EUROPEAN PHARMACOPOEIA 11.

0 Metformin hydrochloride

04/2020:0931 Impurity F. Liquid chromatography (2.2.29).

METFORMIN HYDROCHLORIDE
Metformini hydrochloridum
C4H12ClN5 Mr 165.6 5 min before use.
[1115-70-4]
DEFINITION
1,1-Dimethylbiguanide hydrochloride.
Content : 98.5 per cent to 101.0 per cent (dried substance).
CHARACTERS
Appearance : white or almost white crystals.
Solubility : freely soluble in water, slightly soluble in ethanol
(96 per cent), practically insoluble in acetone and in methylene – stationary phase : spherical end-capped octadecylsilyl silica
chloride.
IDENTIFICATION
First identification : B, E.
Second identification : A, C, D, E.
A. Melting point (2.2.14) : 222 °C to 226 °C.
B. Infrared absorption spectrophotometry (2.2.24).
Comparison : metformin hydrochloride CRS.
C. Thin-layer chromatography (2.2.27).
Test solution. Dissolve 20 mg of the substance to be
examined in water R and dilute to 5 mL with the same
solvent.
Reference solution. Dissolve 20 mg of metformin
hydrochloride CRS in water R and dilute to 5 mL with the
same solvent.
Plate : TLC silica gel G plate R.
Mobile phase : glacial acetic acid R, butanol R, water R
(10:40:50 V/V/V) ; use the upper layer.
Application : 5 μL.
Development : over 3/4 of the plate.
Drying : at 100-105 °C for 15 min.
Detection : spray with a mixture of equal volumes of a
100 g/L solution of sodium nitroprusside R, a 100 g/L
solution of potassium ferricyanide R and a 100 g/L solution
of sodium hydroxide R, prepared 20 min before use.
Results : the principal spot in the chromatogram obtained
with the test solution is similar in position, colour and size
to the principal spot in the chromatogram obtained with
the reference solution.
D. Dissolve about 5 mg in water R and dilute to 100 mL with Test solution. Dissolve 50.0 mg of the substance to be
the same solvent. To 2 mL of the solution add 0.25 mL
of strong sodium hydroxide solution R and 0.10 mL of
α-naphthol solution R. Mix and allow to stand in iced water Reference solution (a). Dissolve 20.0 mg of metformin
for 15 min. Add 0.5 mL of sodium hypobromite solution R impurity A CRS in water R and dilute to 100.0 mL with the
and mix. A pink colour develops.
E. It gives reaction (a) of chlorides (2.3.1).
TESTS
Solution S. Dissolve 2.0 g in water R and dilute to 20 mL with to 20.0 mL with the mobile phase.
the same solvent.
Appearance of solution. Solution S is clear (2.2.1) and
colourless (2.2.2, Method II). Heat the solution to 50 °C and
cool to room temperature.
Derivatisation solution. Prepare the solution immediately
before use. Dilute 1 mL of fluorodinitrobenzene R in 100.0 mL
of acetonitrile R.
Blank solution. To 5.0 mL of acetonitrile R add 100 μL of
triethylamine R1 and 1.0 mL of the derivatisation solution.
Shake well and heat at 60 °C for 30 min. After cooling, dilute
to 10.0 mL with acetonitrile R.
Test solution. Prepare the solution immediately before use.
Suspend 10.0 mg of the substance to be examined in 5.0 mL
of acetonitrile R and sonicate for 5 min. Add 100 μL of
triethylamine R1 and 1.0 mL of the derivatisation solution.
Shake well and heat at 60 °C for 30 min. After cooling, dilute
to 10.0 mL with acetonitrile R. Filter or centrifuge at 800 g for
Reference solution. Dilute 1.0 mL of metformin impurity F CRS
in 100.0 mL of acetonitrile R. Dilute 2.5 mL of the solution
to 100.0 mL with acetonitrile R. To 1.0 mL of this solution
add successively 5.0 mL of acetonitrile R, 100 μL of
triethylamine R1 and 1.0 mL of the derivatisation solution.
Shake well and heat at 60 °C for 30 min. After cooling, dilute
to 10.0 mL with acetonitrile R.
Column :
– size : l = 0.125 m, Ø = 3 mm ;
gel for chromatography R1 (5 μm);
– temperature : 30 °C.
Mobile phase :
– mobile phase A : phosphoric acid R, water for
chromatography R (0.1:99.9 V/V) ;
– mobile phase B : acetonitrile R ;
Time Mobile phase A Mobile phase B
(min) (per cent V/V) (per cent V/V)
0 - 10 60 → 45 40 → 55
10 - 11 45 → 25 55 → 75
11 - 15 25 75
Flow rate : 0.7 mL/min.
Detection : spectrophotometer at 380 nm.
Injection : 5 μL.
Identification of impurities : use the chromatograms obtained
with the blank solution and the reference solution to identify
the peak due to the impurity F derivative.
Retention time : impurity F derivative = about 4 min.
System suitability : reference solution :
– resolution : minimum 3.0 between the peak due to the
impurity F derivative and the nearby eluting peaks due to
the derivatisation reagent.
Limit :
– impurity F : not more than the area of the corresponding
peak in the chromatogram obtained with the reference
solution (0.05 per cent).
Related substances. Liquid chromatography (2.2.29).
examined in the mobile phase and dilute to 10.0 mL with the
mobile phase.
same solvent. Dilute 1.0 mL of the solution to 200.0 mL with
the mobile phase.
Reference solution (b). Dilute 1.0 mL of the test solution to
50.0 mL with the mobile phase. Dilute 1.0 mL of this solution
Reference solution (c). Dissolve 10 mg of melamine R
(impurity D) in about 90 mL of water R. Add 5 mL of the test
solution and dilute to 100 mL with water R. Dilute 1 mL of
this solution to 50 mL with the mobile phase.

