VALIDATION OF SHELF LIFE FOR 70% V/V IPA

VALIDATION TEST

Validation of diluted disinfectant storage conditions are done by following methods

      ·  Surface swab test Method
      ·  Settle Plate Method

1. Prepare 70% v/v solution of Isopropyl Alcohol.

2. Carry out the sterility of diluted IPA solution by Membrane filtration method.

3. Store the disinfectant solution at room temperature, and analyze the sample on the day of preparation, after 24 hrs, 48 hrs, 72 hrs and 96 hours by Surface swab Test, Settle Plate Method.

SURFACE SWAB METHOD

Remove following culture slant from the refrigerator and allow it to attain room temperature:-

      ·  Staphylococcus aureus
      ·  Escherichia coli
      ·  Pseudomonas aeruginosa
      ·  Salmonella, and
      ·  Wild culture

1. Inoculate loop full of the culture from each slant separately into 50 ml of sterile Soyabean casein digest medium and incubate at 32.5 ± 2.5°C for 24 – 48 hrs.

2. Transfer 1.0 ml of the broth culture into 9.0 ml of sterile saline solution (0.9% sodium chloride solution) to obtain a test dilution of 10-1.

3. Transfer 1.0 ml of the 10-1 dilution into 9.0 ml of sterile saline solution to give 10-2 dilution.

4. Similarly serially dilute the culture suspension to obtain dilution of 10-3, 10-4, 10-5  and 10-6.

5. Plate 1.0 ml of the culture suspension from dilution 10-3, 10-4, 10-5 and 10-6 in duplicate into sterile petri dishes.

6. Pour approximately 15-20 ml of sterile Soyabean Casein Digest Agar cooled to about 45°C in each plate. Incubate at 32.5 ± 2.5°C for 24 to 48 hours.

7. Count the number of colonies on each plate and Select the dilution, which gives a count of not less than 10-5.

8. Apply 1.0 ml of each culture suspension containing cell concentration not less than 10-5 cfu/ml, separately on the floor approx 25 cm2 area, and allow to air dry.

9. After drying, take a surface swab as per latest SOP for Swab Testing, and carry out the determination of total aerobic count per cm2 withing 4 hours of sampling.

10. Immediately clean the floor with IPA 70% v/v solution and allow to stand for 30 minutes to facilitate the action of disinfectant solution on the challenge test organisms.

11. After 30 minutes, take a swab and detect the bacterial count as per SOP for swab testing.

12. Repeat the same procedure, step no. 9 to 12 to determine the efficacy of 70% IPA solution after 24 hrs, 48 hrs, 72 hrs and 96 hours at room temperature.

SETTLE PLATE METHOD

1. Expose the pre-incubated sterile Soybean casein digest agar plate for 2 hours.

2. After plate exposure time, immediately clean the floor with IPA 70% v/v solution.

3. After proper cleaning, spray the area with IPA 70 % v/v solution and immediately close the room 30 minutes to facilitate the action of disinfectant solution.

4. After 30 minutes of contact time, immediately expose the plate as per latest SOP for plate exposure and detect the bacterial count per location per 2 hours.

5. Repeat the same procedure, step no. 1 to 4 to determine the efficacy of IPA 70% v/v solution after 24 hrs, 48 hrs, 72 hrs and 96 hours at room temperature

ACCEPTANCE CRITERIA

Diluted disinfectant solution, which is stored at room temperature, is effective when the test result of surface swab and Settle plate shows 90% reduction of the challenged microorganisms.

*  10-3 or 10-1 means 10 to power -1

HOLD TIME STUDY PROTOCOL FOR STERILISED GARMENTS FOR THEIR STERILITY

PROCEDURE

1. Prepare a Dacron bag contains 1 pair of garments used in  sterility testing area, and place 10 cut pieces (6x5 cm) of old dress in-between the dress.

2. Perform the sterilization of the bag as per the SOP for sterilization of dresses.

3. After sterilization place the Dacron bag in Garment cubicle for Hold time study.

4. Collect one cut piece from the dress bag aseptically and direct immerse into the sterilized Soyabean Casein Digest medium and check for the sterility, this sample shall be treated as initial sample. (0 Hour)

5. Similarly collect the dress pieces from the hold bag at regular intervals of 24 hr, 48 hr, 72 hr, 96 hr, and 120 hr and perform the sterility test.

6. Incubate the samples at the specified temperature for 14 days (20-25°C for 7 days and 30-35°C for 7 days).
Note: 0 Hrs starts when the sterilized garments placed in Garment cubicle after sterilization.

ACCEPTANCE CRITERIA

This study is carried out to establish the hold time of sterilized garments after sterilization.
Microbial determination : No growth should be observed in sterility test.

