Meloxicam EUROPEAN PHARMACOPOEIA 9.

– CID gas pressure : 2.7 × 10-3 mbar.

– MRM mode parameters :
Substance MRM channels Collision A. N,N-dimethylmethanaminium,
energy (eV) for the
interval 0-25 min
Meldonium 147.00 → 59.00 18.0

Impurity A 59.97 → 44.98 18.0

Impurity B 74.99 → 59.04 15.0
B. 1,1,1-trimethylhydrazin-1-ium,

Impurity F 115.19 → 71.92 19.0

Impurity C 161.19 → 59.00 23.0

Impurity D 175.19 → 58.07 23.0 C. 2-(3-methoxy-3-oxopropyl)-1,1,1-trimethylhydrazin-1-
Impurity E 189.26 → 58.01 22.0 ium,

Injection : 20 μL of the test solution and reference solutions (b)

and (c).
Relative retention with reference to meldonium
(retention time = about 9 min) : impurity F = about 0.7 ; D. 2-(3-ethoxy-3-oxopropyl)-1,1,1-trimethylhydrazin-1-ium,
impurity B = about 0.8 ; impurity A = about 0.9 ;
impurity C = about 1.5 ; impurity D = about 1.8 ;
impurity E = about 2.0.
System suitability : reference solution (b) :
E. 1,1,1-trimethyl-2-[3-(1-methylethoxy)-3-
– signal-to-noise ratio : impurity A = minimum 10 ; oxopropyl]hydrazin-1-ium,
impurity B = minimum 50 ; impurity C = minimum 200 ;
impurity D = minimum 1000 ; impurity E = minimum 1000 ;
impurity F = minimum 50 ;
– repeatability : maximum relative standard deviation of
10 per cent after 6 injections. F. 1,1-dimethyl-4,5-dihydro-1H-pyrazol-1-ium-3-olate.
Calculation of percentage contents :
– for impurities A, B, C, D, E and F (MRM mode), use the 01/2017:2373
concentration of the corresponding impurity in reference
solution (b) ;
– for impurities other than A, B, C, D, E and F (complete
spectrum mode), use the concentration of meldonium in
reference solution (c). MELOXICAM
Limits :
– impurities A, B, C, D, E, F : for each impurity, maximum
Meloxicamum
0.15 per cent ;
– unspecified impurities : for each impurity, maximum
0.10 per cent ;
– total : maximum 0.3 per cent ;
– reporting threshold : 0.05 per cent.
Chlorides (2.4.4): maximum 100 ppm.
Dilute 5 mL of solution S to 30 mL with distilled water R. C14H13N3O4S2 Mr 351.4
[71125-38-7]
Sulfates (2.4.13) : maximum 100 ppm.
Dilute 10 mL of solution S to 20 mL with distilled water R. DEFINITION
Water (2.5.12) : 19.7 per cent to 21.0 per cent, determined on 4-Hydroxy-2-methyl-N-(5-methylthiazol-2-yl)-2H-1,2-
0.120 g. benzothiazine-3-carboxamide 1,1-dioxide.
Content : 99.0 per cent to 101.0 per cent (dried substance).
Sulfated ash (2.4.14) : maximum 0.1 per cent, determined on
1.0 g. CHARACTERS
Appearance : pale yellow powder.
ASSAY
Solubility : practically insoluble in water, soluble in
Dissolve 0.100 g in 40 mL of glacial acetic acid R. Titrate dimethylformamide, very slightly soluble in ethanol (96 per
with 0.1 M perchloric acid, determining the end-point cent).
potentiometrically (2.2.20). It shows polymorphism (5.9).
1 mL of 0.1 M perchloric acid is equivalent to 14.62 mg of
C6H14N2O2. IDENTIFICATION
Infrared absorption spectrophotometry (2.2.24).
STORAGE Comparison : meloxicam CRS.
In an airtight container. If the spectra obtained in the solid state show differences,
dissolve the substance to be examined and the reference
IMPURITIES substance separately in acetone R, evaporate to dryness and
Specified impurities : A, B, C, D, E, F. record new spectra using the residues.

