EUROPEAN PHARMACOPOEIA 6.

0 Water, purified

7. Using Table 1927.-2, determine the conductivity limit Aluminium (2.4.17) : maximum 10 ppb, if intended for use in
at the measured pH value in step 6. If the measured the manufacture of dialysis solutions.
conductivity in step 4 under stage 2 is not greater than Prescribed solution. To 400 ml of the water to be examined
the conductivity requirements for the pH determined, add 10 ml of acetate buffer solution pH 6.0 R and 100 ml of
the water to be examined meets the requirements of the distilled water R.
test for conductivity. If either the measured conductivity
is greater than this value or the pH is outside the range Reference solution. Mix 2 ml of aluminium standard
of 5.0-7.0, the water to be examined does not meet the solution (2 ppm Al) R, 10 ml of acetate buffer solution
requirements of the test for conductivity. pH 6.0 R and 98 ml of distilled water R.
Blank solution. Mix 10 ml of acetate buffer solution
In order to ensure the appropriate quality of the water, pH 6.0 R and 100 ml of distilled water R.
validated procedures and in-process monitoring of the
electrical conductivity and regular microbial monitoring are Heavy metals (2.4.8) : maximum 0.1 ppm.
applied. Heat 200 ml in a glass evaporating dish on a water-bath until
the volume is reduced to 20 ml. 12 ml of the concentrated
Highly purified water is stored in bulk and distributed in solution complies with limit test A. Prepare the standard
conditions designed to prevent growth of micro-organisms using 10 ml of lead standard solution (1 ppm Pb) R.
and to avoid any other contamination.
Bacterial endotoxins (2.6.14) : less than 0.25 IU/ml.
Table 1927.-2. – Stage 3 - pH and conductivity requirements
(for atmosphere and temperature equilibrated samples) LABELLING
pH Conductivity
The label states, where applicable, that the substance is
(µS·cm− 1)
suitable for use in the manufacture of dialysis solutions.
5.0 4.7
5.1 4.1
01/2008:0008
5.2 3.6
5.3 3.3 WATER, PURIFIED
5.4 3.0
5.5 2.8 Aqua purificata
5.6 2.6
H 2O Mr 18.02
5.7 2.5 [7732-18-5]
5.8 2.4
DEFINITION
5.9 2.4
Water for the preparation of medicines other than those
6.0 2.4 that are required to be both sterile and apyrogenic, unless
6.1 2.4 otherwise justified and authorised.
6.2 2.5
Purified water in bulk
6.3 2.4
PRODUCTION
6.4 2.3
Purified water in bulk is prepared by distillation, by ion
6.5 2.2 exchange, by reverse osmosis or by any other suitable
6.6 2.1 method from water that complies with the regulations on
water intended for human consumption laid down by the
6.7 2.6
competent authority.
6.8 3.1 During production and subsequent storage, appropriate
6.9 3.8 measures are taken to ensure that the total viable aerobic
count is adequately controlled and monitored. Appropriate
7.0 4.6
alert and action limits are set so as to detect adverse trends.
Under normal conditions, an appropriate action limit is a
total viable aerobic count (2.6.12) of 100 micro-organisms
CHARACTERS per millilitre, determined by membrane filtration, using agar
Appearance : clear and colourless liquid. medium S and incubating at 30-35 °C for 5 days. The size of
the sample is to be chosen in relation to the expected result.
TESTS In addition, the test for total organic carbon (2.2.44) with
a limit of 0.5 mg/l or alternatively the following test for
Nitrates : maximum 0.2 ppm. oxidisable substances is carried out : to 100 ml add 10 ml
Place 5 ml in a test-tube immersed in iced water, add 0.4 ml of dilute sulphuric acid R and 0.1 ml of 0.02 M potassium
of a 100 g/l solution of potassium chloride R, 0.1 ml of permanganate and boil for 5 min ; the solution remains
diphenylamine solution R and, dropwise with shaking, faintly pink.
5 ml of nitrogen-free sulphuric acid R. Transfer the tube Conductivity. Determine the conductivity off-line or in-line
to a water-bath at 50 °C. After 15 min, any blue colour in under the following conditions.
the solution is not more intense than that in a reference
solution prepared at the same time in the same manner EQUIPMENT
using a mixture of 4.5 ml of nitrate-free water R and 0.5 ml Conductivity cell :
of nitrate standard solution (2 ppm NO3) R. — electrodes of a suitable material such as stainless steel ;

General Notices (1) apply to all monographs and other texts 3213
Water, purified EUROPEAN PHARMACOPOEIA 6.0

