.
4The five-stage impinger is available from Copley Instruments, plc, Notting-
ham, UK. The inhaler should be connected to the impactor via the induction Procedure—Proceed as directed in the General Procedure
port, shown in Fig. 4 and Fig. 4a. under Apparatus 2, except to use Apparatus 4.

Table 6. Critical Dimensions for Apparatus 5 and 6

Description Dimension (mm)
Preseparator (dimension a—see Figure 9d) 12.80 ± 0.05
Stage 11 Nozzle diameter 14.30 ± 0.05
MOC 1 approximately 0.070
Cup Depth (Dimension b—see Figure 9b) 14.625 ± 0.10
Collection cup surface roughness 0.5 to 2 µm
Stage 1 Nozzle to seal body distance2—dimension c 0 ± 1.18
Stage 2 Nozzle to seal body distance2—dimension c 5.236 ± 0.736
MOC Nozzle to seal body distance2—dimension c 14.429 – 14.571
1See Figure 9c.
2See Figure 9b.
Apparatus 5 for Dry Powder Inhalers— 7 are 8.06, 4.46, 2.82, 1.66, 0.94, 0.55 and 0.34 µm, re-
Design—The design and assembly of Apparatus 55 are spectively. The apparatus contains a terminal micro-orifice
shown in Figures 9, 9a, 9b, 9c, and 9d. The induction port, collector (MOC) that for most formulations may eliminate
used to connect the device to an inhaler, is shown in Figure the need for a final filter as determined by method valida-
4a. The device is a cascade impactor with seven stages and tion. The MOC is an impactor nozzle plate and collection
a micro-orifice collector (MOC). Over the design flow-rate cup. The nozzle plate contains, nominally, 4032 jets, each
range of 30 to 100 L per minute, the 50% efficiency cut-off approximately 70 µm in diameter. Most particles not cap-
diameters of the stages (D50 values) range between 0.24 µm tured on Stage 7 of the impactor will be captured on the
to 11.7 µm, evenly spaced on a logarithmic scale. In the cup surface below the MOC. (For impactors operated at 60
design flow-rate range, there are always at least five stages L per minute, the MOC is capable of collecting 80% of
with D50 values between 0.5 µm and 6.5 µm. The collection 0.14-µm particles). For formulations with a significant frac-
efficiency curves for each stage are sharp and minimize tion of particles not captured by the MOC, there is an op-
overlap between stages. Material may be aluminum, stain- tional filter holder that can replace the MOC or be placed
less steel, or other suitable material. downstream of the MOC containing a suitable after-filter
The impactor layout has removable impaction cups with (glass fiber is often suitable).
all the cups in one plane (Figures 9–9c). There are three Procedure—Assemble the apparatus with the preseparator
main sections to the impactor: the bottom frame that holds (Figure 9d), unless experiments have shown that its omission
the impaction cups, the seal body that holds the jets, and does not result in increased interstage drug losses (>5%) or
the lid that contains the interstage passageways (shown in particle re-entrainment, in which case the preseparator may
Figures 9–9b). Multiple nozzles are used at all but the first be omitted.
stage (Figure 9c). The flow passes through the impactor in a Place cups into the apertures in the cup tray. To ensure
saw-tooth pattern. efficient particle capture, coat the particle collection surface
Stage mensuration is performed periodically together with of each stage with glycerol, silicone oil, or other suitable
confirmation of other dimensions critical to the effective op- liquid typically deposited from a volatile solvent, unless it
eration of the impactor. Critical dimensions are provided be- has been demonstrated to be unnecessary. Insert the cup
low in Table 6. tray into the bottom frame, and lower into place. Close the
In routine operation, the seal body and lid are held to- impactor lid with the seal body attached, and operate the
gether as a single assembly. The impaction cups are accessi- handle to lock the impactor together so that the system is
ble when this assembly is opened at the end of an inhaler airtight.
test. The cups are held in a support tray, so that all cups The preseparator may be assembled as follows: assemble
can be removed from the impactor simultaneously by lifting the preseparator insert into the preseparator base; fit the
out the tray. preseparator base to the impactor inlet; add 15 mL of the
An induction port with internal dimensions identical to solvent used for sample recovery to the central cup of the
those defined in Figure 4a is connected to the impactor in- preseparator insert; place the preseparator body on top of
let. When necessary, with dry powder inhalers, a this assembly; and close the two catches. [Caution—Some
preseparator can be added to avoid overloading the first solvents form flammable vapor-air mixtures that may be ig-
stage. This preseparator connects between the induction nited during passage through a vacuum pump. Take appropri-
port and the impactor. A suitable mouthpiece adapter is ate precautions (e.g., alternative solvents, use of vapor traps,
used to provide an airtight seal between the inhaler and the minimal pump operating times, etc.) to ensure operator safety
induction port. during testing.]
At a volumetric airflow rate of 60 L per minute (the as- Connect an induction port with internal dimensions as de-
signed reference flow rate for cutoff-diameter calculations, fined in Figure 4a either to the impactor inlet or to the
Qn), the cutoff-aerodynamic diameters D50,Qn of Stages 1 to preseparator inlet atop the cascade impactor (Figure 9d).
5The cascade impactor is available as the Next Generation Pharmaceutical
Place a suitable mouthpiece adapter in position at the end
Impactor from MSP Corporation, Minneapolis, MN. of the induction port so that the mouthpiece end of the

