EUROPEAN PHARMACOPOEIA 5.0 2.2.5.

Relative density

Table 2.2.3.-2. – pH of reference buffer solutions at various temperatures

Temperature Potassium Potassium Potassium Potassium Potassium Potassium Disodium Sodium Calcium
(°C) tetraoxalate hydrogen dihydrogen hydrogen dihydrogen dihydrogen tetraborate carbonate hydroxide,
0.05 M tartrate citrate phthalate phosphate phosphate 0.01 M 0.025 M saturated
saturated 0.05 M 0.05 M 0.025 M 0.0087 M + at 25°C
at 25 °C + + sodium
disodium disodium bicarbonate
hydrogen hydrogen 0.025 M
phosphate phosphate
0.025 M 0.0303 M

C4H3KO8,2H2O C4H5KO6 C6H7KO7 C8H5KO4 KH2PO4+ KH2PO4+ Na2B4O7, Na2CO3+ Ca(OH)2

Na2HPO4 Na2HPO4 10H2O NaHCO3

15 1.67 3.80 4.00 6.90 7.45 9.28 10.12 12.81

20 1.68 3.79 4.00 6.88 7.43 9.23 10.06 12.63

25 1.68 3.56 3.78 4.01 6.87 7.41 9.18 10.01 12.45

30 1.68 3.55 3.77 4.02 6.85 7.40 9.14 9.97 12.29

35 1.69 3.55 3.76 4.02 6.84 7.39 9.10 9.93 12.13

+ 0.001 − 0.0014 − 0.0022 + 0.0012 − 0.0028 −0.0028 −0.0082 − 0.0096 −0.034

(1) pH variation per degree Celsius.

Potassium dihydrogen phosphate 0.025 M + disodium Table 2.2.4.-1

hydrogen phosphate 0.025 M. Dissolve 3.39 g of KH2PO4 Reaction pH Indicator Colour
and 3.53 g of Na2HPO4, both previously dried for 2 h at
120 ± 2 °C, in carbon dioxide-free water R and dilute to Alkaline >8 Litmus paper red R Blue
1000.0 ml with the same solvent. Thymol blue Grey or violet-blue
solution R (0.05 ml)
Potassium dihydrogen phosphate 0.0087 M + disodium
hydrogen phosphate 0.0303 M. Dissolve 1.18 g of KH2PO4 Slightly alkaline 8.0 – 10.0 Phenolphthalein Colourless or pink
solution R (0.05 ml)
and 4.30 g of Na2HPO4, both previously dried for 2 h at
120 ± 2 °C, in carbon dioxide-free water R and dilute to Thymol blue Grey
solution R (0.05 ml)
1000.0 ml with the same solvent.
Strongly alkaline > 10 Phenolphthalein Red
Disodium tetraborate 0.01 M. Dissolve 3.80 g of paper R
Na2B4O7,10H2O in carbon dioxide-free water R and dilute Thymol blue Violet-blue
to 1000.0 ml with the same solvent. Store protected from solution R (0.05 ml)
atmospheric carbon dioxide. Neutral 6.0 – 8.0 Methyl red solution R Yellow
Sodium carbonate 0.025 M + sodium hydrogen carbonate Phenol red
0.025 M. Dissolve 2.64 g of Na2CO3 and 2.09 g of NaHCO3 solution R (0.05 ml)
in carbon dioxide-free water R and dilute to 1000.0 ml with Neutral to methyl 4.5 – 6.0 Methyl red solution R Orange-red
the same solvent. Store protected from atmospheric carbon red
dioxide. Neutral to < 8.0 Phenolphthalein Colourless ; pink or red
Calcium hydroxide, saturated at 25 °C. Shake an excess of phenolphtalein solution R (0.05 ml) after adding 0.05 ml
of 0.1 M base
calcium hydroxide R with carbon dioxide-free water R and
decant at 25 °C. Store protected from atmospheric carbon Acid <6 Methyl red solution R Orange or red
dioxide. Bromothymol blue Yellow
solution R1
STORAGE Faintly acid 4.0 – 6.0 Methyl red solution R Orange
Store buffer solutions in suitable chemically resistant, tight Bromocresol green Green or blue
containers, such as type I glass bottles or plastic containers solution R
suitable for aqueous solutions. Strongly acid <4 Congo red paper R Green or blue

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01/2005:20204
2.2.5. RELATIVE DENSITY
2.2.4. RELATIONSHIP BETWEEN
The relative density of a substance is the ratio of the
REACTION OF SOLUTION, mass of a certain volume of a substance at temperature t1 to
APPROXIMATE pH AND COLOUR the mass of an equal volume of water at temperature t2.
OF CERTAIN INDICATORS Unless otherwise indicated, the relative density is
used. Relative density is also commonly expressed as .
To 10 ml of the solution to be examined, add 0.1 ml of Density ρ20, defined as the mass of a unit volume of
the indicator solution, unless otherwise prescribed in the substance at 20 °C may also be used, expressed in
Table 2.2.4.-1. kilograms per cubic metre or grams per cubic centimetre