— cell constant : within 2 per cent of the given value solution prepared at the same time in the same manner
determined using a certified reference solution with a using a mixture of 4.5 ml of nitrate-free water R and 0.5 ml
conductivity less than 1500 µS·cm− 1. of nitrate standard solution (2 ppm NO3) R.
Conductometer : resolution 0.1 µS·cm− 1 on the lowest range. Aluminium (2.4.17) : maximum 10 ppb, if intended for use in
System calibration (conductivity cell and conductometer) : the manufacture of dialysis solutions.
— against one or more suitable certified standard solutions ; Prescribed solution. To 400 ml of the water to be examined
— accuracy : within 3 per cent of the measured conductivity add 10 ml of acetate buffer solution pH 6.0 R and 100 ml of
plus 0.1 µS·cm− 1. distilled water R.
Conductometer calibration : by means of precision resistors Reference solution. Mix 2 ml of aluminium standard
or equivalent devices, after disconnecting the conductivity solution (2 ppm Al) R, 10 ml of acetate buffer solution
cell, for all ranges used for conductivity measurement and pH 6.0 R and 98 ml of distilled water R.
cell calibration (with an accuracy within 0.1 per cent of the Blank solution. Mix 10 ml of acetate buffer solution
stated value, traceable to the official standard). pH 6.0 R and 100 ml of distilled water R.
If in-line conductivity cells cannot be dismantled, system Heavy metals (2.4.8) : maximum 0.1 ppm.
calibration may be performed against a calibrated Heat 200 ml in a glass evaporating dish on a water-bath until
conductivity cell placed close to the cell to be calibrated in the volume is reduced to 20 ml. 12 ml of the concentrated
the water flow. solution complies with limit test A. Prepare the standard
PROCEDURE using 10 ml of lead standard solution (1 ppm Pb) R.
Measure the conductivity without temperature Bacterial endotoxins (2.6.14) : less than 0.25 IU/ml, if
compensation, recording simultaneously the temperature. intended for use in the manufacture of dialysis solutions
Temperature-compensated measurement may be performed without a further appropriate procedure for removal of
after suitable validation. bacterial endotoxins.
The water to be examined meets the requirements if the
measured conductivity at the recorded temperature is not LABELLING
greater than the value in Table 0008.-1. The label states, where applicable, that the substance is
Table 0008.-1. – Temperature and conductivity suitable for use in the manufacture of dialysis solutions.
requirements
Temperature Conductivity Purified water in containers
(°C) (µS·cm− 1)
0 2.4 DEFINITION
10 3.6 Purified water in bulk that has been filled and stored in
conditions designed to assure the required microbiological
20 4.3 quality. It is free from any added substances.
25 5.1
CHARACTERS
30 5.4
Appearance : clear and colourless liquid.
40 6.5
TESTS
50 7.1
It complies with the tests prescribed in the section on
60 8.1 Purified water in bulk and with the following additional tests.
70 9.1 Acidity or alkalinity. To 10 ml, freshly boiled and cooled in a
75 9.7 borosilicate glass flask, add 0.05 ml of methyl red solution R.
The solution is not coloured red.
80 9.7
To 10 ml add 0.1 ml of bromothymol blue solution R1. The
90 9.7 solution is not coloured blue.
100 10.2 Oxidisable substances. To 100 ml add 10 ml of dilute
sulphuric acid R and 0.1 ml of 0.02 M potassium
For temperatures not listed in Table 0008.-1, calculate the permanganate and boil for 5 min. The solution remains
maximal permitted conductivity by interpolation between faintly pink.
the next lower and next higher data points in the table.
Chlorides. To 10 ml add 1 ml of dilute nitric acid R and
Purified water in bulk is stored and distributed in conditions 0.2 ml of silver nitrate solution R2. The solution shows no
designed to prevent growth of micro-organisms and to avoid change in appearance for at least 15 min.
any other contamination.
Sulphates. To 10 ml add 0.1 ml of dilute hydrochloric acid R
CHARACTERS and 0.1 ml of barium chloride solution R1. The solution
Appearance : clear and colourless liquid. shows no change in appearance for at least 1 h.
Ammonium : maximum 0.2 ppm.
TESTS
To 20 ml add 1 ml of alkaline potassium tetraiodomercurate
Nitrates : maximum 0.2 ppm. solution R. After 5 min, examine the solution down the
Place 5 ml in a test-tube immersed in iced water, add 0.4 ml vertical axis of the tube. The solution is not more intensely
of a 100 g/l solution of potassium chloride R, 0.1 ml of coloured than a standard prepared at the same time by
diphenylamine solution R and, dropwise with shaking, adding 1 ml of alkaline potassium tetraiodomercurate
5 ml of nitrogen-free sulphuric acid R. Transfer the tube solution R to a mixture of 4 ml of ammonium standard
to a water-bath at 50 °C. After 15 min, any blue colour in solution (1 ppm NH4) R and 16 ml of ammonium-free
the solution is not more intense than that in a reference water R.

