Ipecacuanha tincture, standardised
cent of total alkaloids, calculated as emetine (C29H40N2O4 ;
Mr 480.7) with reference to the dried drug. The principal
alkaloids are emetine and cephaëline.
CHARACTERS
Ipecacuanha root has a slight odour.
It has the macroscopic and microscopic characters described
under identification tests A and B.
IDENTIFICATION
A. C. ipecacuanha. The root occurs as somewhat tortuous
pieces, from dark reddish-brown to very dark brown,
seldom more than 15 cm long or 6 mm thick, closely
annulated externally, having rounded ridges completely
encircling the root ; the fracture is short in the bark and
splintery in the wood. The transversely cut surface shows
a wide greyish bark and a small uniformly dense wood.
The rhizome occurs as short lengths usually attached to
roots, cylindrical, up to 2 mm in diameter, finely wrinkled
longitudinally and with pith occupying approximately
one-sixth of the whole diameter.
C. acuminata. The root in general resembles the root of
C. ipecacuanha, but differs in the following particulars :
it is often up to 9 mm thick ; the external surface is
greyish-brown or reddish-brown with transverse ridges at
intervals of usually 1 mm to 3 mm, the ridges being about
0.5 mm to 1 mm wide, extending about half-way round
the circumference and fading at the extremities into the
general surface level.
B. Reduce to a powder (355). The powder is light grey to
yellowish-brown. Examine under a microscope, using
chloral hydrate solution R. The powder shows the
following diagnostic characters : parenchymatous cells,
raphides of calcium oxalate up to 80 µm in length either in
bundles or scattered throughout the powder ; fragments
of tracheids and vessels usually 10 µm to 20 µm in
diameter, with bordered pits ; larger vessels and sclereids
from the rhizome. Examine under a microscope using
a 50 per cent V/V solution of glycerol R. The powder
shows simple or two- to eight-compound starch granules
contained in parenchymatous cells, the simple granules
being up to 15 µm in diameter in C. ipecacuanha and up
to 22 µm in diameter in C. acuminata.
C. Examine by thin-layer chromatography (2.2.27), using a
suitable silica gel as the coating substance.
Test solution. To 0.1 g of the powdered drug (180) in a
test-tube add 0.05 ml of concentrated ammonia R and
5 ml of ether R and stir the mixture vigorously with a
glass rod. Allow to stand for 30 min and filter.
Reference solution. Dissolve 2.5 mg of emetine
hydrochloride CRS and 3 mg of cephaëline
hydrochloride CRS in methanol R and dilute to 20 ml
with the same solvent.
Apply separately to the plate as bands 10 µl of each
solution. Develop over a path of 10 cm using a mixture
of 2 volumes of concentrated ammonia R, 15 volumes
of methanol R, 18 volumes of ethyl acetate R and
65 volumes of toluene R. Allow the plate to dry in air.
Spray with a 5 g/l solution of iodine R in alcohol R
and heat at 60 °C for 10 min. Examine in daylight. The
chromatograms obtained with the test solution and with
the reference solution show in the lower part a yellow
zone corresponding to emetine and below it a light
brown zone corresponding to cephaëline. Examine in
ultraviolet light at 365 nm. The zone corresponding to
emetine shows an intense yellow fluorescence and that
1830 See the information section on general monographs (cover pages)
EUROPEAN PHARMACOPOEIA 5.
corresponding to cephaëline a light blue fluorescence.
The chromatogram obtained with the test solution shows
also faint fluorescent zones.
With C. acuminata the principal zones in the
chromatogram obtained with the test solution are similar
in position, fluorescence and size to the zones in the
chromatogram obtained with the reference solution.
With C. ipecacuanha the only difference is that the
zone corresponding to cephaëline in the chromatogram
obtained with the test solution is much smaller than the
corresponding zone in the chromatogram obtained with
the reference solution.
TESTS
Foreign matter (2.8.2). It complies with the test for foreign
matter.
Loss on drying (2.2.32). Not more than 10.0 per cent,
determined on 1.000 g of powdered drug (180) by drying in
an oven at 100-105 °C.
Total ash (2.4.16). Not more than 5.0 per cent.
Ash insoluble in hydrochloric acid (2.8.1). Not more than
3.0 per cent.
ASSAY
To 7.5 g of the powdered drug (180) in a dry flask, add
100 ml of ether R and shake for 5 min. Add 5 ml of dilute
ammonia R1, shake for 1 h, add 5 ml of water R and shake
vigorously. Decant the ether layer into a flask through a plug
of cotton. Wash the residue in the flask with 2 quantities,
each of 25 ml, of ether R, decanting each portion through
the same plug of cotton. Combine the ether solutions and
eliminate the ether by distillation. Dissolve the residue in
2 ml of alcohol (90 per cent V/V) R, evaporate the alcohol to
dryness and heat at 100 °C for 5 min. Dissolve the residue in
5 ml of previously neutralised alcohol (90 per cent V/V) R,
warming on a water-bath, add 15.0 ml of 0.1 M hydrochloric
acid and titrate the excess acid with 0.1 M sodium hydroxide
using 0.5 ml of methyl red mixed solution R as indicator.
1 ml of 0.1 M hydrochloric acid is equivalent to 24.03 mg of
total alkaloids, calculated as emetine.
STORAGE
Store protected from light and moisture.
01/2005:1530
IPECACUANHA TINCTURE,
STANDARDISED
Ipecacuanhae tinctura normata
DEFINITION
Tincture produced from Ipecacuanha root (0094).
Content : 0.18 per cent (m/m) to 0.22 per cent (m/m) of
total alkaloids, calculated as emetine (C29H40N2O4 ; Mr 480.7).
PRODUCTION
The tincture is produced by a suitable procedure from the
herbal drug and ethanol of suitable strength.
CHARACTERS
Appearance : yellowish-brown liquid.
IDENTIFICATION
Thin-layer chromatography (2.2.27).
EUROPEAN PHARMACOPOEIA 5.0 Ipratropium bromide

