Appendix XIX B V-A615
Coloured glass Is obtained by the addition of small
Appendix IX
A. Containers Introduction
(Ph. Eur. general texts 3.2)
ThisAppendixprovides requirements) guidance and information
on containers for pharmaceutical use. Additional guidance is
provided in a numberof British Standards. Attentionis drawn in
particular to British Standards 1679-5: 1973) 1679-6: 1994)
1679-7: 1968 and 1679-8: 1992. The expression 'tamper-evident
container' means a closed container fitted with a device that
reveals irreversibly whether the container has been opened,
whereas) the expression. 'tamper-proof container'means a closed
container in which access. to the contents is prevented undernormal
conditions of use. The two terms areconsidered to be synonymous
by the European Pharmacopoeia Commission.
A container for pharmaceutical use is an article that contains
or is intended to contain a product and is, or may be, in
direct contact with it. The closure is a part of the container.
The container (see General Notices section 1.3) is so
designed that the contents may be removed in a manner
appropriate to the intended use of the preparation.
It provides ayarying degree of protection depending on the
nature of the product and the hazards of the environment,
and minimises the loss of constituents. The container does
not interact physically. or chemically with the contents in a
way that alters their quality beyond the limits tolerated by
official requirements.
Single-dose container A single-dose container holds a
quantity of the preparation intended for total or partial use
on 1 occasion only.
Multidose container A multidose container holds a
quantity of the preparation suitable for 2 or more doses.
Well-closed container A well-closed container protects
the contents from contamination with extraneous solids and
liquids and from loss of contents under ordinary conditions
of handling, storage and transport.
Airtight container An airtight container is impermeable to
solids, liquids and gases under ordinary conditions of
handling, storage and transport. If the container is intended
to be opened on more than 1 occasion, it must be so
designed that it remains airtight after re-closure.
Sealed container A sealed container is a container closed
by fusion of the material of the container.
Tamper-proof container A tamper-proof container is a
closed container fitted with a device that reveals irreversibly
whether the container has been opened.
Child-proof container A container that is fitted with a
closure that prevents opening by children.
B. Glass Containers for Pharmaceutical
Use
(Ph. Eur. method 3.2.1)
Glass containers for pharmaceutical use are glass articles
intended to come into direct contact with pharmaceutical
preparations.
Colourless glass Is highly transparent in the visible
spectrum.
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amounts of metal oxides, chosen according to the desired
spectral absorbance.
Neutral glass Is a borosilicate glass containing significant
amounts of boric oxide, aluminium oxide, alkali metal oxides
and/or alkaline earth oxides in the glass network. Due to its
composition, neutral glass has a high hydrolytic resistance
and a high thermal shock resistance.
Soda-lime-silica glass Is a silica glass containing alkali
metal oxides, mainly sodium oxide, and alkaline earth oxides,
mainlycalcium oxide, in the glass network. Due to its
composition, soda-lime-silica glass has only a moderate
hydrolytic resistance.
The hYdrolytic stability of glass containers for pharmaceutical
use is expressed by the resistanceto the release of soluble
mineral substances into water under the prescribed
conditions of contact between the inner surface of the
container or glass grains and water. The hydrolytic resistance
is evaluated by titrating released alkali reacting ions.
According to their hydrolytic resistance, glass containers are
classified as follows:
- type I glass containers: neutral glass, with a high
hydrolytic resistance due to the chemical composition of
the glass itself;
- type ITglass containers: usually of soda-lime-silica glass
with a high hydrolytic resistance resulting from suitable
treatment of the inner surface;
- type ill glass containers: usually of soda-lime-silica glass
with only moderate hydrolytic resistance.
The following italicised statements constitute general
recommendations concerning the type of glass container that
may be used for different types of pharmaceutical
preparations. The manufacturer of a pharmaceutical product
is responsible for ensuring the suitability of the chosen
container.
Type I glass containers aresuitable for mostpreparations whether
or not for parenteral administration.
Type II glass containers are suitable for mostacidic and neutral,
aqueous preparations whether or notfor parenteral administration.
Type III glass containers are in general suitable for non-aqueous
preparations for parenteral administration, for powders for
parenteral administration (except for freeze-dried preparations)
and for preparations notfor parenteral administration.
Glass containers with a hydrolytic resistance higher than that
recommended above for a particular type of preparation may
generally also be used.
The container chosen for a given preparation shall be such
that the glass material does not release substances in
quantities sufficient to affect the stability of the preparation
or to present a risk of toxicity. In justified cases, further
detailed information may be necessary to assess the impact
on chronic use and for vulnerable patient groups.
Preparations for parenteral administration are normally
presented in colourless glass, but coloured glass may be used
for substances known to be light-sensitive. Colourless or
coloured glass is used for the other pharmaceutical
preparations. It is recommended that all glass containers for
liquid preparations and for powders for parenteral
administration permit the visual inspection of the contents.
The inner surface of glass containers may be specially treated
to improve hydrolytic resistance, to confer water-repellancy,
etc. The outer surface may also be treated, for example to
reduce friction and to improve resistance to abrasion.
V-A616 Appendix XIX B
The outer treatment is such that it does not contaminate the
inner surface of the container.
Except for type I glass containers, glass containers for
pharmaceutical preparations are not to be re-used.
Containers for human blood and blood components must
not be re-used.
PRODUCTION
When glass containers for pharmaceutical use are
manufactured under stressed conditions (e.g. temperature-
time profile) and/or are placed in contact with particularly
aggressive pharmaceutical preparations, they may undergo
delamination, i.e the separation of the inner glass surface into
thin layers called lamellae or flakes. Glass delamination may
be the result of a chemical attack that occurs according to
well-known glass corrosion mechanisms, such as dissolution
by hydrolysis and ion exchange (leaching) as a function of
the pH. The process of interaction between the glass surface
and the pharmaceutical preparation requires incubation time,
and flaking may only become visible a number of months
after filling.
Several risk factors are known to increase the propensity of a
glass to delaminate. The chemical composition of the
pharmaceutical preparation, the presence of buffers like
citrate or phosphate, which are known to corrode glass, and
the ionic strength of the liquid medium may all strongly
favour delamination. The manufacturing process of the
container, chemical treatments of the inner surface, and
terminal sterilisation and processing at the pharmaceutical
filling lines are other important risk factors to be considered.
It is recommended that the user of the container assesses the
compatibility of the glass container and the pharmaceutical
preparation on a case-by-case basis, considering for example
the dosage form, properties of the formulation and glass
quality.
The propensity to delamination of glass containers from
different sources can be assessed and ranked by exposing the
container to accelerated degradation testing, carried out at
specified temperatures for a short time and using the
solutions associated with the actual pharmaceutical
preparation as extractants. The presence of particles in the
extraction solution, the occurrence of phase separation on the
inner surface, and the steep increase of silica concentration in
the extraction solution are all indicators of a potential
propensity for delamination. Accelerated degradation testing
can be used as a predictive tool to select the most
appropriate container for the intended preparation, but the
full compatibility of the active substance with the glass
leachate can only be assessed by a stability test under normal
conditions of use.
TESTS
Glass containers for pharmaceutical use comply with the
relevant test or tests for hydrolytic resistance. When glass
containers have non-glass components, the tests apply only to
the glass part of the container.
To define the quality of glass containers according to the
intended use, one or more of the following tests are
necessary.
Tests for hydrolytic resistance are carried out to define the
type of glass (I, II or III) and to control its hydrolytic
resistance.
In addition, containers foraqueous parenteral preparations
are tested for arsenic release and coloured glass containers
are tested for spectral transmission.
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2020
HYDROLYTIC RESISTANCE
Table 3.2.1.-1. - Types of glass
Type of container Test to be performed
Type I and type II glass containers (to Test A (surface test)
distinguish from type III glass
containers)
Type I glass containers (to distinguish Test B (glass grains test) or test C
from type II and type III glass (etching test)
containers)
Type I and type II glass containers (if Tests A and B, or tests A and C
there are doubts whether the high
hydrolytic resistance is due to the
chemical composition or to the surface
treatment)
The test is carried out by titration of the extraction solutions
obtained under the conditions described for tests A, B
and C. Test C is performed if there are uncertainties whether
the container is type I or type II.
EQUIPMENT
- An autoclave or steam steriliser capable of withstanding a
pressure of 2.5 x 105 N/m 2 (equivalent to
0.25 MPa = 2.5 bar) or more and capable of carrying out
the heating cycle described under Autoclaving process.
Preferably it is equipped with a constant-pressure
regulator or other suitable means in order to maintain the
temperature at 121 ± 1°C. The autoclave vessel is
equipped with a heating device, a thermometer integrated
in the autoclave, a pressure gauge, a vent cock (for
manually operated autoclaves only) and a tray of
sufficient capacity to accommodate, above the water level,
the number of containers needed to carry out the test.
The autoclave has the possibilityto connect a calibrated
resistance thermometer or a calibrated thermocouple from
the inner chamber to an external measuring device to
allow a temperature measurement independent from the
autoclave system.
The autoclave vessel and all ancillary equipment must be washed
thoroughly with waterR before use.
- A calibrated resistance thermometer or calibrated
thermocouple connected to a suitable temperature
measuring device.
- Burettes with a suitable capacity.
- One-mark volumetric flasks, with a capacity of 1000 mL.
- Pipettes and beakers.
- Conical flasks with capacities of 100 mL and 250 mL.
- A water-bath.
- Metal foil (e.g. aluminium, stainless steel).
Flasks and beakers must already have been used for the test
or have been filled with water R and kept in an autoclave at
121°C for at least 1 h before being used.
DETERMINATION OF THE FILLING VOLUME
The filling volume is the volume of water to be introduced
into the container for the purpose of the test. For vials and
bottles the filling volume is 90 per cent of the brimful
capacity. For ampoules it is the volume up to the height of
the shoulder.
Vials and bottles
Select, at random, 6 containers from the sample lot, or 3 if
their capacity exceeds 100 mL, and remove any debris or
dust. Weigh the empty containers with an accuracy of 0.1 g.
Place the containers on a horizontal surface and fill them
with waterR until about the rim edge, avoiding overflow and
introduction of air bubbles. Adjust the liquid levels to the
brimful line. Weigh the filled containers to obtain the mass of

the water expressed to 2 decimal places for containers having
a nominal volume less than or equal to 30 mL, and
expressed to 1 decimal place for containers having a nominal
volume greater than 30 rnL. Calculate the mean value of the
brimful capacity in millilitres and multiply it by 0.9. This
volume, expressed to 1 decimal place, is the filling volume
for the particular container lot.
