EUROPEAN PHARMACOPOEIA 11.

0 Apomorphine hydrochloride hemihydrate

percentage of propidium iodide-positive cells, without gating Solubility : sparingly soluble in water and in ethanol (96 per
but excluding debris. Analyse at least 3000 cells for each of the cent), practically insoluble in toluene.
test and reference solutions.
Use the percentages of dead cells to estimate the potency as the IDENTIFICATION
concentration in milligrams per millilitre of the preparation First identification : B, D.
to be examined necessary to induce 50 per cent of cytotoxicity Second identification : A, C, D.
by fitting a sigmoidal dose response curve to the data obtained A. Ultraviolet and visible absorption spectrophotometry
with the test and the reference preparations and by using a (2.2.25).
4-parameter logistic model (see, for example, chapter 5.3) and
suitable software. The test is not valid unless the percentage of Test solution. Dissolve 10.0 mg in a 10.3 g/L solution of
propidium iodide-positive cells at the lower asymptote of the hydrochloric acid R and dilute to 100.0 mL with the same
curve is less then 15 per cent and the percentage of propidium acid solution. Dilute 10.0 mL of the solution to 100.0 mL
iodide-positive cells at the upper asymptote of the curve is at with a 10.3 g/L solution of hydrochloric acid R.
least 80 per cent. Spectral range : 230-350 nm
The estimated activity is 70 per cent to 130 per cent of the Absorption maximum : at 273 nm.
activity approved for the particular product. Shoulder : at 300-310 nm.
The confidence limits (P = 0.95) are not less than 80 per cent Specific absorbance at the absorption maximum : 530 to 570.
and not more than 125 per cent of the estimated potency. B. Infrared absorption spectrophotometry (2.2.24).
STORAGE Comparison : apomorphine hydrochloride hemihydrate CRS.
Protected from light at the temperature stated on the label. C. To 5 mL of solution S (see Tests) add a few millilitres of
Expiry date. The expiry date is calculated from the beginning sodium hydrogen carbonate solution R until a permanent,
of the assay. white precipitate is formed. The precipitate slowly becomes
greenish. Add 0.25 mL of 0.05 M iodine and shake. The
LABELLING precipitate becomes greyish-green. Collect the precipitate.
The label states : The precipitate dissolves in methylene chloride R giving a
– for liquid preparations, the volume of the preparation in violet-blue solution and in ethanol (96 per cent) R giving
the container and the protein content, a blue solution.
– for freeze-dried preparations : D. To 2 mL of solution S (see Tests) add 0.1 mL of nitric
acid R. Mix and filter. The filtrate gives reaction (a) of
– the name and the volume of the reconstitution liquid
chlorides (2.3.1).
to be added,
– the quantity of protein in the container, TESTS
– that the immunoserum is to be used immediately Solution S. Dissolve 0.25 g without heating in carbon
after reconstitution, dioxide-free water R and dilute to 25 mL with the same solvent.
– the time required for complete dissolution, Appearance of solution. Solution S is clear (2.2.1) and not
– the animal species of origin, more intensely coloured than reference solution BY5 or GY5
– the name and amount of stabiliser, where applicable, (2.2.2, Method II).
– the dilution to be made before use of the product. pH (2.2.3): 4.0 to 5.0 for solution S.
Specific optical rotation (2.2.7) : − 52 to − 48 (dried
01/2020:0136 substance).
Dissolve 0.25 g in a 2.06 g/L solution of hydrochloric acid R
and dilute to 25.0 mL with the same acid solution.
Related substances. Liquid chromatography (2.2.29).
APOMORPHINE HYDROCHLORIDE Test solution. Dissolve 50.0 mg of the substance to be
examined in a 1 per cent V/V solution of glacial acetic acid R
HEMIHYDRATE and dilute to 20.0 mL with the same solution.
Reference solution (a). Dilute 1.0 mL of the test solution to
Apomorphini hydrochloridum 100.0 mL with a 1 per cent V/V solution of glacial acetic
hemihydricum acid R. Dilute 1.0 mL of this solution to 10.0 mL with a 1 per
cent V/V solution of glacial acetic acid R.
Reference solution (b). Dissolve 12.5 mg of apomorphine
impurity B CRS in a 1 per cent V/V solution of glacial acetic
acid R and dilute to 10.0 mL with the same solution.
Reference solution (c). Dilute 2.0 mL of reference solution (b)
to 10.0 mL with a 1 per cent V/V solution of glacial acetic
acid R. Dilute 2.0 mL of this solution to 100.0 mL with a 1 per
C17H18ClNO2,½H2O Mr 312.8 cent V/V solution of glacial acetic acid R.
[41372-20-7] Reference solution (d). Dissolve 25 mg of boldine R in a 1 per
cent V/V solution of glacial acetic acid R and dilute to 10 mL
DEFINITION
with the same solution. To 1 mL of this solution add 1 mL of
(6aR)-6-Methyl-5,6,6a,7-tetrahydro-4H-diben- the test solution and dilute to 10 mL with a 1 per cent V/V
zo[de,g]quinoline-10,11-diol hydrochloride hemihydrate. solution of glacial acetic acid R.
Content : 98.5 per cent to 101.5 per cent (dried substance). Column :
CHARACTERS – size : l = 0.15 m, Ø = 4.6 mm ;
Appearance : white or slightly yellowish-brown or green-tinged – stationary phase : end-capped octadecylsilyl silica gel for
greyish, crystalline powder or crystals ; on exposure to air and chromatography R (5 μm) ;
light, the green tinge becomes more pronounced. – temperature : 35 °C.

