Category: Limit Tests

Title: Testing of Packing Materials

 

1. Objective: To provide general guideline, procedure, instructions for the Testing of Packing Materials used in packaging of Pharmaceuticals Products.

 2. Procedure: The following tests may be used to confirm the identity and quality of packing materials

 3. Definitions:-

      Grammage (GSM):-

Grammage represents the mass of the sample item in gram per square meter. It is expressed as gm/m²   

  Heat Seal Lacquers (HSL):-

Heat Seal Lacquers are generally a premium grade resins used for coating on aluminium foil to be sealed against PVC/PVDC blister substrate. A sealability of aluminium lidding foils against formable polymer materials of blister packages is usually achieved by a coating of aluminium with certain grammages of heat seal lacquers.

 

Hard/Soft Temper (Printed Aluminium foil):-

The temper of a material is best described as the hardness or elasticity of a metal or in our case of foil. The two extremes on the scale are labelled simply as hard tempered or soft tempered foil.

The following tests may be used to confirm the identity and quality of packing materials.

 

• Description

Take a sample of the packing material as per procedure mentioned in SOP/Specification. Check visually and compare it with Specification and approved specimen.

 Material/Artwork code

Take a sample of the packing material as per procedure mentioned in SOP/Specification. Check visually material/artwork code mentioned on the sample/COA and compare with approved specimen artwork code.

• Width

Take a sample of the packing material as per procedure mentioned in SOP/Specification. Measure the width with a calibrated measuring scale or Tape at various points (generally 5 points).

• Thickness

For Printed 5 ply Corrugated Box:-

Take a piece of sample and measure the thickness at various points (generally 5 points) using calibrated Vernier Calliper or screw gauge.

 

For Plain aluminium foil/ PVC film:-

Take a piece of sample about 20 cm long and Measure the thickness at various points (generally 5 points) using calibrated micro meter screw gauge.

 

For Printed Aluminium Foil:-

Take the Average Grammage of foil without HSL coating and Divide it with 2.70 and 1000 and note down reading.

 

Thickness =         Avg. Grammage of Alu. foil without HSL coating

2.70 x 1000

The density of aluminium is 2.70 grams per cubic centimetre

 

     Avg. Grammage of foil without HSL coating =

(Average of Total Grammage – Average Grammage of HSL coating)

• Internal Dimension ( For Printed 5 ply Corrugated Box/ Printed Carton)

Take one sample and Check the internal dimension of sample for Length, Breadth/width and Height using calibrated measuring scale or Tape which is suitable.

 

• Grammage (GSM)

1. Total Grammage (GSM) For PVC film, Printed Aluminium foil, Printed Carton, Plain aluminium foil :-

Cut a 5 pieces of sample in 5 x 5 cm (Length x Width) or any other suitable size as specified and weigh it on a digital balance. Weigh individual sample in gram. Note down the weight and calculate the GSM as follows:

 

Total Grammage (GSM) =    Weight (in g) x 100 x 100

Length x Width (in cm)

 

2. Grammage (GSM) for Printed 5 ply Corrugated Box  :-

Cut a piece of sample of suitable size (about 10 x 10 cm) and immersed in hot water (temperature about 60-70°C⁾ till 5 layers of ply get separate (about 5-10 minutes). In hot water, the stickiness/glue will get dissolve and will help to separate 5 layers. Then remove the sample from water. Take out 5 separated layers and label it as 1, 2,3,4,5 for identification. Then dry the layers in oven at about 60°C or any suitable temperature till the dryness. After drying, Cool and flatten the separated layers of corrugated ply by hand to remove wrinkles. Then cut those 5 separated layers in a piece of 5 cm x 5 cm (Length x Width) each. Weigh individual layer/piece in gm. Note down the weight and calculate the GSM as follows:

GSM =        Weight (in g) x 100 x 100

Length x Width (in cm)

 

3. Grammage (GSM) for poly and Aluminium Layers:-

 For Plain Aluminium foil with poly/ Printed aluminium foil with poly:-

Take sample pieces (5 x 5 cm) of previously weighed for Grammage mentioned under test (a) and introduced it in a beaker containing Concentrated Nitric Acid  till it fully dipped, for about 10 minutes. Nitric Acid will help to separate poly layer from Aluminium foil. Ensure that layers are just separated. Decant the Nitric Acid and rinse the layers with water. Then remove the sample from water. Take out 2 separated layers and wipe with tissue paper or Acetone. Dry it in current of air or in oven below 60°C, cool and re-weigh the individual poly layer and aluminium layer in gm. Note down the weight and calculate the GSM of poly layer and Aluminium layer as follows:

 

GSM for poly layer :        Weight of poly film (in g) x 100 x100

Length x Width (in cm)

 

GSM for Alu. layer :        Weight of Aluminium foil (in g) x 100 x100

Length x Width (in cm)

4. Grammage (GSM) for Aluminium foil without HSL Coating:-

Take sample pieces (5 x 5 cm) of previously weighed for Grammage mentioned under test (a) and introduced it in a beaker containing Ethyl Acetate till it fully dipped. Keep it for about 10 minutes. Ethyl Acetate will help to dissolve HSL layer from Aluminium foil. Decant the Ethyl Acetate. Then remove the sample from Ethyl Acetate. Wipe it with tissue paper. Dry it in current of air or in oven below 60°C, cool and re-weigh the individual aluminium foil in gm. Note down the weight and calculate the GSM for Aluminium foil as follows:

 

GSM for Aluminium foil  (Without HSL Coating) :

=   Weight of Alu. foil after treatment (in g) x 100 x 100

Length x Width (in cm)

 

 5. Grammage (GSM) for HSL Coating:-

       Subtract the Grammage of Aluminium (without HSL Coating) from the Total Grammage of foil.

Grammage of HSL Coating;

= [Total Grammage of foil – Grammage of Aluminium (without HSL)]

 

6. Self-Adhesive Tape:-

Cut a piece of suitable size (e.g.10 x 6 cm; Length x Width) and weigh it on a digital balance. Weigh sample in gram. Note down the weight and calculate the GSM as follows:

 

Grammage (GSM) =    Weight (in g) x 100 x 100

Length x Width (in cm)

 

• Colour Distributions)
• a) Amber/ colour distribution for PVC film:-

Take a sample piece of the packing material as per procedure mentioned in SOP/Specification. Check visually for the physical appearance for colour distribution and compare it with Specification and approved specimen.

 

b) Colour distribution for Printed Aluminium foil/ Printed Carton/ Plain aluminium foil / Printed aluminium foil with poly:-

Take a sample piece of the packing material as per procedure mentioned in SOP/Specification.  Check visually for the physical appearance for colour distribution and compare it with Specification and approved specimen.

 

c) Colour distribution for Self-Adhesive Tape:-

Take a sample piece of the packing material as per procedure mentioned in SOP/Specification.  Check visually for the physical appearance for colour distribution and compare it with Specification and approved specimen.

 

• Delivery Specification:-

For PVC film:-

Check PVC film visually at the time of sampling for its winding. It shall be wound round annual cylindrical cores of 70 mm diameter and width equal of the film. PVC shall be outwards; skewness of winding shall not exceeding 1 mm on any side. At the time of delivery, Rolls should be placed in a box designed to prevent damage to the rolls. Each roll and each box shall be labeled with -film type, width, net weight, name and address of Manufacturer.