2994 See the information section on general monographs (cover pages)

EUROPEAN PHARMACOPOEIA 9.0 Meloxicam

TESTS – disregard limit : 0.3 times the area of the principal peak in
Related substances. Liquid chromatography (2.2.29). the chromatogram obtained with reference solution (a) at
the same wavelength (0.03 per cent).
Test solution. Dissolve 40 mg of the substance to be examined
in a mixture of 5 mL of methanol R and 0.3 mL of 1 M sodium Loss on drying (2.2.32) : maximum 0.5 per cent, determined
hydroxide and dilute to 20.0 mL with methanol R. on 1.000 g by drying in an oven at 105 °C for 4 h.
Reference solution (a). Dilute 2.0 mL of the test solution to Sulfated ash (2.4.14): maximum 0.1 per cent, determined on
100.0 mL with methanol R. Dilute 5.0 mL of this solution to 1.0 g.
100.0 mL with methanol R.
Reference solution (b). Dissolve 2 mg of the substance to ASSAY
be examined, 2 mg of meloxicam impurity A CRS, 2 mg of In order to avoid overheating during the titration, mix
meloxicam impurity B CRS, 2 mg of meloxicam impurity C CRS thoroughly throughout and stop the titration immediately after
and 2 mg of meloxicam impurity D CRS in a mixture of 5 mL the end-point has been reached.
of methanol R and 0.3 mL of 1 M sodium hydroxide and dilute Dissolve 0.250 g in a mixture of 5 mL of anhydrous formic
to 25 mL with methanol R. If complete dissolution cannot acid R and 50 mL of anhydrous acetic acid R. Titrate with 0.1 M
be achieved, filter the solution through a membrane filter perchloric acid, determining the end-point potentiometrically
(nominal pore size 0.45 μm) before injection. (2.2.20).
Column : 1 mL of 0.1 M perchloric acid is equivalent to 35.14 mg
– size : l = 0.15 m, Ø = 4.6 mm ; of C14H13N3O4S2.
– stationary phase : end-capped octadecylsilyl silica gel for STORAGE
chromatography R (5 μm) ;
Protected from light.
– temperature : 45 °C.
Mobile phase : IMPURITIES
– mobile phase A : 1 g/L solution of potassium dihydrogen Specified impurities : A, B, C, D.
phosphate R adjusted to pH 6.0 with 1 M sodium hydroxide ; Other detectable impurities (the following substances would,
if present at a sufficient level, be detected by one or other of
– mobile phase B : methanol R ;
the tests in the monograph. They are limited by the general
Time Mobile phase A Mobile phase B acceptance criterion for other/unspecified impurities and/or
(min) (per cent V/V) (per cent V/V) by the general monograph Substances for pharmaceutical use
0-2 60 40 (2034). It is therefore not necessary to identify these impurities
for demonstration of compliance. See also 5.10. Control of
2 - 10 60 → 30 40 → 70 impurities in substances for pharmaceutical use) : E, F.
10 - 15 30 70

Flow rate : 1.0 mL/min.

Detection : spectrophotometer at 260 nm and 350 nm.
Injection : 10 μL.
Relative retention with reference to meloxicam
(retention time = about 7 min): impurity B = about 0.5 ; A. ethyl 4-hydroxy-2-methyl-2H-1,2-benzothiazine-3-
impurity A = about 1.4 ; impurity C = about 1.7 ; carboxylate 1,1-dioxide,
impurity D = about 1.9.
System suitability : reference solution (b) :
– resolution : minimum 3.0 between the peaks due to
meloxicam and impurity A at 350 nm ; minimum 3.0
between the peaks due to impurity B and meloxicam at
260 nm. B. 5-methylthiazol-2-amine,
Limits :
– correction factor : for the calculation of content, multiply
the peak area of impurity A by 2.0 ;
– impurity A at 350 nm : not more than the area of the
principal peak in the chromatogram obtained with
reference solution (a) at 350 nm (0.1 per cent) ;
– impurity B at 260 nm : not more than the area of the
principal peak in the chromatogram obtained with C. N-[(2Z)-3,5-dimethylthiazol-2(3H)-ylidene]-4-hydroxy-2-
reference solution (a) at 350 nm (0.1 per cent) ; methyl-2H-1,2-benzothiazine-3-carboxamide 1,1-dioxide,
– impurities C, D at 350 nm : for each impurity, not more
than 0.5 times the area of the principal peak in the
chromatogram obtained with reference solution (a) at
350 nm (0.05 per cent) ;
– unspecified impurities : for each impurity, at the wavelength
giving the higher value for the impurity, not more than the
area of the principal peak in the chromatogram obtained
with reference solution (a) at the same wavelength (0.10 per D. N-[(2Z)-3-ethyl-5-methylthiazol-2(3H)-ylidene]-4-
cent) ; hydroxy-2-methyl-2H-1,2-benzothiazine-3-carboxamide
– total : not more than 0.3 per cent ; 1,1-dioxide,

General Notices (1) apply to all monographs and other texts 2995
Melphalan EUROPEAN PHARMACOPOEIA 9.0

