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Sunday, December 28, 2008

TOTAL MICROBIAL COUNT [BACTERIA, FUNGI/MOLDS AND YEAST]

OBJECTIVE
Test for Total Microbial Count is provided to determine compliance with the requirements given in individual monograph / specifications.
PRINCIPLE
Total microbial count is the estimation of the number of viable aerobic micro-organisms present in the pharmaceutical articles of all kinds, from raw materials to the finished forms.
BUFFERS AND MEDIA
1. pH 7.2 Phosphate Buffer
Stock Solution
Dissolve 34 g of Monobasic Potassium Phosphate in about 500 mL of water contained in a 1000 mL volumetric flask. Adjust to pH 7.2 ± 0.1 by the addition of 4 % w/v aqueous solution of Sodium Hydroxide (about 175 mL), add water to volume, and mix. Dispense and sterilize. Store under refrigeration.
For use, dilute the Stock Solution with water in the ration of 1 to 800 , and sterilize in an autoclave at 121 OC , 15 lb pressure for about 15 min.
2. Fluid Soyabean Casein Digest Medium
Pancreatic Digest of Casein 17.0 g
Papacy Digest of Soybean Meal 3.0 g
Sodium Chloride 5.0 g
Dibasic Potassium Phosphate 2.5 g
Dextrose (C6H12O6. H2O) 2.5 g
Distilled Water 1000 mL
Final pH after Sterilization 7.3 ± 0.2
Dissolve the solids in the water, warming slightly to effect solution. Cool to room temperature and adjust pH. 7.3 ± 0.2 and Filter, if necessary. Distribute the media into suitable containers and sterilize in an autoclave at 121 OC for about 15min.
3. Fluid Casein Digest-Soy Lecithin-Polysorbate 20 Medium
Pancreatic Digest of Casein 20.0 g
Soy Lecithin 5.0 g
Polysorbate 20 40 mL
Water 960 mL
Dissolve the pancreatic digest of casein and soy lecithin in 960 mL of water, heating in a water batch at 48 - 50 OC for about 30 min. to effect solution. Add 40 mL of Polysorbate 20. Mix, and dispense as desired and sterilize in an autoclave at 121 OC for about 15min..
4. Sabouraud Glucose Agar with Antibiotics
Peptones (meat and Casein) 10.0 g
D- Glucose Monohydrate 40.0 g
Agar 15.0 g
Water 1000 mL
Adjust the pH to 5.4 +2. Sterilize by heating in an autoclave at 121 OC for 15 min. Immediately before use, add 0.10 g of Benzylpenicillin Sodium and 0.10 g of Tetracycline per liter of medium as sterile solutions or, alternatively, add 50 mg of Chloramphenicol per liter of medium.
5. Peptone Water
(Buffered Sodium Chloride - Peptone Solution pH 7.0)
Potassium Dihydrogen Orthophosphate 3.56 g
Disodium Hydrogen Orthophosphate 7.23 g
Sodium Chloride 4.30 g
Peptone(meat and Casein) 1.0 g
Water 1000 mL
0.1 to 1.0 % w/v Polysorbate 20 or Polysorbate 80 may be added. Sterilize by heating in an autoclave at 121 OC for 15 min.
6. Potato Dextrose Agar Medium
Cook 300 g of peeled and diced potatoes in 500 mL of water prepared by distillation, filter through cheesecloth, add water prepared by distillation to make 1000 mL and add the following :
Agar 15 mg
Glucose 20 mg
pH after sterilization 5.6 ± 0.2
Dissolve by heating, and sterilize.
For use, just prior to pouring the plates, adjust the melted and cooled medium to 45 OC with Sterile Tartaric Acid solution (1 in 10) to a pH of 3.5 ± 0.1 Do not reheat the pH 3.5 medium.
All the above media should be incubated for 24 hours at 37 OC before use. Any contaminated media should be discarded.
Instead of preparing media , one can also use dehydrated media of Hi media / Difco and rehydrate the required quantity as per instructions on the bottle label, dispense in required quantities and sterilize.