Ampoules
Place at least 6 dry ampoules on a flat, horizontal surface and
fill them with water R from a burette, until the water reaches
point A, where the body of the ampoule declines to the
shoulder (see Figure 3.2.1.-1). Read the capacities (expressed
to 2 decimal places) and calculate the mean value..This
volume, expressed to 1 decimal place, is the filling volume
for the particular ampoule lot. The filling volume may also
be determined by weighing.
A Autoclaving process
I! .
~)~".".,.
,..
Figure 3.2.1.-1. - Filling volume of ampoules (up to point A)
Syringes and cartridges
Select 6 syringes or cartridges. Close the small opening
(mouth of cartridges and needle and/or Luer cone of
syringes) using an inert material (e.g. a tip cap) or any other
suitable means to prevent water leakage. Determine the mean
brimful volume in accordance with the procedure described
under Vials and bottles and multiply it by 0.9. This volume,
expressed to 1 decimal place, is the filling volume for the
particular container lot.
TEST A. HYDROLYTIC RESISTANCE OF THE INNER SURFACES OF
GLASS CONTAINERS (SURFACE TEST)
The determination is carried out on unused containers.
The volumes of the test liquid necessary are indicated in
Table 3.2.1.-2.
Table 3.2.1.-2. - Volume of test liquidand numberof titrations
Filling volume (mL) Volwne of test liquid Number oftitrations
for 1 titration (mL)
Up to3 25.0 1 rate so that steam issues vigorously from the vent cock after
Above 3 and up to 30 50.0 2 20-30 min, and maintain a vigorous evolution of steam for a
Above 30 and up to 100 100.0 2 further 10 min.
Above 100 100.0 3 Close the vent cock, follow the temperature increase on the
Cleaning
Remove any debris or dust. Shortly before the test, fill each
container to the brim with zuater R and allow to stand, filled
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Appendix XIX B
with water, for 20 ± 5 min. Empty the containers, carefully
rinse twice with waterR and once with waterRl and allow to
drain.
Closed ampoules are not rinsed before testing. Closed
ampoules may be warmed on a water-bath or in an oven at
about 40 DC for approximately 2 min before opening to avoid
underpressure when opening.
Filling
Fill the containers with waterRl up to the filling volume.
Loosely cap each container with an inert material, for
example with.inverted beakers of such a size that the.bottoms
of the beakers fit snugly down on the rims of the sample.
Ampoules and vials capped with dean alurniniumfoil are
further examples. Place syringes iand ca1tridgesina beaker
andcoverthe .beakerwith clean aluminium foil.
Containers of a volume of 2 mL or less, in which the water is
not sufficiently retained during the autoclaving process, may
be closed in a suitable way, e.g. with a stopper or plug of
inert material, such as silicone, and fixed using a plunger or a
stable fixing or clamping device.
Place the samples, gathered in groups in glass dishes or in
beakers or other suitable holders, on the rack in the autoclave
containing water R at room temperature. Ensure that they are
held above the level of the water in the autoclave.
Reference thermal cycle
The autoclave is run in such a way that the temperature in
the containers to be tested follows a thermal cycle with the
following characteristics: temperature raised from room
temperature to 100 DC within 20-30 min; temperature
maintained at 100 ± 1 DC for 10 ± 1 min; temperature in
the containers raised from 100 DC to 121 DC within
20-22 min; temperature maintained at 121 ± 1 DC for
60 ± 1 min; temperature cooled to 100 DC within
40-44 min.
Autoclave calibration
Before being used for the first time, the autoclave and the
temperature measuring system are calibrated to ensure that
the autoclave settings are suitable to guarantee that the
temperature inside the containers is 121 ± 1 DC.
NOTE: significant differences may be observed between the
temperature measured in the autoclave chamber and inside the
containers.
Take a set of containers of intermediate capacity (10 mL for
instance) and fill them with water Rl. Select a sufficient
number of containers to fill completely the tray within the
autoclave chamber. Insert the end of the calibrated resistance
thermometer or calibrated thermocouple into one of the
containers through a hole in the closure having approximately
the same diameter as the probe and connect it to the external
measuring device. If the container is too small to insert a
thermocouple, place the thermocouple in a similar container
of suitable size filled with water Rl . Close the autoclave door
or lid securely and run the autoclave to achieve the target
thermal cycle in the containers. Where a manual autoclave is
run, leave the vent cock open. Heat the autoclave at a regular
calibrated thermocouple measuring device by comparison
with readings taken from the autoclave thermometer and
adjust the autoclave settings accordingly in order to match
V-A618 Appendix XIX B 2020

the target thermal cycle. Keep the temperature ramp as Table 3.2.1.-3. - Limit values in the testfor surface hydrolytic
smooth as possible. resistance
Using the calibrated thermocouple measuring device, ensure Ma."'Idmum volume of 0.01 M Hel per
that deviations from the holding temperature of 121 ± 1 DC 100 mL of test solution (mL)
are within the tolerance. When cooling down, vent to prevent Filling volume (mL) Types I and IT glass Type ill glass
the formation of a vacuum. For safety reasons (boiling containers containers
retardation) do not open the autoclave before the water in Up to 0.5 3.0 30.0
the containers has reached a temperature of 95 DC. Remove Above 0.5 and up to 1 2.0 20.0
the hot samples from the autoclave and cool cautiously to Above 1 and up to 2 1.8 17.6
room temperature within 30 min. Above 2 and up to 3 1.6 16.1
Record the autoclave settings used to carry out the thermal Above 3 and up to 5 1.3 13.2
cycle and use these settings for routine autoclave runs. Above 5 and up to 10 1.0 10.2
Above 10 and up to 20 0.80 8.1
At regular intervals verify the validation of the calibration.
Above 20 and up to 50 0.60 6.1
Establish a re-calibration plan based on quality assurance
Above 50 and up to 100 0.50 4.8
criteria, recalibrate as appropriate and keep records.
Above 100 and up to 200 0.40 3.8
Routine autoclave runs Above 200 and up to 500 0.30 2.9
Use the autoclave settings established during the calibration Above 500 0.20 2.2
stage and follow the same thermal cycle described above.
Container sets of different capacity can be tested during the - a set of 3 square-mesh sieves of stainless steel, mounted
same run. Keep the glass load very similar in size and mass on frames of the same material and consisting of the
to the load used during the calibration stage. The use of the following:
calibrated thermocouple is no longer necessary provided the (a) sieve no. 710;
calibration is proved to be valid over a defined time span.
(b) sieve no. 425;
At the end of the cycle, remove the hot samples from the
(c) sieve no. 300;
autoclave and cool them cautiously to room temperature
- a mechanical sieve-shaker or a sieving machine may be
within 30 min.
used to sieve the grains;
NOTE: depending on the type or size of the autoclave the heat - a permanent magnet;
transfer and thus the resulting thermal cycle in the containers may - metal foil (e.g. aluminium, stainless steel);
vary with the total load of the autoclave. It may therefore be - a hot-air oven, capable of maintaining a temperature of
necessary to adjust the autoclave load. 140 ± 5°C;
Method - a balance, capable of weighing up to 500 g with an
Carry out the titration within 1 h of removal of the accuracy of 0.005 g;
containers from the autoclave. Combine the liquids obtained - a desiccator;
from the containers and mix. Introduce the prescribed - an ultrasonic bath.
volume (Table 3.2.1.-2) into a conical flask (test solution). Method
Place the same volume of waterR1 into a 2n d similar flask as Rinse the containers to be tested with waterR and dry in the
a blank. Add to each flask 0.05 mL of methyl redsolution R oven. Wrap at least 3 of the glass articles in clean paper and
for each 25 mL of liquid. Titrate the blank with 0.01 M crush to produce 2 samples of about 100 g each, in pieces
hydrochloric acid. Titrate the test solution with the same acid not more than 30 mm across.
until the colour of the resulting solution is the same as that
Where a mortar, pestle and hammer are used, place in the
obtained for the blank. Subtract the value found for the
mortar 30-40 g of the pieces 10-30 mm across taken from
blank titration from that found for the test solution and
1 of the samples, insert the pestle and strike it heavily, once
express the results in millilitres of 0.01 M hydrochloric acid per
only, with the hammer. Transfer the contents of the mortar
100 mL. Express titration values of less than 1.0 mL to
to sieve (a), the coarsest of the set. Repeat the operation until
2 decimal places and titration values of more than or equal to
all fragments have been transferred to the sieve. Shake the set
1.0 mL to 1 decimal place.
of sieves for a short time by hand and remove the glass that
Limits remains on sieves (a) and (b). Submit these portions to
The results, or the average of the results if more than 1 further fracture, repeating the operation until about 109 of
titration is performed, is not greater than the values stated in glass remains on sieve (a). Reject this portion and the portion
Table 3.2.1.-3. that passes through sieve (c). Reassemble the set of sieves
TEST B. HYDROLYTIC RESISTANCE OF GIASS GRAINS (GIASS and shake for 5 min. Transfer to a weighing bottle those
GRAINS TEST) glass grains that pass through sieve (b) and are retained on
Check that the articles as received have been annealed to a sieve (c).
commercially acceptable quality. Where a ball mill is used, place in the ball mill beaker about
The test may be performed on the canes used for the 50 g of the pieces 10-30 rom across taken from 1 of the
manufacture of tubing glass containers or on the containers. samples, add the balls and crush thin-walled glass (wall
Equipment thickness up to 1.5 mm) for up to 2 min and thick-walled
- a mortar, pestle (see Figure 3.2.1.-2) and hammer made glass (wall thickness greater than 1.5 rom) for up to 5 min.
of tempered, magnetic steel; Transfer the grains to sieve (a), sieve for about 30 sand
- as an alternative to the mortar, pestle and hammer, a ball collect the grains retained on sieve (c). Transfer the glass
mill can be used; the ball mill is made of agate, zirconia from sieves (a) and (b) into the ball mill and crush and sieve
or stainless steel with a volume of 250 mL; 2 balls with a again as indicated above. Combine the grains retained on
diameter of 40 rom or 3 balls with a diameter of 30 rom sieve (c);
are suitable;

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048
~I
Figure 3.2.1.-2. - Mortar and pestle apparatus for glass grains
method (dimensions in millimetres)
Repeat the crushing and sieving procedure with the other
glass sample and thus 2 samples of grains, each of which
shall be in excess of 109, are obtained. Spread each sample
on a piece of clean glazed paper and remove any iron
particles by passing the magnet over them. Transfer each
sample into a beaker for cleaning. Add to the grains in each
beaker 30 mL of acetone R and scour the grains by suitable
means, such as a rubber- or plastic-coated glass rod. After
scouring the grains, allow to settle and decant as much
acetone as possible. Add another 30 mL of acetone R, swirl,
allow to settle and decant again, and add 30 mL of acetone R.
