EUROPEAN PHARMACOPOEIA 9.0 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.

PREPARATION OF REFERENCE BUFFER SOLUTIONS 01/2016:20204

Potassium tetraoxalate 0.05 M. Dissolve 12.61 g of
C4H3KO8,2H2O in carbon dioxide-free water R and dilute to
1000.0 mL with the same solvent.
Potassium hydrogen tartrate, saturated at 25 °C. Shake an 2.2.4. APPROXIMATE pH OF
excess of C4H5KO6 vigorously with carbon dioxide-free water R
at 25 °C. Filter or decant. Prepare immediately before use. SOLUTIONS
Potassium dihydrogen citrate 0.05 M. Dissolve 11.41 g Determine the approximate pH using a pH indicator strip R.
of C6H7KO7 in carbon dioxide-free water R and dilute to Alternatively, pH indicators such as those described in
1000.0 mL with the same solvent. Prepare immediately before Table 2.2.4.-1 can be used.
use.
Potassium hydrogen phthalate 0.05 M. Dissolve 10.13 g of
C8H5KO4, previously dried for 1 h at 110 ± 2 °C, in carbon
dioxide-free water R and dilute to 1000.0 mL with the same Table 2.2.4.-1
solvent. Reaction pH Indicator
Potassium dihydrogen phosphate 0.025 M + Disodium
Alkaline >8 Red litmus paper R
hydrogen phosphate 0.025 M. Dissolve 3.39 g of KH2PO4 and
3.53 g of Na2HPO4, both previously dried for 2 h at 120 ± 2 °C, Slightly alkaline 8 – 10 Phenolphthalein solution R
in carbon dioxide-free water R and dilute to 1000.0 mL with
the same solvent. Thymol blue solution R

Potassium dihydrogen phosphate 0.0087 M + Disodium Strongly alkaline > 10 Phenolphthalein paper R
hydrogen phosphate 0.0303 M. Dissolve 1.18 g of KH2PO4
and 4.30 g of Na2HPO4, both previously dried for 2 h at Thymol blue solution R
120 ± 2 °C, in carbon dioxide-free water R and dilute to
Neutral 6–8 Methyl red solution R
1000.0 mL with the same solvent.
Disodium tetraborate 0.01 M. Dissolve 3.80 g of Phenol red solution R
Na2B4O7,10H2O in carbon dioxide-free water R and dilute Acid <6 Methyl red solution R
to 1000.0 mL with the same solvent. Store protected from
atmospheric carbon dioxide. Bromothymol blue solution R1
Sodium carbonate 0.025 M + Sodium hydrogen carbonate Slightly acid 4–6 Methyl red solution R
0.025 M. Dissolve 2.64 g of Na2CO3 and 2.09 g of NaHCO3
in carbon dioxide-free water R and dilute to 1000.0 mL with Bromocresol green solution R
the same solvent. Store protected from atmospheric carbon
dioxide. Strongly acid <4 Congo red paper R

Calcium hydroxide, saturated at 25 °C. Shake an excess of

calcium hydroxide R with carbon dioxide-free water R and 01/2008:20205
decant at 25 °C. Store protected from atmospheric carbon
dioxide.

STORAGE OF BUFFER SOLUTIONS 2.2.5. RELATIVE DENSITY

Store buffer solutions in suitable chemically-resistant, airtight The relative density of a substance is the ratio of the mass
containers, such as type I glass bottles or plastic containers of a certain volume of a substance at temperature t1 to the
suitable for aqueous solutions. mass of an equal volume of water at temperature t2.

