EUROPEAN PHARMACOPOEIA 10.

1 Lisinopril dihydrate

Reference solution (a). Dissolve 5 mg of lisinopril for system

suitability A CRS (containing impurities A and E) in mobile
phase A and dilute to 1 mL with mobile phase A.
Reference solution (b). Dilute 1.0 mL of the test solution to
100.0 mL with mobile phase A. Dilute 1.0 mL of this solution
to 10.0 mL with mobile phase A.
Reference solution (c). Dissolve the contents of a vial of
U. 17-hydroxy-19-nor-17α-pregn-4-en-20-yn-3-one lisinopril impurity F CRS in 1 mL of mobile phase A.
(norethisterone), Reference solution (d). Dissolve 5 mg of lisinopril for peak
identification CRS (containing impurity G) in mobile phase A
and dilute to 1 mL with mobile phase A.
Column :
– size : l = 0.25 m, Ø = 4.6 mm ;
– stationary phase : base-deactivated end-capped octadecylsilyl
silica gel for chromatography R (5 μm) ;
V. 13-ethyl-3-methoxy-18,19-dinor-17α-pregna-1,3,5(10)- – temperature : 50 °C.
trien-20-yn-17-ol, Mobile phase :
– mobile phase A : mix 3 volumes of acetonitrile R1 and
97 volumes of a 3.12 g/L solution of sodium dihydrogen
phosphate R previously adjusted to pH 3.8 with dilute
phosphoric acid R ;
– mobile phase B : mix 20.5 volumes of acetonitrile R1 and
79.5 volumes of a 3.12 g/L solution of sodium dihydrogen
phosphate R previously adjusted to pH 3.5 with dilute
phosphoric acid R ;
W. 13-ethyl-17-hydroxy-18,19-dinor-17α-pregna-5,7,9-trien-
20-yn-3-one. Time Mobile phase A Mobile phase B
(min) (per cent V/V) (per cent V/V)
0-2 100 0
04/2020:1120
2 - 37 100 → 0 0 → 100

37 - 62 0 100

Flow rate : 1.6 mL/min.

LISINOPRIL DIHYDRATE Detection : spectrophotometer at 210 nm.
Injection : 50 μL.
Lisinoprilum dihydricum Identification of impurities : use the chromatogram
supplied with lisinopril for system suitability A CRS and the
chromatogram obtained with reference solution (a) to identify
the peaks due to impurities A and E ; use the chromatogram
obtained with reference solution (c) to identify the peak due to
impurity F ; use the chromatogram supplied with lisinopril for
peak identification CRS and the chromatogram obtained with
reference solution (d) to identify the peak due to impurity G.
C21H31N3O5,2H2O Mr 441.5 Relative retention with reference to lisinopril (retention
[83915-83-7] time = about 14 min) : impurity A = about 0.7 ;
DEFINITION impurity E = about 1.2 ; impurity F = about 1.9 ;
impurity G = about 2.9.
(2S)-1-[(2S)-6-Amino-2-[[(1S)-1-carboxy-3-phenylpropyl]-
amino]hexanoyl]pyrrolidine-2-carboxylic acid dihydrate. System suitability :
Content : 98.5 per cent to 101.5 per cent (anhydrous substance). – resolution : minimum 1.5 between the peaks due to
lisinopril and impurity E in the chromatogram obtained
CHARACTERS with reference solution (a) ;
Appearance : white or almost white, crystalline powder. – signal-to-noise ratio : minimum 45 for the principal peak in
Solubility : soluble in water, practically insoluble in anhydrous the chromatogram obtained with reference solution (b).
ethanol and in heptane. Calculation of percentage contents :
IDENTIFICATION – correction factor : multiply the peak area of impurity F by
2.1 ;
A. Specific optical rotation (2.2.7) : − 47 to − 43 (anhydrous
substance). – for each impurity, use the concentration of lisinopril
dihydrate in reference solution (b).
Dissolve 0.5 g in zinc acetate solution R and dilute to
50.0 mL with the same solvent. Limits :
B. Infrared absorption spectrophotometry (2.2.24). – impurities A, E, F : for each impurity, maximum 0.2 per
cent ;
Comparison : lisinopril dihydrate CRS.
– impurity G : maximum 0.15 per cent ;
TESTS – unspecified impurities : for each impurity, maximum
Related substances. Liquid chromatography (2.2.29). 0.10 per cent ;
Test solution. Dissolve 50.0 mg of the substance to be – total : maximum 0.5 per cent ;
examined in mobile phase A and dilute to 10.0 mL with – reporting threshold : 0.05 per cent ; disregard any peak with
mobile phase A. a retention time less than 3 min.

General Notices (1) apply to all monographs and other texts 4443
Lisinopril dihydrate EUROPEAN PHARMACOPOEIA 10.1

Water (2.5.12) : 8.0 per cent to 9.5 per cent, determined on

0.200 g.
Sulfated ash (2.4.14) : maximum 0.1 per cent, determined on
1.0 g.
ASSAY
Dissolve 0.350 g in 50 mL of water R. Titrate with 0.1 M sodium F. (2S)-1-[(2S)-6-amino-2-[[(1S)-1-carboxy-3-cyclohexyl-
hydroxide, determining the end-point potentiometrically propyl]amino]hexanoyl]pyrrolidine-2-carboxylic acid
(2.2.20) at the 1st point of inflexion. (cyclohexyl analogue),
1 mL of 0.1 M sodium hydroxide is equivalent to 40.55 mg
of C21H31N3O5.
IMPURITIES
Specified impurities : A, E, F, G.
Other detectable impurities (the following substances would,
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.
Control of impurities in substances for pharmaceutical use) : B,
C, D, H, I, J.
G. (2S)-1-[(2S)-6-amino-2-[[(2S)-1-[[(5S)-5-[[(1S)-
1-carboxy-3-phenylpropyl]amino]-6-[(2S)-2-
carboxypyrrolidin-1-yl]-6-oxohexyl]amino]-1-oxo-
4-phenylbutan-2-yl]amino]hexanoyl]pyrrolidine-2-
carboxylic acid (lisinopril dimer),
A. (2RS)-2-amino-4-phenylbutanoic acid,

B. 4-methylbenzenesulfonic acid,

H. (2S)-6-amino-2-[[(1S)-1-carboxy-3-phenylpropyl]amino]-
hexanoic acid,

C. (2S)-2-[(3S,8aS)-3-(4-aminobutyl)-1,4-dioxohexahydro-
pyrrolo[1,2-a]pyrazin-2(1H)-yl]-4-phenylbutanoic acid
(S,S,S-diketopiperazine),

I. (2S)-1-[(2S)-2,6-bis[[(1S)-1-carboxy-3-phenylpropyl]-
amino]hexanoyl]pyrrolidine-2-carboxylic acid,

D. (2S)-2-[(3S,8aR)-3-(4-aminobutyl)-1,4-dioxohexahydro-
pyrrolo[1,2-a]pyrazin-2(1H)-yl]-4-phenylbutanoic acid
(R,S,S-diketopiperazine),

J. (2S)-1-[(2S)-6-[(2S)-6-amino-2-[[(1S)-1-carboxy-3-
E. (2S)-1-[(2S)-6-amino-2-[[(1R)-1-carboxy-3-phenyl- phenylpropyl]amino]hexanamido]-2-[[(1S)-1-carboxy-3-
propyl]amino]hexanoyl]pyrrolidine-2-carboxylic acid phenylpropyl]amino]hexanoyl]pyrrolidine-2-carboxylic
(lisinopril R,S,S-isomer), acid.

4444 See the information section on general monographs (cover pages)