Fill the bath of the ultrasonic vessel with water at room
temperature, then place the beaker in the rack and immerse
it until the level of the acetone is at the level of the water;
apply the ultrasound for 1 min. Swirl the beaker, allow to
settle and decant the acetone as completely as possible, add
30 mL of acetone R and repeat the ultrasonic cleaning
operation. If a fine turbidity persists, repeat the ultrasonic
cleaning and acetone washing until the solution remains
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clear. Swirl and decant the acetone then dry the grains, first
by putting the beaker on a warm plate to remove excess
acetone and then by heating at 140°C for 20 min in the
drying oven.' Transfer the dried grains from each beaker into
separate weighing bottles, insert the stoppers and cool in the
desiccator. Weigh 10.00 g of the cleaned and dried grains
into 2 separate conical flasks. Add 50 mL of waterRl into
each by means of a pipette (test solutions). Pipette 50 mL of
water Rl into a 3rd conical flask as a blank. Distribute the
grains evenly over the flat bases of the flasks by gentle
shaking. Close the flasks with neutral glass dishes or
aluminium foil rinsed with water R, or with inverted beakers
so that the inner surface of the beakers fit snugly down onto
the top rims of the flasks.Placea113 flasks in the rack in the
autoclave containing the water at room temperature, and
ensure that they are held above the level of the water in the
vessel. Carry out theautoclaving procedure in a similar
manner to that described under test A, but maintain the
temperature of 121 ± 1 °C only for 30 ± 1 min. Do not
open the autoclave until it has cooled to 95°C. Remove the
hot samples from the autoclave and cool the flasks in running
tap water as soon as possible, avoiding thermal shock.
To each of the 3 flasks add 0.05 mL of methylredsolution R.
Titrate the blank solution immediately with 0.02 M
hydrochloric acid then titrate the test solutions with the same
acid until the colour matches that obtained with the blank
solution. Subtract the titration volume for the blank solution
from that for the test solution.
NOTE: where necessary to obtain a sharp end-point, theclear
solution is to be decanted into a separate 250 mL flask. Rinse the
grains with 3 quantities, each of 15 ml.; of water Rl by swirling
and add the washings to the main solution. Add o. as mL of
methylredsolution R. Titrate and calculate as described below.
In this case also add 45 mL of water Rl and 0.05 mL of methyl
redsolution R to the blank solution.
Calculate the mean value of the results in millilitres of
0.02 M hydrochloric acid per gram of the sample and if
required its equivalent in alkali extracted, calculated as
micrograms of sodium oxide per gram of glass grains.
1 mL of 0.02 M hydrochloric acid is equivalent to 620 /lg of
sodium oxide.
Repeat the test if the highest and lowest observed values
differ by more than 20 per cent.
Limits
Type I glass containers require not more than 0.1 mL of
0.02 M hydrochloric acid per gram of glass, type II and
type III glass containers require not more than 0.85 mL of
0.02 M hydrochloric acid per gram of glass.
TEST C. TO DETERMINE WHETHER THE CONTAINERS HAVE BEEN
SURFACE-TREATED (ETCHING TEST)
If there are uncertainties whether a container has been
surface-treated, and!or to distinguish between type I and
type II glass containers, test C is used in addition to test A.
Alternatively, tests A and B may be used. Test C may be
carried out either on unused samples or on samples
previously used in test A.
Vials and bottles
The volumes of test liquid required are shown in
Table 3.2.1.-2.
Rinse the containers twice with water R, fill to the brimful
point with a mixture of 1 volume of hydrofluoric acidRand
9 volumes of hydrochloric acidR and allow to stand for
10 min. Empty the containers and rinse carefully 5 times
with waterR. Immediately before the test, rinse once again
with waterR. Submit the containers thus prepared to the
V-A620 Appendix XIX B 2020

same autoclaving and determination procedure as described SPECTRAL TRANSMISSION FOR COLOURED
in Test A for surface hydrolytic resistance. If the results are GLASS CONTAINERS
considerably higher than those obtained from the original Equipment
surfaces (by about a factor of 5 to 10), the samples have A UV-Vis spectrophotometer, equipped with a photo diode
been surface-treated. detector or equipped with a photomultiplier tube coupled
Ampoules, cartridges and syringes with an integrating sphere.
NOTE: ampoules, cartridges and syringes madefrom glass tubing Preparation of the specimen
are not normally subjected to internal surface treatment because Break the glass container or cut it with a circular saw fitted
theirh£gh chemical resistance is dependent upon the chemical with a wet abrasive wheel, such as a carborundum or a
composition of the glass as a material. bonded-diamond wheel. Select sections representative of the
Apply the test method as described above for vials and wall thickness and trim them as suitable for mounting in a
bottles. If the ampoules are not surface-treated, the new spectrophotometer. If the specimen is too small to cover the
values are slightly lower than those obtained in previous tests. opening in the specimen holder, mask the uncovered portion
Distinction between type I and type II glass containers with opaque paper or tape, provided that the length of the
The results obtained in Test C are compared to those specimen is greater than that of the slit. Before placing in the
obtained in Test A. The interpretation of the result is shown holder, wash, dry and wipe the specimen with a lens tissue.
in Table 3.2.1.-4. Mount the specimen with the aid of wax, or by other
convenient means, taking care to avoid leaving fingerprints or
Table 3.2.1.-4. - Distinction between typeI and type II glass other marks.
containers Method
Type I Type II Place the specimen in the spectrophotometer with its
The values are closely similar to The values greatly exceed those found in cylindrical axis parallel to the slit and in such a way that the
those found in the test for surface the test for surface hydrolytic resistance light beam is perpendicular to the surface of the section and
hydrolytic resistance for type I glass and are similar to but not larger than that the losses due to reflection are at a minimum. Measure
containers. those for type III glass containers.
the transmission of the specimen with reference to air in the
spectral region of 290-450 nm, continuously or at intervals of
ARSENIC 20nm.
The test applie: to glass containers for aqueous parenteral
Limits
preparations.
The observed spectral transmission for coloured glass
Hydride generation atomic absorption spectrometry (2.2.23, containers for preparations that are not for parenteral
Method 1). administration does not exceed 10 per cent at any
Test solution Use the extraction solution obtained from wavelength in the range of 290-450 nm, irrespective of the
containers of types I and II, after autoclaving at 121°C for type and the capacity of the glass container. The observed
1 h as described under Test A for surface hydrolytic spectral transmission in coloured glass containers for
resistance. Transfer 10.0 rnL to a 100 mL volumetric flask. parenteral preparations does not exceed the limits given in
Add 10 rnL of hydrochloric add R and 5 mL of a 200 gIL Table 3.2.1.-5.
solution of potassium iodide R. Heat on a water-bath at 80°C
for 20 min, allow to cool and dilute to 100.0 mL with Table 3.2.1.-5. - Limits of spectral transmission for coloured glass
waterR. containers for parenteral preparations
Reference solutions Prepare the reference solutions using Maximum percentage of spectral
transmission at any wavelength between
arsenic standardsolution (1 ppm As) R. Add 10 mL of
290 nm and 450 nm
hydrochloric add Rand 5 mL of a 200 gIL solution of
Nominal volume (mL) Flame-sealed Containers with
potassium iodide R. Heat on a water-bath at 80°C for 20 min, containers closures
allow to cool and dilute to 100.0 rnL with waterR.
Up to I 50 25
The concentration range of the reference solutions is typically
Above I and up to 2 45 20
0.005-0.015 ppm of As.
Above 2 and up to 5 40 15
Acid reservoir Hydrochloric acidR. Above 5 and up to 10 35 13
Reducing reservoir Sodium tetrahydroborate reducing Above 10 and up to 20 30 12
solution R. Above 20 25 10
Use a hydride generation device to introduce the test solution
into the cuvette of the spectrometer. Establish and
standardise instrumental operating conditions according to ANNEX - TEST FOR SURFACE
the manufacturer's instructions, optimise the uptake rate of HYDROLYfIC RESISTANCE-
the peristaltic pump, then connect it to the acid reservoir, the
reducing reservoir and the test solution.
DETERMINATION BY FLAME
SPECTROMETRY
Source Hollow-cathode lamp.
Wavelength 193.7 nm. The surface hydrolytic resistance of glass of types I and II may be
determined by analysis of the leaching solution by flame
Atomisation device Air-acetylene flame.
spectrometry. A numberof elements that, when present as oxides in
Limit Maximum 0.1 ppm of As. glass, contribute to the alkalinity of the solution, are determined
and used to express an alkaliequivalent. The spectrometric method
has the advantage of allowing the use of a much smaller sample of
extractso that it can be applied to small individual containers.
This enables an eoaluation of the uniformity of the containers in a

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2020 1

given batch where this is critical. The results of this measurement extraction solutions of this container type need not be
are not equivalent to those of titrimetry and the 2 methods cannot analysed for these ions. Aspirate the extraction solution from
be considered interchangeable. A correlation between the 2 is each sample directly into the flame of the atomic absorption
dependent on the type of glass and the size and shape of the or atomic emission instrument and determine the
container. The titrimetric method is the reference method of the approximate concentrations of sodium oxide (and potassium
Pharmacopoeia; the spectrometric methodmay be used in justified oxide and calcium oxide, if present) by reference to
and authorised cases. calibration graphs produced from the reference solutions of
A methodsuitable for this type of analysis is shown below. suitable concentration.
The determination is carried out on unused containers. FINAL ANALYSIS
The number of containers to be examined is indicated If dilution is unnecessary, add to each container a volume of
in Table 3.2.1.-6. thespectrochemical buffer solution equivalent to 5 per cent
of the filling volume, mix well and determine sodium oxide,
Table 3.2.1.-6. __ Number of containers to be examinedfor the calcium oxide and potassium oxide, by reference to
spectrometric method calibration graphs. For the determination of.thecalcium
Filling volume (mL) Number of containers Additional containers oxideconcentration by flame speetrometry,a nitrous
to be measured for preliminary oxide/acetylene flame is used.
separately measurements
If dilution is necessary, determine sodium oxide, calcium
Up to 2 20 2 oxide and potassium oxide, if present, following the
Above 2 and up to 5 15 2 procedures as described above. The solutions shall contain
Above 5 and up to 30 10 2 5 per cent V/V of the spectrochemical buffer solution.
