Pharmeuropa 25.

31 July 2013

147

Reference: PNPH/Exp. P4/T (11) 36 ANP

XXXX:2631

ROSUVASTATIN CALCIUM

Rosuvastatinum calcicum
F

C44H54CaF 2N6012S2
[14 7098-20-2]

M, 1001

DEFINITION
Ca lei um bis[ (3R ,5 S, 6E)-7 -[4-( 4-fl uorophenyl )-2-[ methyl( methylsu lfonyl )ami no ]-6-(propan-2yl)pyrimidin-5-yl]-3,5-dihydroxyhept-6-enoate].
Content: 97.0 per cent to 102.0 per cent (anhydrous substance).

CHARACTERS
Appearance: white or almost white, hygroscopic powder.
Solubility: slightly soluble in water, freely soluble in methylene chloride, practically insoluble in
anhydrous ethanol.

IDENTIFICATION

A. Infrared absorption spectrophotometry (2.2.24).

Comparison: rosuvastatin calcium CRS.

B. Enantiomeric purity (see Tests).

Results: the principal peak in the chromatogram obtained with the test solution is similar in
retention time to the principal peak in the chromatogram obtained with reference solution (c).

C. It gives reaction (b) of calcium (2.3.1).

TESTS
Related substances. Liquid chromatography (2.2.29). Carry out the test protected from light.
Test solution. Dissolve 35.0 mg of the substance to be examined in 12 ml of acetonitrile for
chromatography Rand dilute to 50.0 ml with water for chromatography R.
Reference solution (a). Dissolve 35.0 mg of rosuvastatin calcium CRS in 12 ml of acetonitrile for
chromatography Rand dilute to 50.0 ml with water for chromatography R.
Reference solution (b). To 1.0 ml of the test solution add 24 ml of acetonitrile for
chromatography Rand dilute to 100.0 ml with water for chromatography R. To 2.0 ml of this
solution add 2 ml of acetonitrile for chromatography Rand dilute to 10.0 ml with water for
chromatography R.
Reference solution (c). In order to prepare impurity Bin situ, dissolve 10 mg of the substance
to be examined in 10 ml of a 1 per cent \IN solution of trifluoroacetic acid R in acetonitrile
for chromatography R. Stopper and heat at 40 oc for 1 h. Cool, add 20 ml of water for
chromatography Rand adjust to pH 6-8 with a 42 g/L solution of sodium hydroxide R. Dilute
to 50 ml with water for chromatography R.
Reference solution (a). Dissolve 5 mg of rosuvastatin impurity A CRS in 10 ml of acetonitrile for
chromatography Rand dilute to 20.0 ml with water for chromatography R.

PAIPHIExp. P41T (11) 36 ANP

 Pharmeuropa 25.3 I July 2013

148

Reference solution (e). In order to prepare impurity C in situ, heat 0.25 g of the substance to be
examined at 50 oc for 7 days in amber glassware with a porous coveri57J. Dissolve 50 mg in 11 ml
of acetonitrile for chromatography R, add 3.0 ml of reference solution (c) and 1.0 ml of reference
solution (d). Dilute to 50.0 ml with water for chromatography R.
Column:
- size: I= 0.15 m, 0 = 3.0 mm;
- stationary phase: base-deactivated end-capped octadecylsilyl silica gel for chromatography R
(3 1Jm)I5BJ;
- temperature: 40

oc.

Mobile phase:
- mobile phase A: 1 per cent \IN solution of trifluoroacetic acid R, acetonitrile for
chromatography R, water for chromatography R (1 :29:70 \I!\.IN);
- mobile phase 8: 1 per cent \IN solution of trifluoroacetic acid R, water for chromatography R,
acetonitrile for chromatography R (1 :24:75 \I!\.IN);
Time

Mobile phase A

Mobile phase B
(per cent 11/V)

(min)

(per cent 11/V)

0-30

100

30-50

100--+ 60

0--+ 40

50-60

60--+ 0

40--+ 100

60- 75

100

Flow rate: 0.75 ml/min.

