FLUOCINOLONE ACETONIDE IF 2010

- mobile phase: a mixture of450 rnl of acetonitrile and500 Identification

rnl of water, allowed to equilibrate, the volume adjusted
to 1000 rnl with water and mixed again, A. Determine by thin-layer chromatography (2.4.17), coating
- flow rate. 1 rnl per minute, the plate with silica gel GF254.
- spectrophotometer set at 238 nm, Mobile phase. A mixture of60 volumes of n-hexane, 40 volumes
- injection volume. 20 fll. of chloroform, 10 volumes of methanol and 1 volume of
Adjust the sensitivity of the system so that the height of the triethylamine.
principal peak in the chromatogram obtained with reference Test solution. Disperse, by shaking a quantity of the cream
solution (b) is not less than 50 per cent of the full scale of the containing 0.25 mg of Fluocinolone Acetonide in 2 rnl of
recorder. chloroform, add 10 rnl of methanol, shake vigorously, cool in
ice for 15 minutes, centrifuge at 3000 rpm for 15 minutes, decant
Inject reference solution (a). The retention times are:
the clear supernatant liquid, evaporate to dryness on a water-
triamcinolone acetonide about 8.5 minutes and fluocinolone
bath ina current of nitrogen and dissolve the residue in 1 rnl of
acetonide about 10 minutes. The test is not valid unless the
chloroform.
resolution between the peaks corresponding to triamcinolone
acetonide and fluocinolone acetonide is not less than 3.0. ~eterence solution. A 0.025 per cent w/v solution of
fluocinolone acetonide RS in chloroform.
Inject the test solution and reference solution (b). Continue
the chromatography for 4 times the retention time of Apply to the plate 5 fll of each solution. After development,
fluocinolone acetonide. In the chromatogram obtained with dry the plate in air until the odour of the solvent is no longer
the test solution the area of any peak other than the principal detectable, heat at 105° for 5 minutes and spray whilst hot
peak is not greater than the area of the principal peak in the with blue tetrazolium solution. The principal spot in the
chromatogram obtained with reference solution (b) (0.5 per chromatogram obtained with the test solution corresponds to
cent) and the sum of the areas of all such peaks is not greater that in the chromatogram obtained with the reference solution.
than~.5tim~stheareaofthep~cipalpeakinthechromatogram B. In the Assay, the principal peak in the chromatogram
obtamed WIth reference solutlOn (b) (2.5 per cent). Ignore any obtained with the test solution (a) has the same retention time
peak ~ue to the solvent ~d ~y peak ,:"ith an area less than as that of the peak due to Fluocinolone Acetonide in the
.. O:05=trmes=thatof=the=pnnclpal=pealc=m=the=chromatogram chromafogram ol5tamed Witlflnereference solutibn;==== =========
u
c

obtained with reference solution (b) (0.05 per cent).

Losson tIryilig (2.4.19). Not mote than 1.0petcent,detertflitfed Tests
on 1.0 g by drying in an oven at 105° for 3 hours. Other tests. Complies with the tests stated under Creams.
Assay. Weigh accurately about 50 mg, dissolve in ethanol, Assay. Determine by liquid chromatography (2.4.14).
add sufficient ethanol to produce 50.0 rnl and mix. Dilute
For creams containing 0.025 per cent to 0.2 per cent w/w of
2.0 rnl of this solution to 100.0 rnl with ethanol. Measure the
fluocinolone acetonide:
absorbance of the resulting solution at th~ maximum at about
238 nm. Calculate the content of C24H30F206 taking 355 as the Test solution (a). Weigh accurately a quantity of the cream
specific absorbance at 238 nm. containing about 2.5 mg of Fluocinolone Acetonide, add 60 rnl
_ ....•. ' ....• __ ..... • '., .• _....., .'0 _ _ ,.• " _ _ ., ._, • •. " _ _ , _ , _ , .. . "._. __ ~ ~ __ . . ,_._• • __ ._. _

ofasolution prepared by adding 80mlofmethanolto20rnlof

Storage. Store protected from light.
a 25 per cent w/v solution of lithium chloride and disperse by
shaking vigorously. Add 100 rnl of cyclohexane, shake gently
for 2 minutes and separate the lower, aqueous methanolic
layer, taking care to exclude any solid matter that separates at
Fluocinolone Cream the interface. Repeat the extraction using a further 25 rnl of the
lithium chloride solution. To the combined extracts add a
Fluocinolone Acetonide Cream solution containing 11 g of alum in 214 rnl of water followed
FluoeirioloneCream·coutaiiis Fluocinolone Acetonidein a by50rnl of chloroform, shake vigorously for about 3 minutes,
suitable base. allow the layers to separate and fIlter the chloroform extract
through filter paper (such as Whatman No 1), previously
Fluocinolone Cream contains not less than 90.0 per cent and moistened with chloroform, again excluding any solid matter
not more than 110.0 per cent of the stated amount of at the interface. Repeat the extraction with 50- and lO-rnl
fluocinolone acetonide, C24H30F206. quantities of chloroform, filtering the extracts as before.
Usual streugths. 0.0025 per cent; 0.00625 per cent; 0.01 per Evaporate the combined extracts to dryness on a water-bath
cent; 0.025 per cent; 0.2 per cent w/w. in a current of nitrogen, dissolve the residue in 5rnl of