For Printed Aluminium foil:-

Check packing material visually at the time of sampling for its winding. It shall be wound round annual cylindrical cores of 100 mm diameter and width equal to width of the foil. (Aluminium cores should be used.)

Aluminium shall be outwards; skewness of winding shall not exceed 1 mm on any side.

It shall be deliver as rolls on Aluminium cores. At the time of delivery, Rolls should be placed in a box designed to prevent damage to the rolls. Each roll and each box shall be labeled with -foil type, width, net weight and name and address of Manufacturer.

 

 For Plain aluminium foil/ Printed aluminium foil with poly:-

Check packing material visually at the time of sampling for its winding. It shall be wound round annual cylindrical cores of 70-80 mm diameter and width equal to width of the Aluminium foil. It shall be outwards; skewness winding shall not exceed 1 mm on any side. Each roll and each box shall be labeled with -foil type, width, net weight, name and address of Manufacturer.

 

For Printed Carton:- 

Check packing material visually at the time of sampling for properly tied in bundle. Each bundle should contain 50 numbers of cartons.

 

For Printed 5 ply Corrugated Box:- 

Check packing material visually at the time of sampling for properly tied in bundle. It shall be 10-   20 numbers bundled tied with strapping tape.

 

• Physical Inspection of Packing Materials

For PVC film:-

Sample the packing material as per specification/SOP and Check visually. Material shall be smooth of uniform thickness, shall be free from wrinkles, folds, cuts, tear or pinholes, edges shall be straight and parallel without curling or folding.

 

For Printed Aluminium foil with HSL Coating:-

Sample the packing material as per specification/SOP and Check visually. It shall be natural Aluminium coloured foil. Material shall be smooth of uniform thickness, free from wrinkles, folds, cuts, tear or pinholes, edges shall be straight and parallel without curling or folding. HSL coating shall be transparent, colourless of uniform thickness, without bubbles, tear, folds or pinholes and completely covering one side of the aluminium foil. The lamination/lacquer shall not be peel off under normal handling conditions and after sealing.

 

For Plain aluminium foil, Printed aluminium foil with poly:-

Sample packing material as per specification/SOP and Check visually. Material shall be smooth of uniform thickness, free from wrinkles, folds, cuts, tear or pinholes, edges shall be straight and parallel without curling or folding. Poly/LDPE coating shall be uniform thickness, without bubbles, tear, folds or pinholes and completely covering one side of the aluminium foil. The lamination/lacquer shall not be peel off under normal handling conditions and after sealing.

 

• Design/ Printing/ Printing legibility design

a)Design for Printed Carton:-

Sample the packing material as per specification/SOP and Check visually. Printing shall be neat, sharp, and legible without smudging, fading or colour migration during handling and storage.

The printed matter shall be symmetric with respect to the edges of the carton. It should meet its acceptance criteria as per Approved Specimen.

 

b) Printing (Printed Aluminium foil/ Printed aluminium foil with poly):-

Sample the packing material as per specification/SOP and Check visually. Printing shall be neat, sharp, and legible without smudging, fading or colour migration during handling and storage.

The printed matter shall be symmetric with respect to the edges of the foil and shall not be affected in anyway by the cutting. It should meet its acceptance criteria as per Approved Specimen.

 

c)Printing legibility design (Self-Adhesive Tape)

Sample the packing material as per specification/SOP and Check visually. Printing shall be neat, sharp, and legible without smudging, fading or colour migration during handling and storage. Legibility of printed matter and designed should be as per approved specimen.

 

• Punching / Cutting / Pasting

For Printed Carton/ Printed 5 ply Corrugated Box:-

Sample the packing material as per specification/SOP. Check visually and note down the observations for proper staple or proper pasting.

 

• A) Determination of Bursting strength for Printed Carton / Printed 5 ply Corrugated Box:-

Place the representative sample of the material to be analysed over the diaphragm between the upper and lower clamping plates. Lower the tripod plate applying sufficient clamping pressure to prevent slip of the sample between lower and upper clamping plates during analysis. Select the GAUGE-I by moving the lever towards left side (in case bursting strength of the sample is below 8.0 kg/cm²) or Select the GAUGE-II by moving the lever towards right side (bursting strength of the sample is above 8.0 kg/cm²). Set the needle pointer of the gauge at zero position. Press the “Push for burst” button and keep it pressed so that plunger will start rotating in clock wise direction at constant speed till sample bursts. Release pressed push button as soon as the sample bursts. Record the bursting strength reading at which the needle pointer on the gauge stops when the sample bursts.

 

B) Determination of Bursting Factor (Printed 5 ply Corrugated Box)

Calculate Bursting factor by using below formula. Put the bursting strength and Total Grammage (summation of all 5 ply GSM) of the shipper in calculation to find out bursting factor.

 

Bursting Factor (BF) =    Bursting strength x 1000

Total GSM of all 5 ply

 

• Number of Corrugation or Flutes /Meter for 5 ply Corrugated Box:-

Sample the packing material about 1 meter and Check visually. Count the number of corrugation/flutes per meter. Note down the observation. It should comply with the specification.

 

• No of Stitches & Pattern for Printed 5 ply Corrugated Box:-

Sample the packing material as per specification/SOP and Check visually. One side of the shipper shall be stapled in 2 numbers on top and bottom and after every 2.5 cm, two staple.

 

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Title: Microbial Limit Test For Material And Product

 

  Objective: Determination of Microbiological limit test for materials and products as per IP/BP/USP.

2.    Principle:    This test determines whether materials are free from microbial contamination and precautions to be taken to avoid contamination which adversely affect the material or product quality.

3. Procedure:

Total Aerobic Viable Counts (TAVC)
Method :	By Pour Plate Method
Solution A (1:10 dilution):
Aseptically transfer 10 g of sample (10 ml in case of liquid) or (number of tablets equivalent to 10 g weight to 90/100 ml of Buffered sodium chloride peptone solution pH 7.0 or Soyabean casein digest medium. A surface active agent such as 0.1% w/v solution of polysorbate 80 may be added to assist the Suspension of poorly wettable substances (Non fatty samples or which are insoluble in water). If the product is fatty, then homogenise the 10 g sample with 5g polysorbate 80. If necessary, heat to not more than 400C.  Mix carefully and add 85 ml of Buffered sodium chloride peptone solution pH 7.0 or any other suitable diluent. Adjust the volume to 100 ml with same diluent. If the product is containing antibiotic properties carry out dilutions as follow. Aseptically transfer 10 g of sample (10 ml in case of liquid/10 g in case of ointment) or (number of tablets equivalent to 10 g weight) to 40 ml of Buffered sodium chloride peptone solution pH 7.0 or soyabean casein digest medium. A surface active agents such as 0.1% w/v solution of polysorbate 80 may be added to assist the suspension of poorly wettable substances. Transfer 5 ml of homogenate and dilute to 10 ml with same solvent. If necessary heat the solution to not more than 40°C with intermittent shaking but not more than 30 mins.
4 Total Aerobic Microbial Counts (TAMC)
Transfer 1 ml of Solution A i.e 1:10 dilution separately into 2 sterile Petri dishes. Pour about 15 to 20 ml of Soyabean casein digest agar, cooled at about 45°C. Swirl the plate to mix sample and medium. Allow the plates to solidify at room temperature and incubate the plates in inverted position at 30°C to 35°C for 3 to 5 days. After completion of the incubation period count the number of colonies and express the average for the two plates in terms of the number of micro organisms per ml or gram of sample.
Interpretation of Results
If no microbial colonies are recovered from the dishes representing the initial 1:10 dilution of the sample, express the results as “less than 10 CFU per ml or gram of sample”. Calculation (TAMC) CFU/ml = Average number of colonies x dilution factor. The total aerobic viable count (TAMC) is considered to be equal to the number of CFU found using Soyabean casein digest agar; if colonies of fungi are detected on this medium they are counted as a part of TAMC.
Total Yeast & Mould Count (TYMC) Or Total Fungal Count (TFC)
Transfer 1ml of Solution A i.e. 1:10 dilution separately into 2 sterile Petri dishes. Pour about 15 to 20 ml of Sabourauds dextrose agar with or without antibiotic. Cooled at about 45°C. Swirl the plate to mix sample and medium. Allow the plates to solidify at room temperature and incubate the plates in inverted position at 20°C to 25°C for 5 to 7 days. After completion of the incubation period count the number of colonies and express the average for the two plates in terms of the number of microorganisms per ml or gram of sample.

