Salicylates%20(Asrpirin);

1
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• Salicylates (Asrpirin)
• Arylalcanoic acids (Indometacin, Diclofenac)
• 2-Arylpropionic acids (Ibuprofen, Ketoprofen)
• N-Arylanthranilic acids (fenamic acids)
• Pyrazolidine derivatives (Metamizole)
• Oxicams (Piroxicam)
• Sulphonanilides (Nimesulide)
• COX-2 Inhibitors (Celecoxib)
• ????? (Omega-3 fatty acids).

2
NSAIDs
Most NSAIDs act as non-selective inhibitors of
the enzyme cyclooxygenase, inhibiting both the
cyclooxygenase-1 (COX-1) and cyclooxygenase-2
(COX-2) isoenzymes. Cyclooxygenase catalyses the
formation of prostaglandins and thromboxane from
arachidonic acid (itself derived from the
cellular phospholipid bilayer by phospholipase
A2). Prostaglandins act (among other things) as
messenger molecules in the process of
inflammation. This mechanism of action was
elucidated by John Vane, who later received a
Nobel Prize for this work.
3
Indometacin
Indometacin contains not less than 98.5 per cent
and not more than the equivalent of 100.5 per
cent of 1-(4-chlorobenzoyl)- 5 -
methoxy-2-methylindol-3-yl acetic acid,
calculated with reference to the dried
substance. A white or yellow, crystalline
powder, practically insoluble in water, sparingly
soluble in alcohol.
4
Indometacin
IDENTIFICATION First identification A, C.
Second identification A, B, D, E. A. Melting
point (2.2.14) 158C to 162C. B. Dissolve 25
mg in a mixture of 1 volume of 1M hydrochloric
acid and 9 volumes of methanol R and dilute to
100.0 ml with the same mixture of solvents.
Dilute 10.0 ml of the solution to 100.0 ml with
a mixture of 1 volume of 1M hydrochloric acid and
9 volumes of methanol R. Examined between 300 nm
and 350 nm (2.2.25), the solution shows an
absorption maximum at 318 nm. The specific
absorbance at the maximum is 170 to 190. C.
Examine by infrared absorption spectrophotometry
(2.2.24), comparing with the spectrum obtained
with indometacin CRS. Examine the substances in
the solid state without recrystallisation.
5
D. Dissolve 0.1 g in 10 ml of alcohol R, heating
slightly if necessary. To 0.1 ml of the solution
add 2 ml of a freshly prepared mixture of 1
volume of a 250 g/l solution of hydroxylamine
hydrochloride R and 3 volumes of dilute sodium
hydroxide solution R. Add 2 ml of dilute
hydrochloric acid R and 1 ml of ferric chloride
solution R2 and mix. A violet-pink colour
develops.
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Indometacin
7
Indometacin

• E. To 0.5 ml of the solution in alcohol prepared
  in identification test D, add 0.5 ml of
  dimethylaminobenzaldehyde solution R2. A
  precipitate is formed that dissolves on shaking.
  Heat on a water- bath. A bluish-green colour is
  produced. Continue to heat for 5 min and cool in
  iced water for 2 min. A precipitate is formed and
  the colour changes to light greyish-green. Add 3
  ml of alcohol R. The solution is clear and
  violet-pink in colour.