AboveBuand up to 100 5 1 Concentration values less than 1.0 ug/ml, are expressed to
Above:.lOQ 3 1 2 decimal places, values greater than or equal to 1.0 ug/ml;
to 1 decimal place. Correct the result.for the buffer addition
InstrtIctions. on determination of the filling volume, cleaning and for any dilution.
of t:he~RI1tainers, filling and heating are given above under DETERMINATION
Hydfbl§ric resistance and Test A. Determine the mean value of the concentration of individual
SOLUTIONS oxides found in the samples tested, in micrograms of the
Spectrochemical buffer solution Dissolve 80 g of caesium oxide per millilitre of the extraction solution, and calculate
chloride R in-about 300 mL of water Rl, add 10mL of the sum of the individual oxides, expressed as micrograms of
6 M hydrochloric acid R, dilute to 1.0 L with water Rl and sodium oxide per millilitre of the extraction solution, using
mix. the following mass conversion factors:
Stock solutions: - 1 ug of potassium oxide corresponds to 0.658 ug of
=
- sodium oxide, c(NazO) 1 mg/mL; sodium oxide;
=
- potassium oxide, c(KzO) 1 mg/mL; - 1 Ilg of calcium oxide corresponds to 1.105 ug of sodium
=
- calcium oxide, c(CaO) 1 mg/mL. oxide.
Commercially available stock solutions may also be used. Limits
Standard solutions Prepare standard solutions by diluting The mean value is not greater than the value given in
the stock solutions with water Rl to obtain concentrations Table 3~2.1.-7.
suitable for establishing the reference solutions in an
Table 3.2.1.-7. - Limit values in the testfor surface hydrolytic
appropriate manner, e.g, with concentrations of 20 ug/ml, of
resistance by flame spectrometry, for type I and type II glass
sodium oxide, potassium oxide and calcium oxide,
containers
respectively. Commercially available standard solutions may
also be used. Filling volume (mL) Limit values for the concentration of
oxides, expressed as sodium oxide
Reference solutions Prepare the reference solutions for (~Lg/mL)
establishing the calibration graph (set of calibration solutions) Up to 0.5 7.50
by diluting suitable concentrated standard solutions with Above 0.5 and up to 1 5.00
water Rl, so that the normal working ranges of the specific Above 1 and up to 2 4.50
elements are covered, taking into account the instrument Above 2 and up to 3 4.10
used for the measurement. Typical concentration ranges of Above 3 and up to 5 3.20
the reference solutions are: Above 5 and up to 10 2.50
- for determination by atomic emission spectrometry of Above 10 and up to 20 2.00
sodium oxide and potassium oxide: up to 10 Ilg/mL; Above 20 and up to 50 1.50
- for determination by atomic absorption spectrometry of Above 50 and up to 100 1.20
sodium oxide and potassium oxide: up to 3 ug/ml.; Above 100 and up to 200 1.00
- for determination by atomic absorption spectrometry of Above 200 and up to 500 0.75
calcium oxide: up to 7 ~lg/mL. Above 500 0.50
Use reference solutions containing 5 per cent V/V of the
spectrochemical buffer solution.
METHOD
Carry out preliminary measurements of the potassium oxide
and calcium oxide concentrations on one of the extraction
solutions. If, for one container type, the concentration of
potassium oxide is less than 0.2 ug/ml, and the concentration
of calcium oxide is less than 0.1 ug/ml., the remaining

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V-A622 Appendix XIX C
c. Plastic Containers and Closures
(Plastic Containers and Closures for Pharmaceutical Use,
Ph. Bur. method 3.2.2)
A plastic container for pharmaceutical use is a plastic article
which contains or is intended to contain a pharmaceutical
product and is, or may be, in direct contact with it.
The closure is a part of the container.
Plastic containers and closures for pharmaceutical use are
made of materials in which may be included certain
additives; these materials do not include in their composition
any substance that can be extracted by the contents in such
quantities as to alter the efficacy or the stability of the
product or to present a risk of toxicity.
The most commonly used polymers are polyethylene (with
and without additives), polypropylene, poly(vinyl chloride),
poly(ethylene terephthalate) and poly(ethylene-vinyl acetate).
The nature and amount of the additives are determined by
the type of the polymer, the process used to convert the
polymer into the container and the intended purpose of the
container. Additives may consist of antioxidants, stabilisers,
plasticisers, lubricants, colouring matter and impact
modifiers. Antistatic agents and mould-release agents may be
used only for containers for preparations for oral use or for
external use for which they are authorised. Acceptable
additives are indicated in the type specification for each
material described in the Pharmacopoeia. Other additives
may be used provided they are approved in each case by the
competent authority responsible for the licensing for sale of
the preparation.
For selection of a suitable plastic container, it is necessary to
know the full manufacturing formula of the plastic, including
all materials added during formation of the container so that
the potential hazards can be assessed. In justified cases,
further detailed information may be necessary to assess the
impact on chronic use and for vulnerable patient groups.
The plastic container chosen for any particular preparation
should be such that:
- the ingredients of the preparation in contact with the
plastic material are not significantly adsorbed on its
surface and do not significantly migrate into or through
the plastic,
- the plastic material does not release substances in
quantities sufficient to affect the stability of the
preparation or to present a risk of toxicity.
U sing material or materials selected to satisfy these criteria, a
number of identical type samples of the container are made
by a well-defined procedure and submitted to practical
testing in conditions that reproduce those of the intended
use, including, where appropriate, sterilisation. In order to
confirm the compatibility of the container and the contents
and to ensure that there are no changes detrimental to the
quality of the preparation, various tests are carried out such
as verification of the absence of changes in physical
characteristics, assessment of any loss or gain through
permeation, detection of pH changes, assessment of changes
caused by light, chemical tests and, where appropriate,
biological tests.
The method of manufacture is such as to ensure
reproducibility for subsequent bulk manufacture and the
conditions of manufacture are chosen so as to preclude the
possibility of contamination with other plastic materials or
their ingredients. The manufacturer of the product must
ensure that containers made in production are similar in
every respect to the type samples.
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2020
For the results of the testing on type samples to remain valid,
it is important that:
- there is no change in the composition of the material as
defined for the type samples,
- there is no change in the manufacturing process as
defined for the type samples, especially as regards the
temperatures to which the plastic material is exposed
during conversion or subsequent procedures such as
sterilisation,
- scrap material is not used.
Recycling of excess material of well-defined nature and
proportions may be permitted after appropriate validation.
Subject to satisfactory testing for compatibility of each
different combination of container and contents, the
materials described in the Pharmacopoeia are recognised as
being suitable for the specific purposes indicated, as defined
above.
Plastic Containers for Aqueous Solutions for
Parenteral Infusions
(plastic Containers for Aqueous Solutions for Infusion, Ph. Bur.
method3.2.2.1)
DEFINITION
Plastic containers for aqueous solutions for infusion are
manufactured from one or more polymers, if necessary with
additives. The containers described in this section are not
necessarily suitable for emulsions. The polymers most
commonly used are polyethylene, polypropylene and
poly(vinyl chloride). The specifications of this text are to be
read in conjunction with section 3.2.2. Plastic containers and
closures for pharmaceutical use.
The containers may be bags or bottles. They have a site
suitable for the attachment of an infusion set designed to
ensure a secure connection. They may have a site that allows
an injection to be made at the time of use. They usually have
a part that allows them to be suspended and which will
withstand the tension occurring during use. The containers
must withstand the sterilisation conditions to which they will
be submitted. The design of the container and the method of
sterilisation chosen are such that all parts of the containers
that may be in contact with the infusion are sterilised.
The containers are impermeable to micro-organisms after
closure. The containers are such that after :filling they are
resistant to damage from accidental freezing which may occur
during transport of the final preparation. The containers are
and remain sufficientlytransparent to allow the appearance
of the contents to be examined at any time, unless otherwise
justified and authorised.
The empty containers display no defects that may lead to
leakage and the :filled and closed containers show no leakage.
For satisfactory storage of some preparations, the container
has to be enclosed in a protective envelope. The initial
evaluation of storage has then to be carried out using the
container enclosed in the envelope.
TESTS
Solution S
Use solution S within 4 h ofpreparation Fill a
container to its nominal capacity with waterR and close it, if
possible using the usual means of closure; otherwise close
using a sheet of pure aluminium. Heat in an autoclave so
that a temperature of 121 ± 2 °C is reached within 20 min
to 30 min and maintain at this temperature for 30 min.
. If heating at 121°C leads to deterioration of the container,
heat at 100°C for 2 h.

Blank Prepare a blank by heating water R in a borosilicate-
glass flask closed by a sheet of pure aluminium at the
temperature and for the time used for the preparation of
solution S.
Appearance of solution 8
Solution S is clear (2.2.1) and colourless (2.2.2, Method II).
Acidity or alkalinity
To a volume of solution S' corresponding to 4 per cent of the
nominal capacity of the container add 0.1 mL of
phenolphthalein solution R. The solution is colourless.
Add 0.4. mL of 0.01 M sodium hydroxide. The solution is
pink. Add 0.8 mL of 0.01 M hydrochloric acid and 0.1 mL of
methyl red solution R. The solution is orange-red or red.
Absorbance. (2.2.25)
Measure the absorbance of solution S from 230 nm to
360 nm, using the blank (see solution S) as the
compensation liquid. At these wavelengths, the absorbance is
not greater than 0.20.
Reducing substances
To '.20.0 mL of solution S add 1 mL of dilute sulfuric acidR
and 20.0 mL of 0.002 M potassium permanganate. Boil for
3 min. Cool immediately. Add 1 g of potassium iodide R and
titrate:immediately with 0.01 M sodium thiosulfate, using
0.f5,;mL of starch solution R as indicator. Carry out a titration
uS,ipg.,20.0 mLofthe blank. The difference between the
titration volumes is not greater than 1.5 mL.
Transparency
Fill a container previously used for the preparation of
solution S with a volume equal to the nominal capacity of
the primary opalescent suspension (2.2.1) diluted 1 in 200
for a container made from polyethylene or polypropylene and
1 in 400 for other containers. The cloudiness of the
suspension is perceptible when viewed through the container
and compared with a similar container filled with waterR.
LABELLING
The label accompanying a batch of empty containers
includes a statement of:
- the name and address of the manufacturer,
- a batch number which enables the history of the container
and of the plastic material of which it is manufactured to
be traced.
D. Containers for Blood and Blood
Components
1. Sterile Plastic Containers for Human Blood and
Blood Components
(ph. Bur. method3.2.3)
Plastic containers for the collection, storage, processing and
administration of blood and its components are
manufactured from one or more polymers, if necessary with
additives. The composition and the conditions of
manufacture of the containers are registered by the
appropriate competent authorities in accordance with the
relevant national legislation and international agreements.