 

Interpretation of Results
If no microbial colonies are recovered from the dishes representing the 1:10 dilution of the sample, express the results as “less than 10 CFU per ml or gram of sample”. Calculation (TFC /TYMC) CFU/ml = Average number of colonies x dilution factor. The total Fungal Count (TFC) or Total Yeast & Mould Count (TYMC) is considered to be equal to the number of CFU found using Sabourds Dextrose agar with antibiotic. If colonies of bacteria are detected on Sabourds Dextrose agar without antibiotic medium they are counted as a part of TFC/TYMC.
Test for Specified Microorganisms:
Preparation of Solution B:
Transfer 10 ml of Solution A in 90/100 ml of Sterile Soyabean Casein Digest Medium, mix and incubate at 30°C to 35°C for 18-24 hours (Solution B)
Test for Escherichia coli
Primary Test
Inoculate about 1ml from Solution B to 100 ml of MacConkey broth. Incubate at 42°C to 44°C for 24 to 48 hours.
Selection and Subculture:	Take a loopful from MacConkey broth and streak on MacConkey agar. Incubate at 30°C to 35°C for 18 to 72 hours.
Interpretation:	Growth of colonies indicates the possible presence of E. coli. Confirm the results by performing the identification tests or Confirmatory test. (Pink, non – mucoid colonies indicates the presence of E. coli.)

 

Identification Tests / Confirmatory test:
Indole test:	Inoculate the few colonies obtained on MacConkey agar in 5 ml of peptone Water and incubate at 42.0°C to 44.0°C for 24 hours. Add 0.5 ml of Kovac’s reagent into peptone water tubes. Shake well and allow to stand for one minute. If red colour ring is produced in reagent layer, Indole is present Confirms the presence of E. coli.
Acid and Gas Production:	Inoculate the few colonies grown on MacConkey agar in 5ml of MacConkey broth containing inverted durhams tube and incubate at 42.0°C to 44.0°C for 24 hours. Change in color of the broth from purple to yellow and presence of gas in Durhams tube confirms the presence of E. coli.
Test for Salmonella
Preparation of Solution C
Aseptically transfer 10 g of sample (10 ml in case of liquid/10 g in case of ointment) or (number of tablets equivalent to 10 g weight) to 90/100 ml of Soyabean casein digest medium (Solution C). A surface active agents such as 0.1% w/v solution of polysorbate 80 may be added to assist the Suspension of poorly wettable substances. Mix and incubate at 30°C to 35°C for 18 to24 hours.
Primary Test
Inoculate about 0.1 ml from Solution C to 10 ml of Rappaport vassilliadis salmonella Enrichment broth. Incubate at 30°C to 35°C for 18 to 24 hours.
Selection and Subculture:	Take a loopful from Rappaport vassilliadis salmonella enrichment broth and streak on a Xylose Lysine Deoxycholate agar. Incubate at 30°C to 35°C for 18 to 48 hours.

 

Interpretation:			Growth of colonies indicates the possible presence of Salmonella. Confirm the results by performing the identification tests or confirmatory test. (Well developed Red colonies with or without black center indicates the presence of salmonella.)
Identification Tests / Confirmatory test
Inoculate the few colonies to triple sugar iron agar by inoculating surface of slope first and then Stabbing butt using deep inoculation and incubate at 30°C to 35°C for 24 to 48 hours. If tubes have alkaline (pink) slants and acidic (yellow) butts with or without production of hydrogen sulfide gas confirms the presence of Salmonella.
Test for Shigella boydii
Preparation of Solution C( Use same sample as used for Salmonella)
Aseptically transfer 10 g of sample (10 ml in case of liquid/10 g in case of ointment) or (number of tablets equivalent to 10 g weight) to 90/100 ml of Soyabean casein digest medium (Solution C). A surface active agents such as 0.1% w/v solution of polysorbate 80 may be added to assist the Suspension of poorly wettable substances. Mix and incubate at 30°C to 35°C for 18 to 24 hours.
Primary Test
Inoculate about 0.1 ml from Solution C to 10 ml of GN Broth (Medium 11). Incubate at 30°C to 35°C for 18 to 24 hours.
Selection and Subculture:		Take a loopful from GN Broth (Medium 11) and streak on a Xylose Lysine Deoxycholate agar. Incubate at 30°C to 35°C for 18 to 48 hours.
Interpretation:		Growth of colonies indicates the possible presence of Shigella. Confirm the results by performing the identification tests or confirmatory test. (Well developed Red colonies without black center indicates the presence of shigella.)
Identification Tests / Confirmatory test
Inoculate  few colonies to Triple sugar iron agar by inoculating surface of slope first and then Stabbing butt using deep inoculation and incubate at 30°C to 35°C for 24 to 48 hours. If tubes have alkaline (pink) slants and acidic (yellow) butts without production of hydrogen sulfide gas confirms the presence of Shigella boydii.
Test for Pseudomonas aeruginosa
Selection and Subculture:			Streak a loopful of Solution B on Cetrimide agar. Incubate the plates between 30°C to 35oC for 18 to 72 hours.
Interpretation			Growth of colonies indicates the possible presence of Psuedomonas aeruginosa. Confirm the results by performing the identification tests. (Growth of Greenish colour colonies indicates the presence of P. aeruginosa.)
Identification Tests / Confirmatory test
Oxidase test:	place 2 or 3 drops of freshly prepared 1%w/v solution of N, N, N’, N’, tetraethyl- 4-phenylene-diamine dihydrochloride on filter paper and smear with colonies obtained on cetrimide agar. Pink colour develops which change to purple.
Test for Staphylococcus aureus
Selection and Subculture:	Streak a loopful of Solution B on Mannitol Salt Agar. Incubate the plate at 30°C to 35°C for 18 to 72 hours.
Interpretation:	Growth of colonies indicates the possible presence of Staphylococcus aureus. Confirm the results by performing the identification tests or confirmatory test (Yellow or white colonies surrounded by yellow zone indicates the presence of S. aureus. )
Identification Tests / confirmatory Test
Coagulase test:	Transfer individual colonies to tubes containing 0.5 ml coagulase plasma. Incubate the tubes in a water bath at 37°C, examining the tubes after 3 hours and subsequently at suitable interval up to 24 hours. The formation of clot confirms the presence of S. aureus.
Test for bile tolerant gram negative bacteria (Quantitative Test)
Transfer 10 g of the sample in 90 ml of sterile soyabean casein digest medium(0.1g/ml),(Solution D) Homogenize this homogenate and incubate at 20°C to 25OC for 2 to 5 hours but not more than 5 hours to resuscitate the organisms. Transfer 1ml of 0.1g/ml solution in 9 ml of soyabean casein digest medium (0.01g/ml). Further transfer 1ml of 0.01 g/ml Solution in 9 ml of soyabean casein digest medium(0.001g/ml).
Primary test	Inoculate 9 ml of Enterobacteria Enrichment broth–Mossel  with each 1ml Dilutions containing 0.1 g, 0.01 g, 0.001g of the sample. Incubate these tubes at 30°C to 35°C for 24 to 48 hours.
Selection and Subculture:	Subculture from each of these three tubes onto violet red bile glucose agar plates and incubate at 30°C to 35°C for 18 to 24 hours.
Interpretation:	Growth of red or reddish colonies of gram negative bacteria gives a positive result. Note the smallest quantity of product, which gives a positive result, and the largest quantity that gives a negative result. Determine the probable number of bacteria per gram of product from the given table.
Results for each quantity of products						Probable number of bacteria / g or ml of product
0.1 g or  0.1 ml				0.01 g or  0.01 ml	0.001 g or 0.001 ml
+				+	+	More than 103
+				+	–	Less than 103 and more than 102
+				–	–	Less than 102 and more than 10
–				–	–	Less than 10
Controls
Simultaneously perform a negative and positive control.
Refer SOP for Serial dilution preparation of Microbial cultures.
Inoculate NMT 100 CFU/0.1 ml of the specific culture for enumeration media & for Selective media by using surface spread technique for solid media & observe the colony character & follow as per SOP for Identification and Purity Verification of Master Microbial Cultures. For liquid media inoculate with NMT 100 CFU/0.1 ml of specific culture & check for turbidity or colour change.
Interpretation:
Positive Control
Growth obtained on solid media must not differ by a factor greater than 2 from the calculated value For standardized inoculums. In case of solid selective media growth of typical colonies should observe. in case of Liquid selective media typical reactions should occur by showing colour change.
Negative Control
Should not show any growth on solid media and turbidity or colour change in case of liquid media.