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Indometacin
9
Indometacin
10
Indometacin
ASSAY Dissolve 0.300 g in 75 ml of acetone R,
through which nitrogen R, free from carbon
dioxide, has been passed for 15 min. Maintain a
constant stream of nitrogen through the solution.
Add 0.1 ml of phenolphthalein solution R.
Titrate with 0.1 M sodium hydroxide. Carry out a
blank titration. 1 ml of 0.1 M sodium hydroxide
is equivalent to 35.78 mg of C19H16ClNO4.
STORAGE Store in a well-closed container,
protected from light. IMPURITIES A.
4-chlorobenzoic acid.
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Sodium diclofenac
Sodium diclofenac contains not less than 99.0 per
cent and not more than the equivalent of 101.0
per cent of sodium 2-(2,6- dichlorophenyl)aminop
henylacetate, calculated with reference to the
dried substance. A white or slightly yellowish,
crystalline powder, slightly hygroscopic,
sparingly soluble in water, freely soluble in
methanol, soluble in alcohol, slightly soluble
in acetone, practically insoluble in ether.
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Sodium diclofenac
???????
13
Sodium diclofenac
ASSAY Dissolve 0.250 g in 30 ml of glacial
acetic acid R. Titrate with 0.1M perchloric
acid, determining the end-point
potentiometrically (2.2.20). 1 ml of 0.1 M
perchloric acid is equivalent to 31.81 mg of
C14H10Cl2NNaO2. STORAGE Store in a airtight
container, protected from light.
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????? 3 ??? ??????? ??-?????. ???????
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????? ????????? ? ????? ?????????? ??
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??????????? ?? ???? ?? ???????? ?????????
???????? ? ?????? ????????. ??? ????????????
???????? ???????? ???????HClO4 CH3COOH
CH3COOH2 . ClO4-B CH3COOH BH.
CH3COO-BH. CH3COO- CH3COOH2 . ClO4- BH.
ClO4- 2 CH3COOH?????? ?????? ?? ??? ??? ??
???????????? ??? ???????????? ?? ??????? ???????
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??????????.
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15
Ketoprofen
Ketoprofen contains not less than 99.0 per cent
and not more than the equivalent of 100.5 per
cent of (RS)-2-(3-benzoylphenyl) propionic acid,
calculated with reference to the dried
substance. A white or almost white, crystalline
powder, practically insoluble in water, freely
soluble in acetone, in alcohol and in methylene
chloride.
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Ketoprofen
???????
17
Ketoprofen
The chromatographic procedure for Related
substances may be carried out using a
stainless steel column 0.15 m long and 4.6 mm in
internal diameter packed with a spherical
octadecylsilyl silica gel for chromatography R
(5 µm) with a specific surface of 350 m2g-1 and
a pore size of 0.01 µm (10 nm), as mobile
phase at a flow rate of 1 ml per minute a mixture
of 2 volumes of phosphate buffer solution pH 3.5
R, freshly prepared, 43 volumes of acetonitrile
R and 55 volumes of water R, as detector a
spectrophotometer set at 233 nm, a loop
injector. Inject 20 µl of reference solution
(d). The substances are eluted in the following
order ketoprofen and ketoprofen impurity A
(3-acetylbenzophenone). Adjust the sensitivity
of the detector so that the heights of the two
principal peaks in the chromatogram obtained are
not less than 50 per cent of the full scale of
the recorder. The test is not valid unless the
resolution between the peaks corresponding to
ketoprofen and ketoprofen impurity A is at least
7.0.
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Ibuprofen
Ibuprofen contains not less than 98.5 per cent
and not more than the equivalent of 101.0 per
cent of (RS)-2-(4- isobutylphenyl) propionic
acid, calculated with reference to the dried
substance. A white, crystalline powder or
colourless crystals, practically insoluble in
water, freely soluble in acetone, in ether, in
methanol and in methylene chloride. It dissolves
in dilute solutions of alkali hydroxides and
carbonates.
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Ibuprofen
20
Ibuprofen
IDENTIFICATION First identification A, C.
Second identification A, B, D. A. Melting
point (2.2.14) 75C to 78C. B. Dissolve 50.0
mg in a 4 g/l solution of sodium hydroxide R and
dilute to 100.0 ml with the same alkaline
solution. Examined between 240 nm and 300 nm
(2.2.25), using a spectrophotometer with a band
width of 1.0 nm and a scan speed of not more than
50 nm per minute, the solution shows a shoulder
at 258 nm and two absorption maxima, at 264 nm
and 272 nm. The ratio of the absorbance measured
at the maximum at 264 nm to that measured at the
shoulder at 258 nm is 1.20 to 1.30. The ratio of
the absorbance measured at the maximum at 272 nm
to that measured at the shoulder at 258 nm is
1.00 to 1.10. C. Examine by infrared
absorption spectrophotometry (2.2.24), comparing
with the spectrum obtained with ibuprofen CRS.
Examine the substances prepared as discs. D.
Examine by thin-layer chromatography (2.2.27),
using silica gel H R as the coating substance.
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Metamizole sodium
A white or almost white, crystalline powder,
very soluble in water, soluble in alcohol.
Metamizole sodium contains not less than 99.0 and
not more than the equivalent of 100.5 per cent
of sodium (1,5-dimethyl-3-oxo-2-
phenyl-2,3-dihydro-1H -pyrazol-4-yl)-N-
methylaminomethanesulphonate, calculated with
reference to the dried substance.
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Metamizole sodium Impurities
A. R at C4 NHCHO 4-formylamino-1,5-dimethyl-2-
phenyl-1,2- dihydro-3 H-pyrazol-3-one, B. R at
C4 NH2 4-amino-1,5-dimethyl-2-phenyl-1,2-dihydr
o-3H - pyrazol-3-one, C. R at C4 NHCH3
4-methylamino-1,5-dimethyl-2-phenyl-1,2-
dihydro-3 H-pyrazol-3-one, D. R at C4
N(CH3)2 4-dimethylamino-1,5-dimethyl-2-phenyl-1,2
- dihydro-3 H-pyrazol-3-one.
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Metamizole sodium
Related substances Examine by liquid
chromatography (2.2.29). Prepare the solutions
immediately before use. Test solution. Dissolve
50.0 mg of the substance to be examined in
methanol R and dilute to 10.0 ml with the same
solvent. Reference solution (a). Dissolve 10.0
mg of metamizole impurity A CRS in methanol R
and dilute to 20.0 ml with the same solvent.
Reference solution (b). Dilute 1.0 ml of
reference solution (a) to 20.0 ml with methanol
R. Reference solution (c). Dissolve 40 mg of
metamizole sodium CRS in methanol R and dilute
to 20.0 ml with the same solvent. Reference
solution (d). Take 10 ml of reference solution
(c) and boil under a reflux condenser for 10
min. Allow to cool to room temperature and
dilute to 20.0 ml with methanol R. Reference
solution (e). To 6 ml of reference solution (a)
add 1 ml of reference solution (c). 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
base-deactivated octadecylsilyl silica gel for
chromatography R (5 µm), as mobile phase at a
flow rate of 1.0 ml/min a mixture of 28 volumes
of methanol R and 72 volumes of a buffer solution
prepared by adjusting a mixture of 1000 volumes
of a 6.0 g/l solution of sodium dihydrogen
phosphate R and 1 volume of triethylamine R to
pH 7.0 with strong sodium hydroxide solution R,
as detector a spectrophotometer set at 254 nm.
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Metamizole sodium