Preliminary Testing
The methods given herein are invalid unless it is demonstrated that the test specimens to which they are applied to not, of themselves, inhibit the multiplication, under the test conditions, of micro-organisms that may be present. Therefore, prior to doing the tests, inoculate diluted specimens of the substance being examined, with separate viable cultures of Escherichia coli, B. subtilis and Staphylococcus aureus. Add 1 mL of not less than 10 -3 dilution of a 24 hour broth culture of the micro-organism to the first dilution (in buffer solution pH 7.2 , Fluid Soybean-Casein Digest medium, of the test material and following the test procedure. If the organisms fail to grow in the relevant medium the procedure should be modified by
a. increasing the volume of diligent, the quantity of test material remaining the same,
b. incorporating a sufficient quantity of a suitable inactivating agent in the diluents or by,
c. combining the afore-mentioned modifications so as to permit growth in the media.
If inhibitory substances are present in the sample 0.5 % of soy lecithin and 4 % of the Polysorbate 20 may be added to the culture medium.. Alternatively, repeat the test as described in the previous paragraph, using fluid casein digest - soy lecithin - polysorbate 20 medium to demonstrate neutralization of preservatives or other anti-microbial agents in the test material.
PRE - TREATMENT OF THE PREPARATION BEING EXAMINED
Use separate 10 mL or 10 g specimens for testing.
Water - Soluble Products :
Dissolve or dilute 10 g or 10 mL of the preparation being examined, unless otherwise prescribed, in Peptone Water or another suitable medium shown not to have anti-microbial activity under the conditions of the test and adjust the volume to 100 mL with the same medium. If necessary, adjust the pH to about 7.
Non - Fatty Products Insoluble in Water :
Suspend 10 g or 10 mL of the preparation being examined, unless otherwise prescribed, in Peptone Water or another suitable medium shown not to have anti-microbial activity under the conditions of the test and dilute to 100 mL with the same medium. If necessary divide the preparation being examined and homogenize the suspension mechanically.
A suitable surface - active agent such as 0.1 % w/v of Polysorbate 80 may be added to assist the suspension of poorly wettable substances. If necessary, adjust the pH of the suspension to
about 7
Fatty Products :
Homogenize 10 g or 10 mL of the preparation being examined, unless otherwise prescribed with 5 g of Polysorbate 20 or Polysorbate 80. If necessary, heat to not more than 40 OC . Mix carefully while maintaining the temperature in a water bath or in an oven. Add 85 mL of Peptone Water or another suitable medium shown not to have anti-microbial activity n the conditions of the test, heated to not more than 40 OC if necessary. Maintain this temperature for the shortest time necessary for formation of an emulsion and in any case for not more than 30 min. If necessary, adjust the pH of the emulsion to about 7.
For a Fluid Specimen in Aerosol Form :
Chill the container in an Alcohol-dry ice mixture for approximately 1 h, cut open the container, allow it to reach room temperature, permit the propellant to escape, or warm to drive off the propellant if feasible and transfer the quantity of test material required for the procedures specified in one of the two preceding paragraphs, as appropriate. Where 10 g or 10 mL of the specimen, whichever is applicable, cannot be obtained from 10 containers in aerosol form, transfer the entire contents from 10 chilled containers to the culture medium, permit the propellant to escape, and proceed with the test on the residues.
Effectiveness of Culture Media and Validity of the Counting Methods :
When necessary, operate as follows. Grow the following test strains separately in tubes containing casein soybean digest broth at 30 - 35 OC for 18 - 24 hours or, for Candida albicans, at 20 -25 OC for 48 hours.
Staphylococcus aureus such as NCIMB 8625 (ATCC 6538, CIP 530156) or NCIMB 9518 (ATCC 6538, CIP 4038)
Bacillus Subtilis, such as NCIMB 8054 ( ATCC 6633, CIP 52.62)
Candida albicans, such as ATCC 2091 (CIP 1180.79) or ATCC 10231 (NCPF 3179, CIP 48.72)
Dilute portions of each of the cultures using buffered sodium chloride - peptone solution pH 7.0 to make test suspensions containing about 100 viable micro-organisms per milliliter. Use the suspension of each of the micro-organisms separately as a control of the counting methods in the presence and absence of the preparation being examined if necessary.
When testing the method, a count for any of the test organisms differing by not more than a factor of 10 from the calculated value for the inoculum should be obtained. To test the sterility of the medium and of the diligent and the aseptic performance of the test, carry out the total viable aerobic count method using sterile buffered sodium chloride - peptone solution pH 7.0 as the test preparation. There should be no growth of micro-organisms.
PROCEDURE
As per IP
1. Dissolve or suspend 10 g of the substance being examined in sufficient buffer solution, pH 7.2, Fluid Soybean - Casein Digest Medium, or Fluid Casein Digest-Soy lecithin - Polysorbate 20 medium to produce 100 mL.