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Title: Determination of Unsaponifiable Matter

 

1.      Objective: Determination of Unsaponifiable Matter

2.      Principle:  The unsaponifiable matter consists of substances present in oils and fats which are not saponifiable by alkali hydroxides and are determined by extraction with an organic solvent of a solution of the saponified substance under examination .

 

3. Procedure:

3.1 Method

Unless otherwise specified in the individual monograph, introduce about 5 g of the substance under examination, accurately weighed, into a 250-ml flask fitted with a reflux condenser. Add a solution of 2 g of potassium hydroxide in 40 ml of ethanol (95 per cent) and heat on a water bath for 1 hour, shaking frequently. Transfer the contents of the flask to a separating funnel with the aid of 100 ml of hot water and, while the liquid is still warm, shake very carefully with three quantities, each of 100 ml, of peroxide-free ether. Combine the ether extracts in a second separating funnel containing 40 ml of water, swirl gently for a few minutes, allow to separate and reject the lower layer. Wash the extract with two quantities each of 40 ml, of water and with three quantities, each of 40 ml, of a 3 per cent w/v solution of potassium hydroxide, each treatment being followed by a washing with 40 ml of water.
Finally, wash the ether layer with successive quantities, each of 40 ml, of water until the aqueous layer is not alkaline to phenolphthalein solution. Transfer the ether layer to a weighed flask, washing out the separating funnel with peroxide-free ether. Distil off the ether and add to the residue 6 ml of acetone. Remove the solvent completely from the flask with the aid of a gentle current of air. Dry at 100°C to 105°C for 30 minutes. Cool in a desiccator and weigh the residue.
Calculate the unsaponifiable matter as per cent w/w.

Calculation :

Percent of Unsaponifiable matter w/w :         W   x  100

Y

Weight of Sample  : W

Weight of residue  : Y

 

Dissolve the residue in 20 ml of ethanol (95 per cent), previously neutralised to phenolphthalein solution and titrate with 0.1 M ethanolic potassium hydroxide. If the volume of 0.1 M ethanolic potassium hydroxide exceeds 0.2 ml, the amount weighed cannot be taken as the unsaponifiable matter and the test must be repeated.

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Title: Determination of Nitrogen

 

1.      Objective: Determination of nitrogen in organic samples.

2.      Principle:  The decomposition of nitrogen in organic samples by boiling with concentrated sulfuric acid result is an ammonium sulfate solution, adding excess base to the acid digestion mixture to convert NH₄+ to NH₃, followed by boiling and condensation of the NH₃ gas in a receiving solution. The amount of nitrogen in a sample can be calculated from the quantified amount of ammonia ions in the receiving solution.

3. Procedure:

3.1         Use method E for substances containing 2 mg or less of nitrogen.

Method A

Weigh accurately the quantity of the substance under examination specified in the individual monograph or a quantity equivalent to about 35 mg of nitrogen into a 200-ml long-necked flask and add 3 g of anhydrous sodium sulphate, 0.3 g of nitrogen-free mercuric oxide and 20 ml of nitrogen free sulphuric acid, unless otherwise specified in the monograph. Heat the mixture over a small flame until colourless and boil gently for a further 2 hours, unless otherwise directed in the monograph, care being taken to prevent the upper part of the flask from getting overheated. Cool, cautiously dilute to about 75 ml with water and add a piece of granulated zinc and a solution containing 1.5 g of sodium hydroxide per ml of the sulphuric acid used and 2 g of sodium thiosulphate in 25 ml of water. Ensure that before distillation the mixture is strongly alkaline by increasing, if necessary, the quantity of sodium hydroxide. Immediately connect the flask to a distillation apparatus, mix the contents, distill the liberated ammonia into 50.0 ml of 0.1 M sulphuric acid and titrate the excess of acid with 0.1 M sodium hydroxide using methyl red-methylene blue solution as indicator. Repeat the operation without the substance under examination. The difference between the titrations represents the ammonia liberated by the substance under examination.

1 ml of 0.1 M sulphuric acid is equivalent to 0.002802 g of N.

Method B

Weigh accurately the quantity of the substance under examination specified in the monograph or a quantity equivalent to about 35 mg of nitrogen into a 200-ml long necked flask, add 20 ml of nitrogen-free sulphuric acid, unless otherwise specified in the monograph, and heat for 15 minutes. Add 3 g of anhydrous sodium sulphate and 0.3 g of nitrogen free mercuric oxide and complete Method A, beginning at the words “Heat the mixture …”.