inhaler, when inserted, lines up along the horizontal axis of Table 7. Cutoff Aerodynamic Diameter for Stages of
the induction port. The front face of the inhaler mouthpiece Apparatus 5 and 6
is flush with the front face of the induction port, producing Use Eq. 2 to calculate D50,Q for flow rates, Q, in the range 30
an airtight seal. When attached to the mouthpiece adapter, to 100 L per minute with Qn = 60 L per minute.
the inhaler should be positioned in the same orientation as
intended for use. Connect the apparatus to a flow system Stage D50,Qn x
according to the scheme specified in Figure 5. 1 8.06 0.54
Unless otherwise prescribed, conduct the test at the flow 2 4.46 0.52
rate used in the test for Delivered-Dose Uniformity drawing 4 3 2.82 0.50
L of air from the mouthpiece of the inhaler and through the 4 1.66 0.47
apparatus. Connect a flowmeter to the induction port. Use a 5 0.94 0.53
flowmeter calibrated for the volumetric flow leaving the
meter, or calculate the volumetric flow leaving the meter 6 0.55 0.60
(Qout) using the ideal gas law. For a meter calibrated for the 7 0.34 0.67
entering volumetric flow (Qin), use the formula:
Qout = QinP0/(P0 – ∆P)
where P0 is the atmospheric pressure and ∆P is the pressure
drop over the meter. Adjust the flow control valve to
achieve steady flow through the system at the required rate,
Qout (±5%). Ensure that critical flow occurs in the flow-con-
trol valve by the procedure described for Apparatus 2. Adjust
the timer controlling the operation of the two-way solenoid
valve so that it opens the valve for the same duration, T, as
used during testing for Delivered-Dose Uniformity.
Prime or load the dry powder inhaler with powder for
inhalation according to the labeled instructions. With the
vacuum pump running and the two-way solenoid valve
closed, insert the inhaler mouthpiece, held horizontally, into
the induction port mouthpiece adapter. Discharge the pow-
der into the apparatus by activating the timer and opening
the two-way solenoid valve for the required duration,
T(±5%). After the two-way solenoid valve has closed, re-
move the inhaler from the mouthpiece adapter. If additional
doses are required for the sample, reload the inhaler accord-
ing to the labeled instructions, reinsert the mouthpiece into
the mouthpiece adapter, and repeat the operation until the
required number of doses have been discharged. After dis-
charge of the last dose, switch off the vacuum pump.
Dismantle the apparatus, and recover drug for analysis as
follows: remove the induction port and mouthpiece adapter
from the preseparator and extract the drug into an aliquot Fig. 9. Apparatus 5 (shown with the preseparator in place).
of solvent; if used, remove the preseparator from the im-
pactor, without spilling the solvent into the impactor; and
recover the active ingredient from all inner surfaces. Apparatus 6 for Metered-Dose Inhalers—
Open the impactor by releasing the handle and lifting the Design—Apparatus 6 is identical to Apparatus 5 (Figures 9-
lid. Remove the cup tray, with the collection cups, and re- 9d), except that the preseparator is not to be used. Use this
cover the active ingredient from each cup into an aliquot of apparatus at a flow rate of 30 L per minute (±5%), unless
solvent. Using the method of analysis specified in the indi- otherwise prescribed in the individual monograph.
vidual monograph, determine the mass of drug contained in
each of the aliquots of solvent. Procedure—Assemble the apparatus without the
Determine the cutoff diameters of each of the individual preseparator. Place cups into the apertures in the cup tray.
stages of the impactor, at the value of Q = Qout employed in To ensure efficient particle capture, coat the particle collec-
the test by the formula: tion surface of each stage with glycerol, silicone oil, or other
suitable liquid typically deposited from a volatile solvent, un-
D50,Q = D50,Qn (Qn/Q)X, (Eq. 2) less it has been demonstrated to be unnecessary. Insert the
cup tray into the bottom frame, and lower into place. Close
where D50,Q is the cutoff diameter at the flow rate, Qem- the impactor lid with seal body attached, and operate the
ployed in the test, and the subscript, n, refers to the nomi- handle to lock the impactor together so that the system is
nal or reference value for Qn = 60 L of air per minute (see airtight. Connect an induction port with internal dimensions
Table 7). The values for the exponent, x, are listed in Table as defined in Figure 4a to the impactor inlet. Use a mouth-
7. Thus, when Q = 40 L of air per minute, the cutoff diame- piece adapter that ensures that the tip of the inhaler mouth-
ter of Stage 2 is given by the formula: piece is flush with the open end of the induction port. Turn
on the vacuum pump to draw air through the cascade im-
D50,40LPM = 4.46 µm × (60/40)0.52 = 5.51 µm. pactor, and calibrate the airflow through the system with an
appropriate flowmeter attached to the open end of the in-
Analyze the data as directed under Data Analysis. duction port. Adjust the flow-control valve on the vacuum
pump to achieve steady flow through the system at the
required rate, and ensure that the airflow through the sys-
tem is within ±5% of this flow rate. Unless otherwise pre-
scribed in the patient instructions, shake the inhaler for 5
seconds, and discharge one delivery to waste. With the vac-
uum pump running, insert the mouthpiece into the mouth-
piece adapter, and immediately fire the minimum recom-