General Notices (1) apply to all monographs and other texts 1981
Aprepitant EUROPEAN PHARMACOPOEIA 11.0

Mobile phase :
– mobile phase A : 1.1 g/L solution of sodium
octanesulfonate R, adjusted to pH 2.2 with a 50 per
cent m/m solution of phosphoric acid R ;
– mobile phase B : acetonitrile R ;
Time Mobile phase A Mobile phase B A. (6aR)-10-methoxy-6-methyl-5,6,6a,7-tetrahydro-4H-
(min) (per cent V/V) (per cent V/V) dibenzo[de,g]quinolin-11-ol (apocodeine),
0-2 85 15

2 - 32 85 → 68 15 → 32

32 - 37 68 32

Flow rate : 1.5 mL/min.

Detection : spectrophotometer at 280 nm.
Injection : 10 μL. B. 7,8-didehydro-4,5α-epoxy-17-methylmorphinan-3,6α-diol
Identification of impurities : use the chromatogram obtained (morphine),
with reference solution (b) to identify the peak due to
impurity B.
Relative retention with reference to apomorphine
(retention time = about 18 min): impurity B = about 0.4 ;
boldine = about 0.9.
System suitability : reference solution (d) :
– resolution : minimum 2.5 between the peaks due to boldine
and apomorphine.
Calculation of percentage contents : C. (6aR)-9-[7,8-didehydro-4,5α-epoxy-3-hydroxy-
17-methylmorphinan-6α-yl]-6-methyl-5,6,6a,7-
– for impurity B, use the concentration of impurity B in
tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol
reference solution (c) ;
(morphine-apomorphine dimer).
– for impurities other than B, use the concentration of
apomorphine hydrochloride hemihydrate in reference
solution (a). 01/2017:2757
Limits :
– impurity B : maximum 0.15 per cent ;
– unspecified impurities : for each impurity, maximum
0.10 per cent ; APREPITANT
– total : maximum 0.5 per cent ;
– reporting threshold : 0.05 per cent. Aprepitantum
Loss on drying (2.2.32): 2.5 per cent to 4.2 per cent,
determined on 1.000 g by drying in an oven at 105 °C for 2 h.
Sulfated ash (2.4.14) : maximum 0.1 per cent, determined on
1.0 g.

ASSAY
Dissolve 0.250 g in a mixture of 5.0 mL of 0.01 M hydrochloric
acid and 50 mL of ethanol (96 per cent) R. Carry out a
potentiometric titration (2.2.20), using 0.1 M sodium
hydroxide. Read the volume added between the first 2 points C23H21F7N4O3 Mr 534.4
of inflexion. [170729-80-3]
1 mL of 0.1 M sodium hydroxide is equivalent to 30.38 mg of DEFINITION
C17H18ClNO2. 5-[[(2R,3S)-2-[(1R)-1-[3,5-Bis(trifluoromethyl)phenyl]-
ethoxy]-3-(4-fluorophenyl)morpholin-4-yl]methyl]-1,2-
STORAGE dihydro-3H-1,2,4-triazol-3-one.
In an airtight container, protected from light. Content : 98.0 per cent to 102.0 per cent (anhydrous substance).

IMPURITIES CHARACTERS
Specified impurities : B. Appearance : white or almost white powder.
Solubility : very slightly soluble in water, sparingly soluble in
Other detectable impurities (the following substances would, anhydrous ethanol, practically insoluble in heptane.
if present at a sufficient level, be detected by one or other of
the tests in the monograph. They are limited by the general It shows polymorphism (5.9).
acceptance criterion for other/unspecified impurities and/or IDENTIFICATION
by the general monograph Substances for pharmaceutical use
(2034). It is therefore not necessary to identify these impurities A. Specific optical rotation (see Tests).
for demonstration of compliance. See also 5.10. Control of B. Infrared absorption spectrophotometry (2.2.24).
impurities in substances for pharmaceutical use) : A, C. Comparison : aprepitant CRS.

1982 See the information section on general monographs (cover pages)