STP FOR FLUCONAZOLE CAPSULES

• Description: Visual
• Average Weight: Check weight of 20 Capsules at randomly and calculate the average weight by formula :  wt of 20 tablets (gm) x 1000 / 20. found avg. wt in mg
• Average Fill: Check weight of 20 Capsules at randomly. And calculate the average weight by above formula and subtract empty weight of capsule from average weight.
• Uniformity of weight:

Weigh 20 capsules selected at random and calculate the average weight. Not more than two of the individual weights deviate from the average weight by more than the percentage shown in table.

Capsules were weighed individually and the percentage of deviation of its weight from the average weight was determined for each capsule. Formula for calculate the percentage of deviation = (experimental weight – theoretical weight)    x 100%

Theoretical weight

 Average weight of capsules Percentage deviation 300 mg or less 10.0 % More than 300 mg 7.5%
• Identification Test:

In the assay, the principal peak in the chromatogram obtained with the test solution corresponds to the peak in the chromatogram obtained with the reference solution.

6.0)      Disintegration Time

Unless otherwise stated in the individual monograph, introduce one capsule into each tube and add a disc to each tube. The assembly is suspended in the liquid medium in a suitable vessel, preferably a 1-litre beaker. The volume of liquid is such that the wire mesh at its highest point is at least 15mm below the surface of the liquid, and at its lower point is at least 25mm above the bottom of the beaker. At no time should the top of the basket-rack assembly become submerged. There is a thermostatic arrangement for heating the liquid and maintaining the temperature at 37±2°.

If 1 or 2 capsules fail to disintegrate, repeat the test on 12 additional capsules; not less than 16 of the total of 18 capsules tested disintegrate.

If the capsules adhere to the disc and the preparation under examination fails to comply, repeat the test omitting the disc. The preparation complies with the test if all the capsules in the repeat test disintegrate.

7.0)      Dimension of Capsule:

Length and diameter are determined by vernier in mm

8.0)      Dissolution:

Medium: 900ml of 0.1 M HCl.

Time: 45 minutes

Speed: 50 rpm

Limit: NLT 85% of the stated amount of Fluconazole.

Withdraw a suitable volume of the medium and filter through a membrane filter disc with an average pore diameter not more than 1.0 µm, rejecting the first few ml of the filtrate.

Test solution: Dilute the filtrate, if necessary, with the dissolution medium.

Reference solution: Dissolve a weighed quantity of fluconazole reference standard in the dissolution medium and dilute with the dissolution medium to obtain a solution having a known concentration similar to the expected concentration of the test solution.

Chromatographic system:

• A stainless steel column 25cmx4.6mm, packed with octadecylsilane bonded to porous silica (5µm),
• Mobile phase: a mixture of 60 volumes of a buffer solution prepared by dissolving 1.36g of potassium dihydrogen orthophosphate in 1000ml of water and 40 volumes of methanol, adjusted to pH 3.6 with O.P.A.
• Flow rate: 1ml per minute
• Spectrophotometer set at 261nm,
• Injection volume: 20µl

9.0)      Leak test:

The apparatus is used to test for the integrity of packed strips, blisters and Alu-Alu Blister pack containing tablets. Ensure apparatus bath is filled with purified water upto mark indicated and add 0.5% crystal violet solution in water. Samples are placed into the desiccators and the lid is placed in position. The pump starts to produce a vacuum 15inHg inside the desiccators and the vacuum is held for 1 minute. The sample remains at the required vacuum level for given time interval buzzer will sound after time is over and will cut off the vacuum pump. As the package is immersed in a colored dye solution the venting of the desiccators will allow any holes to be penetrated by the dye and the contents of the flexible packaging will also be stained with the same coloring material.

Examine all the strips for any leakage by opening the pockets manually. If anyone pocket shows evidence of leakage, reject the sample, stop the Blister / Strip machine and immediately take corrective action.

10.0)    Related substance: Determined by liquid chromatography.

Note: store and inject the solutions at 4°.

Test solution: Weigh a quantity of the powder containing 100mg of fluconazole, disperse in 100ml of the mobile phase and filter.