Fig. 9a. Components of Apparatus 5.
Fig. 9b. Layout of interstage passageways of Apparatus 5.
Fig. 9c. Nozzle configuration of Apparatus 5.
mended dose into the cascade impactor. Keep the valve suitable solvent, and dilute quantitatively to an appropriate
depressed for a duration sufficient to ensure that the dose volume.
has been completely discharged. If additional sprays are re- Dismantle the apparatus, and recover the drug for analysis
quired for the sample, shake the inhaler, reinsert it into the as follows: remove the induction port and mouthpiece
mouthpiece adapter, and immediately fire the next mini- adapter from the apparatus, and recover the deposited drug
mum recommended dose. into an aliquot of solvent; open the impactor by releasing
Repeat until the required number of doses have been dis- the handle and lifting the lid; remove the cup tray, with the
charged. The number of minimum recommended doses dis- collection cups; and extract the active ingredient in each
charged must be sufficient to ensure an accurate and pre- cup into an aliquot of solvent. Using the method of analysis
cise determination of Aerodynamic Size Distribution. [NOTE— specified in the individual monograph, determine the quan-
The number of minimum recommended doses is typically tity of active ingredient contained in each of the aliquots of
not greater than 10.] After the last dose has been dis- solvent.
charged, remove the inhaler from the mouthpiece adapter. Determine the cutoff diameters of each of the individual
Rinse the mouthpiece adapter and induction port with a stages of the impactor, at the value of Q employed in the