Detection: spectrophotometer at 242 nm.
lr,jection: 10 iJL of the test solution and reference solutions (b) and (e).
Identification of impurities: use the chromatogram obtained with reference solution (e) to identify
the peaks due to impurities A, B and C.
Relative retention with reference to rosuvastatin (retention time =about 25 min):
impurity A = about 0.9; impurity B = about 1.1 ; impurity C = about 1.5.
System suitability: reference solution (e):
- resolution: minimum 2.0 between the peaks due to rosuvastatin and impurity B.
Calculation of percentage contents:
- correction factor: multiply the peak area of impurity C by 2.1 ;
- for each impurity, use the concentration of rosuvastatin in reference solution (b).
Limits:
- impurity C: maximum 0.6 per cent;
- impurity 8: maximum 0.5 per cent;
- impurity A: maximum 0.2 per cent;
- unspecified impurities: for each impurity, maximum 0.10 per cent;
- total: maximum 1.2 per cent;
- reporting threshold: 0.05 per cent.
(57) Place a small vial containing the substance inside a small beaker and then cover the beaker with filter paper secured
with an elastic band.
(58) lnertsil ODS-3 is suitable.

PAIPHIExp. P41T (11) 36 ANP

 Pharmeuropa 25.31 July 2013

149

The following chromatogram is shown for information but will not be published in the European
Pharmacopoeia.
2

10

20

30

40

50

1. impurity A

3. impurity B

5. impurity E

2. rosuvastatin

4. impurity C

6. impurity J

ro

7. impurity F

Figure 2631.-1. -Chromatogram for the test for related substances of rosuvastatin calcium:
reference solution (e) spiked with impurities E, F and J
Enantiomeric purity. Liquid chromatography (2.2.29). Prepare the solutions immediately before
use and protect them from light throughout the test.
Solvent mixture: acetonitrile for chromatography R, water for chromatography R (25:75 11/V).
Test solution. Dissolve 25.0 mg of the substance to be examined in 6 ml of acetonitrile for
chromatography Rand dilute to 25.0 ml with water for chromatography R.
Reference solution (a). Dilute 1.0 ml of the test solution to 100.0 ml with the solvent mixture.
Dilute 1.0 ml of this solution to 10.0 ml with the solvent mixture.
Reference solution (b). Dissolve 5 mg of rosuvastatin impurity G CRS in a mixture of 12 ml
of acetonitrile for chromatography Rand 10 ml of water for chromatography R with the aid of
ultrasound and dilute to 50.0 ml with water for chromatography R.
Reference solution (c). To 25 mg of the substance to be examined add 1.0 ml of reference
solution (b), 6 ml of acetonitrile for chromatography Rand dissolve with the aid of ultrasound;
dilute to 25 ml with water for chromatography R.
Column:
- size: I= 0.15 m, 0

=4.6 mm;

- stationary phase: silica gel OJ for chiral separations R (5 1Jm)I59J;

- temperature: 35

oc.

Mobile phase: acetonitrile for chromatography R, 0.1 per cent \IN solution of trifluoroacetic
acid R (25:75 11/V).
Flow rate: 0.5 ml/min.
(59) Chiralcel OJ-RH is suitable.

PA/PH/Exp. P4/T (11) 36 ANP

oo

m~

 Pharmeuropa 25.3 I July 2013

150

Detection: spectrophotometer at 254 nm.

lr,jection: 10 iJL of the test solution and reference solutions (a) and (c).
Run time: 3 times the retention time of rosuvastatin.
Identification of impurities: use the chromatogram supplied with rosuvastatin impurity G CRS and
the chromatogram obtained with reference solution (c) to identify the peak due to impurity G.
Relative retention with reference to rosuvastatin (retention time= about 25 min): impurity G =about
0.9.
System suitability: reference solution (c):
- resolution: minimum 1.5 between the peaks due to impurity G and rosuvastatin.
Calculation of percentage content:
- use the concentration of rosuvastatin in reference solution (a).
Limit:
- impurity G: maximum 0.1 per cent.
The following chromatogram is shown for information but will not be published in the European
Pharmacopoeia.

.A
I

A.l

/"'

10

20

30

40

50

60

70

1. impurity G

2. rosuvastatin

Figure 2631.-2. -Chromatogram for the test for enantiomeric purity of rosuvastatin calcium:
reference solution (c)

Chlorides: maximum 0.2 per cent.

Dissolve 0.15 g in 60 ml of water R by heating to boiling while stirring. Add 4 ml of dilute nitric
acid R, allow to cool to room temperature and titrate with 0.01 M silver nitrate, determining the
end-point potentiometrically (2.2.20) using a silver indicator electrode and a silver-silver chloride
reference electrode.
1 .0 ml of 0. 01 M silver nitrate is equivalent to 0.3545 mg of Cl.