2. Perform the test for the absence of inhibitory (anti-microbial) properties as described under Preliminary Testing before the determination of Total Microbial Count.
3. Add the substance being examined, to the medium not more than 1 hour after preparing the appropriate dilution’s for inoculation.
A. Plate Method
This method is applicable for substances that are sufficiently soluble or translucent.
1. Dilute further, if necessary, the sample liquid prepared as described above, so that 1 mL will be expected to yield between 30 to 300 colonies.
2. Pipette 1 mL of the final dilution into each of two sterile petri dishes.
3. Immediately add to each dish 15 to 20 mL of Soybean-Casein digest agar medium that has previously been melted and cooled to about 45 OC.
4. Cover the petridishes, mix the sample with the agar by tilting or rotating the dishes, and allow the contents to solidify to room temperature.
5. Invert the petri dishes, and incubate for 48 to 72 hours at 37 OC.
6. After incubation examine the plates of growth, count the number of colonies, and express the average for the two plates in terms of number of micro-organisms per g of the substance.
7. If no colonies are recovered from the dishes representing the initial 1 : 10 dilution of the substance, express the results as “less than 10 micro-organisms per g of substance”.
B. Multiple - Tube Method
This method is applicable for substances that are insoluble or not translucent.
1. Into each of the fourteen test tubes of similar size place 9 mL of sterile Fluid Soyabean casein digest medium.
2. Arrange 12 of the tubes in four sets of three tubes each. Put aside one set of three tubes to serve as the controls.
3. Into each of three tubes of one set [“100” ] and into fourth tube (A) pipette 1 mL of the Solution of suspension of the specimen, and mix.
4. From tube A pipette 1 mL of its contents into the one remaining tube (B) not included in a set, and mix.
5. These two tubes contain 100 mg (or 100 µl) and 10 mg (10 µl) of the specimen respectively.
6. Into each of the second set [“10”] of three tubes pipette 1 mL from tube A and into each tube of the third set [“1”] pipette 1 mL from tube B.
7. Discard the unused contents of tubes A and B.
8. Close well, and incubate all of the tubes.
9. Following the incubation period, examine the tubes for growth; the three control tubes remain clean and the observations in the tubes containing the specimen, when interpreted by reference to Table I. indicate the most probable number of micro-organisms per g or per mL of specimen.
As per BP
Determine the total viable aerobic count of the preparations being examined by the membrane filtration method, the plate count method or the serial dilution method as prescribed. Suitable degrees of dilution should be used so that the number of colony forming units is within the limits suggested for the method to be used.
A. Membrane Filtration
Use membrane filters having a normal pore size not greater than 0.45 µm and 50 nm in diameter the effectiveness of which in retaining bacteria has been established. For example, Cellulose nitrate filters are used for aqueous, oily and weakly alcoholic solutions and Cellulose acetate filters for strongly alcoholic solutions.
1. The filtration apparatus and membrane are sterilized by appropriate means and are designed so that the solution being examined can be introduced and filtered under aseptic conditions and so as to permit the removal of the membrane for transfer to the culture medium.
2. Transfer 10 mL or a quantity to each of two membrane filters and filter immediately.
3. If necessary, dilute the pretreated preparation so that a colony count of 10 to 100 may be expected.
4. Wash each membrane by filtering through it three or more successive quantities , each of approximately 100 mL, of a suitable liquid such as buffered Sodium Chloride - peptone pH 7.0. For fatty substances, this liquid may contain a suitable surface active agent such as polysorbate 20 or polysorbate 80. Transfer one of the membrane filters, intended primarily for the enumerate of bacteria, to the surface of the plate of casein soybean digest agar and the other, intended primarily for the surface of a plate of sabouraud glucose agar with antibiotics.
5. Incubate the plates for 5 days, unless a more reliable count is obtained in a shorter time, at 30 - 35OC in the test intended to detect bacteria and at 20 - 25 OC in the test intended to detect fungi.
6. Count the number of colonies that are formed. Calculate the number of micro-organisms per gram or per milliliter of the preparation being examined, if necessary counting bacteria and fungi separately.
B. Plate Count
i. For Bacteria
1. Using petri dishes 9 to 10 cm in diameter, add to each dish a mixture of 1 mL of the pretreated preparation and about 15 mL of liquefied casein soyabean digest agar at not more than 45 OC .
2. Alternatively, spread the pretreated preparation on the surface of the solidified medium in a petri dish of the same diameter.
3. If necessary, dilute the pre-treated preparation as described above so that a colony count of not more than 300 may be expected.
