Aciclovir EUROPEAN PHARMACOPOEIA 11.

Test solution. Dissolve 50.0 mg of the substance to be Sulfated ash (2.4.14): maximum 0.1 per cent, determined on
examined in mobile phase A and dilute to 50.0 mL with 1.0 g.
mobile phase A. Bacterial endotoxins (2.6.14) : less than 25 IU/g, if intended
Reference solution (a). Dilute 1.0 mL of the test solution to for use in the manufacture of parenteral preparations without
100.0 mL with mobile phase A. Dilute 1.0 mL of this solution a further appropriate procedure for the removal of bacterial
to 10.0 mL with mobile phase A. endotoxins.
Reference solution (b). Dissolve 20.0 mg of tyrosine CRS
(impurity A) in 2 mL of a 40 g/L solution of sodium ASSAY
hydroxide R and dilute to 20.0 mL with water R. Dilute 1.0 mL Dissolve 0.180 g in 50 mL of carbon dioxide-free water R.
of this solution to 10.0 mL with water R. Titrate with 0.1 M sodium hydroxide, determining the
Reference solution (c). Dilute 1.0 mL of reference solution (b) end-point potentiometrically (2.2.20).
to 10.0 mL with mobile phase A. 1 mL of 0.1 M sodium hydroxide is equivalent to 22.32 mg of
Reference solution (d). Dilute 1.0 mL of reference solution (b) C11H13NO4.
to 20.0 mL with the test solution. STORAGE
Column : Protected from light. If the substance is sterile, store in a
– size : l = 0.15 m, Ø = 3 mm ; sterile, airtight, tamper-evident container.
– stationary phase : spherical octadecylsilyl silica gel for IMPURITIES
chromatography R (3 μm) ;
– temperature : 40 °C. Specified impurities : A.
Mobile phase : Other detectable impurities (the following substances would,
if present at a sufficient level, be detected by one or other of
– mobile phase A : mix 1.0 mL of phosphoric acid R and the tests in the monograph. They are limited by the general
1000 mL of water for chromatography R ; acceptance criterion for other/unspecified impurities and/or
– mobile phase B : acetonitrile R1 ; by the general monograph Substances for pharmaceutical
Time Mobile phase A Mobile phase B use (2034). It is therefore not necessary to identify these
(min) (per cent V/V) (per cent V/V) impurities for demonstration of compliance. See also 5.10.
0-2 97 3
Control of impurities in substances for pharmaceutical use) : B.

2 - 15 97 → 62 3 → 38

Flow rate : 0.7 mL/min.

Detection : spectrophotometer at 219 nm.
A. (2S)-2-amino-3-(4-hydroxyphenyl)propanoic acid
Injection : 2 μL of the test solution and reference solutions (a), (tyrosine),
(c) and (d).
Relative retention with reference to N-acetyltyrosine (retention
time = about 6 min) : impurity A = about 0.5.
System suitability : reference solution (d) :
– resolution : minimum 5.0 between the principal peak and
the peak due to impurity A. B. (2S)-2-(acetylamino)-3-[4-(acetoxy)phenyl]propanoic acid
Limits : (diacetyltyrosine).
– impurity A : not more than 0.8 times the area of the
corresponding peak in the chromatogram obtained with 04/2021:0968
reference solution (c) (0.8 per cent) ;
– unspecified impurities : for each impurity, not more than the
area of the principal peak in the chromatogram obtained
with reference solution (a) (0.10 per cent) ;
– total : maximum 1.0 per cent ; ACICLOVIR
– disregard limit : 0.5 times the area of the principal peak in
the chromatogram obtained with reference solution (a) Aciclovirum
(0.05 per cent).
Chlorides (2.4.4): maximum 200 ppm.
Dilute 10 mL of solution S to 15 mL with water R.
Sulfates (2.4.13) : maximum 200 ppm.
Dissolve 1.0 g in distilled water R and dilute to 20 mL with
the same solvent. C8H11N5O3 Mr 225.2
Ammonium (2.4.1, Method B) : maximum 200 ppm, [59277-89-3]
determined on 0.100 g.
DEFINITION
Prepare the standard using 0.2 mL of ammonium standard
solution (100 ppm NH4) R. 2-Amino-9-[(2-hydroxyethoxy)methyl]-1,9-dihydro-6H-
purin-6-one.
Iron (2.4.9) : maximum 20 ppm.
Content : 98.5 per cent to 101.0 per cent (anhydrous substance).
In a separating funnel, dissolve 0.5 g in 10 mL of dilute
hydrochloric acid R. Shake with 3 quantities, each of 10 mL, of CHARACTERS
methyl isobutyl ketone R1, shaking for 3 min each time. To the Appearance : white or almost white, crystalline powder.
combined organic layers add 10 mL of water R and shake for Solubility : slightly soluble in water, very slightly soluble
3 min. The aqueous layer complies with the test. in ethanol (96 per cent), practically insoluble in heptane.
Loss on drying (2.2.32) : maximum 0.5 per cent, determined It dissolves in dilute solutions of mineral acids and alkali
on 1.000 g by drying in an oven at 105 °C. hydroxides.