CONCLUSION

After complete evaluation of the hold time study for sterilized garments used for sterility testing a final hold time study summary report shall be prepared which should essentially contain discussion and conclusion which clearly determine the hold time period for sterilized garments.

VALIDATION PROTOCOL FOR HOLD TIME STUDY OF SWAB TEST SAMPLES

PROCEDURE

1.  Prepare 0.9% saline solution and dispense 10 ml quantity in test tube and put one sterile cotton swab in it and sterilize in autoclave at 15 lbs pressure and 121°C for 15 minutes or use pre sterilized swab tubes and fill with 10 ml sterile 0.9% saline solution.

2. Swab 5x5 cm2 area using parallel  overlapping stroke with slow rotation of swab.

3. Repeat the sampling with crossing first stroke at 90 degree angle. After taking the swab put the swab stick back into 0.9% saline solution tube.

4. After taking swab write the location and date of swab with marker on tube.

5. Bring the tubes to quality control laboratory and gently vortex for 1 minute. Perform the bio-burden testing using 1 ml of solution by membrane filtration.

6. Mark and incubate the plates at 22.5 ± 2.5 °C for 3 days followed by 32.5 ± 2.5 °C for 2 days inverted position.

7. Store the tubes containing the remaining solution with swab stick at 2-8 °C.

8. After 24 hours take the swab tubes and gently vortex for 1 minute.

9. Perform the bio-burden testing again using 1 ml solution from the tubes.

10. Mark and incubate the plates at 22.5 ± 2.5 °C for 3 days followed by 32.5 ± 2.5 °C for 2 days inverted position.

11. After incubation count and observe the plates of both studies for number of colonies on colony counter or under light source with the help of marker.

ACCEPTANCE CRITERIA

There shall be no increase in bio-burden on holding at 2-8°C for 24 hrs.

CONCLUSION

After complete evaluation of the hold time study for swab a final hold time study summary report shall be prepared which should essentially contain discussion and conclusion which clearly determine the hold time period for swab test samples.

VALIDATION OF PURE STEAM

Pure steam is used in various operations in pharmaceuticals but its use in sterilization is very common in pharmaceutical sterile manufacturing.
Pure steam system should be qualified. A WHO guide to good manufacturing practice (GMP) requirements clearly says to perform the performance qualification of pure steam. At the time of performance qualification of pure steam generation system, sample shall be taken from each steam user point and analyzed for three consecutive days. Purified water system must be qualified before starting the qualification of pure steam.

SAMPLING OF PURE STEAM

Sampling for Bacterial Endotoxin Test and chemical tests should be done separately. Depyrogenated tubes or bottles should be used for taking sample for bacterial endotoxin test. Allow the steam to drain for minimum one minute. Open the cap of bottle and fill the bottle with steam condensate by holding the bottle in the holder. Gloves should wear into the hands while sampling the pure steam. Tighten the cap of the bottle and mark with the sampling information. It sample is not analyzed within 2 hours of sampling, store the sample at 2-8 °C.

ANALYSIS OF PURE STEAM

Pure steam should be analyzed for following tests:-

1. Non-Condensable gases:- Non condensable gases are air and carbon dioxide those do not condense with the steam. These are generated due to their presence in the purified water that continuously circulates in the water distribution system. Non condensable gases should not be more than 3.5%.

2. Steam Dryness value:- Dry steam has more energy than the wet steam. Wet steam has water with it and does not have heat energy as dry steam. Dryness of steam is determined by the latent heat. Dryness of the pure steam should not be less than 90%. High moisture content can cause the loss in energy of steam and that may cause the longer sterilization time.

3.  pH:- Steam condensate is analyzed for pH value at 25 °C . It should be between 5-7.

4. Conductivity:- Conductivity should be tested with calibrated conductivity meter at 20 °C. Conductivity should not be more than 1.3 µS/cm.

5. Microorgansims:- Steam condensate is tested for microbial contamination using pore plate method. There should not any microbial contamination in steam condensate.

6. Endotoxin Test:- Determine the endotoxin in the pure steam condensate and it should not be more than 0.25 EU/ml as in water for injection.

LEAK TEST FOR AMPOULES

Ampoules are small glass containers that contain a sterile medicinal liquid intended for parentral use. Presence of capillary pores or tiny cracks can cause microbes or other dangerous contaminants to enter the ampoules or may lead to the leakage of contents to outside. This may lead to contamination of the sterile contents and also spoilage of appearance of the package.
Changes in temperature during storage can cause expansion and contraction of the ampoule and its contents, thereby accentuating interchange if an opening exists. Leaker test for ampoules is intended to detect incompletely sealed ampoules so that they can be discarded in order to maintain the sterile conditions of the medicines.Tip seals are more likely to be incompletely closed than pull seals.Open capillaries or cracks at the point of seal result in LEAKERS.