1 ml of 0.1 M sulphuric acid is equivalent to 0.002802 g of N.

Method C

Weigh accurately the quantity of the substance under examination specified in the monograph or a quantity equivalent to about 15 mg of nitrogen into a 200-ml long necked flask and add 1 g of a powdered mixture of 10 parts of anhydrous sodium sulphate or potassium sulphate and 1 part of cupric sulphate. Add 10 ml of nitrogen-free sulphuric acid, mix, and carefully add 1 ml of hydrogen peroxide solution (100 vol) carefully down the wall of the flask. Heat until the solution becomes clear green in colour or almost colourless for 30 minutes. Cool, carefully add 20 ml of water, cool again and connect the flask to a distillation apparatus. Add 50 ml of 10 M sodium hydroxide and distil immediately by passing steam through the flask. Collect the distillate in 25.0 ml of 0.1 M hydrochloric acid and titrate the excess of acid with 0.1 M sodium hydroxide using methyl red-methylene blue solution as indicator. Repeat the operation using 25 mg of anhydrous dextrose in place of the substance under examination. The difference between the titrations represents the ammonia liberated by the substance under examination.

1 ml of 0.1 M hydrochloric acid is equivalent to 0.001401 g of N.

 

Method D

(When nitrates and nitrites are present)
Weigh accurately the quantity of the substance under examination specified in the monograph or a quantity equivalent to about 15 mg of nitrogen into a 200-ml long necked flask, add 10 ml of nitrogen-free sulphuric acid in which 0.2 g of salicylic acid has been previously dissolved and mix. Allow the mixture to stand for 30 minutes with frequent shaking and add 1 g of a powdered mixture of 10 parts of anhydrous sodium sulphate or potassium sulphate and 1 part of cupric sulphate, mix and carefully add 1 ml of hydrogen peroxide solution (100 vol) down the wall of the flask. Complete Method C beginning at the words “Heat until the solution ….”.

1 ml of 0.1 M hydrochloric acid is equivalent to 0.001401 g of N.

 

Method E

Apparatus:

A unit of the type generally known as semi-micro Kjeldahl apparatus.

Method

Weigh accurately a quantity of the substance under examination equivalent to about 2 mg of nitrogen into the digestion flask of the apparatus. Add 1 g of a powdered mixture of 10 parts of anhydrous sodium sulphate or potassium sulphate and 1 part of cupric sulphate and wash down any adhering material from the neck of the flask with water. Add 7 ml of nitrogen-free sulphuric acid and 1 ml of hydrogen peroxide solution (100 vol) carefully down the wall of the flask. (Do not add hydrogen peroxide during the digestion). Heat until the solution has a clear blue colour and the sides of the flask are free from carbonaceous matter. Cool, add carefully 20 ml of water, cool the solution and arrange for steam distillation. Add through the funnel 30 ml of 10 M sodium hydroxide, rinse the funnel with 10 ml of water, tightly close the apparatus and begin the distillation with steam immediately.

Collect the distillate in 25 ml of 0.01 M sulphuric acid, continue the distillation until the distillate measures about 100 ml. Titrate the distillate with 0.01 M sodium hydroxide using methyl red-methylene blue solution as indicator. Repeat the operation without the substance under examination. The difference between the titrations represents the ammonia liberated by the substance under examination.

1 ml of 0.01 M sulphuric acid is equivalent to 0.0002802 g of N.

 

Method F

(Determination of Protein in Blood Products)
For dried blood products prepare a solution of the preparation as directed in the monograph.

To a volume expected to contain about 0.1 g of protein add sufficient saline solution to produce 20 ml. To 2 ml of the resulting solution, in a 75-ml boiling tube, add 2 ml of a solution containing 75.0 per cent v/v of nitrogen-free sulphuric acid, 4.5 per cent w/v of potassium sulphate and 0.5 per cent w/v of copper (II) sulphate, mix and loosely stopper the tube. Heat gradually to boiling, boil vigorously for 5 hours and cool. If the solution is not clear add 0.25 ml of hydrogen peroxide solution (20 vol), continue heating until a clear solution is produced and cool. During heating, take precautions to ensure that the upper part of the tube is not overheated.

Transfer the solution to a distillation apparatus using three 3-ml quantities of water, add 10 ml of 10M sodium hydroxide and distil rapidly for 4 minutes, collecting the distillate in a mixture of 5 ml of a saturated solution of boric acid and 5 ml of water and keeping the tip of the condenser below the level of the acid. Lower the collection flask so that the condenser can drain freely and continue the distillation for a further 1 minute. Titrate with 0.02 M hydrochloric acid using methyl red mixed solution as indicator (V₁ ml).

To a further volume of the preparation under examination, or of the solution prepared from it, expected to contain about 0.1 g of protein, add 12 ml of saline solution, 2 ml of a 7.5 per cent w/v solution of sodium molybdate and 2 ml of a mixture of 1 volume of nitrogen-free sulphuric acid and 30 volumes of water. Shake, allow to stand for 15 minutes, add sufficient water to produce 20 ml, shake again and centrifuge. Using 2 ml of the resulting clear supernatant liquid repeat the procedure described above beginning at the words ‘in a 75-ml boiling tube … ‘ (V₂ ml). Calculate the protein content in mg per ml of the preparation under examination, using the expression 6.25 x 0.280 (V₁-V₂) and taking into account the initial dilution.

 

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Title: Determination of Friability of uncoated Tablets

 

1.      Objective: Determination of Friability of uncoated Tablets

2.      Principle:    This test is applicable to compressed tablets and is intended to determine the physical strength of tablets.

 3. Procedure:

3.1 Apparatus:

It consists of a drum of transparent synthetic polymer with polished internal surfaces and subject to minimum static build-up. It has a diameter of 283-291 mm and a depth of 36-40 mm (fig.); one side of the drum is removable. A curved projection with an inner radius of 75.5 mm to 85.5 mm and extending from the middle of the drum to the outer wall enables the tumbling of the tablets at each turn of the drum. The outer diameter of the central ring is 24.5 mm to 25.5 mm. The drum is attached to the horizontal axis of a device that rotates at 25 ± 1 rpm. It should be ensured that with every turn of the drum the tablets roll or slide and fall onto the drum wall or onto each other.

Method

For tablets with an average weight of 0.65 g or less take a sample of whole tablets corresponding to about 6.5 g and for tablets with an average weight of more than 0.65 g take a sample of 10 whole tablets.

Dedust the tablets carefully and weigh accurately the required number of tablets. Place the tablets in the drum and rotate it 100 times. Remove the tablets, remove any loose dust from them and weigh them accurately. The test is run only once unless the results are difficult to interpret or if the weight loss is greater than the targeted value, in which case, the test is repeated twice and the mean of the three tests is determined.

A maximum loss of weight (from a single test or from the mean of the three tests) not greater than 1.0 per cent is acceptable for most tablets.

If obviously cracked, chipped or broken tablets are present in the sample after tumbling, the sample fails the test. If the size or shape of the tablet causes irregular tumbling, adjust the drum base so that it forms an angle of about 10° with the horizontal and the tablets do not bind together when lying next to each other, which prevents them from falling freely.