Test solution. To 2.0 ml of the tincture to be examined add water R. Titrate the excess acid with 0.02 M sodium
2 ml of water R and 0.1 ml of concentrated ammonia R. Add hydroxide using 0.15 ml of methyl red mixed solution R
10 ml of ether R and shake. Separate the ether layer, dry it as indicator.
over about 2 g of anhydrous sodium sulphate R and filter. Perform a blank assay replacing the tincture to be examined
Reference solution. Dissolve 2.5 mg of emetine with 10.0 ml of alcohol of the strength stated on the label.
hydrochloride CRS and 3 mg of cephaëline 1 ml of 0.02 M hydrochloric acid is equivalent to 4.807 mg
hydrochloride CRS in methanol R and dilute to of total alkaloids, calculated as emetine.
10 ml with the same solvent.
Plate : TLC silica gel plate R.
Mobile phase : concentrated ammonia R, methanol R, ethyl 01/2005:0919
acetate R, toluene R (2:15:18:65 V/V/V/V).
Application : 10 µl, as bands. IPRATROPIUM BROMIDE
Development : over a path of 10 cm.
Drying : in air. Ipratropii bromidum
Detection A : spray the plate with a 5 g/l solution of iodine R
in alcohol R and heat at 60 °C for 10 min. Examine in
daylight.
Results A : see below the sequence of the zones present in
the chromatograms obtained with the reference solution and
the test solution.
Top of the plate
_______ _______
_______ _______ C20H30BrNO3,H2O Mr 430.4

Emetine : a yellow zone A yellow zone (emetine) DEFINITION

Cephaëline : a light brown zone A light brown zone (cephaëline)
(1R,3r,5S,8r)-3-[[(2RS)-3-Hydroxy-2-phenylpropanoyl]oxy]-
8-methyl-8-(1-methylethyl)-8-azoniabicyclo[3.2.1]octane
Reference solution Test solution bromide monohydrate.
Detection B : examine the plate in ultraviolet light at 365 nm. Content : 99.0 per cent to 100.5 per cent (anhydrous
substance).
Results B : see below the sequence of the zones present in the
chromatograms obtained with the reference solution and the CHARACTERS
test solution. Furthermore, other faint fluorescent zones are Appearance : white or almost white, crystalline powder.
present in the chromatogram obtained with the test solution.
Solubility : soluble in water, freely soluble in methanol,
Top of the plate slightly soluble in alcohol.
_______ _______ mp : about 230 °C, with decomposition.
_______ _______
IDENTIFICATION
Emetine : an intense yellow An intense yellow fluorescent zone First identification : A, E.
fluorescent zone (emetine)
Second identification : B, C, D, E.
Cephaëline : a light blue or A light blue or orange-yellow
orange-yellow fluorescent zone fluorescent zone (cephaëline) A. Infrared absorption spectrophotometry (2.2.24).
Reference solution Test solution Comparison : ipratropium bromide CRS.
B. Examine the chromatograms obtained in the test for
With a tincture from Cephaelis acuminata root the zones of impurity A.
emetine and cephaëline in the chromatogram obtained with
the test solution are similar in size. Results : the principal spot in the chromatogram obtained
with the test solution is similar in position, colour and
With a tincture from Cephaelis ipecacuanha root the zone size to the principal spot in the chromatogram obtained
of emetine is much larger than the zone of cephaëline in the with reference solution (a).
chromatogram obtained with the test solution.
C. To 5 ml of solution S (see Tests), add 2 ml of dilute
TESTS sodium hydroxide solution R. No precipitate is formed.
Ethanol (2.9.10) : 95 per cent to 105 per cent of the quantity D. To about 1 mg add 0.2 ml of nitric acid R and evaporate
stated on the label. to dryness on a water-bath. Dissolve the residue in 2 ml
of acetone R and add 0.1 ml of a 30 g/l solution of
ASSAY potassium hydroxide R in methanol R. A violet colour
Transfer 10.00 g of the tincture to be examined to a develops.
chromatography column about 0.2 m long and about 15 mm E. It gives reaction (a) of bromides (2.3.1).
in internal diameter, filled with 8 g of basic aluminium
oxide R. After infiltration into the aluminium oxide layer TESTS
rinse the internal wall of the column with 3 quantities, each Solution S. Dissolve 0.50 g in carbon dioxide-free water R
of 2 ml, of alcohol (70 per cent V/V) R. Elute in portions, and dilute to 50.0 ml with the same solvent.
with 40 ml of alcohol (70 per cent V/V) R. Avoid whirling or
drying of the surface of the aluminium oxide layer. Collect Appearance of solution. Solution S is clear (2.2.1) and not
the eluate in a 100 ml flask. Evaporate the eluate on a more intensely coloured than reference solution GY7 (2.2.2,
water-bath to about 10 ml. Allow to cool. Add 10.0 ml of Method II).
0.02 M hydrochloric acid and 20 ml of carbon dioxide-free pH (2.2.3) : 5.0 to 7.5 for solution S.

General Notices (1) apply to all monographs and other texts 1831