1362
IP 2010
chlorofonn, transfer to a lO-ml volumetric flask with the aid of
chloroform and add sufficient chlorofonnto produce 10,0 mI.
Test solution (b). Prepare in the same manner as test solution
(a) but adding 1.0 mI of a 0.05 per cent w/v solution of
phenacetin (internal standard) to the chloroform solution
before dilution to 10.0 mI.
Reference solution. A solution containing 0.025 per .cent w/v
of fluocinolone acetonide RS and 0.005 per cent w/v of
phenacetin in chloroform.
For creams containing 0.01 per cent w/w offluocinolone
acetonide:
Test solution (a). Prepare as described above but using a
quantity of the cream containing about 1 mg of Fluocinolone
Acetonide.
Test solution (b). Prepare in the same manner as test solution
(a) but adding 1.0 mI of a 0.02 per cent w/v solution of
phenacetin (internal standard) to the chlorofonn solution
before diluting to 10.0 ml. "
Reference solution. A solution containing 0.01 per cent w/v of
fluocinolone acetonide RS and 0.002 per cent w/v of
phenacetin in chlorofonn.
For creams containing 0.00625 per cent w/w offluocinolone
acetonide:
Test solution (a). Prepare as described above but ijsing a
quantity of the cream containing about 0.625 mg of
Fluocinolone Acetonide.
Test solution (b). Prepare in the same manner as test solution
(a) but adding 1.0 mI of a 0.0125 per cent w/v solution of
phenacetin (internal standard)· to the chloroform solution
before diluting to 10.0 mI.
Reference solution. A solution containing 0.00625 per cent
w/v offluocinolone acetonide RS and 0.00125 per cent w/v of
phenacetin in chlorofonn.
For creams containing 0.0025 per cent w/w offluocinolone
acetonide:
Test solution (a). Prepare as described above but using a
quantity ofthe cream containing about 0.25 mg ofFluocinolone
Acetonide.
Test solution (b). Prepare in the same manner as test solution
(a) but adding 1.0 inlof a 0.005 per cent w/v solution of
phenacetin (internal standard) to the chloroform solution
before diluting to 10.0 ml. . A. Determine by infrared absorption spectrophotometry (2.4.6).
Reference solution. A solution containing 0.0025 per cent w/v
of fluocinolone acetonide RS and 0.0005 per cent w/v of
phenacetin in chloroform.
Chromatographic system
a stainless steel column 25 cm x 4.6 mm, packed with
porous silica particles (5 flIIl),
1363
FLUORESCEIN SODIUM
mobile phase: a mixture of 58 volumes of n-hexane,
40 volumes of chlorofonn, 2 volumes of methanol and
0.1 volume of glacial acetic acid,
flow rate. 1.8 mI per minute,
spectrophotometer set at 243 nm,
injection volume. 20,n.
The assay is not valid unless the resolution between the peaks
due to fluocinolone acetonide and phenacetin is more than 2,
and the capacity factors of fluocinolone acetonide and
phenacetin are about 3 and 2, respectively. If these conditions
are not achieved, adjust the concentration of methanol and
chloroform in the mobile phase. Repeat the adjustment of
chloroform and methanol concentration until correct values
for both resolution and capacity factors have been obtained.
Calculate the content of CzJf3oFz06 in the cream.
Storage. Store at a temperature not exceeding 30°.
Fluorescein Sodium
Soluble Fluorescein
NaO o
COONa
Mol. Wt. 376.3
Fluorescin Sodium is disodium 2-(3-oxo-6-oxido-3H-
xanthen-9-yl)benzoate.
Fluorescein Sodium contains not less than 98.5 per cent and
not more than 100.5 per cent of C2oHlQNazOs, calculated on the
dried basis.
Category. Diagnostic aid (dye for detection of corneal lesions
and foreign bodies).
Description. An orange-red powder; odourless or almost
odourless; hygroscopic.
Identification
Compare the spectrum with that obtained with fluorescein
sodium RS or with the reference spectrum of fluorescein
sodium.
B. A solution is strongly fluorescent, even in extreme dilutions.
The fluorescence disappears when the solution is made acidic
and reappears when it is made alkaline.