Reference solution (a): A 0.1 per cent w/v solution of fluconazole reference standard in the mobile phase.

Reference solution (b): Dilute 1ml of reference solution (a) to 100ml with mobile phase.

Chromatographic system:

• A stainless steel column 15cmx4.6mm, packed with octadecylsilane bonded to porous silica (5µm)
• Mobile phase: a mixture of 80 volumes of water and 20 volumes of acetonitrile.
• Flow rate: 0.5ml per minute
• Spectrophotometer set at 260nm,
• Injection volume: 50µl

 Name Relative retention time Correction factor Fluconazole impurity A1 0.49 0.08 Fluconazole impurity B2 0.81 1.29 Fluconazole impurity C3 0.86 0.05 Fluconazole 1.0 –

Inject reference solution (b). The test is not valid unless the theoretical plates are not less than 1000 and the tailing factor is not more than 2.0.

Inject reference solution (b) and the test solution. Run the chromatogram four times the retention time of the principal peak. In the chromatogram obtained with the test solution, the area of any secondary peak is not more than the area of the peak in the chromatogram obtained with reference solution (b) (1.0 per cent) and the sum of areas of all the secondary peaks is not more than twice the area of the peak in the chromatogram obtained with reference solution (b) (2.0 per cent).

11.0)    Assay: Determined by liquid chromatography.

Test solution: Weigh a quantity of the powder containing 50mg of fluconazole, add 70ml of the mobile phase, sonicate for 15 minutes, and dilute to 100ml with the mobile phase and filter.

Reference solution: A 0.05 per cent w/v solution of fluconazole reference standard in mobile phase.

Chromatographic system:

• A stainless steel column 25cmx4.6mm, packed with octadecylsilane bonded to porous silica (5µm)
• Mobile phase: a mixture of 60 volumes of a buffer solution prepared by dissolving 1.36g of potassium dihydrogen orthophosphate in 1000ml of water and 40 volumes of methanol, adjusted to pH 3.6 with O.P.A.
• Flow rate: 1ml per minute
• Spectrophotometer set at 261nm,
• Injection volume: 20µl

Inject the reference solution. The test is not valid unless the theoretical plate is not less than 1000, tailing factor is not more than 2.0 and the relative standard deviation of replicate injections is not more than 2.0 per cent.

Inject the reference solution and the test solution.

Calculate the content of Fluconazole in the capsules.

Acceptance criteria: 90.0%-110.0%

Alternative Method: By UV Spectrophotometer

Test Solution: Weigh 20 capsules and remove powder from the capsules. Weight sample equivalent to 200mg of Fluconazole 100ml volumetric flask add 20ml 0.1 M HCl into it and sonicate to dissolve the test sample. Make up to mark with 0.1 M HCl. Filter and dilute 5ml of this solution to 25ml in 0.1 M HCl.

Reference Solution: Weigh 40mg of reference standard of Fluconazole in 100ml of volumetric flask. Add 20ml 0.1 M HCl into it and sonicate to dissolve. Make up to mark with 0.1 M HCl.

Check Absorbance of test as well as reference standard at about 261nm in UV Spectrophotometer.

Calculate the content of Fluconazole in the capsules.

Formula:

Abs. of Test      STD Wt.(mg)   5             100                 Potency

—————–X————–X———X——————-X———–X Average wt.

Abs. of STD        100              25      Test Wt.(mg)            100

12.0)    MICROBIOLOGICAL PURITY

Perform the test according to requirements of IP,

Total aerobic Microbial count (TAMC): NMT 103 CFU/g

Total combined yeasts/Moulds count (TYMC): NMT 102 CFU/g

Pathogens: in 1gm drug.

Escherichia Coli – Should be absent

Pseudomonas aeroginosa – Should be absent

Salmonella – Should be absent

Staphylococcus aureus– Should be absent

Abbreviations:

Wt.: Weight

mg: Miligram

ml: Milileter

STD: Standard

inHg: Inch of Mercury

rpm: Rounds per minute

CFU: Colony forming unit

HCl: Hydrochloric acid

O.P.A.: orthophosphoric acid