EUROPEAN PHARMACOPOEIA 6.0 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 at 0.05 M 0.05 M 0.025 M 0.0087 M + at 25°C
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.

Sodium carbonate 0.025 M + sodium hydrogen carbonate Reaction pH Indicator Colour

0.025 M. Dissolve 2.64 g of Na2CO3 and 2.09 g of NaHCO3 Neutral 6.0 – 8.0 Methyl red solution R Yellow
in carbon dioxide-free water R and dilute to 1000.0 ml with
the same solvent. Store protected from atmospheric carbon Phenol red
solution R (0.05 ml)
dioxide.
Neutral to methyl 4.5 – 6.0 Methyl red solution R Orange-red
Calcium hydroxide, saturated at 25 °C. Shake an excess of red
calcium hydroxide R with carbon dioxide-free water R and Neutral to < 8.0 Phenolphthalein Colourless ; pink or red
decant at 25 °C. Store protected from atmospheric carbon phenolphtalein solution R (0.05 ml) after adding 0.05 ml
dioxide. of 0.1 M base
Acid <6 Methyl red solution R Orange or red
STORAGE Bromothymol blue Yellow
solution R1
Store buffer solutions in suitable chemically resistant, tight Faintly acid 4.0 – 6.0 Methyl red solution R Orange
containers, such as type I glass bottles or plastic containers
suitable for aqueous solutions. Bromocresol green Green or blue
solution R
Strongly acid <4 Congo red paper R Green or blue

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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
the mass of an equal volume of water at temperature t2.
APPROXIMATE pH AND COLOUR Unless otherwise indicated, the relative density is
OF CERTAIN INDICATORS used. Relative density is also commonly expressed as .
Density ρ20, defined as the mass of a unit volume of
To 10 ml of the solution to be examined, add 0.1 ml of the substance at 20 °C may also be used, expressed in
the indicator solution, unless otherwise prescribed in kilograms per cubic metre or grams per cubic centimetre
Table 2.2.4.-1. (1 kg·m− 3 = 10− 3 g·cm− 3). These quantities are related by the
following equations where density is expressed in grams per
Table 2.2.4.-1 cubic centimetre :
Reaction pH Indicator Colour
Alkaline >8 Litmus paper red R Blue
Thymol blue Grey or violet-blue
solution R (0.05 ml)
Slightly alkaline 8.0 – 10.0 Phenolphthalein Colourless or pink
solution R (0.05 ml)
Thymol blue Grey Relative density or density are measured with the precision
solution R (0.05 ml)
to the number of decimals prescribed in the monograph
Strongly alkaline > 10 Phenolphthalein Red using a density bottle (solids or liquids), a hydrostatic
paper R
balance (solids), a hydrometer (liquids) or a digital density
Thymol blue Violet-blue
solution R (0.05 ml) meter with an oscillating transducer (liquids and gases).

