## STP FOR ITRACONAZOLE CAPSULES

Description: Visual,

Average Weight: Check weight of 20 Capsules at randomly and calculate the average weight by formula :       wt of 20 Capsule (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:

The retention time of the major peak of the sample solution corresponds to that of the standard dilution, as obtained in the assay.

The UV spectrum of the major peak of the sample solution corresponds to that of the standard solution, as obtained in the assay.

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.

Dimension of Capsule:

Length and diameter are determined by vernier in mm

Dissolution:

Test 1.

Apparatus: Paddle; with a three-prong sinker

Medium: 900ml 0.25% (w/v) sodium lauryl sulfate in 0.1 N hydrochloric acid

Time: 45 minutes

Speed: 75 rpm

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

Standard stock solution: 0.55mg/ml of USP Itraconazole reference standard in 40% glacial acetic acid. Sonicate, if necessary, to dissolve.

Standard solution: 0.02mg/ml of USP Itraconazole reference standard in medium from standard stock solution.

Test solution: A filtered portion of the solution under test suitably diluted with medium, to obtain a

concentration similar to that of the standard solution.

Instrumental conditions

Mode: UV

Analytical wavelength: 260nm

Relative standard deviation: NMT 2.0% for 5 replicate injections

Calculate the percentage of the labeled amount of itraconazole released:

Result= (Av/As) X (Cs/L) X V X D X 100

Av : absorbance of the Test solution

As : absorbance of the standard solution

Cs : concentration of USP Itraconzole

L : Label claim  (mg/capsule)

V : Volume of medium, 900ml

D : Dilution factor for the test solution, if applicable

Test 2.

If the product complies with this test, the labeling indicates that it meets USP dissolution test 2. All solutions containing itraconazole  should be stored in low-actinic or amber glassware and protected from light.

Medium: 900ml simulated gastric fluid without enzymes, desecrated

Time: 60 minutes

Speed: 100 rpm

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

Standard stock solution: 0.55mg/ml of USP Itraconazole reference standard in methanol prepared as follows. Transfer suitable amount of USP Itraconazole reference standard to a suitable volumetric flask and add about 80% of the flask volume of methanol. Heat the solution to 65° in a water bath, with intermittent stirring, until dissolved. Dilute with methanol to final volume.

Standard solution: 0.022mg/ml of USP Itraconazole reference standard in medium from standard stock solution.

Test solution: Pass a portion of the solution under test through a suitable filter and dilute eith medium, if necessary.

Instrumental conditions

Mode: UV

Analytical wavelength: 255nm

Relative standard deviation: NMT 2.0% for 5 replicate injections

Calculate the percentage of the labeled amount of itraconazole released:

Result= (Av/As) X (Cs/L) X V X D X 100

Av : absorbance of the Test solution

As : absorbance of the standard solution

Cs : concentration of USP Itraconzole

L : Label claim  (mg/capsule)

V : Volume of medium, 900ml

D : Dilution factor for the test solution, if applicable

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.

Related substance: Determined by liquid chromatography.

Diluent: methanol and tetrahydrofuran(50:50)

Solution A: 5.8g/L of monobasic ammonium phosphate in water. Adjust with phosphoric acid to a pH of 2.0. Pass through a suitable filter of 0.45 µm pore size.

Solution B: Acetonitrile and tetrahydrofuran (90:10)

Mobile phase: see table below:

 Time (min) Solution A (%) Solution B (%) 0 60 40 25 45 55 40 45 55 42 60 40 50 60 40

System suitability solution: 5mg/ml of USP itraconazole system suitability mixture reference standard in diluent. Sonicate, if necessary, to dissolve.

Standard solution: 0.025 mg/ml of USP Itraconazole reference standard in diluent. Sonicate, if necessary, to dissolve.

Test solution: Nominal 5mg/ml of itraconazole in diluent prepared as follows. Combine the contents of NLT 20 capsules and transfer a portion nominally equivalent to 500mg of itraconazole to 100ml flask. Add about 70ml of diluent and sonicate for 30 min with intermittent shaking. Dilute with dilent to volume. Pass through a suitable filter of 0.45µm pore size.

