2016
the area of any secondary peak is not greater than 0.5 times
the area of the principal peak in the chromatogram obtained
with solution (2) (0.5%);
the sum of the areas of all the secondary peaks is not greater
than the area of the principal peak in the chromatogram
obtained with solution (2 ) ( 1.0 %).
Disregard any peak due to mobile phase A and any peak with
an area less than the area of the principal peak in the
chromatogram obtained with reference solution (4) (0.05%).
Uniformity o f content
Tablets containing less than 2 mg and or less than 2% w/w
of Dexamethasone comply with the requirements stated
under Tablets using the following method of analysis. Carry
out the method for liquid chromatography, Appendix HI D,
using the following solutions.
(1) To one tablet, add sufficient methanol (50%) to produce
a solution containing 0.0025% w/v of Dexamethasone, shake
for 10 minutes and filter through glass-fibre filter (Whatman
GF/C is suitable).
(2) 0.0025% w/v of dexamethasone BPCRS in methanol
(50%).
C H R O M A T O G R A P H IC C O N D IT IO N S
(a) Use a stainless steel column (20 cm x 4.6 mm) packed
with octadecylsUyl silica gel for chromatography (5 (im)
(Spherisorb ODS 1 is suitable).
(b) Use isocratic elution and the mobile phase described
below.
(c) Use a flow rate of 1.4 mL per minute.
(d) Use an ambient column temperature.
(e) Use a detection wavelength of 238 nm.
(f) Inject 20 (jL of each solution.
M OBILE PHASE
47 volumes of methanol and 53 volumes of water.
D E T E R M I N A T IO N O F C O N T E N T
Calculate the content of C 22H 29FO 5 in each tablet using the
declared content of C 22H 29FO 5 in dexamethasone BPCRS.
ASSAY
For ta b lets containing less than 2 m g and/or less than
2 % w /w o f D exam ethasone
Use the average of the 10 individual results obtained in the
test for Uniformity of content.
For tablets containing 2 m g o r m ore an d 2 % w /w o f
D exam ethasone
Weigh and powder 20 tablets. Carry out the method for
liquid chromatography, Appendix HI D, using the following
solutions.
(1) To a quantity of the powdered tablets containing 2.5 mg
of Dexamethasone add 20 mL of methanol (50%), shake for
20 minutes and filter through glass-fibre filter (Whatman
GF/C is suitable).
(2) 0.0125% w/v of dexamethasone BPCRS in methanol
(50%).
C H R O M A T O G R A P H IC C O N D IT IO N S
The chromatographic conditions described under Uniformity
of content may be used.
D E T E R M I N A T IO N O F C O N T E N T
Calculate the content of C 22H 29FO 5 in the tablets using the
declared content of C 22H 29FO 5 in dexamethasone BPCRS.
STORAGE
Dexamethasone Tablets should be protected from light.
Dexamethasone Preparations III-443
Dexamethasone and Neomycin Ear
Spray
Action and use
Glucocorticoid.
D EFIN ITIO N
Dexamethasone and Neomycin Ear Spray is an emulsion
containing Dexamethasone in microfine powder and Neomycin
Sulfate in a suitable vehicle in a suitable metered-dose
container. It may contain acetic acid.
The ear spray complies with the requirements stated under Ear
Preparations and with the following requirements.
Content of dexamethasone, C22H29FO5
80.0 to 120 .0 % of the amount stated to be delivered by
actuation of the valve.
Shake the ear spray vigorously before carrying out the following
tests.
IDENTIFICATION
A. In the Assay for dexamethasone, the chromatogram
obtained with solution ( 1) shows a peak with the same
retention time as the principal peak in the chromatogram
obtained with solution (2).
B. Carry out the method for thin-layer chromatography,
Appendix HI A, using the following solutions.
(1) Discharge the container a sufficient number of times to
obtain a suitable quantity and dilute with methanol, if
necessary, to produce a solution containing 0 . 1 % w/v of
Dexamethasone.
