EUROPEAN PHARMACOPOEIA 9.0
Determination of primary aromatic amino-nitrogen
0.5 mL of starch solution R1 and continue the titration, with
constant shaking especially near the end-point, to liberate all of acid (n1 mL of 0.5 M hydrochloric acid). Carry out a blank test
the iodine from the solvent layer. Add the sodium thiosulfate under the same conditions (n2 mL of 0.5 M hydrochloric acid).
solution dropwise until the blue colour just disappears.
Depending on the volume of 0.01 M sodium thiosulfate used,
it may be necessary to titrate with 0.1 M sodium thiosulfate.
NOTE : there is a 15 s to 30 s delay in neutralising the starch
indicator for peroxide values of 70 and greater, due to the
tendency of trimethylpentane to float on the surface of the
aqueous medium and the time necessary to adequately mix the
solvent and the aqueous titrant, thus liberating the last traces
of iodine. It is recommended to use 0.1 M sodium thiosulfate
for peroxide values greater than 150. A small amount (0.5 per
cent to 1.0 per cent m/m) of high HLB emulsifier (for example 2.5.7. UNSAPONIFIABLE MATTER
polysorbate 60) may be added to the mixture to retard the
phase separation and decrease the time lag in the liberation
of iodine.
Carry out a blank determination (V0 mL). If the result of
the blank determination exceeds 0.1 mL of titration reagent,
replace the reagents and repeat the determination.
c to a temperature below 25 °C and transfer the contents of
= concentration of the sodium thiosulfate solution,
in moles per litre.
01/2008:20506 water R ; repeat this procedure 3 times. Wash the ether phase
2.5.6. SAPONIFICATION VALUE
The saponification value IS is the number that expresses in
milligrams the quantity of potassium hydroxide required to
neutralise the free acids and to saponify the esters present in
1 g of the substance.
Unless otherwise prescribed, use the quantities indicated in
Table 2.5.6.-1 for the determination.
Table 2.5.6.-1
Presumed value IS Quantity of sample (g)
<3 20
3 to 10 12 to 15
10 to 40 8 to 12
40 to 60 5 to 8
60 to 100 3 to 5
100 to 200 2.5 to 3
200 to 300 1 to 2
300 to 400 0.5 to 1
Introduce the prescribed quantity of the substance to be
examined (m g) into a 250 mL borosilicate glass flask fitted
with a reflux condenser. Add 25.0 mL of 0.5 M alcoholic
potassium hydroxide and a few glass beads. Attach the
condenser and heat under reflux for 30 min, unless otherwise
prescribed. Add 1 mL of phenolphthalein solution R1 and
General Notices (1) apply to all monographs and other texts
2.5.8.
titrate immediately (while still hot) with 0.5 M hydrochloric
01/2008:20507
The term “unsaponifiable matter” is applied to the substances
non-volatile at 100-105 °C obtained by extraction with an
organic solvent from the substance to be examined after it has
been saponified. The result is calculated as per cent m/m.
Use ungreased ground-glass glassware.
Introduce the prescribed quantity of the substance to be
examined (m g) into a 250 mL flask fitted with a reflux
condenser. Add 50 mL of 2 M alcoholic potassium hydroxide R
and heat on a water-bath for 1 h, swirling frequently. Cool
the flask to a separating funnel with the aid of 100 mL of
water R. Shake the liquid carefully with 3 quantities, each of
100 mL, of peroxide-free ether R. Combine the ether layers
in another separating funnel containing 40 mL of water R,
shake gently for a few minutes, allow to separate and reject
the aqueous phase. Wash the ether phase with 2 quantities,
each of 40 mL, of water R then wash successively with 40 mL
of a 30 g/L solution of potassium hydroxide R and 40 mL of
several times, each with 40 mL of water R, until the aqueous
phase is no longer alkaline to phenolphthalein. Transfer the
ether phase to a tared flask, washing the separating funnel
with peroxide-free ether R.
Distil off the ether with suitable precautions and add 6 mL of
acetone R to the residue. Carefully remove the solvent in a
current of air. Dry to constant mass at 100-105 °C. Allow to
cool in a desiccator and weigh (a g).
Dissolve the residue in 20 mL of alcohol R, previously
neutralised to phenolphthalein solution R and titrate with
0.1 M ethanolic sodium hydroxide. If the volume of 0.1 M
ethanolic sodium hydroxide used is greater than 0.2 mL, the
separation of the layers has been incomplete ; the residue
weighed cannot be considered as “unsaponifiable matter”. In
case of doubt, the test must be repeated.
01/2008:20508
2.5.8. DETERMINATION OF PRIMARY
AROMATIC AMINO-NITROGEN
Dissolve the prescribed quantity of the substance to be
examined in 50 mL of dilute hydrochloric acid R or in another
prescribed solvent and add 3 g of potassium bromide R. Cool
in ice-water and titrate by slowly adding 0.1 M sodium nitrite
with constant stirring.
Determine the end-point electrometrically or by the use of
the prescribed indicator.
163
2.5.9. Determination of nitrogen by sulfuric acid digestion
01/2008:20509 otherwise prescribed, carefully unstopper the flask. Wash the
ground parts and the walls of the flask, as well as the sample
carrier, with water R. Combine the combustion products and
the washings and proceed as prescribed in the monograph.