Table 8. Table of Mass Summaries for Analyses of Metered-Dose Inhalers and Dry Powder Inhalers
Mass Apparatus 1 Apparatus 2 Apparatus 3a Apparatus 4b Apparatus 5d Apparatus 6d
Mouthpiece Ai — AiAi — Ai — Ai — Ai — Ai —
adapter
Preseparator — — — — AP — — — AP — — —
Stage 0 of A0 B0 — — A0 B0 — — — — — —
impactor
Stage 1 of A1 B1 A1 — A1 B1 A1 — A1 B1 A1 B1
impactor/
impinger
Stage 2 of A2 B2 A2 B2 A2 B2 A2 B2 A2 B2 A2 B2
impactor/
impinger
impactor/
impinger
impactor/
impinger
Stage 5 of A5 B5 A5 B5 A5 B5 — — A5 B5 A5 B5
impactor/
impinger
Stage 6 of A6 B6 — — A6 B6 — — A6 B6 A6 B6
impactor/
impinger
Stage 7 of A7 B7 — — A7 B7 — — A7 B7 A7 B7
impactor/
impinger
Filter AF BF AF BF AF BF AF BF AF BF AF BF
Sums of Masses ΣAc ΣBc ΣAc ΣBc ΣAc ΣBc ΣAc ΣBc ΣAc ΣBc ΣAc ΣBc
aStages 6 and 7 are omitted from Apparatus 3 at airflow rates >60 L per minute.
bStage 5 of Apparatus 4 is the filter stage (see Figure 8).
cΣA is the total drug mass recovered from the apparatus; ΣB is the mass of drug recovered from the impactor (Apparatus 1, 3, 5 and 6) or from the
impactor stages beneath the uppermost stage (Apparatus 2 and 4).
dFor Apparatus 5 and 6, values for the drug masses AF and BF refer to collections from the MOC, and/or the after-filter if used.
test by using Eq. 2 with values obtained from Table 7. Thus,

when Q = 30 L of air per minute, the cutoff diameter of
Stage 2 is given by the formula:
D50,30LPM = 4.46 µm × (60/30)0.52 = 6.40 µm.
To analyze the data, proceed as directed under Data
Analysis.
Data Analysis
This section describes the data analysis required to define
the Aerodynamic Size Distribution of the drug output from
the test inhaler, after the use of Apparatus 1, 2, 3, 4, 5, or 6.
Enter the data collected from Apparatus 1, 2, 3, 4, 5, or 6 in
the table of mass summaries as shown in Table 8. Perform
only those calculations specified in the individual Fig. 9d. Pre-separator layout for Apparatus 5.
monograph.
CALCULATIONS
Fine Particle Dose and Fine Particle Fraction—Calculate

the total mass, ΣA, of drug delivered from the mouthpiece
of the inhaler into the apparatus. Then calculate the total
mass, R, of drug found on the stages of the apparatus and
the filter that captured the drug in the fine particle size