(2 ) 0 . 1 % w/v of dexamethasone BPCRS in methanol.
(3) 0.1% w/v of each of dexamethasone BPCRS and
betamethasone BPCRS in methanol.
C H R O M A T O G R A P H IC C O N D IT IO N S
(a) Use a TLC silica gel F254 plate (Merck silica gel 60 F 254
plates are suitable).
(b) Use the mobile phase as described below.
(c) Apply 5 |iL of each solution.
(d) Develop the plate to 15 cm.
(e) After removal of the plate, allow it to dry in air and
examine under ultraviolet light (254 nm) (detection method
A). Spray with alcoholic solution of sulfuric acid. Heat at 120°
for 10 minutes or until the spots appear. Allow to cool.
Examine the chromatograms in daylight and under ultraviolet
light (365 nm) (detection method B).
M OBILE PHASE
5 volumes of butan-2-ol saturated with water, 10 volumes of
toluene and 85 volumes of ether.
S Y S T E M S U I T A B IL I T Y
The test is not valid unless the chromatogram obtained with
solution (3) shows two spots which may, however, not be
completely separated.
C O N F IR M A T IO N
Method A The principal spot in the chromatogram obtained
with solution ( 1) is similar in position and size to the
principal spot in the chromatogram obtained with
solution (2 ).
Method B The principal spot in the chromatogram obtained
with solution ( 1) is similar in position, colour in daylight,
fluorescence in ultraviolet light at 365 nm and size to the
principal spot in the chromatogram obtained with
solution (2 ).
III-444 Dexamethasone Preparations 2016

C. In the test for Neomycin C, the principal spot in the
chromatogram obtained with solution ( 1) is similar in
position, colour and size to the principal spot in the
chromatogram obtained with solution (4).
Dexamethasone and Neomycin Ear Spray containing acetic acid
complies with the following additional test
D. Discharge the container a sufficient number of times to
produce 0.2 g. The solution yields reaction A characteristic
of acetates, Appendix VI.
TESTS
Acidity
pH, 2.0 to 3.0, Appendix V L.
Neamine
Carry out the method for thin-layer chromatography,
Appendix HI A, using the following solutions.
(1) Discharge the container a sufficient number of times to
obtain a suitable quantity (0.5% w/w of neomycin sulfate is
suitable).
(2) 0.01% w/v of neamine EPCRS in water.
(3) Mix 1 volume of solution (1) and 1 volume of
solution ( 2).
C H R O M A T O G R A P H IC C O N D IT IO N S
(a) Use a TLC silica gel plate.
(b) Use the mobile phase as described below.
(c) Apply 5 |xL of each solution, as 5-mm bands.
(d) Develop the plate to 15 cm.
(e) After removal of the plate, dry it at 100° to 105° for
10 minutes, spray with ninkydrin and stannous chloride reagent,
heat at 110 ° for 15 minutes, spray with the same reagent and
heat at 110° for 15 minutes.
M O B ILE PHASE
10 volumes of dichloromethane, 20 volumes of 13.5m ammonia
and 30 volumes of methanol
S Y S T E M S U I T A B IL I T Y
The test is not valid unless the chromatogram obtained with
solution (3) shows two clearly separated principal spots.
L IM IT S
In the chromatogram obtained with solution ( 1) any spot
corresponding to neamine is not more intense than the spot
in the chromatogram obtained with solution (2) (2%).
Neomycin C
Carry out the method for thin-layer chromatography,
Appendix IH A, using the following solutions.
( 1) Discharge the container a sufficient number of times to
obtain a suitable quantity (0.5% w/w of neomycin sulfate is
suitable).
(2 ) 0.075% w/v of framycetin sulfate EPCRS in water.
(3) Dilute 1 volume of solution (2) to 5 volumes with water.
(4) 0.5% w/v of neomycin sulfate EPCRS in water.
M OBILE PHASE
20 volumes of methanol and 80 volumes of a 2 0 % w/v
solution of sodium chloride.