Water (2.5.32): maximum 6.0 per cent, determined on 20.0 mg.

PAIPHIExp. P41T (11) 36 ANP

min

 Pharmeuropa 25.31 July 2013

151

ASSAY
Liquid chromatography (2.2.29) as described in the test for related substances with the following
modification.

lr,jection: test solution and reference solution (a).

Calculate the percentage content of C44 H54 CaF 2 N6 0 12 S 2 taking into account the assigned content
of rosuvastatin calcium CRS.
STORAGE
In an airtight container protected from light at a temperature of 2 octo 8 oc.
IMPURITIES

Specified impurities: A, B, C, 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): 0, E, F, H, I, J.

A. (3R ,5S, 6E)-7 -[ 4-( 4-fl uorophenyl )-2-[[(2-hyd roxy-2-methyl propyl )sulfonyl]( methyl )ami no ]-6(propan-2-yl )pyri midi n-5-yl]-3, 5-d ihyd roxyhept -6-enoic acid,

and enantiomer

B. (3RS,5RS,6E)-7 -[ 4-( 4-fluorophenyl)-2-[methyl( methylsulfonyl)amino ]-6-(propan-2yl )pyri midi n-5-yl]-3, 5-d ihyd roxyhept -6-eno ic acid,

C. (3R ,6E)-7 -[ 4-( 4-fl uorophenyl )-2-[ methyl( methylsu lfonyl )ami no ]-6-(pro pan-2-yl )pyri midi n-5-yl]3-hydroxy-5-oxohept-6-enoic acid,

PAIPHIExp. P41T (11) 36 ANP

 Pharmeuropa 25.3 I July 2013

152

D. N-[4-(4-fluorophenyi)-5-[(E)-2-[(2S,4R)-4-hydroxy-6-oxotetrahydro-2H-pyran-2-yl]ethenyl]-6(propan-2-yl)pyrimidin-2-yi]-N-methylmethanesulfonamide,
F

E. (3R,5S,6E)-7 -[ 4-( 4-fluorophenyl)-2-[([2-[4-( 4-fluorophenyl)-2-[methyl( methylsulfonyl)amino ]-6(propan-2-yl)pyrimidin-5-yl]-2-hydroxyethyl]sulfonyl)(methyl)amino]-6-(propan-2-yl)pyrimidin5-yl]-3,5-dihydroxyhept-6-enoic acid,

F

F. 1 ,1-dimethylethyl [(4R,6S)-6-[(E)-2-[4-(4-fluorophenyl)-2-[methyl(methylsulfonyl)amino]-6(propan-2-yl)pyrimidin-5-yl]ethenyl]-2,2-dimethyl-1 ,3-dioxan-4-yl]acetate,

F

H
OH

G. (3S,5R,6E)-7 -[ 4-( 4-fluorophenyl)-2-[methyl( methylsulfonyl)amino ]-6-(propan-2-yl)pyrimid in-5yl]-3, 5-d ihyd roxyhept-6-eno ic acid,
F

OH
H
and epimer at

H. (3R ,5R S)-5-[8-fl uoro-2-[ methyl( methylsu lfonyl )ami no ]-4-(propan-2-yl )-5, 6dihydrobenzo[h]quinazolin-6-yl]-3,5-dihydroxypentanoic acid,

PA/PH/Exp. P4/T (11) 36 ANP

 Pharmeuropa 25.31 July 2013

153

OH
and epi mer at

I. (3S,5RS)-5-[8-fluoro-2-[methyl(methylsulfonyl)amino]-4-(propan-2-yl)-5,6dihydrobenzo[h]quinazolin-6-yl]-3,5-dihydroxypentanoic acid,
F

J. (3R,5S,6E)-7 -[ 4-( 4-fluorophenyi)-2-[([(E)-2-[4-( 4-fluorophenyl)-2-[methyl( methylsulfonyl)amino]-6-(propan-2-yl)pyrimidin-5-yl]ethenyl]sulfonyl)(methyl)amino]-6-(propan-2-yl)pyrim id in-5-yl]-3, 5-d ihyd roxyhept -6-e noic acid.

PA/PH/Exp. P4/T (11) 36 ANP