range appropriate for the particular drug being tested. The 2 with Table 7. For Apparatus 1, use the cutoff diameters
Fine Particle Dose is calculated by the formula: quoted by the manufacturer. For Apparatus 3, present the
data as cumulative percentages of mass on and below the
R/n stated stage, and avoid assigning values to stage cutoff
diameters.
where R is as stated above, and n is the number of doses Repeat the calculation for each of the stages in the im-
discharged during the test. The Fine Particle Fraction that pactor or impinger stack, in reverse numerical order (largest
would be delivered from the inhaler is then calculated by to smallest stage number). For each stage, calculate the cu-
the formula: mulative percentage of mass less than the stated aerody-
namic diameter by adding the percentage of the mass on
R/ΣA. that stage to the total percentage from the stages below
and entering the value opposite the effective cutoff diame-
Cumulative Percentage (Cum%) of Drug Mass Less ter of the stage above it in the stack. Thus, the percentage
Than Stated Aerodynamic Diameter—Construct Table 9 by of drug on the filter can be seen to have aerodynamic diam-
dividing the mass of drug on the filter stage by ΣB (see eters less than the cutoff diameter of the stage above the
Table 8). Multiply the quotient by 100, and enter this num- filter, and the percentage on the filter plus the percentage
ber as a percentage opposite the effective cutoff diameter of on the stage above have diameters less than the cutoff di-
the stage immediately above it in the impactor or impinger ameter of the stage above that, and so on. Repeat the cal-
stack. For Apparatus 2 or 4, use Equation 1 to calculate the culation for each of the remaining stages in reverse numeri-
stage cutoff diameters, D50,Q, at the airflow rate, Q, em- cal order (see Table 9).
ployed during the test. For Apparatus 5 and 6, use Equation
Table 9. Cumulative Percentage (Cum%) of Mass Less than the Stated Aerodynamic Diameter
Apparatus 1 Apparatus 2 Apparatus 3a Apparatus 4b Apparatus 5 Apparatus 6
Mass Cum%c D50d Cum%c D50,Qd Cum%c D50,Qe Cum%c D50,Qd Cum%c D50,Qd Cum%c D50,Qd
Filter 0.4 0.625 0.4 1.7 0.34 0.34
Stage 7 b 0.7 — — b 0.7 — — b 0.55 b 0.55
Stage 6 c 1.1 — — c 1.1 — — c 0.94 c 0.94
Stage 5 d 2.1 b 1.25 d 2.1 — — d 1.66 d 1.66
Stage 4 e 3.3 c 2.5 e 3.3 b 3.1 e 2.82 e 2.82
Stage 3 f 4.7 d 5.0 f 4.7 c 6.8 f 4.46 f 4.46
Stage 2 g 5.8 100 10.0 g 5.8 100 13.0 g 8.06 g 8.06
Stage 1 h 9.0 — — h 9.0 — — — —— — —
Stage 0 100 — — — 100 — — — 100 — 100 —
aStages 6 and 7 are omitted from Apparatus 3 at flow rates >60 L per minute; thus, values for b and c should be omitted for Apparatus 3, where
necessary.
bThe filter stage in Apparatus 4 is Stage 5 (see Figure 8).
c [(mass on stage / ΣB)×100] % + (total% of ΣB from stages below).
dThe 50% cutoff diameter of the stage immediately above that indicated (e.g., for Stage 4, enter the cutoff diameter for Stage 3; for Apparatus 2
or 4, calculate as D50,Q from Eq. 1; for Apparatus 5 or 6, calculate as D50,Q from Eq. 2 using Table 7). Values entered in the Table are correct for
Apparatus 1, 2, 4, 5, and 6 only when used at 28.3, 60.0, 60.0, 60.0, and 60.0 L per minute, respectively.
eThe D
50 values are only valid at a flow rate of 28.3 L per minute.
If necessary, and where appropriate, plot the percentage

of mass less than the stated aerodynamic diameters, versus
the aerodynamic diameter, D50,Q, on log probability paper.
Calculate the GSD by the equation:
Use these data and/or plot to determine values for MMAD

and GSD etc., as appropriate and when necessary (see Fig-
ure 10).
Fig. 10. Plot of cumulative percentage of mass less than

stated aerodynamic diameter (probability scale) versus aero-
dynamic diameter (log scale).