4. Prepare at least two such petri dishes using the same dilution and incubate at 30 - 35OC for unless a more reliable count is obtained in a shorter time.
5. Count the number of colonies that are formed.
6. Calculate the results using plates with the greatest number of colonies but taking 300 colonies per plate as the maximum consistent with good evaluation.
ii. For Fungi
1. Using petri dishes 9 - 10 cm in diameter, add to each dish a mixture of 1 mL of the pre-treated preparation and about 15 mL of liquefied Sabouraud glucose agar with antibiotics at not more than 45 OC.
2. Alternatively, spread the pretreated preparation on the surface of the solidified medium in a petri dish of the same diameter. If necessary, diluted the pretreated preparation as described above so that a colony count of not more than 100 may be expected.
3. Prepare atleast two such plates using the same dilution and incubate at 20 - 25 OC for 5 days, unless a more reliable count is obtained in a shorter time.
4. Count the colonies that are formed. Calculate the results using plates with not more than 100 colonies.
Serial Dilution
1. Prepare a series of 12 tubes each containing 9 - 10 mL of caseing soyabean digest broth.
2. To each of the first three tubes add 1 mL of the preparation diluted, dissolved or homogenized in the proportion 1 in 10 , as described above.
3. To the next three tubes add 1 mL of a 2 in 100 dilution of the preparation and to the next three tubes add 1 mL of a 1 in 100 dilution of the preparation.
4. To the last three tubes add 1 mL of the diligent.
5. Incubate the tubes at 30 - 35 OC for at least 5 days.
6. The last three tubes should show no microbial growth.
7. If the reading of the results is difficult or uncertain owing to the nature of the preparation being examined, sub-culture on a liquid or solid medium and read the results after a further period of incubation.
8. Determine the most probable number of micro-organisms per gram or per milliliter of the preparation being examined from Table I.
As per USP
1. For specimens that are sufficiently soluble or translucent to permit use of the plate Method, otherwise, use Multiple-tube Method.
2. With either method, first dissolve or suspend 10 g of the specimen if it is a solid, or 10 mL , accurately measured if the specimen is a liquid, in pH 7.2 Phosphate Buffer, Fluid Soyabean-Casein Digest Medium, or Fluid Casein Digest-soy Lecithin - Polysorbate 20 Medium to make 100 mL
3. For viscous specimens that cannot be pipetted at this initial 1:10 dilution, dilute the specimen until a suspension is obtained i.e., 1:50 or 1: 100 etc., that can be pipetted.
4. Perform the test for absence of inhibitory (anti-microbial) properties as described under Preparatory Testing before the determination of Total Aerobic Microbial Count.
5. Add the specimen to the medium not more than 1 h after preparing the appropriate dilution’s for inoculation.
A. Plate Method
1. Dilute further, if necessary, the fluid so that 1mL will be expected to yield between 30 and 300 colonies.
2. Pipette 1 mL of the final dilution onto each of two sterile petridishes.
3. Promptly add to each dish 15 - 20 mL of Soyabean Casein Digest Agar Medium when testing for bacteria and add 15 - 20 mL Sabouraud Dextrose Agar medium or Potato Dextrose Agar medium for molds and yeasts,. that previously has been melted and cooled to approx. 45 OC.
4. Cover the petridishes, mix the sample with the agar by tilting or rotating the dishes, and allow the contents to solidify at room temperature.
5. Invert the petridishes, and incubate for 48 - 72 hours at 37 OC for bacteria and incubate for 5 - 7 days at 20 - 25 OC for molds and yeasts.
6. Following incubation, examine the plates for growth, count the number of colonies, and express the average for the two plates in terms of the number of micro-organisms per g or per mL of specimen.
7. If no microbial colonies are recovered from the dishes representing the initial 1 : 10 dilution of the specimen, express the results as “less than 10 micro-organisms per g or per mL of specimen.”
B. Multiple Tube Method
1. Place 9 mL of sterile Fluid Soybean casein digest medium into each of fourteen test tubes of similar size.
2. Arrange twelve of the tubes in four sets of the three tubes each.
3. Put aside one set of three tubes to serve as the controls.
4. Into each of three tubes of one set [“100”] and into a fourth tube (A) pipette 1 mL of the solution or suspension of the specimen, and mix.
5. From tube a , pipette 1 mL of its contents into the one remaining tube (B) not included in a set, and mix. These two tubes contain 100 mg (100 µl) and 10 mg (10 µl) of the specimen, respectively.
