1.0) Description: Visual
2.0) Average Weight: Check weight of 20 tablets at randomly and calculate the average weight by formula :
wt of 20 tablets (gm) x 1000 / 20. found avg. wt in mg
3.0) Uniformity of weight:
Weigh 20 tablets 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.
Tablets were weighed individually and the percentage of deviation of its weight from the average weight was determined for each tablet. Formula for calculate the percentage of deviation = (experimental weight – theoretical weight) x 100%
theoretical weight
| Average weight of tablets | Percentage deviation |
| More than 80mg but Less than 250mg | 7.5% |
| 250 mg or More | 5% |
4.0) 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.
5.0) Hardness:
The standard method used for tablet hardness testing is compression testing. The tablet is placed between two jaws that crush the tablet. The machine measures the force applied to the tablet and detects when it fractures. Although compressive force is applied to the tablet, along the diameter of the tablet at right angles to the applied force
6.0) Disintegration Time
Unless otherwise stated in the individual monograph, introduce one tablet 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 tablets fail to disintegrate, repeat the test on 12 additional tablets; not less than 16 of the total of 18 tablets tested disintegrate.
If the tablets 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 tablets in the repeat test disintegrate.
7.0) Dimension of Tablet:
Length, breadth and thickness are determined by vernier in mm
8.0) Dissolution:
Apparatus: Paddle
Medium: 0.01M hydrochloric acid; 900ml
Speed: 50rpm
Time: 45 minutes
Limit: NLT 75 percent of the stated amount of olanzapine in the medium.
Withdraw a suitable volume of the medium and filter. Determine by liquid chromatography
Note: Protect all the solution from light.
Test solution: Use the filtrate, if necessary dilute with dissolution medium.
Reference solution: Weigh 16mg of olanzapine reference standard, dissolve in about 2.5ml of acetonitrile and dilute to 25ml with 0.01M hydrochloric acid. Dilute suitably to get 0.00016 per cent w/v in dissolution medium.
Chromatography System:
- A stainless steel column 25cm X4.6mm, packed with octadecylsilane bonded to porous silica (5µm)
- Mobile Phase: A mixture of 70 volumes of buffer solution prepared by dissolving 3gm of ammonium dihydrogen orthophosphate in 900ml water, add 2ml of triethylamine and dilute to 1000ml with water. Adjust the pH to 2.5 with orthophosphoric acid and 30 volumes of methanol
- Flow rate: 1ml per minute,
- Spectropohtometer set at 260nm,
- Injection volume: 10µl
Inject the reference solution. The test is not valid unless the tailing factor is not more than 2.0 and the column efficiency is not less than 2500 theoretical plates.
Inject the reference solution and the test solution.
Calculate the content of Olanzapine.
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) Assay: Determined by liquid chromatography.
Reference solution: Weigh 25mg of olanzapine reference standard in 50ml volumetric flask. Add 10ml of mobile phase to dissolve the standard. Dilute upto the mark with mobile phase. Dilute 5ml of this solution to 50ml in mobile phase. Filter the solution and inject.
Test preparation: Weigh and powder 20 tablets. Weigh sample containing 5mg of olanzapine in to 100ml volumetric flask. Add 10ml of mobile phase to dissolve the sample. Dilute this upto the mark. Filter and inject the sample.
Chromatography System:
- A stainless steel column 25cm X4.6mm, packed with octadecylsilane bonded to porous silica (5µm)
- Mobile Phase: A mixture of 70 volumes of buffer solution prepared by dissolving 3gm of ammonium dihydrogen orthophosphate in 900ml water, add 2ml of triethylamine and dilute to 1000ml with water. Adjust the pH to 2.5 with orthophosphoric acid and 30 volumes of methanol
- Flow rate: 1ml per minute,
- Spectropohtometer set at 260nm,
- Injection volume: 10µl
Inject the reference solution. The test is not valid unless the tailing factor is not more than 2.0, the column efficiency is not less than 2500 theoratical plates. The relative standard deviation for replicate injections is not more than 2 per cent.
Inject the reference solution and the test solution.
Calculate the content of Olanzapine.
Formula:
Area of Test STD Wt.(mg) 5 100 Potency
—————–X————–X———X—————–X———–X Average weight
Area of STD 50 50 Test Wt.(mg) 100
Alternative method: (By UV spectrophotometer)
Reference solution:Weigh 25mg of olanzapine reference standard in 50ml volumetric flask. Add 10ml of methanol and dissolve the standard. Dilute upto the mark with methanol. Dilute 1ml of this solution to 50ml in 0.1M HCl.
Test preparation: Weigh and powder 20 tablets. Weigh sample containing 10mg of olanzapine in to 100ml volumetric flask. Add 10ml of methanol to dissolve the sample. Dilute this upto the mark. Filter and dilute it further 5ml to 50ml in 0.1M HCl.
Scan the solution by UV Spectrophotometer. It will give absorbance maxima at about 260nm. Note down the reading and calculate the content of olanzapine in tablet.
Formula:
Acceptance criteria: 90.0%-110.0%
Formula:
Area of Test STD Wt.(mg) 1 100 50 Potency
—————–X————–X———X—————–X——–X———–X Average weight
Area of STD 50 50 Test Wt.(mg) 5 100
Acceptance criteria: 90.0%-110.0%
11.0) MICROBIAL ANALYSIS
A.) Total microbial count
B.) Pathogen detection
A.) TOTAL MICROBIAL COUNT
- Pour plate method by serial dilution:
Take 2 test tubes and label them A & B and 6 petriplates (3 for SCDA &3 for SDA) and label the petriplates as 10ˉ¹, 10ˉ², 10ˉ³.