General Notices (1) apply to all monographs and other texts 3359
Methacrylic acid - ethyl acrylate copolymer (1:1) EUROPEAN PHARMACOPOEIA 11.0

Column :
– size : l = 0.25 m, Ø = 4.6 mm ;
– stationary phase : strong cation-exchange silica gel for
chromatography R (10 μm).
Mobile phase : 17 g/L solution of ammonium dihydrogen
phosphate R adjusted to pH 3.0 with phosphoric acid R. C. N2,N2-dimethyl-1,3,5-triazine-2,4,6-triamine
Flow rate : 1.0 mL/min. (N,N-dimethylmelamine),
Detection : spectrophotometer at 218 nm.
Injection : 20 μL.
Run time : twice the retention time of metformin.
Identification of impurities : use the chromatogram obtained
with reference solution (a) to identify the peak due to
impurity A ; use the chromatogram obtained with reference D. 1,3,5-triazine-2,4,6-triamine (melamine),
solution (c) to identify the peak due to impurity D.
Relative retention with reference to metformin
(retention time = about 14 min) : impurity A = about 0.3 ;
impurity D = about 0.4.
System suitability : reference solution (c) : E. 1-methylbiguanide,
– resolution : minimum 10 between the peaks due to
impurity D and metformin.
Limits :
F. N-methylmethanamine (dimethylamine).
– impurity A : not more than the area of the corresponding
peak in the chromatogram obtained with reference
solution (a) (0.02 per cent) ; 01/2019:1128
– 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 : maximum 0.2 per cent ;
– disregard limit : 0.3 times the area of the principal peak in METHACRYLIC ACID - ETHYL
the chromatogram obtained with reference solution (b)
(0.03 per cent); do not disregard the peak due to ACRYLATE COPOLYMER (1:1)
impurity A.
Loss on drying (2.2.32) : maximum 0.5 per cent, determined Acidi methacrylici et ethylis acrylatis
on 1.000 g by drying in an oven at 105 °C for 5 h. polymerisatum 1:1
Sulfated ash (2.4.14) : maximum 0.1 per cent, determined on DEFINITION
1.0 g.
Copolymer of methacrylic acid and ethyl acrylate having a
ASSAY mean relative molecular mass of about 250 000. The ratio of
Dissolve 0.100 g in 4 mL of anhydrous formic acid R. Add carboxylic groups to ester groups is about 1:1. The substance is
80 mL of acetonitrile R. Carry out the titration immediately. in the acid form (type A) or partially neutralised using sodium
Titrate with 0.1 M perchloric acid, determining the end-point hydroxide (type B). It may contain suitable surface-active
potentiometrically (2.2.20). agents such as sodium dodecyl sulfate and polysorbate 80.
Content :
1 mL of 0.1 M perchloric acid is equivalent to 16.56 mg of
C4H12ClN5. – type A : 46.0 per cent to 50.6 per cent of methacrylic
acid units (dried substance) ;
IMPURITIES – type B : 43.0 per cent to 48.0 per cent of methacrylic
Specified impurities : A, F. acid units (dried substance).
Other detectable impurities (the following substances would, CHARACTERS
if present at a sufficient level, be detected by one or other of
the tests in the monograph. They are limited by the general Appearance : white or almost white, free-flowing powder.
acceptance criterion for other/unspecified impurities and/or Solubility : practically insoluble in water (type A) or dispersible
by the general monograph Substances for pharmaceutical use in water (type B), freely soluble in anhydrous ethanol,
(2034). It is therefore not necessary to identify these impurities practically insoluble in ethyl acetate. It is freely soluble in a
for demonstration of compliance. See also 5.10. Control of 40 g/L solution of sodium hydroxide.
impurities in substances for pharmaceutical use): B, C, D, E.
IDENTIFICATION
A. Infrared absorption spectrophotometry (2.2.24).
Preparation : dissolve 0.1 g of the substance to be examined
in 1 mL of ethanol (90 per cent V/V) R and place 2 drops
A. cyanoguanidine, of the solution on a sodium chloride plate ; dry to allow
the formation of a film and cover with another sodium
chloride plate.
Comparison : methacrylic acid - ethyl acrylate
copolymer (1:1) - type A CRS or methacrylic acid - ethyl
acrylate copolymer (1:1) - type B CRS.
B. It complies with the limits of the assay.
B. (4,6-diamino-1,3,5-triazin-2-yl)guanidine, C. Sulfated ash (see Tests).

3360 See the information section on general monographs (cover pages)