EUROPEAN PHARMACOPOEIA 6.0 Water, purified

7. Using Table 1927.-2, determine the conductivity limitat the measured pH value in step 6. If the measuredconductivity in step 4 under stage 2 is not greater thanthe conductivity requirements for the pH determined,the water to be examined meets the requirements of thetest for conductivity. If either the measured conductivityis greater than this value or the pH is outside the rangeof 5.0-7.0, the water to be examined does not meet therequirements of the test for conductivity.

In order to ensure the appropriate quality of the water,validated procedures and in-process monitoring of theelectrical conductivity and regular microbial monitoring areapplied.

Highly purified water is stored in bulk and distributed inconditions designed to prevent growth of micro-organismsand to avoid any other contamination.

Table 1927.-2. – Stage 3 - pH and conductivity requirements(for atmosphere and temperature equilibrated samples)

pH Conductivity(µS·cm−1)

5.0 4.7

5.1 4.1

5.2 3.6

5.3 3.3

5.4 3.0

5.5 2.8

5.6 2.6

5.7 2.5

5.8 2.4

5.9 2.4

6.0 2.4

6.1 2.4

6.2 2.5

6.3 2.4

6.4 2.3

6.5 2.2

6.6 2.1

6.7 2.6

6.8 3.1

6.9 3.8

7.0 4.6

CHARACTERS

Appearance : clear and colourless liquid.

TESTS

Nitrates : maximum 0.2 ppm.

Place 5 ml in a test-tube immersed in iced water, add 0.4 mlof a 100 g/l solution of potassium chloride R, 0.1 ml ofdiphenylamine solution R and, dropwise with shaking,5 ml of nitrogen-free sulphuric acid R. Transfer the tubeto a water-bath at 50 °C. After 15 min, any blue colour inthe solution is not more intense than that in a referencesolution prepared at the same time in the same mannerusing a mixture of 4.5 ml of nitrate-free water R and 0.5 mlof nitrate standard solution (2 ppm NO3) R.

Aluminium (2.4.17) : maximum 10 ppb, if intended for use inthe manufacture of dialysis solutions.Prescribed solution. To 400 ml of the water to be examinedadd 10 ml of acetate buffer solution pH 6.0 R and 100 ml ofdistilled water R.Reference solution. Mix 2 ml of aluminium standardsolution (2 ppm Al) R, 10 ml of acetate buffer solutionpH 6.0 R and 98 ml of distilled water R.Blank solution. Mix 10 ml of acetate buffer solutionpH 6.0 R and 100 ml of distilled water R.

Heavy metals (2.4.8) : maximum 0.1 ppm.Heat 200 ml in a glass evaporating dish on a water-bath untilthe volume is reduced to 20 ml. 12 ml of the concentratedsolution complies with limit test A. Prepare the standardusing 10 ml of lead standard solution (1 ppm Pb) R.

Bacterial endotoxins (2.6.14) : less than 0.25 IU/ml.

LABELLINGThe label states, where applicable, that the substance issuitable for use in the manufacture of dialysis solutions.

01/2008:0008

WATER, PURIFIED

Aqua purificata

H2O Mr 18.02[7732-18-5]

DEFINITIONWater for the preparation of medicines other than thosethat are required to be both sterile and apyrogenic, unlessotherwise justified and authorised.

Purified water in bulkPRODUCTIONPurified water in bulk is prepared by distillation, by ionexchange, by reverse osmosis or by any other suitablemethod from water that complies with the regulations onwater intended for human consumption laid down by thecompetent authority.During production and subsequent storage, appropriatemeasures are taken to ensure that the total viable aerobiccount is adequately controlled and monitored. Appropriatealert and action limits are set so as to detect adverse trends.Under normal conditions, an appropriate action limit is atotal viable aerobic count (2.6.12) of 100 micro-organismsper millilitre, determined by membrane filtration, using agarmedium S and incubating at 30-35 °C for 5 days. The size ofthe sample is to be chosen in relation to the expected result.In addition, the test for total organic carbon (2.2.44) witha limit of 0.5 mg/l or alternatively the following test foroxidisable substances is carried out : to 100 ml add 10 mlof dilute sulphuric acid R and 0.1 ml of 0.02 M potassiumpermanganate and boil for 5 min ; the solution remainsfaintly pink.

Conductivity. Determine the conductivity off-line or in-lineunder the following conditions.

EQUIPMENTConductivity cell :— 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 valuedetermined using a certified reference solution with aconductivity less than 1500 µS·cm−1.