1878 See the information section on general monographs (cover pages)

EUROPEAN PHARMACOPOEIA 11.0
IDENTIFICATION
Infrared absorption spectrophotometry (2.2.24).
Comparison : aciclovir CRS.
TESTS
Appearance of solution. The solution is clear (2.2.1) and not
more intensely coloured than reference solution Y7 (2.2.2,
Method II).
Dissolve 0.25 g in a 4 g/L solution of sodium hydroxide R and
dilute to 25 mL with the same solvent.
Related substances. Liquid chromatography (2.2.29). Prepare
the solutions immediately before use.
Solvent mixture : dimethyl sulfoxide R, water R (20:80 V/V).
Phosphate buffer solution pH 2.5. Dissolve 3.48 g of
dipotassium hydrogen phosphate R in 1000 mL of water for
chromatography R and adjust to pH 2.5 with phosphoric acid R.
Phosphate buffer solution pH 3.1. Dissolve 3.48 g of
dipotassium hydrogen phosphate R in 1000 mL of water for
chromatography R and adjust to pH 3.1 with phosphoric acid R.
Test solution. Dissolve 25 mg of the substance to be examined
in 5.0 mL of dimethyl sulfoxide R and dilute to 25.0 mL with
water R.
Reference solution (a). Dissolve 5 mg of aciclovir for system
suitability A CRS (containing impurities B, J, K, N, O and P) in
1 mL of dimethyl sulfoxide R and dilute to 5 mL with water R.
Reference solution (b). Dilute 1.0 mL of the test solution to
100.0 mL with the solvent mixture. Dilute 1.0 mL of this
solution to 10.0 mL with the solvent mixture.
Reference solution (c). Dissolve the contents of a vial of
aciclovir for impurity C identification CRS in 200 μL of
dimethyl sulfoxide R and dilute to 1 mL with water R.
Reference solution (d). Dissolve the contents of a vial of
aciclovir for impurity G identification CRS in 1 mL of reference
solution (a).
Column :
– size : l = 0.25 m, Ø = 4.6 mm ;
– stationary phase : end-capped octadecylsilyl silica gel for
chromatography R (5 μm).
Mobile phase :
– mobile phase A : acetonitrile R, phosphate buffer solution
pH 3.1 (1:99 V/V) ;
– mobile phase B : acetonitrile R, phosphate buffer solution
pH 2.5 (50:50 V/V) ;
Time Mobile phase A Mobile phase B
(min) (per cent V/V) (per cent V/V)
0-5 100 0 Other detectable impurities (the following substances would,
5 - 27 100 → 80 0 → 20
27 - 40 80 20
Flow rate : 1.0 mL/min.
Detection : spectrophotometer at 254 nm.
Injection : 10 μL of the test solution and reference solutions (b), Control of impurities in substances for pharmaceutical use) : A,
(c) and (d).
Identification of impurities : use the chromatogram supplied
with aciclovir for impurity C identification CRS and the
chromatogram obtained with reference solution (c) to identify
the peak due to impurity C ; use the chromatograms supplied
with aciclovir for system suitability A CRS and aciclovir
for impurity G identification CRS and the chromatogram
obtained with reference solution (d) to identify the peaks due
to impurities B, G, J, K, N, O and P.
Relative retention with reference to aciclovir (retention
time = about 13 min) : impurity B = about 0.4 ;
impurity P = about 0.7 ; impurity C = about 0.9 ;
impurity N = about 1.37 ; impurities O and Q = about 1.42 ;
impurity J = about 1.62 ; impurities K and R = about 2.5 ;
impurity G = about 2.6.
General Notices (1) apply to all monographs and other texts
Aciclovir
System suitability :
– resolution : minimum 1.5 between the peaks due to
impurity C and aciclovir in the chromatogram obtained
with reference solution (c) ; minimum 1.5 between the
peaks due to impurities K and G in the chromatogram
obtained with reference solution (d).
Limits :
– correction factor : for the calculation of content, multiply
the peak area of impurity C by 2.2 ;
– impurity B : not more than 7 times the area of the principal
peak in the chromatogram obtained with reference
solution (b) (0.7 per cent);
– impurity J : not more than twice the area of the principal
peak in the chromatogram obtained with reference
