Aciclovir

EUROPEAN PHARMACOPOEIA 8.0

Loss on drying (2.2.32): maximum 0.5 per cent, determined

on 1.000 g by drying in an oven at 105 C.
Sulfated ash (2.4.14) : maximum 0.1 per cent, determined on
1.0 g.
Bacterial endotoxins (2.6.14) : less than 25 IU/g, if intended
for use in the manufacture of parenteral preparations without
a further appropriate procedure for the removal of bacterial
endotoxins.

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
ASSAY
dilute to 25 mL with the same solvent.
Dissolve 0.180 g in 50 mL of carbon dioxide-free water R.
Related substances. Liquid chromatography (2.2.29). Prepare
Titrate with 0.1 M sodium hydroxide, determining the
the solutions immediately before use.
end-point potentiometrically (2.2.20).
Solvent mixture : dimethyl sulfoxide R, water R (20:80 V/V).
1 mL of 0.1 M sodium hydroxide is equivalent to 22.32 mg
of C11H13NO4.
Phosphate buffer solution pH 2.5. Dissolve 3.48 g of
dipotassium hydrogen phosphate R in 1000 mL of water R and
STORAGE
adjust to pH 2.5 with phosphoric acid R.
Protected from light. If the substance is sterile, store in a
Phosphate buffer solution pH 3.1. Dissolve 3.48 g of
sterile, airtight, tamper-proof container.
dipotassium hydrogen phosphate R in 1000 mL of water R and
adjust to pH 3.1 with phosphoric acid R.
IMPURITIES
Test solution. Dissolve 25 mg of the substance to be examined
Specified impurities : A.
in 5.0 mL of dimethyl sulfoxide R and dilute to 25.0 mL with
Other detectable impurities (the following substances would,
water R.
if present at a sufcient level, be detected by one or other of
Reference solution (a). Dissolve 5 mg of aciclovir for system
the tests in the monograph. They are limited by the general
suitability CRS (containing impurities A, B, J, K, N, O and P) in
acceptance criterion for other/unspecied impurities and/or
1 mL of dimethyl sulfoxide R and dilute to 5.0 mL with water R.
by the general monograph Substances for pharmaceutical
Reference solution (b). Dilute 1.0 mL of the test solution to
use (2034). It is therefore not necessary to identify these
100.0 mL with the solvent mixture. Dilute 1.0 mL of this
impurities for demonstration of compliance. See also 5.10.
Control of impurities in substances for pharmaceutical use) : B. solution to 10.0 mL with the solvent mixture.
Reference solution (c). Dissolve the contents of a vial of
aciclovir for peak identification 1 CRS (containing impurities C
and I) in 200 L of dimethyl sulfoxide R and dilute to 1.0 mL
with water R.
A. (2S)-2-amino-3-(4-hydroxyphenyl)propanoic acid
Reference solution (d). Dissolve the contents of a vial of
(tyrosine),
aciclovir for peak identification 2 CRS (containing impurities F
and G) in 1.0 mL of reference solution (a).
Column :
size : l = 0.25 m, = 4.6 mm ;
stationary phase : end-capped octadecylsilyl silica gel for
chromatography R (5 m).
B. (2S)-2-(acetylamino)-3-[4-(acetoxy)phenyl]propanoic acid
Mobile
phase :
(diacetyltyrosine).
mobile phase A : acetonitrile R, phosphate buffer solution
pH 3.1 (1:99 V/V) ;
01/2014:0968
mobile phase B : acetonitrile R, phosphate buffer solution
pH 2.5 (50:50 V/V);

ACICLOVIR
Aciclovirum

Time
(min)
0-5

Mobile phase A
(per cent V/V)
100

Mobile phase B
(per cent V/V)
0

5 - 27

100 80

0 20

27 - 40

80

20

Flow rate : 1.0 mL/min.

