Sulfanilamide EUROPEAN PHARMACOPOEIA 10.
B. Examine the chromatograms obtained in the test for related STORAGE
substances. The principal spot in the chromatogram Protected from light.
obtained with test solution (b) is similar in position and
size to the principal spot in the chromatogram obtained
with reference solution (a).
01/2017:1571
C. Dissolve 0.5 g in 1 mL of a 40 per cent V/V solution of
sulfuric acid R, heating gently. Continue heating until a
crystalline precipitate appears (about 2 min). Cool and
add 10 mL of dilute sodium hydroxide solution R. Cool
again, add 25 mL of ether R and shake the solution for
5 min. Separate the ether layer, dry over anhydrous sodium SULFANILAMIDE
sulfate R and filter. Evaporate the ether by heating in a
water-bath. An oily residue is obtained which becomes Sulfanilamidum
crystalline on cooling ; if necessary, scratch the wall of the
container with a glass rod. The residue melts (2.2.14) at
102 °C to 106 °C.
D. Dissolve about 5 mg in 10 mL of 1 M hydrochloric acid.
Dilute 1 mL of the solution to 10 mL with water R. The
solution, without further acidification, gives the reaction of
primary aromatic amines (2.3.1). C 6H 8N2O 2S Mr 172.2
[63-74-1]
TESTS DEFINITION
Appearance of solution. Dissolve 1.0 g in a mixture of 10 mL Sulfanilamide contains not less than 99.0 per cent
of 1 M sodium hydroxide and 15 mL of water R. The solution and not more than the equivalent of 101.0 per cent of
is clear (2.2.1) and not more intensely coloured than reference 4-aminobenzenesulfonamide, calculated with reference to the
solution Y4 or BY4 (2.2.2, Method II). dried substance.
Acidity. To 1.25 g, finely powdered, add 25 mL of carbon
CHARACTERS
dioxide-free water R. Heat at 70 °C for 5 min. Cool in iced
water for about 15 min and filter. To 20 mL of the filtrate White or yellowish-white crystals or fine powder, slightly
add 0.1 mL of bromothymol blue solution R1. Not more than soluble in water, freely soluble in acetone, sparingly soluble
0.5 mL of 0.1 M sodium hydroxide is required to change the in alcohol, practically insoluble in methylene chloride. It
colour of the indicator. dissolves in solutions of alkali hydroxides and in dilute
mineral acids.
Related substances. Examine by thin layer chromatography
(2.2.27), using TLC silica gel GF254 plate R. IDENTIFICATION
Test solution (a). Dissolve 0.10 g of the substance to be First identification : B.
examined in acetone R and dilute to 5 mL with the same Second identification : A, C, D.
solvent.
A. Melting point (2.2.14) : 164.5 °C to 166.0 °C.
Test solution (b). Dilute 1 mL of test solution (a) to 10 mL B. Examine by infrared absorption spectrophotometry
with acetone R. (2.2.24), comparing with the spectrum obtained with
Reference solution (a). Dissolve 20 mg of sulfamethoxypyr- sulfanilamide CRS. Examine the substances prepared as
idazine CRS in acetone R and dilute to 10 mL with the same discs.
solvent. C. Examine the chromatograms obtained in the test for
Reference solution (b). Dilute 2.5 mL of test solution (b) to related substances. The principal spot in the chromatogram
50 mL with acetone R. obtained with test solution (a) is similar in position and
size to the principal spot in the chromatogram obtained
Apply separately to the plate 5 μL of each solution. Develop with reference solution (a).
over a path of 15 cm using a mixture of 1 volume of dilute
ammonia R1, 9 volumes of water R, 30 volumes of 2-propanol R D. Dissolve about 5 mg in 10 mL of 1 M hydrochloric acid.
and 50 volumes of ethyl acetate R. Dry the plate at 100-105 °C Dilute 1 mL of the solution to 10 mL with water R. The
and examine in ultraviolet light at 254 nm. Any spot in the solution, without further acidification, gives the reaction of
chromatogram obtained with test solution (a), apart from primary aromatic amines (2.3.1).
the principal spot, is not more intense than the spot in the TESTS
chromatogram obtained with reference solution (b) (0.5 per
cent). Solution S. To 2.5 g add 50 mL of carbon dioxide-free water R.
Heat at about 70 °C for about 5 min. Cool in iced water for
Heavy metals (2.4.8). 1.0 g complies with test D for heavy about 15 min and filter.
metals (20 ppm). Prepare the reference solution using 2 mL
of lead standard solution (10 ppm Pb) R. Acidity. To 20 mL of solution S add 0.1 mL of bromothymol
blue solution R1. Not more than 0.2 mL of 0.1 M sodium
Loss on drying (2.2.32). Not more than 0.5 per cent, hydroxide is required to change the colour of the indicator.
determined on 1.000 g by drying in an oven at 105 °C.
Related substances. Examine by thin-layer chromatography
Sulfated ash (2.4.14). Not more than 0.1 per cent, determined (2.2.27), using a TLC silica gel F254 plate R.
on 1.0 g.
Test solution (a). Dissolve 20 mg of the substance to be
ASSAY examined in 3 mL of a mixture of 2 volumes of concentrated
ammonia R and 48 volumes of methanol R and dilute to 5 mL
Carry out the assay of primary aromatic amino-nitrogen with the same mixture of solvents.
(2.5.8), using 0.2500 g, determining the end-point Test solution (b). Dissolve 0.10 g of the substance to be
electrometrically. examined in 0.5 mL of concentrated ammonia R and dilute to
1 mL of 0.1 M sodium nitrite is equivalent to 28.03 mg of 5 mL with methanol R. If the solution is not clear, heat gently
C11H12N4O3S. until dissolution is complete.