General Notices (1) apply to all monographs and other texts 25

2.2.6. Refractive index EUROPEAN PHARMACOPOEIA 6.0

When the determination is made by weighing, the buoyancy Precision is a function of the repeatability and stability of the
of air is disregarded, which may introduce an error of 1 unit oscillator frequency, which is dependent on the stability of
in the 3rd decimal place. When using a density meter, the the volume, mass and spring constant of the cell.
buoyancy of air has no influence. Density meters are able to achieve measurements with an
Oscillating transducer density meter. The apparatus error of the order of 1 × 10− 3 g·cm− 3 to 1 × 10− 5 g·cm− 3 and a
consists of: repeatability of 1 × 10− 4 g·cm− 3 to 1 × 10− 6 g·cm− 3.
— a U-shaped tube, usually of borosilicate glass, which
contains the liquid to be examined ; 01/2008:20206
— a magneto-electrical or piezo-electrical excitation system
that causes the tube to oscillate as a cantilever oscillator 2.2.6. REFRACTIVE INDEX
at a characteristic frequency depending on the density of
the liquid to be examined ; The refractive index of a medium with reference to air is
— a means of measuring the oscillation period (T), which equal to the ratio of the sine of the angle of incidence of a
may be converted by the apparatus to give a direct beam of light in air to the sine of the angle of refraction of
reading of density, or used to calculate density using the the refracted beam in the given medium.
constants A and B described below. Unless otherwise prescribed, the refractive index is measured
at 20 ± 0.5 °C, with reference to the wavelength of the D-line
The resonant frequency (f) is a function of the spring
of sodium (λ = 589.3 nm) ; the symbol is then .
constant (c) and the mass (m) of the system :
Refractometers normally determine the critical angle. In
such apparatus the essential part is a prism of known
refractive index in contact with the liquid to be examined.
Hence : Calibrate the apparatus using certified reference materials.
When white light is used, the refractometer is provided
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.
M = mass of the tube, The thermometer is graduated at intervals of 0.5 °C or less.
V = inner volume of the tube.
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Introduction of 2 constants and
, leads to the classical equation for the oscillating
transducer : 2.2.7. OPTICAL ROTATION
Optical rotation is the property displayed by chiral substances
of rotating the plane of polarisation of polarised light.
The constants A and B are determined by operating the Optical rotation is considered to be positive (+) for
instrument with the U-tube filled with 2 different samples dextrorotatory substances (i.e. those that rotate the plane
of known density, for example, degassed water R and air. of polarisation in a clockwise direction) and negative (−) for
Control measurements are made daily using degassed laevorotatory substances.
water R. The results displayed for the control measurement The specific optical rotation is the rotation, expressed
using degassed water R shall not deviate from the reference in radians (rad), measured at the temperature t and at
value (ρ20 = 0.998203 g·cm− 3, = 1.000000) by more than the wavelength λ given by a 1 m thickness of liquid or a
its specified error. For example, an instrument specified solution containing 1 kg/m3 of optically active substance.
to ± 0.0001 g·cm− 3 shall display 0.9982 ± 0.0001 g·cm− 3 in For practical reasons the specific optical rotation
order to be suitable for further measurement. Otherwise is normally expressed in milliradians metre squared per
a re-adjustment is necessary. Calibration with certified kilogram (mrad·m2·kg− 1).
reference materials is carried out regularly. Measurements The Pharmacopoeia adopts the following conventional
are made using the same procedure as for calibration. The definitions.
liquid to be examined is equilibrated in a thermostat at 20 °C
before introduction into the tube, if necessary, to avoid the The angle of optical rotation of a neat liquid is the angle
formation of bubbles and to reduce the time required for of rotation α, expressed in degrees (°), of the plane of
measurement. polarisation at the wavelength of the D-line of sodium
(λ = 589.3 nm) measured at 20 °C using a layer of 1 dm ;
Factors affecting accuracy include : for a solution, the method of preparation is prescribed in
— temperature uniformity throughout the tube, the monograph.
— non-linearity over a range of density, The specific optical rotation of a liquid is the angle
of rotation α, expressed in degrees (°), of the plane of
— parasitic resonant effects,
polarisation at the wavelength of the D-line of sodium
— viscosity, whereby solutions with a higher viscosity than (λ = 589.3 nm) measured at 20 °C in the liquid substance to
the calibrant have a density that is apparently higher be examined, calculated with reference to a layer of 1 dm
than the true value. and divided by the density expressed in grams per cubic
The effects of non-linearity and viscosity may be avoided by centimetre.
using calibrants that have density and viscosity close to those The specific optical rotation of a substance in solution
of the liquid to be examined (± 5 per cent for density, ± 50 per is the angle of rotation α, expressed in degrees (°), of the
cent for viscosity). The density meter may have functions for plane of polarisation at the wavelength of the D-line of
automatic viscosity correction and for correction of errors sodium (λ = 589.3 nm) measured at 20 °C in a solution of
arising from temperature changes and non-linearity. the substance to be examined and calculated with reference

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