Ipecacuanha liquid extract, standardised
D. D1, D2, D3 and D4 : 3-[[[[3-(acetylmethylamino)-5-(dim-
ethylcarbamoyl)-2,4,6-triiodobenzoyl]amino]acetyl]ami-
no]-5-[(2-hydroxyethyl)carbamoyl]-2,4,6-triiodobenzoic
acid,
E. 3-[[[[3-[[[[3-(acetylmethylamino)-2,4,6-triiodo-5-(methyl-
carbamoyl)benzoyl]amino]acetyl]amino]-5-[(2-hydroxy-
ethyl)carbamoyl]-2,4,6-triiodobenzoyl]amino]acetyl]ami-
no]-5-[(2-hydroxyethyl)carbamoyl]-2,4,6-triiodobenzoic
acid,
F. specified impurity whose structure is unknown,
G. 3-[[[[3-(acetylmethylamino)-2,4,6-triiodo-5-
(methylcarbamoyl)benzoyl]amino]acetyl]amino]-5-
[[2-(acetyloxy)ethyl]carbamoyl]-2,4,6-triiodobenzoic acid,
H. 3,3′-[[5-(acetylmethylamino)-2,4,6-triiodo-1,3-phe-
nylene]bis(carbonyliminomethylenecarbonylimi-
no)]bis[5-[(2-hydroxyethyl)carbamoyl]-2,4,6-triiodoben-
zoic] acid.
01/2005:1875
IPECACUANHA LIQUID EXTRACT,
STANDARDISED
Ipecacuanhae extractum fluidum
normatum
DEFINITION
Standardised liquid extract produced from Ipecacuanha
root (0094).
1828 See the information section on general monographs (cover pages)
EUROPEAN PHARMACOPOEIA 5.
Content : minimum 1.80 per cent and maximum 2.20 per
cent of total alkaloids, calculated as emetine (C29H40N2O4 ;
Mr 480.7).
PRODUCTION
The extract is produced from the herbal drug and solvent of
suitable strength by an appropriate procedure.
CHARACTERS
Appearance : dark-brown liquid.
IDENTIFICATION
Thin-layer chromatography (2.2.27).
Test solution. Dilute 5.0 ml of the extract to be examined to
50 ml with alcohol (70 per cent V/V) R. To 2.0 ml of this
solution add 2 ml of water R and 0.1 ml of concentrated
ammonia R. Add 10 ml of ether R and shake. Separate the
upper layer, dry it over about 2 g of anhydrous sodium
sulphate R 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
10 ml with the same solvent.
Plate : TLC silica gel plate R.
Mobile phase : concentrated ammonia R, methanol R, ethyl
acetate R, toluene R (2:15:18:65 V/V/V/V).
Application : 10 µl, as bands.
Development : over a path of 10 cm.
Drying : in air.
Detection A : spray with a 5 g/l solution of iodine R in
alcohol R. Heat at 60 °C for 10 min and allow to cool for
30 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. Furthermore, other zones may be present
in the chromatogram obtained with the test solution.
Top of the plate
_______ _______
_______ _______
Emetine : a yellow zone A yellow zone (emetine)
Cephaëline : a light brown zone A light brown zone (cephaëline)
Reference solution Test solution
Detection B : examine in ultraviolet light at 365 nm.
Results B : see below the sequence of the zones present in the
chromatograms obtained with the reference solution and the
test solution. Furthermore, other faint fluorescent zones are
present in the chromatogram obtained with the test solution.
Top of the plate
_______ _______
_______ _______
Emetine : an intense yellow An intense yellow fluorescent zone
fluorescent zone (emetine)
Cephaëline : a light blue or A light blue or orange-yellow
orange-yellow fluorescent zone fluorescent zone (cephaëline)
Reference solution Test solution
With a liquid extract from Cephaelis acuminata root, the
zones of emetine and cephaëline in the chromatogram
obtained with the test solution are of similar size.
With a liquid extract from Cephaelis ipecacuanha root, the
zone of emetine is much larger than the zone of cephaëline
in the chromatogram obtained with the test solution.
EUROPEAN PHARMACOPOEIA 5.0
TESTS
Ethanol (2.9.10) : minimum 95 per cent and maximum
105 per cent of the quantity stated on the label.
ASSAY
Dilute 1.00 g of the extract to be examined to 10 ml with
alcohol (70 per cent V/V) R and transfer to a chromatography
column about 0.2 m long and about 15 mm in internal
diameter, containing 8 g of basic aluminium oxide R, using
a glass rod. After infiltration into the aluminium oxide
layer, rinse the flask, glass rod and internal wall of the
column with 3 quantities, each of 2 ml, of alcohol (70 per
cent V/V) R. Elute in portions with 40 ml of alcohol (70 per
cent V/V) R. Avoid disturbance or drying of the surface of
the aluminium oxide layer. Collect the eluate in a 100 ml
flask. Evaporate the eluate on a water-bath to about 10 ml.
Allow to cool. Add 10.0 ml of 0.02 M hydrochloric acid and
20 ml of carbon dioxide-free water R. Titrate the excess acid
with 0.02 M sodium hydroxide using 0.15 ml of methyl red
mixed solution R as indicator.
Perform a blank assay by repeating the assay but replacing
the extract to be examined with 10.0 ml of alcohol of the
strength stated on the label.
1 ml of 0.02 M hydrochloric acid is equivalent to 4.807 mg
of total alkaloids, calculated as emetine.
01/2005:0093
IPECACUANHA, PREPARED
Ipecacuanhae pulvis normatus
DEFINITION
Prepared ipecacuanha is ipecacuanha root powder (180)
adjusted, if necessary, by the addition of powdered lactose
or ipecacuanha root powder with a lower alkaloidal content
to contain 1.9 per cent to 2.1 per cent of total alkaloids,
calculated as emetine (C29H40N2O4 ; Mr 480.7) with reference
to the dried drug.
CHARACTERS
Light grey to yellowish-brown powder with a slight odour.
IDENTIFICATION
A. 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. Examined in glycerol (85 per
cent) R, it may be seen to contain lactose crystals.
B. Examine by thin-layer chromatography (2.2.27), using a
suitable silica gel as the coating substance.
Test solution. To 0.1 g 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.
General Notices (1) apply to all monographs and other texts
Ipecacuanha root
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
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
Loss on drying (2.2.32). Not more than 5.0 per cent,
determined on 1.000 g by drying in an oven at 100 °C to
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 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 two 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 in an airtight container, protected from light.
01/2005:0094
IPECACUANHA ROOT
Ipecacuanhae radix
DEFINITION
Ipecacuanha root consists of the fragmented and dried
underground organs of Cephaelis ipecacuanha (Brot.) A.
Rich., known as Matto Grosso ipecacuanha, or of Cephaelis
acuminata Karsten, known as Costa Rica ipecacuanha, or of
a mixture of both species. It contains not less than 2.0 per
1829