• When the chromatograms are recorded in the
  prescribed conditions, the substances elute in
  the following order impurity A, metamizole,
  impurity B, impurity C and impurity D. Inject 10
  µl of reference solution (b). Adjust the
  sensitivity of the system so that the height of
  the principal peak in the chromatogram obtained
  is at least 50 per cent of the full scale of the
  recorder.
• Inject 10 µl of reference solution (d). The
  chromatogram shows two principal peaks due to
  metamizole and impurity C.
• Inject 10 µl of reference solution (e). The
  test is not valid unless in the chromatogram
  obtained the resolution between the peaks
  corresponding to impurity A and metamizole is at
  least 2.5.
• Inject 10 µl of the test solution and 10 µl
  of reference solution (b) and continue the
  chromatography for 3.5 times the retention time
  of metamizole. In the chromatogram obtained with
  the test solution the area of any peak
  corresponding to impurity C is not greater than
  the area of the principal peak in the
  chromatogram obtained with reference solution
  (b) (0.5 per cent), the area of any peaks, apart
  from the principal peak and the peak due to
  impurity C is not greater than 0.4 times the
  area of the principal peak in the chromatogram
  obtained with reference solution (b) (0.2 per
  cent). The sum of the areas of all the peaks,
  apart from the principal peak, is not greater
  than the area of the principal peak in the
  chromatogram obtained with reference solution (b)
  (0.5 per cent). Disregard any peak with an area
  less than 0.05 times that of the principal peak
  in the chromatogram obtained with the reference
  solution (b).