 

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Title: Determination of uniformity content

 

1.      Objective: Determination of uniformity content of single- dose preparation as per IP.

 

2.      Principle:    The test for uniformity of content of single-dose preparations is based on the assay of the individual contents of active substance(s) of a number of single-dose units to determine whether the individual contents are within limits set with reference to the average content of the sample.

 

3. Procedure:

3.1 Uniformity content of single- dose preparation:

Determine the content of active ingredient(s) in each of 10 dosage units taken at random using the method given in the monograph or by any other suitable analytical method.

Acceptance limits;

For tablets, powders and suspensions for injection and ophthalmic inserts:

The preparation complies with the test if each individual content is 85 to 115 per cent of the average content. The preparation fails to comply with the test if more than one individual content is outside these limits or if one individual content is outside the limits of 75 to 125 per cent of the average content.

If one individual content is outside the limits of 85 to 115 per cent of the average content but within the limits of 75 to 125 per cent, repeat the determination using another 20 dosage units. The preparation complies with the test if not more than one of the individual contents of the total sample of 30 dosage units is outside 85 to 115 per cent of the average content and none is outside the limits of 75 to 125 per cent of the average content.

For capsules, powders other than for parenteral use, granules, pessaries and suppositories:

The preparation complies with the test if not more than one individual content is outside the limits of 85 to 115 per cent of the average content and none is outside the limits of 75 to 125 per cent of the average content. The preparation fails to comply with the test if more than three individual contents are outside the limits of 85 to 115 per cent of the average content or if one or more individual contents are outside the limits of 75 to 125 per cent of the average content.

If two or three individual contents are outside the limits of 85 to 115 per cent of the average content but within the limits of 75 to 125 per cent, repeat the determination using another 20 dosage units. The preparation complies with the test if not more than three individual contents of the total sample of 30 dosage units are outside the limits of 85 to 115 per cent of the average content and none is outside the limits of 75 to 125 per cent of the average content.

 

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Title: Determination of uniformity weight

 

1.      Objective: Determination of uniformity weight tablets, capsules and powders as per IP.

 

2.      Principle:   The test for uniformity of weight of single-dose preparations is based on the individual single-dose units to determine whether the individual unit weight are within limits set with reference to the average content of the sample.

 3. Procedure:

3.1 Uniformity weight tablets, capsules and powders:

Weigh individually 20 units selected at random or, for single dose preparations in individual containers, the contents of 20 units, 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 the table and none deviates by more than twice that percentage.

 

Dosage        form	Average weight	Percentage deviation	Number of tablets
Uncoated and film coated tablets	80 mg or less	±  10	18
		±  20	2
	More than 80 mg but less than 250 mg	±  7.5	8
		±  15	2
	250 mg or more	±  5	18
		±  10	2
Capsules, granules and powders(single dose)	Less than 300 mg	±  10	18
		±  20	2
	300 mg or more	±  7.5	18
		±  15	2

 

Dosage        form	Average weight	Percentage deviation	Number of tablets
Powders for parenteral use	More than 40 mg	± 10	18
		± 20	2

 

    For capsules 

Weigh an intact capsule. Open it without losing any part of the shell and remove the contents as completely as possible. For soft gelatin capsules, wash the shell with a suitable solvent and keep aside until the odour of the solvent is not perceptible. Weigh the shell. The difference between the weighing gives the weight of the contents. Repeat the procedure with another 19 capsules.

For powders for parenteral use

Remove any adhering labels from a container and wash and dry the outside. Open the container and immediately weigh it along with the contents. Empty it as completely as possible by gentle tapping, rinse it with water and then with ethanol. Dry at 100°C to 105°C for one hour, or if the nature of the contents does not permit drying at this temperature, dry at a lower temperature to constant weight. Cool in a desiccator and weigh. The difference in the weighing gives the weight of the contents of the container. Repeat the procedure with another 19 containers.

 

Weight of 20 unit :  X

Average weight :   X/20 = Y

 

Y= should meet specified criteria

 

 

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Title: Determination of Disintegration Test (DT) for Tablets and Capsules 

 

1.      Objective: Determination of disintegration test for Tablets and Capsules.

2.      Principle:    This test determines whether dosage forms such as tablets, capsules, disintegrate within a prescribed time when placed in a liquid medium under the prescribed experimental conditions.

 

3. Procedure:

3.1 Disintegration Test: DT

For the purpose of this test, disintegration does not imply complete solution of the dosage unit or even of its active constituent. Disintegration is defined as that state in which no residue of the unit under test remains on the screen of the apparatus or, if a residue remains, it consists of fragments of disintegrated parts of tablets component parts such as insoluble coating of the tablets or of capsule shells, or of any melted fatty substance from the pessary or suppository or is a soft mass with no palpable core. If discs have been used with capsules, any residue remaining on the lower surfaces of the discs consists only of fragments of shells.

 For Tablets and Capsules

Apparatus

Figure 1. Diagram for disintegration apparatus A (dimensions are expressed in millimetres).

The apparatus consists of a basket-rack assembly, a 1-litre beaker, a thermostatic arrangement for heating the fluid and a mechanical device for raising and lowering the basket in the immersion fluid at a constant frequency rate.

Basket-rack assembly. The basket-rack assembly is rigid and supports six cylindrical glass tubes, 77.5 ± 2.5 mm long, 21.5 mm in internal diameter and with a wall thickness of about 2 mm (Fig.). The tubes are held vertically by two superimposed transparent plastic plates, 90 ± 2mm in diameter  and 6.75 ± 1.75 mm thick perforated by six holes having the same diameter as the tubes. The holes are equidistant from the centre of the plate and are equally spaced from one another. Attached to the underside of the lower plate is a woven stainless steel wire cloth with a plain square weave with 2.0 ± 0.2 mm mesh apertures and with a wire diameter of 0.615 ± 0.045 mm. The upper plate is covered with a stainless steel disc perforated by six holes, each about 24 ± 2 mm in diameter, which fits over the tubes and holds them between the plastic plates. The holes coincide with those of the upper plastic plate and the upper open ends of the glass tubes. A suitable means is provided to suspend the basket-rack assembly from the raising and lowering device using a point on its axis.

The plates are held rigidly in position and 77.5 mm apart by vertical metal rods at the periphery and a metal rod is also fixed to the centre of the upper plate to enable the assembly to be attached to the device for raising and lowering it smoothly at a constant frequency of between 28 and 32 cycles per minute through a distance of 50 to 60 mm. The time required for the upward stroke is equal to the time required for the downward stroke, and the change in stroke direction should be smooth and not abrupt. There should be no appreciable horizontal motion or movement of the axis from the vertical.

The design of the basket-rack assembly may be somewhat different provided specifications for the glass tubes and the screen mesh size are unchanged.

Discs. A cylindrical disc for each tube, each 20.7 ± 0.15 mm thick in diameter and 9.5 ± 0.15 mm thick, made of transparent plastic with a relative density of 1.18 to 1.20, and pierced with five holes, each 2 mm in diameter, one in the centre and the other four spaced equally on a circle of radius 6 mm from the centre of the disc. Four equally-spaced grooves are cut in the lateral surface of the disc in such a way that at the upper surface of the disc they are 9.5 mm wide and 2.55 mm deep and at the lower surface 1.6 mm square.

Medium

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 25 mm below the surface of the liquid, and at its lower point is at least 25 mm 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°C ± 2°C.