USP 35 Physical Tests / 〈610〉 Alternative Microbial Sampling 253
Add the following: BULK TESTING FOR LOW-CONTENT INDP

Bulk lot testing may be preferable for low-content INDP in
〈610〉 ALTERNATIVE
▲ lieu of finished product testing to allow larger sample sizes
that are representative of the batch, without unduly increas-
MICROBIOLOGICAL SAMPLING ing the risk of inadvertent microbial contamination. Bulk
testing can be performed on the bulk powder or liquid for-
METHODS FOR NONSTERILE mulation just before filling. If bulk testing is performed in
lieu of finished product testing, then manufacturing
INHALED AND NASAL PRODUCTS processes following bulk sampling (e.g., filling and packag-
ing) must be validated in accordance with current good
manufacturing practice (CGMP) for their ability to prevent
microbial contamination. For microbial enumeration tests, at
least 10 g or 10 mL of bulk material, or, for specified micro-
INTRODUCTION organisms tests, 1 g or 1 mL of bulk material may be sam-
pled. For small batch sizes (i.e., less than 1000 g or 1000
Proper microbiological sampling of microbiologically sus- mL), the recommended sample size is 1% of the batch for
ceptible nonsterile products can be difficult because these both microbial enumeration and specified microorganisms
products are often filled into unique primary containers that tests.
are designed to protect the product from inadvertent con-
tamination during storage and use. These unique designs SAMPLING METHODS FOR HIGH-CONTENT
may increase the difficulty of taking an aseptic sample of
sufficient size or volume for microbiological testing. Unless INDP
special approaches are used, products such as inhaled, nasal
liquid, or powder dosage forms can be difficult to sample
without potential exposure to extraneous microbial contami- Dry Powder Inhalers
nation. This general test chapter provides these special ap-
proaches for sampling either low- or high-content inhaled or DPIs have an internal reservoir that contains a sufficient
nasal dosage forms. Alternative sampling approaches may quantity of formulation for multiple doses that are metered
provide better ways to sample containers in an aseptic man- by the device itself during activation by the patient. For
ner. Any alternative methodology should employ aseptic DPIs, appropriate validated procedures should be used to
techniques and should be conducted under environmental sample a nonsterile drug product container.
and other conditions that are appropriate for aseptic
sampling.
Inhalation Aerosols
INHALED OR NASAL DOSAGE FORMS Consider safety issues related to both inhalation of the
drug substance and the potential of a flammability hazard.
Low-content inhaled and nasal drug products (low-con- Avoid contamination of samples by employing aseptic tech-
tent INDP) are products that have a target fill of less than niques whenever necessary.
100 mg of powder or 1 mL of liquid formulation per unit
(primary container). Examples are pre-metered inhalation
powders, more commonly known as dry powder inhalers AUTOMATIC ACTUATION METHOD
(DPIs), and single-dose nasal sprays.
High-content INDP are multidose drug products that have The contents of the inhalation aerosol containers may be
a target fill of more than 100 mg of powder or more than 1 collected by automatically actuating each aerosol container
mL of liquid formulation per unit. Examples are aerosols for and collecting the delivered formulation on a suitable sterile
inhalation and nasal delivery, known as metered-dose inhal- filter.
ers (MDIs); device-metered inhalation powders; and mul-
tidose nasal sprays.
The appropriate sample quantity or volume should be ROOM TEMPERATURE METHOD
based on the test methodology, including any relevant gen-
eral test chapters, such as 〈61〉 Microbiological Examination of Disinfect the outside of the test containers with an ap-
Nonsterile Products: Microbial Enumeration Tests and 〈62〉 Mi- propriate disinfectant, and allow the containers to dry in a
crobiological Examination of Nonsterile Products: Tests for Spec- controlled environment. Empty the contents of the aerosol
ified Microorganisms. Testing may be performed on the un- container into a sterile vessel using a needle apparatus or
packaged bulk dry powder or liquid formulation or the similar device (e.g., icemaker water line tap). If it has been
finished product. If testing is performed on the bulk material demonstrated that the propellant does not inhibit the
alone, then the process leading from the bulk to the fin- growth of microorganisms, the contents of the sterile vessel
ished product should be validated for its ability to prevent may be added directly to the liquid media or buffer for the
microbial contamination. Testing should be performed on test. Otherwise, allow the propellant to evaporate from the
the finished product if this process is not validated. vessel by leaving the vessel at room temperature for several
minutes. Remove any residual gaseous propellant by tilting
the vessel slightly or by allowing a slow stream of microbio-
SAMPLE SIZE DETERMINATION logically inert sterile gas to pass over the surface. For some
less volatile propellants such as chlorofluorocarbon (CFC)
For each microbiological test, sample 10 drug product 11/12 combinations, the vessel may be heated slightly (to
containers or units or a number of units that can provide a temperatures ≤ 45°) to assist with evaporation. After the pro-
minimum of 1 gram of product that are representative of pellant has evaporated, add the liquid media or buffer, and
the batch. For batch sizes smaller than 200 units (e.g., mix the contents to prepare for testing.
batches used in clinical trials), sample size may be reduced Direct expulsion into the broth media or buffer may be
to 1% of the units or 1 unit, whichever is greater. The con- feasible if a needle apparatus that is thin and strong enough
tents of individual containers may be pooled for testing. to puncture the container and to allow slow removal of the
contents is available. In this case, the contents may be ex-