S Y S T E M S U I T A B IL I T Y
The test is not valid unless, in the chromatogram obtained
with solution (4), a spot appears with an Rf value slightly less
than that of the principal spot.
L IM IT S
In the chromatogram obtained with solution (1) the spot
with an Rf value slightly less than that of the principal spot
(neomycin C) is not more intense than the spot in the
chromatogram obtained with solution (2 ) (15%) but is more
intense than the spot in the chromatogram obtained with
solution (3) (3%).
Related substances
D exam ethasone
Carry out the method for liquid chromatography,
Appendix HI D, using the following solutions.
(1) After priming the pump, discharge the container a
sufficient number of times to obtain 2.5 mg of
Dexamethasone, add 1.5 mL of acetonitrile R and 5 m L of
mobile phase A. Mix with the aid of ultrasound, add
sufficient mobile phase A to produce 10 mL and filter
through a 0.45-|jm filter.
(2) Dilute 1 m L of solution (1) to 100 mL with mobile
phase A.
(3) 0.002% w/v of each of methylprednisolone BPCRS and
dexamethasone BPCRS in mobile phase A.
(4) Dilute 1 mL of solution (2) to 20 mL with mobile phase
A.
C H R O M A T O G R A P H IC C O N D IT IO N S
(a) Use a stainless steel column (25 cm x 4.6 mm) packed
with octadecylsUyl silica gel for chromatography (5 Jim) (Hypersil
ODS is suitable).
(b) Use gradient elution and the mobile phase described
below.
(c) Use a flow rate of 2.5 mL per minute.
(d) Use a column temperature of 45°.
(e) Use a detection wavelength of 254 nm.
(f) Inject 20 |iL of each solution.
M OBILE PHASE
Mobile phase A 25% v/v acetonitrile.
Mobile phase B acetonitrile.
Time Mobile phase A Mobile phase B Comment
(min) (per cent V/V) (per cent V/V)
0 100 0 isocratic
15 0-+100 begin linear gradient
1
o0
o

C H R O M A T O G R A P H IC C O N D IT IO N S
40 0 100 end chromatogram,
(a) Use a TLC silica gel plate. return to 100 A
(b) Use the mobile phase as described below.
41 100 0 begin equilibration
(c) Apply 5 nL of each solution, as 5-mm bands. with A
(d) Develop the plate to 15 cm.
46 = 0 100 0 end equilibration, begin
(e) After removal of the plate, dry it at 100° to 105° for next chromatogram
10 minutes, spray with ntnhydrin solution R1 and heat at 100°
to 105° for 10 minutes.
2016
When the chromatograms are recorded under the prescribed
conditions, the retention times are: methylprednisolone,
about 12 minutes; dexamethasone, about 14 minutes.
S Y S T E M S U I T A B IL I T Y
The test is not valid unless:
(a) in the chromatogram obtained with solution (3), the
resolution factor between the peaks corresponding to
methylprednisolone and dexamethasone is at least 1.5 (if
necessary, adjust the concentration of acetonitrile in mobile
phase A);
(b) in the chromatogram obtained with solution (4), the
signal to noise ratio of the peak due to dexamethasone is at
least 10 .
L I M IT S
In the chromatogram obtained with solution (1):
the area of any secondary peak is not greater than 0.5 times
the area of the principal peak in the chromatogram obtained
with solution (2) (0.5%);
the sum of the areas of all the secondary peaks is not greater
than the area of the principal peak in the chromatogram
obtained with solution (2) ( 1%).
Disregard any peak due to mobile phase A and any peak with
an area less than the area of the principal peak in the
chromatogram obtained with reference solution (4) (0.05%).
ASSAY
For dexam ethasone
Carry out the method for liquid chromatography,
Appendix ID D, using the following solutions protected from
light.
( 1) After priming the pump, discharge the container a
sufficient number of times to obtain 1 mg of
Dexamethasone, add 10 mL of methanol, place in an
ultrasonic bath for 10 minutes, cool, dilute to 25 mL with
water, mix and filter through a 0.45-|im PTFE filter.