TEST PROCEDURE:

Leakers are detected by this process in a visible manner. Ampoules are placed in a vacuum chamber, completely submerged in a deeply colored dye solution of about 0.5 to 1% methylene blue.
A negative pressure is applied within the ampoule. Subsequent atmospheric pressure causes the dye to penetrate an opening thus making it visible after the ampoule has been washed.
The vacuum, about 27 inches Hg, should be sharply released after 30 minutes. Detection of leakers is prominent when ampoules are immersed in a bath of dye during autoclaving cycle as this has the advantage of accomplishing both leaker detection and sterilization in one operation.

DISADVANTAGES:

Capillaries of 15 micron or smaller diameter cannot be detected by this test.
Vials and bottles are not subjected to such a leaker test as the rubber closer is not rigid.

LEAKAGE TEST FOR INJECTABLE AND NON INJECTABLE PLASTIC CONTAINERS

Fill about 10 containers with water and fit with an appropriate closure.
Keep these containers in an inverted position at room temperature for about 24 hrs. Check for any leakage from any container.

WATER VAPOUR PERMEABILITY TEST

Take five containers and fill them with nominal volume of water. Heats seal these containers with an aluminium foil - polyehtylene laminate or other seal. Weigh the containers accurately and allow to stand for 14 days at a relative humidity of 55 - 65% and a temperature between 20 to 25@C. Reweigh the containers. The loss in weight in each container should not be more than 0.2%.

VARIOUS METHODS FOR LEAKAGE DETECTION

Above mentioned test are specific for certain types of material. Some other approaches also available to detect the leak in various types of container based upon different mechanisms. Let's see one by one:-

(1) OBSERVATION OF VISUAL DEFECTS (pinholes, capillaries):

This is done by leak test using methylene blue dye what I mentioned for ampoules.

(2) WEIGHT CHANGE:

This calculated by plotting a graph with loss or gain versus time under specifically defined condition (what I mentioned in water vapour permeability test).

(3) PRESSURE VACUUM CHANGES:

By the application of pressure and/or vacuum under defined conditions can help detect any leaks to external atmospheres like gases in the form of bubbles. Detection by visual inspection are limited to around 1 cm3 per minute with the possible detection of pinholes up to 20-25m.
Pressure decay systems are operated to a specified pressure and then monitored for pressure drop by which leaks down to 10-3 cm3/sec.
Pressure increase method can also be used in which leakage from a pack under vacuum is detected as a positive pressure change.
Flexible packs generally intend to extend (or indent) when the pack is under vacuum or pressure in sealed condition. A grossly leaking pack does not show any such movement and a slightly leaking one shows less movement. By assessing these movements the leak can be analyzed with the help of transducers or spring loaded sensors.
The sense of deflection and force changes can be detected with typical instruments like blister testers.
Both these methods are more sensitive than the older conventional vacuum with dye tests and can detect pinholes down to 10m. These tests are also non destructive and the time period is as small as 0.5 to unit second in order to minimize temperature effects.

(4) GASEOUS DETECTION TESTS:

This test makes use of gases associated with the product or a gas which is specifically introduced for the leakage detection process. The commonly used gases are:
Helium uses mass spectrometry which enables leakage to be detected down to 10-12 Pa m3s. This test needs high vacuum that may not be ideal for the component being tested. Other halogens are also used for gas detection.
Oxygen, i.e. Mocon Ox-tran: It uses a stream of dry nitrogen whereby presence of oxygen is detected coulometrically.
Carbon dioxide, e.g. Mocon Permatran C used to detect carbon dioxide in another dry gas using infrared technique.
Moisture vapour, e.g. Mocon Permatran W or Dynamic water vapour tester measures moisture by a photoelectric sensor.
Radio isotope tracer gas, e.g. using krypton 85 where a high sensitivity is reported.

(5) BURST TESTS:

The strength of the seals can be estimated by tensile tests as seal or peel tests or the air pressure that is sufficient to create rupture.
This test is carried out by placing a hypodermic needles arrangement and pressurizing the pack against it at a specific rate until the pack bursts. Depending on the nature of the seal the rupture may arise in the body of the pack or at the seal.
An alternative to this can be done by applying steadily increasing mechanical compression on a pack placed in a jig.

(6) MICROBIAL INTEGRITY:

Different methods have been developed under normal pressure and vacuum in order to check whether highly contaminated liquid, gel or media based material will grow back or penetrate closure systems. Alternative methods of leak detection are more preferable as variable results been observed with this method.

(7) CRACK, PINHOLE, CAPILLARY DETECTION:

The old conventional dye/vacuum immersion tests were used earlier to check the seal efficiency in ampoules. These methods have been replaced by electrical conductivity and capacitance type tests.
Typical equipments include NIKKA DENSOK AMPOULE INSPECTION MACHINE.
This machine employs a high frequency and high voltage. It distinguishes between good glass (a non conductor) and areas of cracks or pinholes where current will flow between the inner and outer glass surfaces.
There are other machines that make use of capacitance and dielectric constants where a material with defects will display a higher dielectric constant.