USP 601 AEROSOLS, NASAL SPRAYS

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37 by aptuit on Fri Dec 07 08:51:21 EST 2012

232 〈591〉 Zinc Determination / Chemical Tests USP 35

〈591〉 ZINC DETERMINATION Physical Tests and

Official from December 1, 2012

USP 35 Physical Tests / 〈601〉 Aerosols 233

Official from December 1, 2012

234 〈601〉 Aerosols / Physical Tests USP 35

Official from December 1, 2012

USP 35 Physical Tests / 〈601〉 Aerosols 235

Fig. 1. Sampling apparatus for pressurized metered-dose inhalers.

Delivered-Dose Uniformity Unless otherwise specified in the individual monograph,

Official from December 1, 2012

236 〈601〉 Aerosols / Physical Tests USP 35

Table 1. Component Specifications for Apparatus B (see Fig. 2)

6PU, UK), or equivalent.

Official from December 1, 2012

USP 35 Physical Tests / 〈601〉 Aerosols 237

Table 1. Component Specifications for Apparatus B (see Fig. 2) (Continued)

6PU, UK), or equivalent.

Official from December 1, 2012

238 〈601〉 Aerosols / Physical Tests USP 35

Delivered-Dose Uniformity over the Entire DRY POWDER INHALERS

Official from December 1, 2012

USP 35 Physical Tests / 〈601〉 Aerosols 239

Because the dimensions of the induction port used to con-

Fig. 4. Apparatus 1: Assembly of induction port and en-

Because the size distributions produced by different im-

Official from December 1, 2012

240 〈601〉 Aerosols / Physical Tests USP 35

Official from December 1, 2012

USP 35 Physical Tests / 〈601〉 Aerosols 241

more than 125% of the average minimum recommended

Official from December 1, 2012

242 〈601〉 Aerosols / Physical Tests USP 35

Official from December 1, 2012

USP 35 Physical Tests / 〈601〉 Aerosols 243

between the inhaler mouthpiece and the induction port.

Official from December 1, 2012

244 〈601〉 Aerosols / Physical Tests USP 35

Table 3. Component Specifications for Figure 5

also Footnote h in Table 1.

Table 4. Component Units of Multistage Liquid Impinger (see Fig. 8)

Official from December 1, 2012

USP 35 Physical Tests / 〈601〉 Aerosols 245

Table 4. Component Units of Multistage Liquid Impinger (see Fig. 8) (Continued)

Fig. 8a. Apparatus 4: Details of jet tube and impaction plate.

Fig. 8. Apparatus 4: Schematic of multistage liquid im-

Fig. 8b. Apparatus 4: Expanded view of Stage 5. (See Table

Under steady flow conditions, at the appropriate volumet-

Official from December 1, 2012

246 〈601〉 Aerosols / Physical Tests USP 35

n.a.: not applicable.

Official from December 1, 2012

USP 35 Physical Tests / 〈601〉 Aerosols 247

Official from December 1, 2012

248 〈601〉 Aerosols / Physical Tests USP 35

Table 6. Critical Dimensions for Apparatus 5 and 6