6. Into each of the second set [“10”] of three tubes pipette 1 mL from the tube A, and into each tube of the third set [“1”] pipette 1 mL from tube B.
7. Discard the unused contents of tubes A and B.
8. Close well, and incubate all of the tubes. Following the incubation period, examine the tubes for growth; the three control tubes remain clear and the observation in the tubes containing the specimen, when interpreted by reference to Table I, indicate the most probable number of micro-organisms per g r per mL of specime
Most Probable Count by Multiple - Tube Method
TABLE - I




Calculations
Take the average count of each dilution and multiply it by the dilution factor. Calculate the average form the readings obtained. This gives the count per gram or per mL of the sample/
Interpretation of Results :
If a limit is prescribed, it is to be interpreted as follows :
10 2 micro-organisms, MAXIMUM LIMIT OF ACCEPTANCE: 5 x 10 2
10 3 micro-organisms, MAXIMUM LIMIT OF ACCEPTANCE : 5 x 10 3
NOTE: Use the method as per the pharmacopoeial status (grade) of the material. In case of In- house specifications, follow the method as per the IP or as specified

ABNORMAL / UNDUE TOXICITY / SAFETY TEST

OBJECTIVE
The test for Abnormal / Undue Toxicity / Safety test is provided to determine compliance with the requirements given in individual monograph / specifications.
PRINCIPLE
This test is intended to detect any unexpected, unacceptable, biological reactivity in a substance. This is an invitro test designed for safety assessment of biologics and biotechnology derived products.
ANIMAL QUARTERS
1. Keep the animals individually in a quiet area with an appropriate uniform temperature allowing a variation of ± 2 OC and uniform humidity and free from disturbance.
2. Carry out the test in a quiet room where there is no risk of disturbance which can excite the animals and in which the room temperature is within 3 OC of that of their living quarters or in which the animals have been kept for at least 18 h before the test.
3. Withhold food from the animals overnight and until the test is completed , with hold water only during the test.
TEST FOR UNDUE TOXICITY ( As per USP )
GENERAL TEST
1. Inject intravenously into each of 5 healthy mice, weighing between 17 - 22 g the quantity of the substance to be examined prescribed in the individual monograph, dissolved in 0.5 mL of water for injection should normally occupy about 15 - 30 sec, unless otherwise prescribed in the individual monograph.
2. The substance passes the test if none of the mice dies within 24 h or within such time as is specified in the individual monograph.
3. If one of the animals dies, repeat the test. The substance passes the test if none of the animals in the second group dies within the time interval specified.
TEST FOR ABNORMAL TOXICITY (As per BP)
Use Method - A unless otherwise directed.
Where under the heading Abnormal Toxicity in an individual monograph it states that the substance being examined is to be dissolved in Albumin solution, this reagent may be replaced by a solution in water for injections containing an appropriate amount of a suitable protein that has been shown to be pyrogen-free and also shown, by a suitable method, to be free from proteolytic activity. The solution is adjusted to an appropriate pH.
METHOD A
1. Inject intravenously into each of five mice the quantity of the substance being examined prescribed in the monograph, dissolve in 0.5 mL of water for injection or Sodium Chloride injection unless otherwise prescribed in the monograph.
2. The injection should normally be administered over a period of 15 to 30 sec., unless otherwise prescribed in the monograph.
3. None of the mice should die within 24 h, or within such time as is specified in the monograph.
4. If one of the animals dies within the specified time, repeat the test. None of the animals in the second group should die within the specified time.
METHOD B
1. Unless otherwise prescribed in the monograph inject one dose but not more than 1.0 mL intraperitoneally into each of 5 mice and one dose but not more than 5.0 mL intraperitoneally into each of two guinea-pigs.
2. None of the animals should die or show of ill health within 7 days.
3. If one of the animals dies or shows signs of ill health within 7 days, repeat the test. None of the animals in the second group should die or shows signs of ill health within 7 days.
SAFETY TEST ( As per USP)
1. Select five healthy mice not previously used for testing, weighing between 17 - 22 g, unless otherwise specified in the individual monograph, and maintained on an adequate balanced diet.
2. Prepare a test solution as directed in the individual monograph. Unless otherwise directed in the individual monograph, inject intravenously a dose of 0.5 mL of the test solution into each of the mice, using a 26 gauge needle of suitable length.