When the composition of the materials of the different parts
of the containers correspond to the appropriate specifications,
their quality is controlled by the methods indicated in those
specifications (see 3.1. Materials usedfor the manufacture of
containers and subsections).
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Appendix XIX D V-A623
Materials other than those described in the Pharmacopoeia
may be used provided that their composition is authorised by
the competent authority and that the containers
manufactured from them comply with the requirements
prescribed for Sterile Plastic Containers for Human Blood
and Blood Components.
In normal conditions of use the materials do not release
monomers, or other substances, in amounts likely to be
harmful nor do they lead to any abnormal modifications of
the blood.
The containers may contain anticoagulant solutions,
depending on their intended use, and are supplied sterile.
Each container is fitted with attachments suitable for the
intended use. The container may be in the form of a
single unit or the collecting container may be connected by
one or more tubes to one or more secondary containers to
allow separation of the blood components to be effected
within a closed system.
The outlets are of a shape and size allowing for adequate
connection of the container with the blood-giving equipment.
The protective coverings on the blood-taking needle and on
the appendages must be such as to ensure the maintenance
of sterility. They must be easily removable but must be
tamper-proof.
The capacity of the containers is related to the nominal
capacity prescribed by the national authorities and to the
appropriate volume of anticoagulant solution. The nominal
capacity is the volume of blood to be collected in the
container. The containers are of a shape such that when
filled they may be centrifuged.
The containers are fitted with a suitable device for
suspending or fixing which does not hinder the collection,
storage, processing or administration of the blood.
The containers are enclosed in sealed, protective envelopes.
CHARACTERS
The container is sufficientlytransparent to allow adequate
visual examination of its contents before and after the taking
of the blood and is sufficientlyflexible to offer minimum
resistance during filling and emptying under normal
conditions of use. The container contains not more than
5 mL of air.
TESTS
Solution 8 1
Fill the container with 100 mL of a sterile, pyrogen-free
9 gIL solution of sodium chloride R. Close the container and
heat it in an autoclave so that the contents are maintained at
110°C for 30 min.
If the container to be examined contains an anticoagulant
solution, first empty it, rinse the container with 250 mL of
waterfor injections R at 20 ± 1 °C and discard the rinsings.
Solution S2
Introduce into the container a volume of waterfor injections R
corresponding to the intended volume of anticoagulant
solution. Close the container and heat it in an autoclave so
that the contents are maintained at 110°C for 30 min. After
cooling, add sufficient waterfor injections R to fill the
container to its nominal capacity.
If the container to be examined contains an anticoagulant
solution, first empty it and rinse it as indicated above.
Resistance to centrifugation
Introduce into the container a volume of waterR, acidified by
the addition of 1 mL of dilute hydrochloric acidR, sufficient to
fill it to its nominal capacity. Envelop the container w,i!J:?
V'-A624 Appendix XIX D
absorbent paper impregnated with a 1 in 5 dilution of
bromophendblue solution R1 or other suitable indicator and
then dried. Centrifuge at 5000 g for 10 min. No leakage
perceptible on the indicator paper and no permanent
distortion occur.
Resistance to stretch
Introdu.ce into the container a volume of water R, acidified by
the addition of 1 mL of dilute hydrochloric acid R, sufficient to
fill it to its nominal capacity. Suspend the container by the
suspending device at the opposite end from the blood-taking
tube and apply along. the axis of this tube an immediate force
of 20 N (2.05 kgf). Maintain the traction for 5 s. Repeat the
test wifu the force applied to each of the parts for filling and
emptying. No break and no deterioration occur.
Leakage
Place the container which has been submitted to the stretch
test between two plates covered with absorbent paper
impregnated with a 1 in 5 dilution of bromophenol blue
solution R1 or other suitable indicator and then dried.
Progressively apply force to the plates to press the container
so that its internal pressure (i.e. the difference between the
applied pressure and atmospheric pressure) reaches 67 kPa
within 1 min. lvlaintain the pressure for 10 min. No signs of
leakage are detectable on the indicator paper or at any point
of attachment (seals, joints, etc.).
Vapour permeability
For a container containing an anticoagulant solution, fill with
a volume of a 9 gIL solution of sodium chloride R equal to the
volume of blood for which the container is intended.
For an empty container, fill with the same mixture of
anticoagulant solution and sodium chloride solution. Close
the container, weigh it and store it at 5 ± 1 DC in an
atmosphere with a relative humidity of (50 ± 5) per cent for
21 days. At the end of this period the loss in mass is not
greater than 1 per cent.
Emptying under pressure
Fill the container with a volume of waterR at 5 ± 1 DC
equal to the nominal capacity. Attach a transfusion set
without an intravenous cannula to one of the connectors.
Compress the container so as to maintain throughout the
emptying an internal pressure (i.e the difference between the
applied pressure and atmospheric pressure) of 40 kPa.
The container empties in less than 2 min.
Speed of filling
Attach the container by means of the blood-taking tube fitted
with the needle to a reservoir containing a suitable solution
having a viscosity equal to that of blood, such as a 335 gIL
solution of sucrose Rat 37 DC. Maintain the internal pressure
of the reservoir (i.e. the difference between the applied
pressure and atmospheric pressure) at 9.3 kPa with the base
of the reservoir and the upper part of the container at the
same level. The volume of liquid which flows into the
container in 8 min is not less than the nominal capacity of
the container.
Resistance to temperature variations
Place the container in a suitable chamber having an initial
temperature of 20-23 DC. Cool it rapidly in a deep-freeze to
-80 DC and maintain it at this temperature for 24 h. Raise
the temperature to 50 DC and maintain for 12 h. Allow to
cool to room temperature. The container complies with the
tests for resistance to centrifugation, resistance to stretch,
leakage, vapour permeability emptying under pressure and
speed of filling prescribed above.
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Transparency
Fill the empty container with a volume equal to its nominal
capacity of the primary opalescent suspension (2.2.1) diluted
so as to have an absorbance (2.2.25) at 640 run of 0.37 to
0.43 (dilution factor about 1 in 16). The cloudiness of the
suspension must be perceptible when viewed through the
bag, as compared with a similar container filled with water R.
Extractable matter
Tests are carried out by methods designed to simulate as far
as possible the conditions of contact between the container
and its contents which occur in conditions of use.
The conditions of contact and the tests to be carried out on
the eluates are prescribed, according to the nature of the
constituent materials, in the particular requirements for each
type of container.
Haemolytic effects in buffered systems
Stock buffer solution Dissolve 90.0 g of sodium chloride R,
34.6 g of disodium hydrogen phosphate dodecahydrate Rand
2.43 g of sodium dihydrogen phosphate R in water R and dilute
to 1000 mL with the same solvent.
Buffer solution A o To 30.0 mL of stock buffer solution
add 10.0 mL of water R.
Buffer solution B o To 30.0 mL of stock buffer solution
add 20.0 mL of water R.
Buffer solution Co To 15.0 mL of stock buffer solution
add 85.0 mL of waterR.
Introduce 1.4 mL of solution Sz into each of three centrifuge
tubes. To tube I add 0.1 mL of buffer solution Ao, to tube II
add 0.1 mL of buffer solution Bo and to tube III add 0.1 mL
of buffer solution Co. To each tube add 0.02 mL of fresh,
heparinised human blood, mix well and warm on a water-
bath at 30 ± 1 DC for 40 min. Use blood collected less than
3 h previously or blood collected into an anticoagulant
citrate-phosphate-dextrose solution (CPD) less than 24 h
previously.
Prepare three solutions containing, respectively:
3.0 mL of buffer solution Ao and 12.0 mL of waterR
(solution AI),
4.0 mL of buffer solution Bo and 11.0 mL of waterR
(solution B I),
4.75 mL of buffer solution Bo and 10.25 mL of waterR
(solution C I).
To tubes I, II and III add, respectively, 1.5 mL of
solution AI' 1.5 mL of solution B I and 1.5 mL of
solution C I. At the same time and in the same manner,
prepare three other tubes, replacing solution Sz by waterR.
Centrifuge simultaneously the tubes to be examined and the
control tubes at exactly 2500 g in the same horizontal
centrifuge for 5 min. After centrifuging, measure the
absorbances (2.2.25) of the liquids at 540 nm using the stock
buffer solution as compensation liquid. Calculate the
haemolytic value as a percentage from the expression:
A exp x 100
A lOO
A JOO absorbance of tube ill,
A exp absorbance of tube I or II or of the corresponding control
tubes.
The solution in tube I gives a haemolytic value not greater
than 10 per cent and the haemolytic value of the solution in
tube II does not differ by more than 10 per cent from that of
the corresponding control tube.
Sterility (2.6.1)
The containers comply with the test for sterility. Introduce
aseptically into the container 100 mL of a sterile 9 gIL
solution of sodium chloride and shake the container to
ensure that the internal surfaces have been entirely wetted.
Filter the contents of the container through a membrane
filter and place the membrane in the appropriate culture
medium, as prescribed in the test for sterility.
Pyrogens (2.6.8)
Solution Sl complies with the test for pyrogens. Inject 10 mL
of the solution per kilogram of the rabbit's mass.
PACKAGING
The. containers are packed. in protective envelopes.
On removalfrom its protective envelope the container shows
no leakage and no growth of micro-organisms. The protective
envelope is sufficiently robust to withstand normal handling.
The protective envelope is sealed in such a manner that it
cannot be opened and re-closed without leaving visible traces
that the seal has been broken.
L{\BELLING
Th'e labelling complies with the relevant nationallegislation and
international agreements. The label states:
----..:,'tile name and address of the manufacturer,
-/(a batch number which enables the history of the container
and of the plastic material of which it is manufactured to
be traced.
A part of the label is reserved for:
- the statement of the blood group, the reference number
and all other information required by national legislation
or international agreements, and an empty space is
provided for the insertion of supplementary labelling.
The label of the protective envelope or the label on the
container, visible through the envelope, states:
- the expiry date,
- that, once withdrawn from its protective envelope, the
container must be used within 10 days.
The ink or other substance used to print the labels or the
writing must not diffuse into the plastic material of the
container and must remain legible up to the time of use.
2. Empty Sterile Containers of Plasticised Poly
(Vinyl Chloride) for Human Blood and Blood
Components
(ph. Eur. method 3.2.4)
DEFINITION
Unless otherwise authorised as described in general chapter
3.2.3. Sterile plastic containers for human blood and blood
components, the nature and composition of the material from
which the containers are made comply with the requirements
in general chapter 3.1.1.1. Materials based on plasticised
poly(vinyl chloride) for containers for human blood and blood
components.
TESTS
They comply with the tests prescribed in general chapter
3.2.3. Sterile plastic containers for human blood and blood
components and with the following tests.