Conductometer : resolution 0.1 µS·cm−1 on the lowest range.System calibration (conductivity cell and conductometer) :— against one or more suitable certified standard solutions ;— accuracy : within 3 per cent of the measured conductivity

plus 0.1 µS·cm−1.Conductometer calibration : by means of precision resistorsor equivalent devices, after disconnecting the conductivitycell, for all ranges used for conductivity measurement andcell calibration (with an accuracy within 0.1 per cent of thestated value, traceable to the official standard).If in-line conductivity cells cannot be dismantled, systemcalibration may be performed against a calibratedconductivity cell placed close to the cell to be calibrated inthe water flow.

PROCEDUREMeasure the conductivity without temperaturecompensation, recording simultaneously the temperature.Temperature-compensated measurement may be performedafter suitable validation.The water to be examined meets the requirements if themeasured conductivity at the recorded temperature is notgreater than the value in Table 0008.-1.

Table 0008.-1. – Temperature and conductivityrequirements

Temperature(°C)

Conductivity(µS·cm−1)

0 2.4

10 3.6

20 4.3

25 5.1

30 5.4

40 6.5

50 7.1

60 8.1

70 9.1

75 9.7

80 9.7

90 9.7

100 10.2

For temperatures not listed in Table 0008.-1, calculate themaximal permitted conductivity by interpolation betweenthe next lower and next higher data points in the table.Purified water in bulk is stored and distributed in conditionsdesigned to prevent growth of micro-organisms and to avoidany other contamination.

CHARACTERSAppearance : clear and colourless liquid.

TESTS

Nitrates : maximum 0.2 ppm.Place 5 ml in a test-tube immersed in iced water, add 0.4 mlof a 100 g/l solution of potassium chloride R, 0.1 ml ofdiphenylamine solution R and, dropwise with shaking,5 ml of nitrogen-free sulphuric acid R. Transfer the tubeto a water-bath at 50 °C. After 15 min, any blue colour inthe solution is not more intense than that in a reference

solution prepared at the same time in the same mannerusing a mixture of 4.5 ml of nitrate-free water R and 0.5 mlof nitrate standard solution (2 ppm NO3) R.

Aluminium (2.4.17) : maximum 10 ppb, if intended for use inthe manufacture of dialysis solutions.Prescribed solution. To 400 ml of the water to be examinedadd 10 ml of acetate buffer solution pH 6.0 R and 100 ml ofdistilled water R.Reference solution. Mix 2 ml of aluminium standardsolution (2 ppm Al) R, 10 ml of acetate buffer solutionpH 6.0 R and 98 ml of distilled water R.Blank solution. Mix 10 ml of acetate buffer solutionpH 6.0 R and 100 ml of distilled water R.

Heavy metals (2.4.8) : maximum 0.1 ppm.Heat 200 ml in a glass evaporating dish on a water-bath untilthe volume is reduced to 20 ml. 12 ml of the concentratedsolution complies with limit test A. Prepare the standardusing 10 ml of lead standard solution (1 ppm Pb) R.

Bacterial endotoxins (2.6.14) : less than 0.25 IU/ml, ifintended for use in the manufacture of dialysis solutionswithout a further appropriate procedure for removal ofbacterial endotoxins.

LABELLINGThe label states, where applicable, that the substance issuitable for use in the manufacture of dialysis solutions.

Purified water in containersDEFINITIONPurified water in bulk that has been filled and stored inconditions designed to assure the required microbiologicalquality. It is free from any added substances.

CHARACTERSAppearance : clear and colourless liquid.

TESTSIt complies with the tests prescribed in the section onPurified water in bulk and with the following additional tests.

Acidity or alkalinity. To 10 ml, freshly boiled and cooled in aborosilicate glass flask, add 0.05 ml ofmethyl red solution R.The solution is not coloured red.To 10 ml add 0.1 ml of bromothymol blue solution R1. Thesolution is not coloured blue.

Oxidisable substances. To 100 ml add 10 ml of dilutesulphuric acid R and 0.1 ml of 0.02 M potassiumpermanganate and boil for 5 min. The solution remainsfaintly pink.

Chlorides. To 10 ml add 1 ml of dilute nitric acid R and0.2 ml of silver nitrate solution R2. The solution shows nochange in appearance for at least 15 min.

Sulphates. To 10 ml add 0.1 ml of dilute hydrochloric acid Rand 0.1 ml of barium chloride solution R1. The solutionshows no change in appearance for at least 1 h.