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

2.2.6. Refractive index EUROPEAN PHARMACOPOEIA 9.0

Unless otherwise indicated, the relative density is used. – parasitic resonant effects,
Relative density is also commonly expressed as . Density ρ20, – viscosity, whereby solutions with a higher viscosity than
defined as the mass of a unit volume of the substance at 20 °C the calibrant have a density that is apparently higher than
may also be used, expressed in kilograms per cubic metre or the true value.
grams per cubic centimetre (1 kg·m− 3 = 10− 3 g·cm− 3). These
The effects of non-linearity and viscosity may be avoided by
quantities are related by the following equations where density
using calibrants that have density and viscosity close to those
is expressed in grams per cubic centimetre :
of the liquid to be examined (± 5 per cent for density, ± 50 per
cent for viscosity). The density meter may have functions for
automatic viscosity correction and for correction of errors
arising from temperature changes and non-linearity.
Precision is a function of the repeatability and stability of the
oscillator frequency, which is dependent on the stability of the
volume, mass and spring constant of the cell.
Relative density or density are measured with the precision Density meters are able to achieve measurements with an
to the number of decimals prescribed in the monograph error of the order of 1 × 10− 3 g·cm− 3 to 1 × 10− 5 g·cm− 3 and a
using a density bottle (solids or liquids), a hydrostatic balance repeatability of 1 × 10− 4 g·cm− 3 to 1 × 10− 6 g·cm− 3.
(solids), a hydrometer (liquids) or a digital density meter
with an oscillating transducer (liquids and gases). When the
determination is made by weighing, the buoyancy of air is 01/2008:20206
disregarded, which may introduce an error of 1 unit in the
3rd decimal place. When using a density meter, the buoyancy
of air has no influence.
Oscillating transducer density meter. The apparatus consists of :
– a U-shaped tube, usually of borosilicate glass, which
2.2.6. REFRACTIVE INDEX
contains the liquid to be examined ; The refractive index of a medium with reference to air is equal
– a magneto-electrical or piezo-electrical excitation system to the ratio of the sine of the angle of incidence of a beam of
that causes the tube to oscillate as a cantilever oscillator light in air to the sine of the angle of refraction of the refracted
at a characteristic frequency depending on the density of beam in the given medium.
the liquid to be examined ; Unless otherwise prescribed, the refractive index is measured
– a means of measuring the oscillation period (T), which may at 20 ± 0.5 °C, with reference to the wavelength of the D-line
be converted by the apparatus to give a direct reading of of sodium (λ = 589.3 nm); the symbol is then .
density, or used to calculate density using the constants A Refractometers normally determine the critical angle. In such
and B described below. apparatus the essential part is a prism of known refractive
The resonant frequency (f) is a function of the spring index in contact with the liquid to be examined.
constant (c) and the mass (m) of the system : 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
Hence : of operation at the temperature prescribed. The thermometer
is graduated at intervals of 0.5 °C or less.

01/2008:20207
M = mass of the tube,
V = inner volume of the tube.
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
The constants A and B are determined by operating the of rotating the plane of polarisation of polarised light.
instrument with the U-tube filled with 2 different samples Optical rotation is considered to be positive (+) for
of known density, for example, degassed water R and air. dextrorotatory substances (i.e. those that rotate the plane of
Control measurements are made daily using degassed water R. polarisation in a clockwise direction) and negative (−) for
The results displayed for the control measurement using laevorotatory substances.
degassed water R shall not deviate from the reference value The specific optical rotation is the rotation, expressed
(ρ20 = 0.998203 g·cm− 3, = 1.000000) by more than its in radians (rad), measured at the temperature t and at the
specified error. For example, an instrument specified to wavelength λ given by a 1 m thickness of liquid or a solution
± 0.0001 g·cm− 3 shall display 0.9982 ± 0.0001 g·cm− 3 in containing 1 kg/m3 of optically active substance. For practical
order to be suitable for further measurement. Otherwise a reasons the specific optical rotation is normally
re-adjustment is necessary. Calibration with certified reference expressed in milliradians metre squared per kilogram
materials is carried out regularly. Measurements are made (mrad·m2·kg− 1).
using the same procedure as for calibration. The liquid to
The Pharmacopoeia adopts the following conventional
be examined is equilibrated in a thermostat at 20 °C before
definitions.
introduction into the tube, if necessary, to avoid the formation
of bubbles and to reduce the time required for measurement. The angle of optical rotation of a neat liquid is the angle of
rotation α, expressed in degrees (°), of the plane of polarisation
Factors affecting accuracy include :
at the wavelength of the D-line of sodium (λ = 589.3 nm)
– temperature uniformity throughout the tube, measured at 20 °C using a layer of 1 dm ; for a solution, the
– non-linearity over a range of density, method of preparation is prescribed in the monograph.

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