(2) Dilute 10 volumes of a 0.010% w/v solution of
dexamethasone BPCRS in methanol to 25 volumes with water.
C H R O M A T O G R A P H IC C O N D IT IO N S
(a) Use a stainless steel column (15 cm x 4.6 mm) packed
with octadecylsUyl silica gel for chromatography (5 pm)
(Spherisorb ODS 2 is suitable).
(b) Use isocratic elution and the mobile phase described
below.
(c) Use a flow rate of 2 mL per minute.
(d) Use an ambient column temperature.
(e) Use a detection wavelength of 254 nm.
(f) Inject 20 |iL of each solution.
M OBILE PHASE
Mix 10 volumes of glacial acetic add, 350 volumes of
acetonitrile and 640 volumes of water and filter (Whatman
GF/F is suitable).
D E T E R M I N A T IO N O F C O N T E N T
Calculate the content of C22H 29FO 5 in the ear spray using
the declared content of C 22H 29FO 5 in dexamethasone BPCRS
F or n eo m ycin su lfa te
Prime the pump and discharge the container a sufficient
number of times to obtain a quantity of the emulsion
containing 3250 IU; dilute to 50 mL with sterile phosphate
buffer pH 8.0. Dilute 10 mL of the resulting solution to
100 m L with the same solvent and carry out the
microbiological assay of antibiotics, Appendix XIV A.
The precision of the assay is such that the fiducial limits of
Dexamethasone Preparations III-445
error are not less than 95% and not more than 105% of the
estimated potency. The upper fiducial limit of error is not
less than 90.0% and the lower fiducial limit of error is not
more than 115.0% of the stated number of IU per m L
STORAGE
Dexamethasone and Neomycin Ear Spray should not be
allowed to freeze.
LABELLING
The strength with respect to Neomycin Sulfate is stated as
the number of IU (Units) per m L
Dexamethasone Sodium Phosphate Eye
Drops, Solution
Action and use
Glucocorticoid.
DEFINITION
Dexamethasone Sodium Phosphate Eye Drops, Solution are
a sterile Solution of Dexamethasone Sodium Phosphate in a
suitable vehicle.
The eye drops comply with the requirements stated under Eye
Preparations and with the following requirements.
Content of dexamethasone sodium phosphate,
C22H28FNa20 8P
95.0 to 105.0% of the stated amount.
IDENTIFICATION
Mix a quantity of the eye drops containing 20 mg of
Dexamethasone Sodium Phosphate with 5 mL of
0.1m sodium hydroxide, add 50 m L of dichloromethane and mix
with the aid of ultrasound for 20 minutes, filter the
dichloromethane layer and evaporate to dryness using a
rotary evaporator. Dry the residue at 105° for 2 hours.
The infrared absorption spectrum of the dried residue,
Appendix II A, is concordant with the reference spectrum of
dexamethasone (R S 089).
TESTS
Alkalinity
pH, 7.0 to 7.5, Appendix V L.
Related substances
Carry out the method for liquid chromatography,
Appendix LEI D, using the following solutions in the mobile
phase.
(1) Disperse a quantity of the eye drops containing 20 mg of
Dexamethasone Sodium Phosphate in 70 mL, mix with the
aid of ultrasound for 10 minutes, dilute to 100 m L and filter.
(2) Dilute 3 volumes of solution (1) to 100 volumes.
(3) 0.02% w/v of dexamethasone impurity standard BPCRS.
(4) Dilute 1 volume of solution (1) to 100 volumes and
dilute 1 volume of this solution to 10 volumes
C H R O M A T O G R A P H IC C O N D IT IO N S
(a) Use a stainless steel column (15 cm x 3.9 mm) packed
with octadecylsUyl silica gd for chromatography R (5 (im)
(Waters Symmetry C18 is suitable).
(b) Use isocratic elution and the mobile phase described
below.
(c) Use a flow rate of 1.5 mL per minute.
(d) Use an ambient column temperature.
(e) Use a detection wavelength of 254 nm.