Ammonium : maximum 0.2 ppm.To 20 ml add 1 ml of alkaline potassium tetraiodomercuratesolution R. After 5 min, examine the solution down thevertical axis of the tube. The solution is not more intenselycoloured than a standard prepared at the same time byadding 1 ml of alkaline potassium tetraiodomercuratesolution R to a mixture of 4 ml of ammonium standardsolution (1 ppm NH4) R and 16 ml of ammonium-freewater 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 ammoniumchloride buffer solution pH 10.0 R, 50 mg of mordantblack 11 triturate R and 0.5 ml of 0.01 M sodium edetate. Apure blue colour is produced.

Residue on evaporation : maximum 0.001 per cent.Evaporate 100 ml on a water-bath and dry in an oven at100-105 °C. The residue weighs a maximum of 1 mg.

Microbial contamination. Total viable aerobic count (2.6.12)not more than 102 micro-organisms per millilitre, determinedby membrane filtration, using agar medium B.

LABELLINGThe label states, where applicable, that the substance issuitable for use in the manufacture of dialysis solutions.

01/2008:0359corrected 6.0

WHEAT STARCH

Tritici amylumDEFINITIONWheat starch is obtained from the caryopsis of Triticumaestivum L. (T. vulgare Vill.).

CHARACTERSAppearance : very fine, white or almost white powder whichcreaks when pressed between the fingers.Solubility : practically insoluble in cold water and in alcohol.Wheat starch does not contain starch grains of any otherorigin. It may contain a minute quantity, if any, of tissuefragments of the original plant.

IDENTIFICATIONA. Examined under a microscope using equal volumes

of glycerol R and water R, it presents large and smallgranules, and, very rarely, intermediate sizes. The largegranules, 10 µm to 60 µm in diameter, are discoid or,more rarely, reniform when seen face-on. The centralhilum and striations are invisible or barely visible and thegranules sometimes show cracks on the edges. Seen inprofile, the granules are elliptical and fusiform and thehilum appears as a slit along the main axis. The smallgranules, rounded or polyhedral, are 2 µm to 10 µm indiameter. Between crossed nicol prisms, the granulesshow 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,add 0.05 ml of iodine solution R1. A dark blue colour isproduced 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 for60 s. Allow to stand for 15 min.

Foreign matter. Examined under a microscope using amixture of equal volumes of glycerol R and water R, notmore than traces of matter other than starch granules arepresent. 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)modified as follows : wash any adhering particles from the

neck into the flask with 25 ml of sulphuric acid R ; continuethe heating until a clear solution is obtained ; add 45 ml ofstrong sodium hydroxide solution R.

Oxidising substances (2.5.30) : maximum 20 ppm, calculatedas H2O2.

Sulphur dioxide (2.5.29) : maximum 50 ppm.

Iron (2.4.9) : maximum 10 ppm.Shake 1.5 g with 15 ml of dilute hydrochloric acid R. Filter.The filtrate complies with the limit test for iron.

Loss on drying (2.2.32) : maximum 15.0 per cent, determinedon 1.000 g by drying in an oven at 130 °C for 90 min.

Sulphated ash (2.4.14) : maximum 0.6 per cent, determinedon 1.0 g.

Microbial contamination. Total viable aerobic count (2.6.12)not more than 103 bacteria and not more than 102 fungi pergram, determined by plate count. It complies with the testfor Escherichia coli (2.6.13).

01/2008:1379

WHEAT-GERM OIL, REFINED

Tritici aestivi oleum raffinatumDEFINITIONFatty oil obtained from the germ of the grain of Triticumaestivum L. by cold expression or by other suitablemechanical means and/or by extraction. It is then refined. Asuitable antioxidant may be added.

CHARACTERSAppearance : 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.

IDENTIFICATIONA. Identification of fatty oils by thin-layer chromatography

(2.3.2). The chromatogram obtained is similar to the typechromatogram 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 themanufacture of parenteral dosage forms : maximum 0.3.

Peroxide value (2.5.5) : maximum 10.0. If intended for use inthe manufacture of parenteral dosage forms : maximum 5.0.

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 foralkaline impurities in fatty oils.

Composition of fatty acids. Gas chromatography (2.4.22,Method C). Use the mixture of calibrating substances inTable 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,— linolenic acid : 3.0 per cent to 10.0 per cent,— eicosenoic acid : maximum 2.0 per cent.

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