Cefaclor

EUROPEAN PHARMACOPOEIA 5.0

CHARACTERS
Appearance : clear, almost colourless or slightly yellow,
viscous, hygroscopic liquid.
Solubility : slightly soluble in light petroleum, miscible with
alcohol and with glacial acetic acid.
Relative density : about 0.958.
Refractive index : about 1.479.
IDENTIFICATION
First identification : D.
Second identification : A, B, C.
A. It complies with the test for optical rotation (see Tests).
B. It complies with the test for hydroxyl value (see Tests).
C. Iodine value (2.5.4) : 82 to 90.
D. It complies with the test for composition of fatty acids
(see Tests).
TESTS
Optical rotation (2.2.7) : + 3.5 to + 6.0.
Specific absorbance (2.2.25) : maximum 1.0, determined at
the absorption maximum at 269 nm 1 nm.
To 1.0 g add alcohol R and dilute to 100.0 ml with the same
solvent.
Acid value (2.5.1) : maximum 2.0.
Dissolve 5.0 g in 25 ml of the prescribed mixture of solvents.
Hydroxyl value (2.5.3, Method A) : minimum 150.
Peroxide value (2.5.5) : maximum 10.0.
Unsaponifiable matter (2.5.7) : maximum 0.8 per cent,
determined on 5.0 g.
Composition of fatty acids. Gas chromatography (2.4.22)
with the following modifications.
Use the mixture of calibrating substances in Table 2.4.22.-3.
Test solution. Introduce 75 mg of the substance to be
examined into a 10 ml centrifuge tube with a screw cap.
Dissolve in 2 ml of 1,1-dimethylethyl methyl ether R1
with shaking and heat gently (50-60 C). Add, when still
warm, 1 ml of a 12 g/l solution of sodium R in anhydrous
methanol R, prepared with the necessary precautions, and
mix vigorously for at least 5 min. Add 5 ml of distilled
water R and mix vigorously for about 30 s. Centrifuge for
15 min at 1500 g. Use the upper layer.
Reference solution. Dissolve 50 mg of methyl
ricinoleate CRS and 50 mg of methyl stearate CRS in
10.0 ml of 1,1-dimethylethyl methyl ether R1.
Column :
material : fused silica,
size : l = 30 m, = 0.25 mm,
stationary phase: macrogol 20 000 R (film thickness
0.25 m).
Carrier gas : helium for chromatography R.
Flow rate : 0.9 ml/min.
Split ratio : 1:100.
Temperature :

Column

Time
(min)

Temperature
(C)

0 - 55

215

Injection port

250

Detector

250

Detection : flame ionisation.

Injection : 1 l.
1198

Calculate the percentage content of each fatty acid by

normalisation.
Correct the area of the peak due to methyl ricinoleate,
multiplying by a factor R calculated using the following
expression :

m1

mass of methyl ricinoleate in the reference

solution,

m2

mass of methyl stearate in the reference solution,

A1

area of the peak due to methyl ricinoleate in

the chromatogram obtained with the reference
solution,

A2

area of the peak due to methyl stearate in the

chromatogram obtained with the reference
solution.

Composition of the fatty-acid fraction of the oil :

palmitic acid : maximum 2.0 per cent,
stearic acid : maximum 2.5 per cent,
oleic acid and isomers (C18:1 equivalent chain length on
macrogol 20 000 : 18.3) : 2.5 per cent to 6.0 per cent,
linoleic acid (C18:2 equivalent chain length on macrogol
20 000 : 18.8) : 2.5 per cent to 7.0 per cent,
linolenic acid (C18:3 equivalent chain length on macrogol
20 000 : 19.2) : maximum 1.0 per cent,
eicosenoic acid (C20:1 equivalent chain length on
macrogol 20 000 : 20.2) : maximum 1.0 per cent,
ricinoleic acid (equivalent chain length on macrogol
20 000 : 23.9) : 85.0 per cent to 92.0 per cent,
any other fatty acid : maximum 1.0 per cent.
Water (2.5.12) : maximum 0.3 per cent, determined on 5.0 g.
STORAGE
In an airtight, well-filled container, protected from light.
LABELLING
The label states the name and concentration of any added
antioxidant.
01/2005:0986
corrected

CEFACLOR
Cefaclorum

C15H14ClN3O4S,H2O

Mr 385.8

DEFINITION
Cefaclor is the monohydrate of (6R,7R)-7-[[(2R)-2amino-2-phenylacetyl]amino]-3-chloro-8-oxo-5-thia-1azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid. It contains not
See the information section on general monographs (cover pages)