(8) THERMAL CONDUCTIVITY:

These make use of a thermistor bridge that is balanced against air and is subsequently upset if another gas leaks into the leak.

(9) CHEMICAL TRACER TESTS:

This test follows the principle of interaction between material i.e. ammonia on one side and hydrochloric acid on the other forms a white cloud of ammonium chloride indicating leakage. Pinhole can be well detected by this method.

(10) THERMOCOUPLE GAUGES:

This technique based on the mechanism of temperature change. These are mainly used to detect a drop in temperature when a solvent type systems escape under vacuum. It is also used to detect the presence of warmer gases.

TABLET COATING PROBLEMS

BLISTERING:- It is the  detachment of film from the tablet substrate as the elasticity of surface film compromised.

Cause- generally occur due to higher temperature during various stages of coating.

CHIPPING: It is generally a edge phenomena in which at the edge of tablet the surface film chipped off.

Cause decrese in rotation speed of pan, poor coating solution.

CRATERING: It is defect in which a defect in film coating leads to craters appearing exposing the tablet surface.

Cause insufficient drying time, high volume of coating solution used

PICKING: It is defect where a surface film is pulled away from the surface when the tablet sticks together and then part due to being detached away from the core. 

Cause over wetting of tablets due to polymer solution, improper drying.

PITTING: It is a defect in which deformation occur or pitts formation occur in the surface of a tablet core without any visible disruption of the film coating.

Cause it generally occur when the temp of core of the tablets generally higher than the melting points of material used in it. 

BLOOMING: It is defect where dullness in coating colour of tablet occur after  prolonged storage at high temperatures.

Causes use of low molecular weight plasticizer, 

BLUSHING: It is defect in which whitish specks or haziness occur  in the film.

Cause occur due to the high caoting temp. Leads to percipitation of polymers on surface. 

COLOUR VARIATION: A defect which involves variation in colour of the film.
Cause poor mixing, uneven spray pattern, migration of dyes during drying etc.

Infilling- it refers to filling of intagliations(distinctive words or symbols on tablet surface)

Cause due to accumulation or settling of foam in intagliations caused due to air spraying 

ORANGE PEEL EFFECT: It is surface defect resulting in appearence of an orange peel having rough and nonglossy texture.

Cause too high spray pressure, poor tablet composition, rapid drying etc

CRACKING/SPLITTING: It is defect in which the cracks in the films occur across the crown of the tablet (cracking) or splits around the edges of the tablet (Splitting).

Causes occur when the internal stress of the film exceeds the tensile strength of fim, use of high molecular weight polymers.

Links 

http://www.pharmainfo.net/tablet-ruling-dosage-form-years/problems-tablet-manufacture-and-related-remedies/tablet-coating-problems

https://www.lfatabletpresses.com/articles/different-tablet-coating-defects-and-remedies

TABLET PROBLEMS

Problems occuring due to process

CAPPING:- It occur when the upper or lower part of the tablet separates horizontally (partially or complete) from the tablet body and comes off as a cap, during ejection of tablet from the compression machine. 

Causes- over drying of granules, lesser amount of binder, improper tooling, air entrapment.

LAMINATION:- ‘Lamination’ is the separation of a tablet into two or more distinct horizontal layers.

Causes- over speed of turret, improper setting of lower punches, over dried granules.

Problems due to excipients

CHIPPING:- ‘Chipping’ is defined as the breaking of tablet edges during the release of tablet from the press or during handling and coating operations.

Cause - punch tips worn, over drying of granules, lower amount of lubricants.

CRACKING:- When the small and fine cracks observed on the upper and lower central surface of tablets, or sometimes on the sidewalls of tablets are referred to as Cracking.

Causes- over dried granules, large size of granules, tablets expansion.

STICKING:- when the tablet material adhere to the die wall is referred as sticking.
Filming is a slow form of sticking and occur due to excessive moisture in the granules.

Cuses- excessive binder, lesser drying of granules

PICKING:- when a small amount of material from a tablet is sticking and  removed off from the tablet-surface by a punch face is called picking.
The problem is more common on the upper punch faces than on the lower ones.

Cause lesser drying of granules, lesser addition of lubricants, binder addition was high embossing letters on punch tips

MOTTLING:- it is the  unequal distribution of colour on a tablet, with light or dark spots present in an uniform surface.

Causemigration of die to the durface of granules during drying improper mixing of die coloured drug used with colourless excipients

DOUBLE IMPRESSION:- ‘Double Impression’ occur due to free rotation of punches, which have a monogram or other engraving on their face.

Cause - occur due to free rotation of punches