3930 See the information section on general monographs (cover pages)
EUROPEAN PHARMACOPOEIA 10.0 Sulfasalazine
Reference solution (a). Dissolve 20 mg of sulfanilamide CRS in Preparation : discs.
3 mL of a mixture of 2 volumes of concentrated ammonia R Comparison : sulfasalazine CRS.
and 48 volumes of methanol R and dilute to 5 mL with the
same mixture of solvents. TESTS
Reference solution (b). Dilute 1.25 mL of test solution (a) to Related substances. Liquid chromatography (2.2.29).
50 mL with a mixture of 2 volumes of concentrated ammonia R Test solution. Dissolve 25.0 mg of the substance to be
and 48 volumes of methanol R. examined in dilute ammonia R3 and dilute to 25.0 mL with
Reference solution (c). Dissolve 20 mg of the substance to the same solvent.
be examined and 20 mg of sulfamerazine CRS in 3 mL of
Reference solution (a). Dilute 1.0 mL of the test solution to
a mixture of 2 volumes of concentrated ammonia R and
100.0 mL with dilute ammonia R3.
48 volumes of methanol R and dilute to 5 mL with the same
mixture of solvents. Reference solution (b). Dissolve 1.0 mg of sulfasalazine
Apply to the plate 5 μL of each solution. Develop over a derivative for resolution CRS in 10.0 mL of reference
path corresponding to two-thirds of the plate height using solution (a). Dilute 1.0 mL of this solution to 10.0 mL with
a mixture of 3 volumes of dilute ammonia R1, 5 volumes of reference solution (a).
water R, 40 volumes of nitromethane R and 50 volumes of Column :
dioxan R. Dry the plate at 100 °C to 105 °C and examine in – size : l = 0.25 m, Ø = 4.6 mm ;
ultraviolet light at 254 nm. Any spot in the chromatogram – stationary phase : octadecylsilyl silica gel for
obtained with test solution (b), apart from the principal chromatography R (5 μm).
spot, is not more intense than the spot in the chromatogram
obtained with reference solution (b) (0.5 per cent). The test Mobile phase :
is not valid unless the chromatogram obtained with reference – mobile phase A : in a 1000 mL volumetric flask dissolve
solution (c) shows two clearly separated principal spots. 1.13 g of sodium dihydrogen phosphate R and 2.5 g of
sodium acetate R in 900 mL of water R ; adjust to pH 4.8
Loss on drying (2.2.32). Not more than 0.5 per cent,
with glacial acetic acid R and dilute to 1000 mL with
determined on 1.000 g by drying in an oven at 105 °C.
water R ;
Sulfated ash (2.4.14). Not more than 0.1 per cent, determined – mobile phase B : mobile phase A, methanol R (10:40 V/V);
on 1.0 g.
Time Mobile phase A Mobile phase B
ASSAY (min) (per cent V/V) (per cent V/V)
Carry out the determination of primary aromatic 0 - 15 60 → 45 40 → 55
amino-nitrogen (2.5.8), using 0.140 g and determining the
15 - 25 45 55
end-point electrometrically.
1 mL of 0.1 M sodium nitrite is equivalent to 17.22 mg of 25 - 60 45 → 0 55 → 100
C6H8N2O2S. 60 - 65 0 100
STORAGE
Flow rate : 1 mL/min.
Store protected from light.
Detection : spectrophotometer at 320 nm.
01/2017:0863 Injection : 20 μL.
corrected 9.2 Relative retention with reference to sulfasalazine :
impurity H = about 0.16 ; impurity I = about 0.28 ;
impurity C = about 0.80 ; impurity F = about 0.85 ;
impurity G = about 1.39 ; impurity E = about 1.63 ;
impurity B = about 1.85 ; impurity D = about 1.90 ;
SULFASALAZINE impurity A = about 2.00.
System suitability : reference solution (b) :
Sulfasalazinum – resolution : minimum 3.0 between the peaks due to
sulfasalazine and sulfasalazine derivative for resolution.
Limits :
– impurities A, B, C, D, E, F, G, I : for each impurity, not more
than the area of the principal peak in the chromatogram
obtained with reference solution (a) (1 per cent);
– total : not more than 4 times the area of the principal peak
in the chromatogram obtained with reference solution (a)
C18H14N4O5S Mr 398.4 (4 per cent) ;
[599-79-1] – disregard limit : 0.05 times the area of the principal peak
in the chromatogram obtained with reference solution (a)
DEFINITION (0.05 per cent) ; disregard any peak with a retention time
2-Hydroxy-5-[2-[4-(pyridin-2-ylsulfamoyl)phenyl]diazenyl]- less than 6 min (due to impurities H and J).
benzoic acid. Impurities H and J. Liquid chromatography (2.2.29).
Content : 97.0 per cent to 101.5 per cent (dried substance). Test solution. Dissolve 25.0 mg of the substance to be
CHARACTERS examined in dilute ammonia R3 and dilute to 25.0 mL with
Appearance : bright yellow or brownish-yellow, fine powder. the same solvent.
Solubility : practically insoluble in water, very slightly soluble Reference solution (a). Dissolve 5.0 mg of salicylic acid R
in ethanol (96 per cent), practically insoluble in methylene (impurity H) and 5.0 mg of sulfapyridine CRS (impurity J)
chloride. It dissolves in dilute solutions of alkali hydroxides. in dilute ammonia R3 and dilute to 10.0 mL with the same
solvent.
IDENTIFICATION Reference solution (b). Dilute 2.0 mL of reference solution (a)
Infrared absorption spectrophotometry (2.2.24). to 100.0 mL with dilute ammonia R3.
General Notices (1) apply to all monographs and other texts 3931