Cefaclor

EUROPEAN PHARMACOPOEIA 5.0

less than 96.0 per cent and not more than the equivalent of
102.0 per cent of C15H14ClN3O4S, calculated with reference
to the anhydrous substance.
CHARACTERS
A white or slightly yellow powder, slightly soluble in water,
practically insoluble in methanol and in methylene chloride.
IDENTIFICATION
Examine by infrared absorption spectrophotometry (2.2.24),
comparing with the spectrum obtained with cefaclor CRS.
TESTS
pH (2.2.3). Suspend 0.250 g in carbon dioxide-free water R
and dilute to 10 ml with the same solvent. The pH of the
suspension is 3.0 to 4.5.
Specific optical rotation (2.2.7). Dissolve 0.250 g in a 10 g/l
solution of hydrochloric acid R and dilute to 25.0 ml with
the same solution. The specific optical rotation is + 101 to
+ 111, calculated with reference to the anhydrous substance.
Related substances. Examine by liquid chromatography
(2.2.29).
Test solution. Dissolve 50.0 mg of the substance to be
examined in 10.0 ml of a 2.7 g/l solution of sodium
dihydrogen phosphate R adjusted to pH 2.5 with phosphoric
acid R.
Reference solution (a). Dissolve 2.5 mg of cefaclor CRS
and 5.0 mg of delta-3-cefaclor CRS in 100.0 ml of a 2.7 g/l
solution of sodium dihydrogen phosphate R adjusted to
pH 2.5 with phosphoric acid R.
Reference solution (b). Dilute 1.0 ml of the test solution
to 100.0 ml with a 2.7 g/l solution of sodium dihydrogen
phosphate R adjusted to pH 2.5 with phosphoric acid R.
The chromatographic procedure may be carried out using :
a stainless steel column 0.25 m long and 4.6 mm in
internal diameter packed with end-capped octadecylsilyl
silica gel for chromatography R (5 m),
as mobile phase at a flow rate of 1.0 ml/min :
Mobile phase A. Adjust a 7.8 g/l solution of sodium
dihydrogen phosphate R to pH 4.0 with phosphoric
acid R,
Mobile phase B. Mix 450 ml of acetonitrile R with 550 ml
of mobile phase A,
as detector a spectrophotometer set at 220 nm,
a 20 l loop injector.
Equilibrate the column with a mixture of 5 volumes of
mobile phase B and 95 volumes of mobile phase A for at least
15 min between each analysis. Inject the solutions. Operate
by gradient elution increasing the concentration of mobile
phase B continuously and linearly by 0.67 per cent V/V per
minute for 30 min (25 per cent V/V). Then increase the
concentration of mobile phase B continuously and linearly
by 5 per cent V/V per minute for 15 min (100 per cent V/V).
Finally elute with mobile phase B for 10 min. Change the
composition to a mixture of 5 volumes of mobile phase B and
95 volumes of mobile phase A to re-equilibrate the column.
Inject reference solution (a). The test is not valid unless
the resolution between the peaks due to cefaclor and
delta-3-cefaclor is at least 2 and the symmetry factor of
the cefaclor peak is at most 1.2. If necessary, adjust the
acetonitrile content of the mobile phase.
General Notices (1) apply to all monographs and other texts

Inject the test solution and reference solution (b). In the

chromatogram obtained with the test solution : the area of
any peak, apart from the principal peak and any peaks due
to the mobile phase, is not greater than 0.5 times the area
of the principal peak in the chromatogram obtained with
reference solution (b) (0.5 per cent) ; and the sum of all such
peaks is not more than twice the area of the principal peak
in the chromatogram obtained with reference solution (b)
(2 per cent). Disregard any peak with an area less than
0.1 times that of the principal peak in the chromatogram
obtained with reference solution (b).
Heavy metals (2.4.8). 1.0 g complies with limit test C
(30 ppm). Prepare the standard using 3 ml of lead standard
solution (10 ppm Pb) R.
Water (2.5.12). 3.0 per cent to 6.5 per cent, determined on
0.200 g by the semi-micro determination of water.
ASSAY
Examine by liquid chromatography (2.2.29).
Test solution. Dissolve 15.0 mg of the substance to be
examined in the mobile phase and dilute to 50.0 ml with the
mobile phase.
Reference solution (a). Dissolve 15.0 mg of cefaclor CRS in
the mobile phase and dilute to 50.0 ml with the mobile phase.
Reference solution (b). Dissolve 15.0 mg of cefaclor CRS
and 15.0 mg of delta-3-cefaclor CRS in the mobile phase and
dilute to 50.0 ml with the mobile phase.
The chromatographic procedure may be carried out using :
a stainless steel column 0.25 m long and 4.6 mm in
internal diameter packed with octadecylsilyl silica gel for
chromatography R (5 m),
as mobile phase at a flow rate of 1.5 ml/min a mixture
prepared by adding 220 ml of methanol R to a mixture of
780 ml of water R, 10 ml of triethylamine R and 1 g of
sodium pentanesulphonate R, adjusted to pH 2.5 with
phosphoric acid R,
as detector a spectrophotometer set at 265 nm,
a 20 l loop injector.
Inject reference solution (b). The assay is not valid unless
the resolution between the peaks due to cefaclor and
delta-3-cefaclor is at least 2.5. Adjust the concentration of
methanol in the mobile phase, if necessary. The assay is not
valid unless the symmetry factor of the cefaclor peak is at
most 1.5. Inject reference solution (a) six times. The assay is
not valid unless the relative standard deviation of the peak
area of cefaclor is at most 1.0 per cent. Inject alternately the
test solution and reference solution (a).
IMPURITIES