2.5.9. DETERMINATION OF
NITROGEN BY SULFURIC ACID
DIGESTION
SEMI-MICRO METHOD
Place a quantity of the substance to be examined (m g)
containing about 2 mg of nitrogen in a combustion flask, add
4 g of a powdered mixture of 100 g of dipotassium sulfate R,
5 g of copper sulfate pentahydrate R and 2.5 g of selenium R,
and three glass beads. Wash any adhering particles from the
neck into the flask with 5 mL of sulfuric acid R, allowing it
to run down the sides of the flask, and mix the contents by
rotation. Close the mouth of the flask loosely, for example by
means of a glass bulb with a short stem, to avoid excessive
loss of sulfuric acid. Heat gradually at first, then increase the
temperature until there is vigorous boiling with condensation
of sulfuric acid in the neck of the flask ; precautions should be
taken to prevent the upper part of the flask from becoming
overheated. Continue the heating for 30 min, unless otherwise
prescribed. Cool, dissolve the solid material by cautiously
adding to the mixture 25 mL of water R, cool again and place
in a steam-distillation apparatus. Add 30 mL of strong sodium
hydroxide solution R and distil immediately by passing steam
through the mixture. Collect about 40 mL of distillate in
20.0 mL of 0.01 M hydrochloric acid and enough water R
to cover the tip of the condenser. Towards the end of the
distillation, lower the receiver so that the tip of the condenser
is above the surface of the acid. Take precautions to prevent
any water on the outer surface of the condenser from reaching
the contents of the receiver. Titrate the distillate with 0.01 M
sodium hydroxide, using methyl red mixed solution R as
indicator (n1 mL of 0.01 M sodium hydroxide).
Repeat the test using about 50 mg of glucose R in place of the
substance to be examined (n2 mL of 0.01 M sodium hydroxide).
01/2008:20510
2.5.10. OXYGEN-FLASK METHOD
Unless otherwise prescribed the combustion flask is a conical
flask of at least 500 mL capacity of borosilicate glass with
a ground-glass stopper fitted with a suitable carrier for the
sample, for example in platinum or platinum-iridium.
Finely grind the substance to be examined, place the
prescribed quantity in the centre of a piece of filter paper
measuring about 30 mm by 40 mm provided with a small strip
about 10 mm wide and 30 mm long. If paper impregnated
with lithium carbonate is prescribed, moisten the centre of
the paper with a saturated solution of lithium carbonate R
and dry in an oven before use. Envelop the substance to be
examined in the paper and place it in the sample carrier.
Introduce into the flask water R or the prescribed solution
designed to absorb the combustion products, displace the air
with oxygen by means of a tube having its end just above the
liquid, moisten the neck of the flask with water R and close
with its stopper. Ignite the paper strip by suitable means with
the usual precautions. Keep the flask firmly closed during the
combustion. Shake the flask vigorously to completely dissolve
the combustion products. Cool and after about 5 min, unless
164
EUROPEAN PHARMACOPOEIA 9.0
01/2008:20511
corrected 8.0
2.5.11. COMPLEXOMETRIC
TITRATIONS
ALUMINIUM
Introduce 20.0 mL of the prescribed solution into a 500 mL
conical flask, add 25.0 mL of 0.1 M sodium edetate and
10 mL of a mixture of equal volumes of a 155 g/L solution of
ammonium acetate R and dilute acetic acid R. Boil for 2 min,
then cool. Add 50 mL of ethanol R and 3 mL of a freshly
prepared 0.25 g/L solution of dithizone R in ethanol R. Titrate
the excess of sodium edetate with 0.1 M zinc sulfate until the
colour changes from greenish-blue to reddish-violet.
1 mL of 0.1 M sodium edetate is equivalent to 2.698 mg of Al.
BISMUTH
Introduce the prescribed solution into a 500 mL conical flask.
Dilute to 250 mL with water R and then, unless otherwise
prescribed, add dropwise, with shaking, concentrated
ammonia R until the mixture becomes cloudy. Add 0.5 mL
of nitric acid R. Heat to about 70 °C until the cloudiness
disappears completely. Add about 50 mg of xylenol orange
triturate R and titrate with 0.1 M sodium edetate until the
colour changes from pinkish-violet to yellow.
1 mL of 0.1 M sodium edetate is equivalent to 20.90 mg of Bi.
CALCIUM
Introduce the prescribed solution into a 500 mL conical
flask, and dilute to 300 mL with water R. Add 6.0 mL of
strong sodium hydroxide solution R and about 200 mg of
calconecarboxylic acid triturate R. Titrate with 0.1 M sodium
edetate until the colour changes from violet to full blue.
1 mL of 0.1 M sodium edetate is equivalent to 4.008 mg of Ca.
MAGNESIUM
Introduce the prescribed solution into a 500 mL conical flask
and dilute to 300 mL with water R. Add 10 mL of ammonium
chloride buffer solution pH 10.0 R and about 50 mg of mordant
black 11 triturate R. Heat to about 40 °C then titrate at this
temperature with 0.1 M sodium edetate until the colour
changes from violet to full blue.
1 mL of 0.1 M sodium edetate is equivalent to 2.431 mg of Mg.
LEAD
Introduce the prescribed solution into a 500 mL conical flask
and dilute to 200 mL with water R. Add about 50 mg of
xylenol orange triturate R and hexamethylenetetramine R until
the solution becomes violet-pink. Titrate with 0.1 M sodium
edetate until the violet-pink colour changes to yellow.
1 mL of 0.1 M sodium edetate is equivalent to 20.72 mg of Pb.
ZINC
Introduce the prescribed solution into a 500 mL conical
flask and dilute to 200 mL with water R. Add about 50 mg
of xylenol orange triturate R and hexamethylenetetramine R
until the solution becomes violet-pink. Add 2 g of
hexamethylenetetramine R in excess. Titrate with 0.1 M
sodium edetate until the violet-pink colour changes to yellow.
1 mL of 0.1 M sodium edetate is equivalent to 6.54 mg of Zn.
See the information section on general monographs (cover pages)