Pour 9ml buffer solution (NaCl / Peptone water) in each test tube.
Crush the tablets in pestle motar and make it in powder form.
Take 99ml of Soyabean Casein Digest Medium (Tryptone Soya Broth) in borosilicate glass bottle and add 1 gm of tablet powder sample to it to make 1:100 dilution and mix well.
Pipette out 1ml of this dilution with the help of sterile micropipette and open the lids of empty sterile petriplate labeled 10ˉ¹ and dispense into the middle of each petriplate aseptically and then close the lid of petriplate. Transfer 1ml of tablet sample sample to the test tube labeled A to make 1:10 dilution and mix well. Take 1ml of sample from test tube A and dispense aseptically into the petriplate labeled 10ˉ². Transfer 1ml of sample from test tube A to test tube B and mix well. Then take 1ml of sample from test tube B and dispense the sample into the petriplate labeled 10ˉ³. Remove the cotton plug from the flask of molten agar (SCDA & SDA) and pass the rim of opened flask through the flame of Bunsen burner. Open the lid of petriplate containing the sample and pour 20-25ml sterile Soyabean Casein Digest Agar (for TAMC) and Sabouraud Dextrose Agar (for TYMC) cooled at 45ºC into the petriplates. Close the lid of petriplates and mix thoroughly by gently swirling the plates & allow solidifying the media at room temperature.
Use aseptic technique throughout the procedure
Invert and incubate the SCDA plates at 30- 35ºC for 3-5 days (for TAMC) and SDA plates at 20-25ºC for 5-7 days (for TYMC). Also incubate the Soyabean Casein Digest Medium containing capsule sample for 24 hours at 30- 35ºC temperature for pathogen detection test.
After incubation examine the SCDA plates for total bacterial count and SDA plates for total yeast and mold count. Take maximum from the two plates and calculate the no. of CFU/ml of water sample.
B.) PATHOGEN DETECTION
- Detection of Escherichia coli:
Streak a loopful culture of enriched pre-incubated Soyabean Casein Digest Medium containing capsule sample on the surface of sterile MacConkey Aagr plate. Incubate the plates at 30-35 for 18-72 hours. After incubation presence of brick red colonies on MacConkey agar indicate the presence of E.Coli. Further confirmation is done by gram’s staining.
- Detection of Salmonella:
Transfer 0.1 ml of enriched Soyabean Casein Digest medium to 10 ml of Rappaport Vassiliadis
Salmonella enrichment broth and incubate at 30-35 for 24-48 hours. Growth of green colonies with blank centre and in 48 hours the colonies become uniformly black colonies surrounded by a dark zone and metallic sheen indicates the possibility of presence of Salmonella.
If subcultured on plate Xylose-Lysine Deoxycholate Agar and incubate at 30-35 for 24-48 hours. Well developed red colonies with or without black centre indicates the possibility of presence of Salmonella. Further confirmation is done by gram’s staining.
- Detection of Pseudomonas aeruginosa:
Streak a loopful culture of pre-incubated Soyabean Casein Digest Medium (containing capsule sample) on the surface of Cetrimide Agar medium and incubate at 30-35 for 18-72 hours. A greenish colored colony indicates the possibility of presence of P.aeruginosa. Carry out the further conformation by gram’s staining. Further confirmation done by oxidase test.
- Detection of Staphylococcus aureus:
Streak a loopful culture of pre-incubated Soyabean Casein Digest medium (containing capsule sample) on the surface of Mannitol Salt Agar plates. Incubate the plates at 30-35 for 18-72 hours. Examine the plates after incubation. If upon examination of incubated plates none of them contains yellow colonies with yellow zones the sample meets the requirements for the absence of Staphylococcus aureus. If growth occurs further confirm by streak the colonies on the surface of Baired Parker agar plates black, shinny colonies surrounded by clear zones indicates the presence of Staphylococcus aureus. Further confirm by gram’s staining.
- Detection of Asperigillus brasiliensis:
Streak a loopful culture of enriched fluid Soyabean Casein Digest medium on the surface of Sabooraud Dextrose Agar plates. Incubate the plates at 20-25 for 18-72 hours.
Growth of black color colonies may indicate the presence of Asperigillus brasiliensis.
Further confirm by microscopic examination.
- Test for Candida albicans:
Streak a loopful culture of enriched fluid Soyabean Casein Digest medium on the surface of Sabooraud Dextrose Agar plates. Incubate the plates at 20-25 for 18-72 hours.
Growth of white colonies may indicate the presence of Candida albicans.
Abbreviations:
Wt.: Weight
mg: Miligram
ml: Milileter
STD: Standard
inHg: Inch of Mercury
RPM: Rounds per minute
E.coli: Escherichia coli
P.: Pseudomonas
TAMC: Total aerobic microbial count
SCDA: Soyabean casein digest medium
TYMC: Total yeast and mold count
SDA: Sabourand dextrose agar
CFU: Colony forming unit
NaCl: Sodium Chloride
HCl: Hydrochloric acid