General Notices (1) apply to all monographs and other texts 27

2.2.6. Refractive index EUROPEAN PHARMACOPOEIA 5.0

(1 kg·m− 3 = 10− 3 g·cm− 3). These quantities are related by the Factors affecting accuracy include :
following equations where density is expressed in grams per — temperature uniformity throughout the tube,
cubic centimetre : — non-linearity over a range of density,
— parasitic resonant effects,
— viscosity, whereby solutions with a higher viscosity than
the calibrant have a density that is apparently higher than
the true value.
The effects of non-linearity and viscosity may be avoided by
using calibrants that have density and viscosity close to those
Relative density or density are measured with the precision of the liquid to be examined (± 5 per cent for density, ± 50 per
to the number of decimals prescribed in the monograph cent for viscosity). The density meter may have functions for
using a density bottle (solids or liquids), a hydrostatic automatic viscosity correction and for correction of errors
balance (solids), a hydrometer (liquids) or a digital density arising from temperature changes and non-linearity.
meter with an oscillating transducer (liquids and gases). Precision is a function of the repeatability and stability of the
When the determination is made by weighing, the buoyancy oscillator frequency, which is dependent on the stability of
of air is disregarded, which may introduce an error of 1 unit the volume, mass and spring constant of the cell.
in the 3rd decimal place. When using a density meter, the Density meters are able to achieve measurements with an
buoyancy of air has no influence. error of the order of 1 × 10− 3 g·cm− 3 to 1 × 10− 5 g·cm− 3 and a
Oscillating transducer density meter. The apparatus repeatability of 1 × 10− 4 g·cm− 3 to 1 × 10− 6 g·cm− 3.
consists of:
— a U-shaped tube, usually of borosilicate glass, which 01/2005:20206
contains the liquid to be examined ;
— a magneto-electrical or piezo-electrical excitation system
2.2.6. REFRACTIVE INDEX
that causes the tube to oscillate as a cantilever oscillator The refractive index of a medium with reference to air is
at a characteristic frequency depending on the density of equal to the ratio of the sine of the angle of incidence of a
the liquid to be examined ; beam of light in air to the sine of the angle of refraction of
— a means of measuring the oscillation period (T), which the refracted beam in the given medium.
may be converted by the apparatus to give a direct Unless otherwise prescribed, the refractive index is measured
reading of density, or used to calculate density using the at 20 ± 0.5 °C, with reference to the wavelength of the D-line
constants A and B described below. of sodium (λ = 589.3 nm) ; the symbol is then .
The resonant frequency (f) is a function of the spring Refractometers normally determine the critical angle. In
constant (c) and the mass (m) of the system : such apparatus the essential part is a prism of known
refractive index in contact with the liquid to be examined.
Calibrate the apparatus using certified reference materials.
When white light is used, the refractometer is provided
Hence : with a compensating system. The apparatus gives readings
accurate to at least the third decimal place and is provided
with a means of operation at the temperature prescribed.
The thermometer is graduated at intervals of 0.5 °C or less.

M = mass of the tube, 01/2005:20207

V = inner volume of the tube.
2.2.7. OPTICAL ROTATION
Introduction of 2 constants and
, leads to the classical equation for the oscillating Optical rotation is the property displayed by chiral substances
transducer : of rotating the plane of polarisation of polarised light.
Optical rotation is considered to be positive (+) for
dextrorotatory substances (i.e. those that rotate the plane
of polarisation in a clockwise direction) and negative (−) for
The constants A and B are determined by operating the laevorotatory substances.
instrument with the U-tube filled with 2 different samples
of known density, for example, degassed water R and air. The specific optical rotation is the rotation, expressed
Control measurements are made daily using degassed in radians (rad), measured at the temperature t and at
water R. The results displayed for the control measurement the wavelength λ given by a 1 m thickness of liquid or a
3
using degassed water R shall not deviate from the reference solution containing 1 kg/m of optically active substance.
value (ρ20 = 0.998203 g·cm− 3, = 1.000000) by more than For practical reasons the specific optical rotation
its specified error. For example, an instrument specified is normally expressed in milliradians metre squared per
to ± 0.0001 g·cm− 3 shall display 0.9982 ± 0.0001 g·cm− 3 in kilogram (mrad·m2·kg− 1).
order to be suitable for further measurement. Otherwise The Pharmacopoeia adopts the following conventional
a re-adjustment is necessary. Calibration with certified definitions.
reference materials is carried out regularly. Measurements The angle of optical rotation of a neat liquid is the angle
are made using the same procedure as for calibration. The of rotation α, expressed in degrees (°), of the plane of
liquid to be examined is equilibrated in a thermostat at 20 °C polarisation at the wavelength of the D-line of sodium
before introduction into the tube, if necessary, to avoid the (λ = 589.3 nm) measured at 20 °C using a layer of 1 dm ;
formation of bubbles and to reduce the time required for for a solution, the method of preparation is prescribed in
measurement. the monograph.

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