C8H11N5O3
Mr 225.2 Detection : spectrophotometer at 254 nm.
Injection : 10 L of the test solution and reference solutions (b),
[59277-89-3]
(c) and (d).
DEFINITION
Identification of impurities : use the chromatogram supplied
2-Amino-9-[(2-hydroxyethoxy)methyl]-1,9-dihydro-6Hwith aciclovir for peak identification 1 CRS and the
purin-6-one.
chromatogram obtained with reference solution (c) to identify
Content : 98.5 per cent to 101.0 per cent (anhydrous substance). the peaks due to impurities C and I ; use the chromatogram
supplied with aciclovir for peak identification 2 CRS and the
CHARACTERS
chromatogram obtained with reference solution (d) to identify
the peaks due to impurities A, B, F, G, J, K, N, O and P.
Appearance : white or almost white, crystalline powder.
Relative retention with reference to aciclovir (retention
Solubility : slightly soluble in water, very slightly soluble
time = about 13 min) : impurity B = about 0.4 ;
in ethanol (96 per cent), practically insoluble in heptane.
impurity P = about 0.7 ; impurity C = about 0.9 ;
It dissolves in dilute solutions of mineral acids and alkali
impurity N = about 1.37 ; impurities O and Q = about 1.42 ;
hydroxides.

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Aciclovir

EUROPEAN PHARMACOPOEIA 8.0

impurity I = about 1.57 ; impurity J = about 1.62 ;

impurity F = about 1.7 ; impurity A = about 1.8 ; impurities K
and R = about 2.5 ; impurity G = about 2.6.
System suitability :
resolution : minimum 1.5 between the peaks due to
A. 2-[(2-amino-6-oxo-1,6-dihydro-9H-purin-9impurity C and aciclovir in the chromatogram obtained
yl)methoxy]ethyl acetate,
with reference solution (c) ; minimum 1.5 between the
peaks due to impurities F and A and 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 I by 1.5 ;

B. 2-amino-1,7-dihydro-6H-purin-6-one (guanine),

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) ;
sum of impurities O and Q : not more than 3 times the area
of the principal peak in the chromatogram obtained with
reference solution (b) (0.3 per cent) ;

C. 2-amino-7-[(2-hydroxyethoxy)methyl]-1,7-dihydro-6Hpurin-6-one,

sum of impurities K and R : not more than twice the area

of the principal peak in the chromatogram obtained with
reference solution (b) (0.2 per cent) ;
impurities A, G, J, N, P : for each impurity, not more than
twice the area of the principal peak in the chromatogram
obtained with reference solution (b) (0.2 per cent) ;

F. N-[9-[(2-hydroxyethoxy)methyl]-6-oxo-6,9-dihydro-1Hpurin-2-yl]acetamide,

impurities C, F, I : for each impurity, not more than the area

of the principal peak in the chromatogram obtained with
reference solution (b) (0.1 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) ;
G. 2-[[2-(acetylamino)-6-oxo-1,6-dihydro-9H-purin-9yl]methoxy]ethyl acetate,
total : not more than 15 times the area of the principal peak
in the chromatogram obtained with reference solution (b)
(1.5 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.

I. 2-amino-7-[[2-[(2-amino-6-oxo-1,6-dihydro-9H-purin-9yl)methoxy]ethoxy]methyl]-1,7-dihydro-6H-purin-6-one,

Bacterial endotoxins (2.6.14, Method D) : less than

0.50 IU/mg, if intended for use in the manufacture of
parenteral preparations without a further appropriate
procedure for the removal of bacterial endotoxins.
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.

J. 9,9-[ethylenebis(oxymethylene)]bis(2-amino-1,9-dihydro6H-purin-6-one),

1 mL of 0.1 M perchloric acid is equivalent to 22.52 mg

of C8H11N5O3.
IMPURITIES
Specified impurities : A, B, C, F, G, I, J, K, N, O, P, Q, R.

K. 2,2-(methylenediimino)bis[9-[(2-hydroxyethoxy)methyl]1,9-dihydro-6H-purin-6-one],

Other detectable impurities (the following substances would,

if present at a sufcient level, be detected by one or other of
the tests in the monograph. They are limited by the general
acceptance criterion for other/unspecied 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. Control of
L. N-(9-acetyl-6-oxo-6,9-dihydro-1H-purin-2-yl)acetamide
impurities in substances for pharmaceutical use) : L, M.
(N2,9-diacetylguanine),
General Notices (1) apply to all monographs and other texts

1483

Acitretin

EUROPEAN PHARMACOPOEIA 8.0

Carry out all operations as rapidly as possible and avoid

exposure to actinic light ; use freshly prepared solutions.