Reference solution
Heat water R in a borosilicate-glass flask in an autoclave at
110 DC for 30 min.
Acidity or alkalinity
To a volume of solution Sz (see 3.2.3) corresponding to
4 per cent of the nominal value of the container add 0.1 mL
of phenolphthalein solution R. The solution remains colourless.
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Add 0.4 mL of O.OiNI sodium hydroxide. The solution is
pink. Add 0.8 mL of O.OINI hydrochloric acid and 0.1 mL of
methyl red solution R. The solution is orange-red or red.
Absorbance (2.2.25)
Maximum 0.30, determined between wavelengths of 230 nm
and 250 nm on solution Sz (see 3.2.3); maximum 0.10,
determined between wavelengths of 251 nm and 360 nm on
solution Sz. Use the reference solution as the compensation
liquid.
Reducing substances
Immediately after preparation of solution Sz (see 3.2.3),
transfer to a borosilicate-glass flask a volume corresponding
to 8 per cent of the nominal value of the container. At the
same time, prepare a blank using an equal volume of the
freshly prepared reference solution in another borosilicate-
glass flask. To each solution add 20.0 mL of 0.002 M
potassium permanganate and 1 mL of dilute sulfuric acid R.
Allow to stand protected from light for 15 min. To each
solution add 0.1 g of potassium iodide R. Allow to stand
protected from light for 5 min and titrate immediately with
0.01 M sodium thiosulfate, using 0.25 mL of starch solution R
as indicator. The difference between the 2 titrations is not
greater than 2.0 mL.
Plastic additives 01, 24, 25, 26 and 27
Gas chromatography (2.2.28) coupled with mass
spectrometry (2.2.43).
Internal standard solution S3 1 mg/mL solution of di-n-
octylphthalate R in tetrahydrofuran for chromatography R.
Internal standard solution S4 5 ug/ml, solution of di-n-
octylphthalate R in anhydrous ethanolR.
Test solution Cut 0.2 g of the material to be examined
into pieces about 0.5 cm in length. Dissolve the pieces in
12.5 mL of internal standard solution S3 using a
polytetrafluoroethylene magnetic stirring bar. Complete
dissolution of the material to be examined is obtained after
stirring for about 20-30 min. The poly(vinyl chloride) is
precipitated as a white powder by adding dropwise 37.5 mL
of anhydrous ethanolR. Centrifuge, then dilute 1.0 mL ofthe
supernatant to 50.0 mL with anhydrous ethanolR. The final
concentration of the internal standard in the test solution is
5/lg/mL.
The stock solutions may be stored at 4 DC for up to 2 weeks.
Stock solution (a) Dissolve 20.0 mg of plastic
additive 01 CRS in internal standard solution S4 and dilute to
20.0 mL with internal standard solution S4.
Stock solution (b) Dissolve 20.0 mg of plastic
additive 24 CRS in internal standard solution S4 and dilute to
20.0 mL with internal standard solution S4.
Stock solution (c) Dissolve 20.0 mg of plastic
additive 25 CRS in internal standard solution S4 and dilute to
20.0 mL with internal standard solution S4.
Stock solution (d) Dissolve 20.0 mg of plastic
additive 26 CRS in internal standard solution S4 and dilute to
20.0 mL with internal standard solution S4.
Stock solution (e) Dissolve 20.0 mg of plastic
additive 27 CRS in internal standard solution S4 and dilute to
20.0 mL with internal standard solution S4.
Reference solutions (al)-(a5) Dilute stock solution (a)
with internal standard solution S4 to obtain 5 reference
solutions containing 10-40 ug/ml, of plastic additive 01 CRS.
Reference solutions (bl)-(b5) Dilute stock solution (b)
with internal standard solution S4 to obtain 5 reference
solutions containing 10-40 ug/ml, of plastic additive 24 CRS.
V-A626 Appendix XIX D 2020

Reference solutions (cl)-(c5) Dilute stock solution (c) after the injection of a standard solution. The ratios in the
with internal standard solution S4 to obtain 5 reference table below are given for information.
solutions containing 10-40 ug/ml, of plastic additive 25 CRS.
Reference solutions (dl)-(d5) Dilute stock solution (d) Substance Ion 1 Ion 2 Ion 3 Ion ratio Ion ratio
[rn/z] [rn/z] [rn/z] 2/1 3/1
with internal standard solution S4 to obtain 5 reference
(%) (%)
solutions containing 10-40 ug/ml, of plastic additive 26 CRS.
Plastic additive 01 149 167 279 50 30
Reference solutions (el)-(e5) Dilute stock solution (e) Plastic additive 24 155 127 299 30 13
with internal standard solution S4 to obtain 5 reference Plastic additive 25 71 213 315 45 20
solutions containing 10-40 ~glrnL of plastic additive 27 CRS. Plastic additive 26 305 193 323 55 20
Column: Plastic additive 27 261 149 167 130 85
- material: fused silica; DnOP (internal 149 279 167 / /
- size: 1 = 30 m, 0 = 0.25 mm; standard)
- stationary phase: poly(dimethyl) (diphenyl)siloxane R (film
thickness 0.25 urn), System suitability:
Carrier gas heliumfor chromatography R. - resolution: if plastic additive 27 is tested, minimum 1.5
Flow rate 1 mUmin. between the peaks due to the internal standard and
Split ratio 1:20. plastic additive 27;
- repeatability: maximum relative standard deviation of
Temperature: 1.0 per cent for the retention time of the peak due to the
plastic additive, determined on 6 injections of a reference
Time Temperature
solution of each plastic additive tested situated in the
(min) CC)
middle of the calibration range (e.g. 20 ug/ml.);
Column o 100
maximum relative standard deviation of 3.0 per cent for
0-3.3 100 ~ 200
the ratio of the area of the peak due to the plastic additive
3.3 - 20 200 ~ 250
to that due to the internal standard, determined on
20 - 22.5 250
6 injections of a reference solution of each plastic additive
22.5 - 23 250 ~ 270
tested situated in the middle of the calibration range
23 - 25 270
(e.g. 20 ug/ml.).
25 - 25.6 270 ~ 320
25.6 - 30.6 320
From the calibration curve obtained with the reference
Injection port 300
solutions, calculate the percentage content of plastic additives
in the material to be examined.
Detection Mass spectrometer as described below; adjust Limits:
the detector settings so as to comply with the system - plastic additive 01: maximum 40 per cent;
suitability criteria: - plastic additive 24: maximum 45 per cent;
- quadrupole mass spectrometer equipped with. an electron - plastic additive 25: maximum 45 per cent;
impact ionisation mode (70 eV); - plastic additive 26: maximum 45 per cent;
- ion source temperature: 230 DC; - plastic additive 27: maximum 45 per cent.
- acquisition system: performed on full-scan (m/z = 40-350) Chlorides (2.4.4)
and on single-ion monitoring (SIM) modes; Maximum 0.4 ppm, determined with solution S2 (see 3.2.3).
- solvent delay: 2.5 min; Prepare the standard using a mixture of 1.2 mLof chloride
- mass spectrometer parameters for the fragmentometric standardsolution (5 ppm Cl) Rand 13.8 mL of waterR.
mode (SIM) set as follows: Residue on evaporation
Evaporate to dryness 100 mL of solution S2 (see 3.2.3) in a
Substance Ion 1 Ion 2 Ion 3
[rn/z]
borosilicate-glass beaker of appropriate capacity, previously
[m/z] [m/z]
heated to 105 DC. Evaporate to dryness in the same
Plastic additive 01 149 167 279
conditions 100 mL of the reference solution (blank test).
Plastic additive 24 155 127 299
Dry to constant mass at 100-105 DC. The residue from
Plastic additive 25 71 213 315
solution S2 weighs a maximum of 3 mg, taking into account
Plastic additive 26 305 193 323
the blank test.
Plastic additive 27 261 149 167
DnOP (internal 149 279 167 STORAGE
standard) See general chapter 3.2.3. Sterile plastic containers for human
blood and blood components.
Injection 1 ~L. LABELLING
Relative retention With reference to di-s-octyl phthalate See general chapter 3.2.3. Sterile plastic containers for human
(retention time = about 22 min): plastic blood and blood components.
additive 01 = about 0.80; plastic additive
= =
24 about 0.95-1.09; plastic additive 27 about 1.02; 3. Sterile Containers of Plasticised Poly(Vinyl
Chloride) for Human Blood Containing
plastic additive 25 = about 1.14; plastic additive
26 = about 1.34. Anticoagulant Solution .
The specificity of the detection is checked by monitoring 3
(Ph. Bur. method3.2.5)
different ions for each substance using a mass spectrometer DEFINITION
in SIM mode. Ion ratios are determined from the peak areas Sterile plastic containers containing an anticoagulant solution
complying with the monograph Anticoagulant and preservative
solutions for human blood (0209) are used for the collection,

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2020
storage and administration of blood. Before filling they
comply with the description and characters given in general
chapter 3.2.4. Empty sterile containers of plasticised poly(vinyl
chloride) for human blood and blood components.
Unless otherwise authorised as described in general chapter
3.2.3. Sterile plastic containers for human blood and blood
components, the nature and composition of the material from
which the containers are made should comply with the
requirements prescribed in general chapter 3.1.1.1. Materials
based on plasticised poly(vinyl chloride) for containers for human
bloodand blood components.
TESTS
They comply.with thetests prescribed in general chapter
3.2.3. Sterile plastic containers for human bloodand blood
components and with the following tests.
Volume of anticoagulant solution
Empty the container, collecting the anticoagulant solution in
a graduated cylinder. The volume does not differ by more
than ± 10 per cent from the stated volume.
Absorbance (2.2.25)
Mb~sure the absorbance of the anticoagulant solution
rerrioved from the container between 250 nm and 350 nm,
using as.the compensation liquid an anticoagulant solution of
th~·;sap.1e composition that has not been in contact with a
pla~ticInaterial.The absorbance at the maximum at 280 nm
is not greater than 0.5.
Plastic additives 01,24, 25, 26 and 27
Gas chromatography (2.2.28) coupled with mass
spectrometry. (2.2.43).
Carefully remove the anticoagulant solution by means of the
flexible transfer tube. Using a funnel fitted to the tube,
completely fill the container with waterR, leave in contact for
1 min while squeezing the container gently, then empty
completely. Repeat the rinsing. The container, emptied and
rinsed in this manner, complies with the test for plastic
additives 01, 24, 25, 26 and 27 prescribed in general chapter
3.2.4. Empty sterile containers of plasticised poly(vinyl chloride)
for human blood and blood components.