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Metamizole sodium

• Dissolve 0.200 g in 10 ml of 0.01 M
  hydrochloric acid previously cooled in iced
  water and titrate immediately, dropwise, with
  0.05 M iodine. Before each addition of 0.05 M
  iodine dissolve the precipitate by swirling. At
  the end of the titration add 2 ml of starch
  solution R and titrate until the blue colour of
  the solution persists for at least 2 min. The
  temperature of the solution during the titration
  must not exceed 10C.
• 1 ml of 0.05Miodine is equivalent to 16.67 mg
  of C13H16N3NaO4S.

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Phenylbutazone contains not less than 99.0 per
cent and not more than the equivalent of 101.0
per cent of 4-butyl-1,2- diphenyl-pyrazolidine-3,5
-dione, calculated with reference to the dried
substance. CHARACTERS A white or almost white,
crystalline powder, practically insoluble in
water, soluble in ether, sparingly soluble in
alcohol. It dissolves in alkaline solutions.
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Phenylbutazone
A. Melting point (2.2.14) 104C to 107C. B.
Dissolve 30.0 mg in 25 ml of methanol R, add 50
ml of 1M sodium hydroxide and dilute to 100.0 ml
with water R. Dilute 5.0 ml of the solution to
250.0 ml with water R. Examined between 240 nm
and 350 nm (2.2.25), the solution shows an
absorption maximum at 264 nm. The specific
absorbance at the maximum is 650 to 700. Use as
the compensation liquid a mixture of 0.5 ml of
methanol R, 1.0 ml of 1M sodium hydroxide and
98.5 ml of water R. C. Examine by infrared
absorption spectrophotometry (2.2.24), comparing
with the spectrum obtained with phenylbutazone
CRS. D. To 0.1 g add 1 ml of glacial acetic acid
R and 2 ml of hydrochloric acid R and heat the
mixture under a reflux condenser for 30 min.
Cool, add 10 ml of water R and filter. To the
filtrate add 3 ml of a 7 g/l solution of sodium
nitrite R. A yellow colour is produced. To 1 ml
of the solution add a solution of 10 mg of b-
naphthol R in 5 ml of sodium carbonate solution
R. A reddish- brown to reddish-violet precipitate
is forme.
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Phenylbutazone
Dissolve 0.500 g in 25 ml of acetone R and add
0.5 ml of bromothymol blue solution R1. Titrate
with 0.1M sodium hydroxide until a blue colour
is obtained which persists for 15 s. Carry out a
blank titration. 1 ml of 0.1 M sodium hydroxide
is equivalent to 30. 84 mg of C19H20N2O2.
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Celecoxib
Pale yellow solid, mp 157 -159 degrees.
4-5-(4-Methylphenyl)-3-(trifluoromethyl)-1H
-pyrazol-1-ylbenzene sulfonamide
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Piroxicam
Piroxicam contains not less than 98.5 per cent
and not more than the equivalent of 101.0 per
cent of 4-hydroxy-2-methyl-N-(pyridin-2-yl)-2H-1,2
-benzothiazine-3-carboxamide 1,1-dioxide,
calculated with reference to the dried
substance.
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Nimesulide

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• ??????????? ??? ????, ????? ????????? ?
  ??????, ?????? ????????? ? ??????
• ??? ?????????? ?????
• ?.?. ?????149?
• pKa6.5