A. (2R)-2-amino-2-phenylacetic acid (phenylglycine),

B. (6R,7R)-7-amino-3-chloro-8-oxo-5-thia-1azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid,
1199

Cefadroxil monohydrate

EUROPEAN PHARMACOPOEIA 5.0

01/2005:0813
corrected

CEFADROXIL MONOHYDRATE
Cefadroxilum monohydricum
C. (6R,7R)-7-[[(2S)-2-amino-2-phenylacetyl]amino]-3-chloro-8oxo-5-thia- 1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid,

C16H17N3O5S,H2O

Mr 381.4

DEFINITION
(6R,7R)-7-[[(2R)-2-Amino-2-(4-hydroxyphenyl)acetyl]amino]-3methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic
acid monohydrate.
Content : 95.0 per cent to 102.0 per cent (anhydrous
substance).

D. (2R,6R,7R)- and (2S,6R,7R)-7-[[(2R)-2-amino2-phenylacetyl]amino]-3-chloro-8-oxo-5-thia1-azabicyclo[4.2.0]oct-3-ene-2-carboxylic acid

(delta-3-cefaclor),

CHARACTERS
Appearance : white or almost white powder.
Solubility : slightly soluble in water, very slightly soluble
in alcohol.
IDENTIFICATION
E. 2-[[(2R)-2-amino-2-phenylacetyl]amino]-2-(5-chloro-4-oxo-3,
Infrared absorption spectrophotometry (2.2.24).
4-dihydro-2H-1,3-thiazin-2-yl)acetic acid,

F. 3-phenylpyrazin-2-ol.

G. (2R,6R,7R)- and (2S,6R,7R)-7-[[(2R)-2-amino-2phenylacetyl]amino]-3-methylene-8-oxo-5-thia-1azabicyclo[4.2.0]octane-2-carboxylic acid (isocefalexine),

H. (6R,7R)-7-[[(2R)-2-[[(2R)-2-amino-2-phenylacetyl]amino]2-phenylacetyl]amino]-3-chloro-8-oxo-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid
(N-phenylglycyl cefaclor).
1200

TESTS
pH (2.2.3) : 4.0 to 6.0.
Suspend 1.0 g in carbon dioxide-free water R and dilute to
20 ml with the same solvent.
Specific optical rotation (2.2.7) : + 165 to + 178 (anhydrous
substance).
Dissolve 0.500 g in water R and dilute to 50.0 ml with the
same solvent.
Absorbance (2.2.25). Dissolve 20.0 mg in phosphate
buffer solution pH 6.0 R and dilute to 100.0 ml with the
same solvent. The absorbance of the solution determined
at 330 nm is not greater than 0.05. Dilute 10.0 ml of
the solution to 100.0 ml with phosphate buffer solution
pH 6.0 R. Examined between 235 nm and 340 nm, the
diluted solution shows an absorption maximum at 264 nm.
The specific absorbance at this maximum is 225 to 250
(anhydrous substance).
Related substances. Liquid chromatography (2.2.29).
Test solution. Dissolve 50.0 mg of the substance to be
examined in mobile phase A and dilute to 50.0 ml with
mobile phase A.
Reference solution (a). Dissolve 10.0 mg of
D--(4-hydroxyphenyl)glycine CRS (impurity A) in
mobile phase A and dilute to 10.0 ml with mobile phase A.
Reference solution (b). Dissolve 10.0 mg of
7-aminodesacetoxycephalosporanic acid CRS
(impurity B) in phosphate buffer solution pH 7.0 R5 and
dilute to 10.0 ml with the same buffer solution.
Reference solution (c). Dilute 1.0 ml of reference solution (a)
and 1.0 ml of reference solution (b) to 100.0 ml with mobile
phase A.

See the information section on general monographs (cover pages)