M. 2-[[2-(acetylamino)-6-oxo-1,6-dihydro-7H-purin-7yl]methoxy]ethyl acetate,
N. unknown structure,
O. unknown structure,

P. 2-amino-9-(2-hydroxyethyl)-1,9-dihydro-6H-purin-6-one,

Q. mixture of 2-amino-9-[[2-(hydroxymethoxy)
ethoxy]methyl]-1,9-dihydro-6H-purin-6-one and
2-amino-9-[[2-(hydroxyethoxy)methoxy]methyl]-1,9dihydro-6H-purin-6-one,

IDENTIFICATION
First identification : B.
Second identification : A, C.
A. Ultraviolet and visible absorption spectrophotometry
(2.2.25).
Test solution. Dissolve 15.0 mg in 10 mL of
tetrahydrofuran R and dilute immediately to 100.0 mL
with the same solvent. Dilute 2.5 mL of this solution to
100.0 mL with tetrahydrofuran R.
Spectral range : 300-400 nm.
Absorption maximum : at 358 nm.
Specific absorbance at the absorption maximum : 1350 to
1475.
B. Infrared absorption spectrophotometry (2.2.24).
Preparation : discs.
Comparison : acitretin CRS.
If the spectra obtained in the solid state show differences,
dissolve the substance to be examined and the reference
substance separately in 2-propanol R heating under reux,
lter, evaporate to dryness and record new spectra using
the residues.
C. Examine the chromatograms obtained in the assay.
Results : the principal peak in the chromatogram obtained
with test solution (b) is similar in retention time to
the principal peak in the chromatogram obtained with
reference solution (a).

TESTS
Related substances. Liquid chromatography (2.2.29).
Maintain the sampler at 4 C.
Test solution (a). Dissolve 25.0 mg of the substance to be
examined in 5 mL of tetrahydrofuran R and dilute immediately
to 100.0 mL with anhydrous ethanol R.
Test solution (b). Dilute 10.0 mL of test solution (a) to 25.0 mL
with anhydrous ethanol R.
R. 9,9-[methylenebis(oxyethyleneoxymethylene)]bis(2Reference solution (a). Dissolve 25.0 mg of acitretin CRS in
amino-1,9-dihydro-6H-purin-6-one).
5 mL of tetrahydrofuran R and dilute immediately to 100.0 mL
with anhydrous ethanol R. Dilute 10.0 mL of this solution to
07/2010:1385 25.0 mL with anhydrous ethanol R.
corrected 7.0 Reference solution (b). Dissolve 1.0 mg of tretinoin CRS in
anhydrous ethanol R and dilute to 20.0 mL with the same
ACITRETIN
solvent. Mix 5.0 mL of this solution with 2.5 mL of reference
solution (a) and dilute to 100.0 mL with anhydrous ethanol R.
Acitretinum
Reference solution (c). Dilute 2.5 mL of reference solution (a)
to 50.0 mL with anhydrous ethanol R. Dilute 3.0 mL of this
solution to 20.0 mL with anhydrous ethanol R.
Column :
size l = 0.25 m, = 4 mm ;
stationary phase : microparticulate octadecylsilyl silica gel
for chromatography R (5 m) with a specic surface area
C21H26O3
Mr 326.4
of 200 m2/g, a pore size of 15 nm and a carbon loading of
[55079-83-9]
20 per cent ;
temperature : 25 C.
DEFINITION
Mobile phase : a 0.3 per cent V/V solution of glacial acetic
(all-E)-9-(4-Methoxy-2,3,6-trimethylphenyl)-3,7acid R in a mixture of 8 volumes of water R and 92 volumes
dimethylnona-2,4,6,8-tetraenoic acid.
of anhydrous ethanol R.
Content : 98.0 per cent to 102.0 per cent (dried substance).
Flow rate : 0.6 mL/min.
CHARACTERS
Detection : spectrophotometer at 360 nm.
Appearance : yellow or greenish-yellow, crystalline powder.
Injection : 10 L of test solution (a) and reference solutions (b)
and (c).
Solubility : practically insoluble in water, sparingly soluble in
tetrahydrofuran, slightly soluble in acetone and in ethanol
Run time : 2.5 times the retention time of acitretin.
(96 per cent), very slightly soluble in cyclohexane.
Retention time : impurity A = about 4.8 min ; tretinoin = about
It is sensitive to air, heat and light, especially in solution.
5.2 min ; acitretin = about 6.2 min ; impurity B = about
10.2 min.
It shows polymorphism.

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See the information section on general monographs (cover pages)