3. Observe the animals over the 48 h. following the injection.
4. If, at the end of 48 h., all of the animals survive and not more than one of the animals shows outward symptoms of a reaction not normally expected of the level of toxicity related to the article, the requirements of this test are met.
5. If one or more animals die or if more than one of the animals shows signs of abnormal or untoward toxicity of the article under test, repeat the test using at least another 10 mice similar to those used in the initial test, but weighing 20 ± 1 g. In either case, if all of the animals survive for 48 h. and show no symptoms of a reaction indicative of an abnormal or undue level of toxicity of the article, the requirements of the test are met.
NOTE:
1. For biologics, perform the test according to the procedures prescribed in the Federal regulations (see Biologics [1041], Section 610.11), using not less than 2 mice similar to those described above but weighing less than 22 g and not less than 2 healthy guinea - pigs weighing less than 400 g.
2. For a liquid product or a freeze-dried product that has been constituted as directed in the labeling, inject a volume of 0.5 mL intraperitoneally into each mouse, and inject a volume of 5.0 mL intaperitoneally into each guinea-pig.
3. For freeze-dried products for which the volume of constitution is not indicated in the label, or for non liquid products other than freeze-dried products, perform the test using the route of administration, test dose, and diligent approved by the Center for Biologics Evaluation and Research (FDA) on the basis of substantial evidence demonstrating that the test variation will assure sensitivity equal to or greater than that of the test described above.
INTERPRETATION :
1. Observe the animals for a minimum observation period of 7 days.
2. If all of the animals survive the test period, do not exhibit any response that is not specific for or expected from the product and that may indicate a difference in such product quality, and weigh no less at the end of, the test period than at the time of injection, the requirements of the test are met.
3. If the article fails to meet the requirements, the test may be repeated as in the initial test, in the one or both species in which the requirements were not met .
4. If the animals fulfill the criteria specified for the initial test, the article meets the requirements of the test. If the article fails to meet the requirements after the first repeat test, and not less than 50 % of the total number of animals of the species in which the requirements of the test were not met in the combined initial and first retests have survived, a second retest may be performed.
5. Use twice the number of animals of the relevant species used in the initial test. If the animals fulfill the criteria specified for the initial test, the requirements of the test are met.
Note: Use the method as per the Pharmacopoeial status (grade) of material. In case of In-house Specifications, follow the method as per IP or as specified.

IDENTIFICATION OF STAPHYLOCOCCUS AUREUS

OBJECTIVE
The test for Staphylococcus aureus is provided to determine compliance with the requirements given in individual monograph/specifications,.
PRINCIPLE
Identification of Staphylococcus aureus is the detection of a pathogenic bacteria which causes infection in human body. It can be identified by using specific, differential media which support growth of only Staphylococcus aureus.
REAGENT AND MEDIA
Fluid Soybean - Casein Digest Medium
Pancreatic digest of Casein 17.0 g
Papai digest of Soybean meal 3.0 g
Sodium Chloride 5.0 g
Dibasic Potassium Phosphate 2.5 g
Dextrose (C6H12O6.H2O) 2.5 g
Distilled Water 1000 mL
Final pH after sterilization 7.3 ± 0.2
Dissolve the solids in the water, warming slightly to effect solution. Cool to room temperature and add, if necessary, sufficient 0.1 N Sodium Hydroxide to give a final pH after sterilization of between 7.1 and 7.5. Filter, if necessary, to clarify. Distribute into suitable containers and sterilize in an autoclave at 121 OC for about 15 min.
Mannitol - Salt Agar Medium
Pancreatic digest of Casein 5.0 g
Peptic digest of animal tissue 5.0 g
Beef Extract 1.0 g
D-Mannitol 10.0 g
Sodium Chloride 75.0 g
Agar 15.0 g
Phenol Red 0.025 g
Water 1000 mL
Final pH after sterilization 7.4 ± 0.2
Mix, then heat with frequent agitation, and boil for 1 min. to effect solution.
Vogel - Johnson Agar Medium
Pancreatic Digest Casein 10.0 g
Yeast extract 5.0 g
Mannitol 10.0 g
Dibasic Potassium Phosphate 5.0 g
Lithium Chloride 5.0 g
Glycine 10.0 g
Agar 16.0 g
Phenol Red 25.0 mg
Water 1000 mL
Boil the solution of solids for 1 min. sterilize, cool to between 45 - 50 OC and add 20 mL of sterile Potassium tellurite solution (1 in 100)
Baird - Parker Agar Medium
Pancreatic digest of Casein 10.0 g
Beef Extract 5.0 g
Yeast Extract 1.0 g
Lithium Chloride 5.0 g
Agar 20.0 g
Glycine 12.0 g
Sodium Pyruvate 10.0 g
Water 950 mL
pH after sterilization 7.2 ± 0.2
Heat to boiling for 1 min., shaking frequently. Adjust the pH so that after sterilization it is 6.6 - 7.0. Sterilize by heating in an autoclave at 121 OC for 15 min., cool to 45 - 50 OC and add 10 mL of sterile 1 % w/v solution Potassium tellurite and 50 mL of egg yolk emulsion.