Chromatographic system:

• A stainless steel column (4.6mm-15-cm,) 3µm packing L1
• Flow rate: 1.5ml per minute,
• Column temperature: 30°
• Spectrophotometer set at 225nm,
• Injection volume: 10µl

Suitability requirements:

Resolution: NLT 1.5 between itraconazole and n-butyl isomer, system suitability solution

Tailing factor: NMT 2.0, standard solution

Relative standrad deviation: NMT 10.0%, standard solution

Calculate the percentage of any individual impurity in the portion of capsules taken:

Result= (rv/rs) X (Cs/Cv)X100

rv: peak response of any individual impurity from the test solution

rs : peak response of itraconazole from the standard solution

Cs : concentration of USP itraconazole reference standard in the standard solution(mg/ml)

Cv : nominal concentration of itraconazole in the test solution (mg/ml)

Acceptance criteria: Any individual unspecified impurity should be NMT 0.2%

Total impurities should be NMT 1.5%

Assay: Determined by liquid chromatography.

Diluent: methanol and tetrahydrofuran(50:50)

Test stock solution: Nominally 1mg/ml of itraconazole in diluent prepared as follows. Transfer an amount nominally equivalent to 100mg of itraconazole, from the contents of NLT 20 capsules, to a 100ml volumetric flask. Add 70ml of diluent, and sonicate for about 30 minutes. Dilute with diluent to volume. Pass through a suitable filter of 0.45µm pore size.

Test solution: Nominall 0.1mg/ml of itraconazole in diluent from Test stock solution.

Reference solution: 0.1mg/ml of USP Itraconazole reference standard in diluent. Sonicate, if necessary, to dissolve.

Chromatographic system:

• A stainless steel column (4.6mmx15-cm), 3µm packing L1
• Mobile phase: Solution A and Solution B(45:55)

Solution A: 5.8g/L of monobasic ammonium phosphate in water. Adjust with phosphoric acid to a pH of 2.0. Pass through a suitable filter of 0.45 µm pore size.

Solution B: Acetonitrile and tetrahydrofuran (90:10)

• Flow rate: 1.5ml per minute,
• Column temperature: 30°
• Spectrophotometer set at 225nm,
• Injection volume: 10µl

Inject reference solution. The test is not valid unless the tailing factor is not more than 2.0. The relative standard deviation for replicate injections is not more than 2.0 per cent.

Inject reference solution and the test solution.

Calculate the percentage of the labeled amount of itraconazole in the portion of capsules taken

Result= (rv/rs) X (Cs/Cv) X100

rv: peak response of itraconazole from the test solution

rs : peak response of itraconazole from the standard solution

Cs : concentration of USP itraconazole reference standard in the standard solution(mg/ml)

Cv : nominal concentration of itraconazole in the test solution (mg/ml)

Alternative Method: By UV Spectrophotometer

Test Solution: Weigh and powder 20 capsules. Weight sample equivalent to 25mg of Itraconazole in 100ml volumetric flask add 10ml 0.1 M methanolic HCl into it and sonicate to dissolve the test sample. Make up to mark with 0.1M methanolic HCl. Filter and dilute 2ml of this solution to 25ml in same solvent.

Reference Solution: Weigh 25mg of reference standard of Itraconazole in 100ml of volumetric flask. Add 10ml 0.1 M methanolic HCl into it and sonicate to dissolve. Make up to mark with 0.1M methanolic HCl. Filter and Dilute 2ml of this solution to 25ml with same solvent.

Check Absorbance of test as well as reference standard at about 262nm in UV Spectrophotometer.

Calculate the content of Itraconazole in capsules.

Formula:

Area of Test      STD Wt.(mg)   2            100             25        Potency

—————–X—————-X——-X—————X——–X———–X Average weight

Area of STD        100              25      Test Wt.(mg)     2           100

Acceptance criteria: 90.0%-110.0%

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

g/L: gram/litre