Method

Unless otherwise stated in the individual monograph, introduce one tablet or capsule into each tube and, if directed in the appropriate general monograph, add a disc to each tube. Suspend the assembly in the beaker containing the specified liquid and operate the apparatus for the specified time. Remove the assembly from the liquid. The tablets or capsules pass the test if all of them have disintegrated.

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

Apparatus

Use the apparatus for tablets and capsules described above.

Method

Put one tablet into each tube, suspend the assembly in the beaker containing 0.1 M hydrochloric acid and operate without the discs for 2 hours, unless otherwise stated in the individual monograph. Remove the assembly from the liquid.

No tablet shows signs of cracks that would allow the escape of the contents or disintegration, apart from fragments of coating.

Replace the liquid in the beaker with mixed phosphate buffer pH 6.8, add a disc to each tube and operate the apparatus for a further 60 minutes. Remove the assembly from the liquid. If the tablet fails to comply because of adherence to the disc, repeat the test on a further 6 tablets without the discs. The tablets pass the test if all six have disintegrated.

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Title: Determination of Dissolution Test (IP)

 

1.      Objective: Determination of Dissolution test for tablets and Capsules as per IP Standard.

2.      Principle:  Dissolution test is done to verify the release of drug in the solution from the tablet because binders, granulation, mixing and coating may affect the release of drug from tablets. 

3. Procedure:

3.1  Dissolution test :

The amount of dissolved active ingredient is known as D in dissolution test. The limit of D may be different in different monographs according to the nature of the formulation and its active ingredients. Dissolution test is done using 6 units or dosage forms. These dosages forms are run for the specified time period, sampled and analyzed for the dissolved amount of active ingredient in percentage. This is first stage of the dissolution and known as S₁ Stage.

In S₁ stage dissolved amount of each unit should not be less than D+5%. It shows that every unit should be above the 5% of the specified limit in the individual monograph.

If any of the unit is found below this limit then we have to analyze the sample in S₂ stage.
Six additional units are tested for the dissolved content. The average of the all 12 units should not be less than D and no unit should be less than D-15%. Average should be equal to or more then D but some units may below the D. If any unit is found below the D-15 or average of all units is less than D then sample is analyzed in S₃ stage.

Twelve more units are analyzed for the dissolved active content. At this stage the average of all 24 units should not be less than D, only two units may below the D-15% and no unit should be less than Q-25%. This stage gives more flexibility to the sample because average of 24 units should be equal to or more then D but two units may up to D-25

Dissolution stages give the flexibility to the sample that is unable to pass the dissolution test. These stages are accepted by all regulatory bodies. Hence, it is a widely accepted test method for the dissolution of solid dosage form

Use Apparatus I unless otherwise directed. All parts of the apparatus that may come into contact with the preparation under examination or with the dissolution medium are chemically inert and do not adsorb, react or interfere with the preparation under examination. All metal parts of the apparatus that may come into contact with the preparation or the dissolution medium must be made from stainless steel, type 316 or equivalent or coated with a suitable material to ensure that such parts do not react or interfere with the preparation under examination or the dissolution medium. No part of the assembly, including the environment in which the assembly is placed, contributes significant motion, agitation or vibration beyond that due to the smoothly rotating element.

An apparatus that permits observation of the preparation under examination and the stirrer during the test is preferable.

Apparatus -1

An assembly consisting of the following:

1. A cylindrical vessel, A, made of borosilicate glass or any other suitable transparent material, with a hemispherical bottom and with a nominal capacity of 1000 ml and an inside diameter of 98-106 mm (Fig.). The vessel has a flanged upper rim and is fitted with a lid that has a number of openings, one of which is central.
2. A motor with a speed regulator capable of maintaining the speed of rotation of the paddle within 4 per cent of that specified in the individual monograph. The motor is fitted with a stirring element which consists of a drive shaft and blade forming a paddle, B (Fig.).

The blade passes through the diameter of the shaft so that the bottom of the blade is flush with the bottom of the shaft. The shaft is positioned so that its axis is within 2 mm of the axis of the vessel and the lower edge of the blade is 23 to 27 mm from the inside bottom of the vessel. The apparatus operates in such a way that the paddle rotates smoothly and without significant wobble.

3. A water-bath set to maintain the dissolution medium at 36.5°C to 37.5°C. The bath liquid is kept in constant and smooth motion during the test. The vessel is securely clamped in the water bath in such a way that the displacement vibration from other equipment, including the water circulation device, is minimized

Apparatus -2

 

The assembly is the same as in Apparatus 1 except that in the stirring element the paddle is replaced by a basket, D (see Figs.). The metallic shaft rotates smoothly and without significant wobble. The basket consists of two components. The top part, with a vent, is attached to the shaft C, it is fitted with three spring clips, or other suitable means, that allow removal of the lower part for introduction of the preparation under examination and that firmly hold the lower part of the basket concentric with the axis of the vessel during rotation. The lower detachable part of the basket is made of welded-steam cloth, with a wire thickness of 0.254 mm diameter and with 0.381 mm square openings, formed into a cylinder with narrow rim of sheet metal around the top and the bottom. The basket may be plated with a 2.5 mm layer of gold for use with acidic media. The distance between the inside bottom of the vessel and the basket is maintained at 23 to 27 mm during the test.

Dissolution medium

Use the dissolution medium specified in the individual monograph. If the medium is a buffered solution, adjust the solution so that its pH is within 0.05 units of the pH specified in the monograph. The dissolution medium should be deaerated prior to testing.

Time

Where a single time specification is given in the monograph, the test may be concluded in a shorter period if the requirement for the minimum amount dissolved is met. If two or more times are specified, specimen are to be withdrawn only at the stated times, within a tolerance of ± 2 per cent.

Method

Conventional and prolonged-release solid dosage forms

Place the stated volume of the dissolution medium, free from dissolved air, into the vessel of the apparatus. Assemble the apparatus and warm the dissolution medium to 36.5°C to 37.5°C. Unless otherwise stated, place one dosage unit in the apparatus, taking care to exclude air bubbles from the surface of the dosage unit. When Apparatus 1 is used, allow the tablet or capsule to sink to the bottom of the vessel prior to the rotation of the paddle. A suitable device such as a wire of glass helix may be used to keep horizontal at the bottom of the vessel tablets or capsules that would otherwise float. When Apparatus 2 is used, place the tablet or capsule in a dry basket at the beginning of each test. Lower the basket into position before rotation.

Operate the apparatus immediately at the speed of rotation specified in the individual monograph. Within the time interval specified, or at each of the times stated, withdraw a specimen from a zone midway between the surface of the dissolution medium and the top of the rotating blade or basket, not less than 10 mm from the wall of the vessel. Except in the case of single sampling, add a volume of dissolution medium equal to the volume of the samples withdrawn. Filter the sample solution promptly through a membrane filter disc with an average pore diameter not greater than 1.0 micron. Discard the first few ml of the filtrate. Perform the analysis as directed in the individual monograph. Repeat the whole operation five times. Where two or more tablets or capsules are directed to be placed together in the apparatus, carry out six replicate tests.