N-(4-Nitro-2-phenoxyphenyl)methanesulphonamide
36
ASSAY Dissolve 0.240 g in 30 ml of previously
neutralised acetone R and add 20 ml of water R.
Titrate with 0.1 M sodium hydroxide, determining
the end-point potentiometrically (2.2.20).
1 ml of 0.1 M sodium hydroxide is equivalent to
30.83 mg of C13H12N2O5S.
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Nimesulide
A R1 SO2-CH3, R2 H, R3 R4 NO2
N-(2,4-dinitro-6-phenoxyphenyl)
methane-sulphonamide B R1 SO2-CH3, R2 R3
R4 H N-(2-phenoxyphenyl) methane-sulphonamide C
R1 R2 R3 R4 H 2-phenoxyaniline D R1
R2 R4 H, R3 NO2 4-nitro-2-phenoxyaniline E
R1 R2 SO2-CH3, R3 R4 H N,N-bis(methylsulph
onyl)-2-phenoxyaniline F R1 R2 SO2-CH3, R3
NO2, R4 H N,N-bis(methylsulphonyl)-4-nitro-2-phe
noxyaniline
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Nimesulide

• C18 3.5 µm (150 x 4.6 mm i.d.) ?????? ? C18 guard
  column (10 x 2.1 mm i.d.)
• ???????? ???? ??????????? 50 mM ????????
  ???????? ?????, ??3(5347)
• ??????? 1.1 ml/min
• UV-???????? ??? 240 nm

• Hypersil BDS-C18 column
• ???????? ???? 1g/l ???????????????????????
  (pH7.6) ??????? (5644)
• UV-???????? ??? 302 nm

• Supersphere Select B, 5µM, 125 x 3mm i.d.
• ???????? ???? 1000g ???????, 650g ???????????,
  1650g ????, 10g ???????????????????????, 4g
  ?????? ????????? ????????. pH5.5 (???? ????????
  ???????? 85)
• ??????? 0.7 ml/min
• UV-???????? ??? 297 nm

• 50mmx4.6mm i.d. monolithic column
• ???????? ???? ??????????? ???????? ?????,
  pH7 10mM (3466, v/v)
• ??????? 4.0 ml/min

39
Omega-3 fatty acids
Omega-3 fatty acids are a family of
polyunsaturated fatty acids which have in common
a carbon-carbon double bond in the ?-3 position.
Alpha-linolenic acid
Eicosapentaenoic acid
Docosahexaenoic acid
40
Omega-3 fatty acids
41
Omega-3 fatty acids
Column 1 dimensions l 0.3 m, Ø 7.8 mm
stationary phase styrene-divinylbenzene
copolymer R (7 µm), with a pore size of 10 nm.
Columns 2 and 3 placed closest to the injector
dimensions l 0.3 m, Ø 7.8 mm
stationary phase styrene-divinylbenzene
copolymer R (7 µm), with a pore size of 50 nm.
Mobile phase tetrahydrofuran R. Flow rate
0.8 ml/min. Detection Differential
refractometer. Injection 40 µl.
42
Styrene (Vinyl benzene) can be copolymerized with
other monomers for example, divinylbenzene for
cross-linking the polystyrene chains
Divinylbenzene
43
Omega-3 fatty acids
(Ph. Eur. method 2.2.30) Size-exclusion
chromatography is a chromatographic technique
which separates molecules in solution according
to their size. With organic mobile phases, the
technique is known as gel-permeation
chromatography and with aqueous mobile phases,
the term gel-filtration chromatography has been
used. The sample is introduced into a column,
which is filled with a gel or a porous particle
packing material, and is carried by the mobile
phase through the column. The size separation
takes place by repeated exchange of the solute
molecules between the solvent of the mobile
phase and the same solvent in the stagnant liquid
phase (stationary phase) within the pores of the
packing material. The pore-size range of the
packing material determines the molecular-size
range within which separation can occur.
Size-exclusion ?????????? ?? ??????
gel-permeation ??? ??????????
gel-filtration ??? ??????????
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