solution (b) (0.2 per cent);
– sum of impurities K and R : not more than 1.5 times the area
of the principal peak in the chromatogram obtained with
reference solution (b) (0.15 per cent);
– sum of impurities O and Q : not more than 1.5 times the
area of the principal peak in the chromatogram obtained
with reference solution (b) ( 0.15 per cent);
– impurities C, N, P : for each impurity, not more than
1.5 times the area of the principal peak in the chromatogram
obtained with reference solution (b) (0.15 per cent) ;
– unspecified impurities : for each impurity, not more than
0.5 times the area of the principal peak in the chromatogram
obtained with reference solution (b) (0.05 per cent) ;
– total : not more than 10 times the area of the principal peak
in the chromatogram obtained with reference solution (b)
(1.0 per cent);
– disregard limit : 0.3 times the area of the principal peak in
the chromatogram obtained with reference solution (b)
(0.03 per cent).
Water (2.5.12) : maximum 6.0 per cent, determined on 0.500 g.
Sulfated ash (2.4.14): maximum 0.1 per cent, determined on
1.0 g.
ASSAY
Dissolve 0.150 g in 60 mL of anhydrous acetic acid R. Titrate
with 0.1 M perchloric acid, determining the end-point
potentiometrically (2.2.20). Carry out a blank titration.
1 mL of 0.1 M perchloric acid is equivalent to 22.52 mg
of C8H11N5O3.
IMPURITIES
Specified impurities : B, C, J, K, N, O, P, Q, R.
if present at a sufficient level, be detected by one or other of
the tests in the monograph. They are limited by the general
acceptance criterion for other/unspecified impurities and/or
by the general monograph Substances for pharmaceutical
use (2034). It is therefore not necessary to identify these
impurities for demonstration of compliance. See also 5.10.
F, G, I, L, M.
A. 2-[(2-amino-6-oxo-1,6-dihydro-9H-purin-9-
yl)methoxy]ethyl acetate,
B. 2-amino-1,7-dihydro-6H-purin-6-one (guanine),
1879
Acitretin
C. 2-amino-7-[(2-hydroxyethoxy)methyl]-1,7-dihydro-6H-
purin-6-one,
F. N-[9-[(2-hydroxyethoxy)methyl]-6-oxo-6,9-dihydro-1H-
purin-2-yl]acetamide,
G. 2-[(2-acetamido-6-oxo-1,6-dihydro-9H-purin-9-
yl)methoxy]ethyl acetate,
I. 2-amino-7-[[2-[(2-amino-6-oxo-1,6-dihydro-9H-purin-9-
yl)methoxy]ethoxy]methyl]-1,7-dihydro-6H-purin-6-one,
J. 9,9′-[ethane-1,2-diylbis(oxymethylene)]bis(2-amino-1,9-
dihydro-6H-purin-6-one),
K. 2,2′-(methylenediazanediyl)bis[9-[(2-hydroxye-
thoxy)methyl]-1,9-dihydro-6H-purin-6-one],
L. N-(9-acetyl-6-oxo-6,9-dihydro-1H-purin-2-yl)acetamide
(N2,9-diacetylguanine),
M. 2-[(2-acetamido-6-oxo-1,6-dihydro-7H-purin-7-
yl)methoxy]ethyl acetate,
N. unknown structure,
O. unknown structure,
1880
EUROPEAN PHARMACOPOEIA 11.0
P. 2-amino-9-(2-hydroxyethyl)-1,9-dihydro-6H-purin-6-one,
Q. mixture of 2-amino-9-[[2-(hydroxyethoxy)
methoxy]methyl]-1,9-dihydro-6H-purin-6-one and
2-amino-9-[[2-(hydroxymethoxy)ethoxy]methyl]-1,9-
dihydro-6H-purin-6-one,
R. 9,9′-[methylenebis(oxyethane-2,1-diyloxymethy-
lene)]bis(2-amino-1,9-dihydro-6H-purin-6-one).
01/2023:1385
ACITRETIN
Acitretinum
C21H26O3 Mr 326.4
[55079-83-9]
DEFINITION
(2E,4E,6E,8E)-9-(4-Methoxy-2,3,6-trimethylphenyl)-3,7-
dimethylnona-2,4,6,8-tetraenoic acid.
Content : 98.0 per cent to 102.0 per cent (dried substance).
CHARACTERS
Appearance : yellow or greenish-yellow, crystalline powder.
Solubility : practically insoluble in water, sparingly soluble in
tetrahydrofuran, slightly soluble in acetone and in ethanol
(96 per cent), very slightly soluble in cyclohexane.
It is sensitive to air, heat and light, especially in solution.
It shows polymorphism (5.9).
Carry out all operations as rapidly as possible and avoid
exposure to actinic light ; use freshly prepared solutions.
IDENTIFICATION
First identification : A.
Second identification : B.
A. Infrared absorption spectrophotometry (2.2.24).
Comparison : acitretin CRS.
See the information section on general monographs (cover pages)