All the above media should be incubated for 24 h. at 37 OC before use. Any contaminated media should be discarded Instead of preparing media, one can also use dehydrated media of Hi media / Difco and rehydrate the required quantity as per instruction son the bottle label, dispense in required quantities and sterilize at 15 psi at 121 OC for 15 min.
PRE-TREATMENT OF THE PREPARATION BEING EXAMINED
Water-Soluble Products
1. Dissolve or dilute 10 g or 10 mL of the preparation being examined, unless otherwise prescribed, in Lactose broth or another suitable medium shown not to have anti-microbial activity under the conditions of the test and adjust the volume to 100 mL with the same medium.
2. If necessary, adjust the pH to about 7.0
Non-Fatty Products Insoluble in Water:
1. Suspend 10 g or 10 mL of the preparation being examined, unless otherwise prescribed, in Lactose broth or another suitable medium shown not to have anti-microbial activity under the conditions of the test and dilute to 100 mL with the same medium.
2. If necessary divide the preparation being examined and homogenize the suspension mechanically.
3. A suitable surface-active agent such as 0.1 % w/v of Polysorbate 80 may be added to assist the suspension of poorly wettable substances.
4. If necessary, adjust the pH of the suspension to about 7.
Fatty Products :
1. Homogenize 10 g or 10 mL of the preparation being examined, unless otherwise prescribed with 5 g of Polysorbate 20 or Polysorbate 80.
2. If necessary, heat to not more than 40 OC
3. Mix carefully while maintaining the temperature in a water bath or in an oven.
4. Add 85 mL of Lactose broth or another suitable medium shown not to have anti-microbial activity in the conditions of the test, heated to not more than 40 OC, if necessary,
5. Maintain this temperature for the shortest necessary for formation of an emulsion and in any case for not more than 30 min.
6 If necessary, adjust the pH of the emulsion to about 7.0
For Fluid Specimen in Aerosol Form:
1. Chill the container in an Alcohol-dry ice mixture for approx. 1 h., cut open the container allow it to reach room temperature, permit the propellant to escape, or warm to drive off the propellant if feasible and transfer the quantity of test material required for the procedures specified in one of the two proceeding paragraphs, as appropriate.
2. Where 10 g or 10 mL of the specimen, whichever is applicable, cannot be obtained from 10 containers in aerosol form, transfer the entire contents from 10 chilled containers to the culture medium, permit the propellant to escape, and proceed with the test on the residues.
PROCEDURE
As per IP’95
Primary Test
1. Place the prescribed quantity in a sterile screw-capped jar containing g 100 mL of fluid soybean-casein digest medium and incubate at 35 - 37 OC for 24 - 48 h.
2. Subculture on a plate containing a layer of Mannitol salt agar medium or Vogen-Johnson agar medium and incubate at 35 - 37 OC for 18 - 24 h.
3. Examine the resulting growth by Gram’s staining and apply the coagulase test. Gram-positive cocci (in cluster) in yellow colonies (in Mannitol salt agar medium) and in colonies, black surrounded by yellow zones (in Vogen - Johnson agar medium), and giving an positive coagulase test indicate the presence of Staphylococcus aureus.
Confirmatory Test :
Coagulase Test:
1. Transfer representative colonies from the agar surfaces of the Mannitol salt agar medium or Vogel-Johnson agar medium to individual tubes, each containing 0.5 mL of mammalian, preferably rabbit or horse, plasma with or without suitable additives. Incubate in a water bath at 37 OC examine the tubes after 24 h.
2. If coagulation in any degree is observed, the test is positive.
As per BP
Primary Test:
1. Inoculate 100 mL of casein digest broth with a quantity of the solution, suspension or emulsions thus obtained containing 1 g or 1 mL of the preparation being examined, after pretreatment as described above.