STORAGE
See general chapter 3.2.3. Sterile plastic containers for human
blood and bloodcomponents.
LABELLING
See general chapter 3.2.3. Sterile plastic containers for human
bloodand blood components.
ED Rubber Closures for Containers for
Aqueous Parenteral Preparations
(Rubber Closures for Containers for Aqueous Parenteral
Preparations for Powders and for Freeze-dried Powders, Ph. Eur.
method 3.2.9)
Rubber closures for containers for aqueous parenteral
preparations, for powders and for freeze-dried powders are
elastomers made of materials obtained by vulcanisation
(cross-linking) of macromolecular organic substances, using
appropriate additives. They cover all types of rubber closures
including stoppers for vials, sealing disks and plunger
stoppers for cartridges, as well as rubber tip caps, needle
shields and plunger stoppers for syringes. The elastomers are
polymers, obtained by chemical synthesis, or are polymers of
natural origin. The choice of the principal components and
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l.IJ1::lenUIX XIX E
of the various additives (for example, vulcanisers,
accelerators, stabilisers, pigments) depends on the properties
required for the finished article. The specifications apply to
rubber closures made from rubber of 1 kind only, to coated
closures, to bi-layer seals and to lubricated closures. Coated
closures consist of a bulk of rubber, bearing on its surface or
part of its surface a layer ofa different polymer. Bi-layer seals
are composed of 2 different layers of rubber, 1 of which
exhibits a higher level of chemicalpurity and is intended for
contact with a pharmaceutical preparation; the other layer
exhibits a higherleveLof elasticity and is intended to improve
self-sealing and fragmentation resistance .ofthe seal.
Lubricated .rubber closures are closures treated with silicone
oil (3.1. 8)..or.other lubricants, for .exarnple•materials
chemicaliYor mechanically bonded to theclosures.
If closuresarelubricated.theyc0rn.ply.in lubricated state. with
the requirements as de.fined.inthis general chapter.
The. specifications do not apply to closures made from
silicone elastomer (which are dealt with in general chapter
3.1.9. Silicone elastomer for closures and tubing).
Rubber closures may be classified in 2 types: type I closures
meet the strictest requirements and are preferred; type IT
closures have mechanical properties suitable for special uses
(for example, multiple piercing) and. cannot meet
requirements as severe as for type I closures because of their
chemical composition.
The closures chosen for use with aparticuIar preparation are
such that:
- the. components of the preparation in contact with the
closures are not adsorbed onto the surface of the closures
and do not migrate into or through the closures to an
extent sufficient to .affect the preparation adversely;
-the closures do not release substances in quantities
sufficient to affect the stability of the preparation or to
present a risk of toxicity;
- the closures are compatible with the preparation for
which they are used throughout its period of validity.
The manufacturer of the preparation must obtain from the
supplier an assurance that the composition of the closure
does not vary and that it is identical to that of the closure
used during compatibility testing. If the supplier informs the
manufacturer of the preparation that changes have been
made to the composition, a risk assessment should be applied
to determine the need to repeat the compatibility testing,
totally or partly, depending on the nature of the changes.
The closures are washed and may be sterilised before use.
CHARACTERS
Rubber closures are elastic. They are translucent or opaque
and have no characteristic colour, the latter depending on the
additives used. They are practically insoluble in
tetrahydrofuran, in which, however, a considerable reversible
swelling may occur. They are homogeneous and practically
free from flash and adventitious materials (for example,
fibres, foreign particles, waste rubber).
IDENTIFICATION
Identification of the type of rubber usedfor the closures is not
within the scope of this specification. The identification tests given
below distinguish between closures made from rubber and those
made from silicone elastomer and plastic materials but do not
differentiate all types of rubber. Other identity tests may be carried
out with the aim of detecting differences in a batch compared with
the closures usedfor compatibility testing. One or more of the
following analytical methods may be applied for thispurpose:
determination of relative density, determination of sulfated ash,
V-A628 Appendix XIX E
determination of sulfurcontent, thin-layer chromatography carried
out on an extract, ultraviolet absorption spectrophotometry of an
extract, infrared absorption spectrophotometry of a pyrolysate or
attenuatedtotal reflectance (ATR).
A. Infrared absorption spectrophotometry (2.2.24).
Examine by attenuated total reflectance (ATR).
If necessary, cut the sample along an appropriate axis and
examine the cut surface. For coated, bi-layer and lubricated
closures, perform the test for each different part of the
closure. This is not required for silicone oil used as lubricant.
Identification of silicone oil can be performed prior to it
being used.
Comparison Type sample.
If direct ATR measurement on the surface is not feasible
(mainly rubber closures filled with carbon black), heat an
appropriate amount of rubber in a heat-resistant test-tube
over an open flame to dry the sample and continue heating
until pyrolysate vapours are condensed near the top edge of
the test-tube. Examine the pyrolysate of the sample by ATR
and compare the spectrum with that obtained with the
pyrolysate of the type sample.
B. Total ash (2.4.16).
If the sample has not been subjected to steam sterilisation,
drying at 100-105 °C can be omitted. Determine the
percentage content of total ash in the sample to be examined
and compare with the percentage content of total ash in the
type sample (Ao). The total ash content falls within the
following ranges depending on the total ash content of the
type sample, or, if not available, in the range defined as the
target for the specific rubber type.
Total ash in the type sample, Ao Limit for total ash in the sample
(per cent) (per cent)
Ao ~ 5.0 CAo -0.75) to CAo + 0.75)
5.0 < Ao ~ 10 CAo -1.0) to CAo + 1.0)
Ao> 10 CAo -2.0) to CAo + 2.0)
In addition to the use of platinum and silica crucibles
described in general chapter 2.4.16, porcelain crucibles may
be used. The sample may be ignited using a microwave oven
instead of a muffle furnace.
TESTS
Solution S
Place a number of uncut closures with a total surface area of
about 100 cm 2 in a wide-necked flask (type I glass, 3.2.1),
add 200 mL of waterR and weigh. Cover the mouth of the
flask with a borosilicate-glass beaker. Heat in an autoclave so
that a temperature of 121 ± 2 °C is reached within
20-30 min and maintain at this temperature for 30 min.
Immerse the temperature probe for the autoclave
programme-control in water in a container comparable to
that used for the sample. Cool to room temperature over
about 30 min. Make up to the original mass with water R.
Shake and decant the solution immediately. Shake solution S
before each test. If using a tightly closed flask (type I glass,
3.2.1) with an inert closure instead of a wide-necked flask
covered with a borosilicate-glass beaker, it is not necessary to
make up to the original mass.
Blank solution Prepare a blank solution in the same
manner using 200 mL of water R.
Appearance of solution S
Solution S is not more intensely coloured than reference
solution GY s (2.2.2, l\IIethod II). For type I closures,
solution S is not more opalescent than reference
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2020
suspension II (2.2.1) and for type II closures, solution S is
not more opalescent than reference suspension III. In case of
nephelometric determination, the limit for type I closures is
6 NTU and the limit for type II closures is 18 NTU.
Acidity or alkalinity
To 20 mL of solution S add 0.1 mL of bromothymol blue
solution R1. Carry out a titration using 20.0 mL of the blank
(see solution S). Not more than 0.3 mL of 0.01 M sodium
hydroxide or 0.8 mL of 0.01 M hydrochloric acid is required to
change the colour of the indicator to blue or yellow,
respectively. If after adding the indicator the solution is
green, it is neutral and no titration is needed.
Absorbance
Carry out the test within 5 h of preparation of solution S. Filter
solution S through a membrane filter (nominal pore size
0.45 urn), rejecting the first few millilitres of filtrate. Measure
the absorbance (2.2.25) of the filtrate at wavelengths from
220-360 nm using the blank (see solution S) as
compensation liquid: absorbance within the 220-360 nm
range does not exceed 0.2 for type I closures or 4.0 for type
II closures. If necessary, dilute the filtrate before
measurement of the absorbance and correct the result for the
dilution.
Reducing substances
Carry out the test within 4 h of preparation of solution S.
To 20.0 mL of solution S add 1 mL of dilute sulfuric acid R
and 20.0 mL of 0.002 M potassium permanganate. Boil for
3 min. Cool. Add 1 g of potassium iodide R and titrate
immediately with 0.01 M sodium thiosulfate, using 0.25 mL of
starch solution R as indicator. Carry out a titration using
20.0 mL of the blank (see solution S). The difference
between the titration volumes is not greater than 3.0 mL for
type I closures and 7.0 mL for type II closures.
Ammonium (2.4.1, MethodA)
Maximum 2 ppm.
Dilute 5 mL of solution S to 14 mL with water R.
Extractable zinc
Maximum 5 ~lg of extractable Zn per millilitre of solution S.
Atomic absorption spectrometry (2.2.23, Method I).
Test solution Use solution S. If results are outside the
calibration range, dilute 10.0 mL of solution S to an
appropriate volume with 0.1 M hydrochloric acid.
Reference solutions Prepare the reference solutions using
zinc standardsolution (10 ppm Zn) R diluted with 0.1 M
hydrochloric acid.
Source Zinc hollow-cathode lamp.
Wavelength 213.9 nm.
Atomisation device Air-acetylene flame.
Extractable heavy metals (2.4.8)
Maximum 2 ppm.
Solution S complies with test A. Prepare the reference
solution using leadstandardsolution (2 ppm Pb) R.
Residue on evaporation
Evaporate 50.0 mL of solution S to dryness on a water-bath
and dry at 100-105 °C. The residue weighs not more than
2.0 mg for type I closures and not more than 4.0 mg for type
II closures.
Volatile sulfides
Place closures, cut if necessary, with a total surface area of
20 ± 2 cm 2 in a 100 mL conical flask and add SO mL of a
20 g/Lsolution of citric acid monohydrate R. Place a piece of
lead acetate paper R over the mouth of the flask and maintain
2020
the paper in position by placing over it an inverted weighing
bottle. Heat in an autoclave at 121 ± 2 °C for 30 min.
Any black stain on the paper is not more intense than that of
a standard, treated in the same manner, prepared by mixing
50 mL of a 20 gIL solution of citric acid monohydrate R and
5.0 mL of a freshly prepared 0.0308 gIL solution of sodium
sulfide R.
The tests for penetrability, fragmentation and self-sealing are
performed on whole closures.
For the tests for penetrability, fragmentation and self-sealing, treat
non-sterilised closures as described for the preparation of solution S
and allow to dry. To perform these 3 tests, use for each
closure a new, lubricated, long-bevel' (bevel angle 12 ± 2°)
hypodermicneedle with an externaldiameter of O.8mm and
pierce the closures with the needle perpendicular to the
surface without rotating the needle.