Reference solution (b). Dilute 10.0 mL of test solution (a) to

100.0 mL with methanol R1.
Reference solution (c). Dilute 1.0 mL of reference solution (b)
to 100.0 mL with methanol R1.
Reference solution (d). Dilute 5.0 mL of reference solution (b)
E. methyl 4-hydroxy-2-methyl-2H-1,2-benzothiazine-3- to 100.0 mL with methanol R1.
carboxylate 1,1-dioxide, Reference solution (e). In order to prepare impurity I in
situ, dissolve 5 mg of melphalan for system suitability CRS
(containing impurities B, D, G, H and J) in methanol R1, dilute
to 5.0 mL with the same solvent and heat at 60 °C for 15 min.
Column :
– size : l = 0.15 m, Ø = 4.6 mm ;
– stationary phase : base-deactivated end-capped octadecylsilyl
F. isopropyl 4-hydroxy-2-methyl-2H-1,2-benzothiazine-3- silica gel for chromatography R (5 μm).
carboxylate 1,1-dioxide. Mobile phase :
– mobile phase A : mixture of 5 volumes of acetonitrile for
07/2012:1698
chromatography R and 95 volumes of water R containing
0.01 per cent V/V of triethylamine R, 0.05 per cent m/m
of ammonium acetate R and 0.05 per cent V/V of glacial
acetic acid R ;
– mobile phase B : mixture of 40 volumes of water R
MELPHALAN containing 0.01 per cent V/V of triethylamine R, 0.05 per
cent m/m of ammonium acetate R and 0.05 per cent V/V
Melphalanum of glacial acetic acid R, and 60 volumes of acetonitrile for
chromatography R ;
Time Mobile phase A Mobile phase B
(min) (per cent V/V) (per cent V/V)
0 - 20 100 → 0 0 → 100
20 - 25 0 100

C13H18Cl2N2O2 Mr 305.2 Flow rate : 1.5 mL/min.

[148-82-3]
DEFINITION
4-[Bis(2-chloroethyl)amino]-L-phenylalanine.
Content : 94.0 per cent to 102.0 per cent (anhydrous and
diethylamine-free substance).
CHARACTERS
Appearance : white or almost white, hygroscopic powder.
Solubility : practically insoluble in water, slightly soluble in
methanol. It dissolves in dilute mineral acids.
IDENTIFICATION
Infrared absorption spectrophotometry (2.2.24).
Comparison : Ph. Eur. reference spectrum of melphalan.
TESTS
Appearance of solution. If intended for use in the
manufacture of parenteral preparations, the solution is clear
(2.2.1) and colourless (2.2.2, Method II).
Dissolve 0.25 g in dilute hydrochloric acid R and dilute to
25 mL with the same acid.
Specific optical rotation (2.2.7) : − 36.0 to − 30.0 (anhydrous – impurity G : not more than twice the area of the principal
and diethylamine-free substance).
Dissolve 0.175 g at 45 °C for 10 min in methanol R and dilute
to 25.0 mL with the same solvent.
Related substances. Liquid chromatography (2.2.29). Use
freshly prepared solutions and protect from light.
Test solution (a). Dissolve 50.0 mg of the substance to be
examined in methanol R1 and dilute to 50.0 mL with the same
solvent.
Test solution (b). Dilute 1.0 mL of test solution (a) to 10.0 mL – impurity I : not more than twice the area of the principal
with methanol R1.
Reference solution (a). Dissolve 50.0 mg of melphalan
hydrochloride CRS in methanol R1 and dilute to 50.0 mL with – unspecified impurities : for each impurity, not more than the
the same solvent. Dilute 1.0 mL of the solution to 10.0 mL
with methanol R1.
Detection : spectrophotometer at 260 nm.
Injection : 20 μL of test solution (a) and reference solutions (c),
(d) and (e).
Identification of impurities : use the chromatogram
supplied with melphalan for system suitability CRS and the
chromatogram obtained with reference solution (e) to identify
the peaks due to impurities B, D, G, H, I and J.
Relative retention with reference to melphalan (retention
time = about 10 min) : impurity B = about 0.3 ;
impurity D = about 0.6 ; impurity I = about 0.8 ;
impurity J = about 1.04 ; impurity G = about 1.4 ;
impurity H = about 1.5.
System suitability : reference solution (e) :
– peak-to-valley ratio : minimum 2.5, where Hp = height
above the baseline of the peak due to impurity J and
Hv = height above the baseline of the lowest point of the
curve separating this peak from the peak due to melphalan.
Limits :
– impurity D : not more than 6 times the area of the principal
peak in the chromatogram obtained with reference
solution (d) (3.0 per cent) ;
peak in the chromatogram obtained with reference
solution (d) (1.0 per cent) ;
– impurities J, H : for each impurity, not more than the area
of the principal peak in the chromatogram obtained with
reference solution (d) (0.5 per cent) ;
– impurity B : not more than 3 times the area of the principal
peak in the chromatogram obtained with reference
solution (c) (0.3 per cent) ;
peak in the chromatogram obtained with reference
solution (c) (0.2 per cent) ;
area of the principal peak in the chromatogram obtained
with reference solution (c) (0.10 per cent) ;

2996 See the information section on general monographs (cover pages)