3214 See the information section on general monographs (cover pages)

EUROPEAN PHARMACOPOEIA 6.0 Wheat-germ oil, refined

Calcium and magnesium. To 100 ml add 2 ml of ammonium neck into the flask with 25 ml of sulphuric acid R ; continue
chloride buffer solution pH 10.0 R, 50 mg of mordant the heating until a clear solution is obtained ; add 45 ml of
black 11 triturate R and 0.5 ml of 0.01 M sodium edetate. A strong sodium hydroxide solution R.
pure blue colour is produced. Oxidising substances (2.5.30) : maximum 20 ppm, calculated
Residue on evaporation : maximum 0.001 per cent. as H2O2.
Evaporate 100 ml on a water-bath and dry in an oven at Sulphur dioxide (2.5.29) : maximum 50 ppm.
100-105 °C. The residue weighs a maximum of 1 mg. Iron (2.4.9) : maximum 10 ppm.
Microbial contamination. Total viable aerobic count (2.6.12) Shake 1.5 g with 15 ml of dilute hydrochloric acid R. Filter.
not more than 102 micro-organisms per millilitre, determined The filtrate complies with the limit test for iron.
by membrane filtration, using agar medium B.
Loss on drying (2.2.32) : maximum 15.0 per cent, determined
LABELLING on 1.000 g by drying in an oven at 130 °C for 90 min.
The label states, where applicable, that the substance is Sulphated ash (2.4.14) : maximum 0.6 per cent, determined
suitable for use in the manufacture of dialysis solutions. on 1.0 g.
Microbial contamination. Total viable aerobic count (2.6.12)
not more than 103 bacteria and not more than 102 fungi per
01/2008:0359 gram, determined by plate count. It complies with the test
corrected 6.0 for Escherichia coli (2.6.13).

WHEAT STARCH 01/2008:1379

Tritici amylum WHEAT-GERM OIL, REFINED

DEFINITION
Wheat starch is obtained from the caryopsis of Triticum
aestivum L. (T. vulgare Vill.).
CHARACTERS
Appearance : very fine, white or almost white powder which
creaks when pressed between the fingers.
Solubility : practically insoluble in cold water and in alcohol.
Wheat starch does not contain starch grains of any other
origin. It may contain a minute quantity, if any, of tissue
fragments of the original plant.
IDENTIFICATION
A. Examined under a microscope using equal volumes
of glycerol R and water R, it presents large and small
granules, and, very rarely, intermediate sizes. The large
granules, 10 µm to 60 µm in diameter, are discoid or,
more rarely, reniform when seen face-on. The central
hilum and striations are invisible or barely visible and the
granules sometimes show cracks on the edges. Seen in
profile, the granules are elliptical and fusiform and the
hilum appears as a slit along the main axis. The small
granules, rounded or polyhedral, are 2 µm to 10 µm in
diameter. Between crossed nicol prisms, the granules
show a distinct black cross intersecting at the hilum.
B. Suspend 1 g in 50 ml of water R, boil for 1 min and cool.
A thin, cloudy mucilage is formed.
C. To 1 ml of the mucilage obtained in identification test B, the manufacture of parenteral dosage forms : maximum 5.0.
add 0.05 ml of iodine solution R1. A dark blue colour is
produced which disappears on heating.
TESTS
pH (2.2.3) : 4.5 to 7.0.
Shake 5.0 g with 25.0 ml of carbon dioxide-free water R for Method C). Use the mixture of calibrating substances in
60 s. Allow to stand for 15 min.
Foreign matter. Examined under a microscope using a
mixture of equal volumes of glycerol R and water R, not
more than traces of matter other than starch granules are
present. No starch grains of any other origin are present.
Total protein : maximum 0.3 per cent of total protein
(corresponding to 0.048 per cent N2, conversion factor :
6.25), determined on 6.0 g by sulphuric acid digestion (2.5.9) — linolenic acid : 3.0 per cent to 10.0 per cent,
modified as follows : wash any adhering particles from the
Tritici aestivi oleum raffinatum
DEFINITION
Fatty oil obtained from the germ of the grain of Triticum
aestivum L. by cold expression or by other suitable
mechanical means and/or by extraction. It is then refined. A
suitable antioxidant may be added.
CHARACTERS
Appearance : clear, light yellow liquid.
Solubility : practically insoluble in water and in alcohol,
miscible with light petroleum (40 °C to 60 °C).
Relative density : about 0.925.
Refractive index : about 1.475.
IDENTIFICATION
A. Identification of fatty oils by thin-layer chromatography
(2.3.2). The chromatogram obtained is similar to the type
chromatogram for wheat-germ oil.
B. It complies with the test for composition of fatty acids
(see Tests).
TESTS
Acid value (2.5.1) : maximum 0.9. If intended for use in the
manufacture of parenteral dosage forms : maximum 0.3.
Peroxide value (2.5.5) : maximum 10.0. If intended for use in
Unsaponifiable matter (2.5.7) : maximum 5.0 per cent,
determined on 5.0 g.
Alkaline impurities (2.4.19). It complies with the test for
alkaline impurities in fatty oils.
Composition of fatty acids. Gas chromatography (2.4.22,
Table 2.4.22.-3.
Composition of the fatty-acid fraction of the oil :
— palmitic acid : 14.0 per cent to 19.0 per cent,
— stearic acid : maximum 2.0 per cent,
— oleic acid : 12.0 per cent to 23.0 per cent,
— linoleic acid : 52.0 per cent to 59.0 per cent,
— eicosenoic acid : maximum 2.0 per cent.

General Notices (1) apply to all monographs and other texts 3215