For each of the tablet or capsule tested, calculate the amount of dissolved active ingredient in solution as a percentage of the stated amount where two or more tablets or capsules are placed together, determine for each test the amount of active ingredient in solution per tablet or capsules and calculate as a percentage of the stated amount.

Acceptance criteria

Conventional-release dosage forms

Unless otherwise specified, the requirements are met if the quantities of active substance dissolved from the dosage units conform to Table 1. If the results do not conform to the requirements at stage S) given in the table, continue testing with additional dosage units through stages S2 and S3 unless the results conform at stage S2′

Where capsule shells interfere with the analysis, remove the contents of not less than 6 capsules as completely as possible, and dissolve the empty capsule shells in the specified volume of the dissolution medium. Perform the analysis as directed in the individual monograph. Make any necessary correction.

Correction factors should not be greater than 25 per cent of the stated amount

Table 1

Level	Samples tested	Acceptance criteria
S1	6	Each value is not less than D + 5%
S2	6	Average value of the 12 dosage units (S1 + S2) is equal to or greater than D and no unit is less than D-15%.
S3	12	Average value of 24 dosage units (S1 + S2 + S3) is equal to or greater than D; not more than 2 units are less than D – 15%; no unit is less than D – 25%.

D = is the amount of dissolved active ingredient specified in the individual monograph, expressed as   percentage of label amount.

 

Prolonged-release dosage forms

Unless otherwise specified, the requirements are met if the quantities of active substance dissolved from the dosage units conform to Table 2. If the results do not conform to the requirements at stage L1 given in the table, continue testing with additional dosage units through stages L2 and L1 unless the results conform at stage L2. The limits embrace each value of D, the amount dissolved at each specified dosing interval. Where more than one range is specified, the acceptance criteria apply to each range.

Table 2

Level	Samples tested	Acceptance criteria
L1	6	No individual value lies outside each of the stated ranges and no individual value is less than the stated amount at the final test time.
L2	6	The average value of the 12 dosage units (L1 + L2) lies within each of the stated ranges and is not less than the stated amount at the final test time; none is more than 10% of the labelled content outside each of the stated ranges; and none is more than 10% of labelled content below the stated amount at the final test time.
L3	12	The average value of the 24 dosage units (L1 + L2 + L3) lies within the stated ranges and is not less than the stated amount at the final test time; not more than 2 of the 24 dosage units are more than 10% of labelled content outside each of the stated ranges; not more than 2 of the 24 dosage units are more than 10% of labelled content below the stated amount at the final test time; and none of the 24 dosage units is more than 20% of labelled content below the stated content at the final test time; none of the units are more than 20% of labelled content outside each of the stated ranges or more than 20% of labelled content below the stated amount at the final test time

 

Modified-release dosage forms. Use method A or Method B.

Method A

Acid stage. Place 750 ml of a.1M hydrochloric acid in the vessel, and assemble the apparatus. Warm the dissolution medium to 36.5°C to 37.5°C. Place one dosage unit in the apparatus, cover the vessel and operate the apparatus at the specified rate. After 2 hours of operation in the acid medium, withdraw an aliquot of the liquid and proceed immediately as directed under Buffer stage. Perform the analysis of the aliquot using a suitable assay method.

Buffer stage

Complete the operations of adding the buffer and adjusting the pH within 5 minutes. With the apparatus operating at the rate specified, add to the medium in the vessel 250 ml of a 0.2 M solution of trisodium phosphate dodecahydrate that has been warmed to 36°C to 37°C. Adjust, if necessary, with 2M hydrochloric acid or 2 M sodium hydroxide to a pH of 6.8 ± 0.05. 2 M hydrochloric acid or 2 M sodium hydroxide to a pH of 6.8 ± 0.05.

Method B

Acid stage. Place 1000 ml of 0.1M hydrochloric acid in the vessel and assemble the apparatus. Warm the dissolution medium to 36°C to 37°C. Place one dosage unit in the apparatus, cover the vessel and operate the apparatus at the specified rate. After 2 hours of operation in the acid medium, withdraw an aliquot of the liquid and proceed immediately as directed under Buffer stage. Perform the analysis of the aliquot using a suitable assay method.

Buffer stage

Use buffer that has previously been warmed to 36°Cto 37°C. Drain the acid from the vessel and add 1000 ml of pH 6.8 phosphate buffer, prepared by mixing 3 volumes of 0.1M hydrochloric acid with 1 volume of 0.2 M solution of trisodium phosphate dodecahydrate and adjusting, if necessary, with 2M hydrochloric acid or 2M sodium hydroxide to a pH of 6.8 ± 0.05. This may also be done by removing from the apparatus the vessel containing the acid and replacing it with another vessel containing the buffer and transferring the dosage unit to the vessel containing the buffer. Continue to operate the apparatus for 45 minutes, or for the specified time. At the end of this period, withdraw an aliquot of the liquid and perform the analysis using a suitable assay method.

Acceptance criteria

Acid stage

Unless otherwise specified, the requirements of this part of the test are met if the quantities, based on the percentage of the labelled content of active substance dissolved from the units tested conform to Table 3. Continue the testing through the 3 levels unless the results of both acid and buffer stages conform at an earlier level.
Table 3

Level	Samples tested	Acceptance criteria
A1	6	No individual value exceeds 10% dissolved
A2	6	Average value of the 12 dosage units (A1 + A2) is not more than 10% dissolved, and no individual value is greater than 25% dissolved
A3	12	Average value of 24 dosage units (A1 + A2 + A3) is not more than 10% dissolved, and no individual value is greater than 25% dissolved.

Buffer stage

Unless otherwise specified, the requirements of this part of the test are met if the quantities, based on the percentage of the labelled content of active substance dissolved from the units tested conform to Table 4. Continue the testing through the 3 levels unless the results of both acid and buffer stages conform at an earlier level. The value of D in Table 4 is 75 per cent dissolved unless otherwise specified. The quantity, D, is the specified total amount of active substance dissolved in both the acid and buffer stages, expressed as a percentage of the labelled content

Table 4

Level	Samples tested	Acceptance criteria
B1	6	No value is less than D + 5%
B2	6	Average value of the 12 dosage units (B1 + B2) is equal to or greater than D, and no unit is less than D – 15%
B3	12	Average value of the 24 dosage units (B1 + B2 + B3) is equal to or greater than D; not more than 2 units are less than D – 15%, and no unit is less than D – 25%.

 

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Title: Determination of solubility

 

1.      Objective: Determination of solubility of powder and liquid.

2.      Principle: The approximate solubility’s of the articles of the Pharmacopoeia are given here primarily as Information; they are not meant to be test for identifying the materials. However they may indirectly help in the primary evaluation of the integrity of an article

 3. Procedure:

3.1    Solubility of powder and liquid:

A test for solubility becomes a test for purity only when where a special quantitative test is given in the individual monograph and is an official requirement.

They have been indicated by descriptive terms in the accompanying table and with reference to a temperature of 15°C to 30°C

Descriptive term	Parts of solvent required for part of solute
Very soluble	Less than 1
Freely soluble	From  1 to 10
soluble	From  10 to 30
Sparingly soluble	From  30 to 100
Slightly soluble	From  100 to 1000
Very Slightly soluble	From  1000 to 10,000
Practically insoluble, or Insoluble	From  10,000 to more

 

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