Penetrability
For closures intended to be pierced by a hypodermic needle,
carry out the following test. Fill 10 suitable vials to the
nominal volume with water R, fit the closures to be examined
and-secure with a cap. The force required for piercing,
determined with an accuracy of ± 0.25 N, is not greater
than 10 N for each closure.
Fr~entation
Fot.el9sures intended to be pierced by a hypodermic needle,
carry.out the following test. If the closures are to be used for
aqueous preparations, introduce in 12 clean vials a volume of
water R corresponding to the nominal volume minus 4 mL,
close the vials with the closures to be examined, secure with
a cap and allow to stand for 16 h. If the closures are to be
used with dry preparations, close 12 clean vials with the
closures to be examined. Using a needle fitted to a clean
syringe, inject into the vial 1 mL of water R and remove
1 mL of air; carry out this operation 4 times for each closure,
piercing the closure each time at a different site. Use a new
needle for each closure and check that the needle is hot
blunted during the test. Pass the liquid in the vials through a
filter with a pore size of 0.5 urn, Count the fragments of
rubber visible to the naked eye. The total number of
fragments does not exceed 5. This limit is based on the
assumption that fragments with a diameter equal to or
greater than 50 urn are visible to the naked eye; in cases of
doubt or dispute, the fragments are examined with a
microscope to verify their nature and size.
Self-sealing test
For closures intended to be used with multidose containers,
carry out the following test. Fill 10 vials matching the design
of the stopper to the nominal volume with waterR, fit the
closures to be examined, secure with a cap and crimp tightly.
Pierce each closure 10 times, piercing the closure each time
at a different site. Immerse the vials upright in a 1 gIL
solution of methylene blue R and reduce the external pressure
by 27 kPa for 10 min. Restore atmospheric pressure and
leave the vials immersed for 30 min. Rinse the outside of the
vials. None of the vials contains any trace of coloured
solution.
1 See ISO 7864) Sterile hypodermic needles jar single use.
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Sets the Transfusion and
Blood Components
(ph. Eur. method 3.2.6)
DEFINITION
Sets for the transfusion of blood and blood components
consist principally of plastic tubing to which are fitted the
parts necessary to enable the set to be used for transfusion in
the appropriate manner. Sets include a closure-piercing
device, a blood filter, a drip chamber, a flow regulator, a
Luer COnnector and, usually, a site that allows an injection to
be made at the time of use. When the sets are to be used
with containers requiring an air-filter, this may be
incorporated in the closure-piercing device or a separate air-
inlet device may be used. The chamber enclosing the blood
filter, the drip chamber and the main tubing are transparent.
The materials chosen and the design of the set are such as to
ensure absence of haemolytic effects. The sets comply with
current standards regarding dimensions and performance.
All parts of the set that may be in contact with blood and
blood components are sterile and pyrogen-free. Each set is
presented in an individual package that maintains the sterility
of the contents. The sets are not to be re-sterilised or
re-used.
Sets for the transfusion of blood and blood components are
manufactured in accordance with the rules of good
manufacturing practice for medical devices and any relevant
national regulations.
TESTS
Carry out the tests on sterilised sets.
Solution S
Make a closed circulation system from 3 sets and a 300 mL
borosilicate-glass vesseL Fit to the vessel a suitable thermostat
device that maintains the temperature of the liquid in the
vessel at 37 ± 1°C. Circulate 250 mL of waterfor
injections R through the system in the direction used for
transfusion for 2 h at a rate of 1 lA1 (for example using a
peristaltic pump applied to as short a piece of suitable
silicone elastomer tubing as possible). Collect the whole of
the solution and allow to cool.
Appearance of solution
Solution S is clear (2.2.1) and colourless (2.2.2, Method II).
Acidity or alkalinity
To 25 mL of solution S add 0.15 mL of BRP indicator
solution R. Not more than 0.5 mL of 0.01 M sodium hydroxide
is required to change the colour of the indicator to blue.
To 25 mL of solution S add 0.2 mL of methyl orange
solution R. Not more than 0.5 mL of 0.01 M hydrochloric acid
is required to reach the beginning of the colour change of the
indicator.
Absorbance (2.2.25)
Maximum 0.30, determined between wavelengths of 230 urn
and 250 urn on solution S; maximum 0.15, determined
between wavelengths of 251 urn and 360 urn on solution S.
Reducing substances
Carry out the test within 4 h ofpreparation of
solution S To 20.0 mL of solution S add 1 mL of dilute
sulfuric acid Rand 20.0 mL of 0.002 M potassium
permanganate. Boil for 3 min and cool immediately. Add 1 g
of potassium iodide R and titrate with 0.01 M sodium thiosulfate
using 0.25 mL of starchsolution R as indicator. Carry out a
blank test using 20 mL of waterfor injections R. .
V-A630 Appendix XIX F
The difference between the titration volumes is not greater
than 2.0 mL.
Ethylene oxide
Gas chromatography (2.2.28).
Column:
- material: stainless steel;
- size: 1 = 1.5 m, 0 = 6.4 mID;
- stationary phase: silanised diatomaceous earth for gas
chromatography R impregnated with macrogol 1500 R (3 g
per 10 g).
Carrier gas heliumfor chromatography R.
Flow rate 20 mIJmin.
Temperature:
- column: 40 "C;
- injection port: 100 "C;
- detector. 150 "C.
Detection Flame ionisation.
Verify the absence of peaks interfering with the ethylene
oxide peak by carrying out the test using an unsterilised set
or using the same chromatographic system with the following
modifications.
Column:
- size: 1= 3 m, 0 = 3.2 mID;
- stationary phase: silanised diatomaceous earth for gas
chromatography R impregnated with
triscyanoethoxypropane R (2 g per 10 g);
- temperature: 60 "C.
Ethylene oxide solution Prepare in a fume cupboard. Place
50.0 mL of dimethylacetamide R in a 50 mL vial, stopper,
secure the stopper and weigh to the nearest 0.1 mg. Fill a
50 mL polyethylene or polypropylene syringe with gaseous
ethylene oxide R, allow the gas to remain in contact with the
syringe for about 3 min, empty the syringe and fill again with
50 mL of gaseous ethylene oxide R. Fit a hypodermic needle
to the syringe and reduce the volume of gas in the syringe
from 50 mL to 25 mL. Inject these 25 mL of ethylene oxide
slowly into the vial, shaking gently and avoiding contact
between the needle and the liquid. Weigh the vial again: the
increase in mass is 45 mg to 60 mg and is used to calculate
the exact concentration of the solution (about 1 gIL).
Test Weigh the set after removing the package. Cut the set
into pieces of maximum dimension 1 em and place the
pieces in a 250-500 mL vial containing 150 mL of
dimethylacetamide R. Close the vial with a suitable stopper
and secure the stopper. Place the vial in an oven at
70 ± 1 "C for 16 h. Remove 1 mL of the hot gas from the
vial and inject it onto the column. From the calibration curve
and the height of the peak obtained, calculate the mass of
ethylene oxide in the vial.
Calibration curve In a series of 7 vials of the same type as
that used for the test and each containing 150 mL of
dimethylacetamide R, place respectively 0 mL, 0.05 mL,
0.10 mL, 0.20 mL, 0.50 mL, 1.00 mL and 2.00 mL of the
ethylene oxide solution, i.e, about 0 ug, 50 !J.g, 100 ug,
200 !J.g, 500 ug, 1000 !J.g and 2000 ug of ethylene oxide.
Stopper the vials, secure the stoppers and place the vials in
an oven at 70 ± 1 "C for 16 h. Inject 1 mL of the hot gas
from each vial onto the column and draw a calibration curve
from the heights of the peaks and the mass of ethylene oxide
in each flask.
Limit If the label states that ethylene oxide has been used
for sterilisation:
- ethylene oxide: maximum 10 ppm.
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2020
Extraneous particles
Fill the set via the normal inlet with a 0.1 gIL solution of
sodium laurilsulfate R, previously filtered through a sintered-
glass filter (16) (2.1.2) and heated to 37 "C. Collect the
liquid via the normal outlet. When examined under suitable
conditions of visibility, the liquid is clear and practically free
from visible particles and filaments (it is assumed that
particles and filaments with a diameter equal to or greater
than 50 um are visible to the naked eye).
Flow rate
Pass through a complete set with the flow regulator fully
open 50 mL of a solution having a viscosity of 3 mf'a-s
(3 cP) (for example a 33 gIL solution of macrogol 4000 R at
20 "C) under a static head of 1 m. The time required for
passage of 50 mL of the solution is not greater than 90 s.
Resistance to pressure
Make tight the extremities of the set and any air-inlet device.
Connect the set to a compressed air outlet fitted with a
pressure regulator. Immerse the set in a tank of water at
20-23 "C. Apply progressively an excess pressure of 100 kPa
and maintain for 1 min. No air bubble escapes from the set.
Transparency
Use as reference suspension the primary opalescent
suspension (2.2.1) diluted 1 in 8 for sets having tubing with
an external diameter less than 5 mID and diluted 1 in 16 for
sets having tubing with an external diameter of 5 mm or
greater. Circulate the reference suspension through the set
and compare with a set from the same batch filled with
water R. The opalescence and presence of bubbles are
discernible.
Residue on evaporation
Evaporate 50.0 mL of solution S to dryness on a water-bath
and dry to constant mass in an oven at 100-105 "C. Carry
out a blank test using 50.0 mL of water for injections R.
The difference between the masses of the residues is not
greater than 1.5 mg.
Sterility (2.6.1)
The sets comply with the test for sterility. If the sets are
stated to be sterile only internally, pass 50 mL of buffered
sodium chloride-peptone solution pH 7.0 (2.6.12) through
the set and use to carry out the test by the membrane
filtration method.
If the sets are stated to be sterile both internally and
externally, open the package with the necessary aseptic
precautions and:
- for the direct inoculation method, place the set or its
components in a suitable container containing a sufficient
quantity of the culture medium to ensure complete
immersion;
- for the membrane filtration method, place the set or its
components in a suitable container containing a sufficient
quantity of buffered sodium chloride-peptone solution
pH 7.0 (2.6.12) to allow total rinsing for 10 min.
Pyrogens (2.6.8)
Connect together 5 sets and pass through the assembly at a
flow rate not exceeding 10 mUmin 250 mL of a sterile,
pyrogen-free 9 gIL solution of sodium chloride R. Collect the
solution aseptically in a pyrogen-free container. The solution
complies with the test for pyrogens. Inject per kilogram of
the rabbit's mass, 10 mL of the solution.
LABELLING
The label states, where applicable, that the set has been
sterilised using ethylene oxide.