2. Mix and incubate at 35 - 37 OC for 24 - 48 h.
3. Subculture on a suitable medium such as Baird-Parker agar.
4. Incubate at 35 - 37 OC for 24 - 48 h.
5. If no growth of micro-organisms is detected, the preparation being examined passes the test.
6. Black colonies of Gram-positive cocci often surrounded by clean zones may indicate the presence of Staphylococcus aureus.
Confirmatory Test:
1. For catalase-positive cocci, confirmation may be obtained, for example, by coagulase and deoxyribonuclease tests.
2. The preparation being examined passes the test if cultures of the type described do not appear or if the Confirmatory biochemical tests are negative.
As per USP
Primary Test
1. To the specimen ad Fluid Soybean-Casein digest medium to make 100 mL mix, and incubate.
2. Examine the medium for growth and if growth is present, use an inoculating loop to streak a portion of the medium on the surface of Vogel-Johnson agar medium (or Baird-Parker agar medium, or Mannitol-Salt agar medium), on petridishes.
3. Cover and invert the dishes and incubate.
4. If, upon examination, none of the plates contains colonies having the characteristics listed in Table I for the media used, the test specimen meets the requirements for freedom from Staphylococcus aureus.
Confirmatory Test
Coagulase Test
1 With the aid of an inoculation loop, transfer representative suspected colonies from agar surfaces of Vogel-Johnson agar medium (or Baird-Parker agar medium, or Mannitol-Salt agar medium) to individual tubes, each containing 0.5 mL of mammalian, preferably rabbit or horse, plasma with or without suitable additives.
2. Incubate in a water bath at 37 OC, examining the tubes after 3 h. and subsequently at suitable intervals up to 24 h.
3. Test positive e and negative controls simultaneously with the unknown specimens.
4. If no coagulation in any degree is observed, the specimen meets the requirements of the test for absence of Staphylococcus aureus.
TABLE I
Morphological Characteristics of Staphylococcus aureus on Selective Agar Media


NOTE:
1. Carry out a control test by repeating the test, adding the prescribed quantity and a volume of broth containing 10 to 50 cells / colonies of Staphylococcus aureus (ATCC 6538; NCTC 10788), prepared from a 24 h. culture in fluid soybean-casein digest medium, to a sterile screw-capped jar containing 100 mL of fluid soybean-casein digest medium.
2. The test is invalid if the results do not indicate that the control contains Staphylococcus aureus.
3. Use the method as per the pharmacopoeial status (grade) of the material. In case of In-house specifications, follow the method as per IP or as specified.

Friday, December 26, 2008

TEST FOR ANTIMICROBIAL PRESERVATIVE EFFECTIVENESS (PRESERVATIVE EFFICACY)

1.0 OBJECTIVE
The test for Anti-microbial preservative - Effectiveness is provided to determine compliance with the requirements given in individual monograph / specifications.
2.0 PRINCIPLE
Preservative efficacy test is basically a challenge test in which the function of a preservative in pharmaceutical preparation use to be checked by challenging it with different microorganisms (bacteria, fungi and yeast.)
Test Organisms
Aspergillus niger IMI 147 007 (ATCC 16404, IP 1431.83)
Candida albicans NCPE 3179 (ATCC 10231, IP 48.72)
Pseudomonas aeruginosa NCIMB 8626 (ATCC 9027, CIP 82.118)
Staphylococcus aureus NCTC 10788 (NCIMB 9518, ATCC 6538, CIP 4.13) .
Single-strain challenges are used and the designated micro-organisms are supplemented by other strains or species that may represent likely contaminants to the preparation. For example, Escherichia coli [NCIMB 8545 (ATCC 8739, CIP 53.126)] is used for all oral preparations and Zygosaccharomyces Rouxii[NCYC 381 (IP 2021.92)] for oral preparations containing a high concentration a sugar.
Medium
a. Soyabean Casein Digest Agar Medium . (or Tryptone Soya Agar)
Pancreatic digest of Casein 17.0 g
Papaic digest of Soyabean meal 3.0 g
Sodium Chloride 5.0 g
Dibasic Potassium Phosphate 2.5 g
Dextrose (C6H12O.H2O) 2.5 g
Distilled water 1000 mL
Final pH after Sterilization 7.3 ± 0.2
Dissolve the solids in the water, warming slightly to effect solution. Cool to room temperature and add, if necessary, sufficient 0.1 N Sodium Hydroxide to give a final pH after sterilization of between 7.1 and 7.5. Filter, if necessary, to clarify, distribute into suitable containers and sterilize in an autoclave at 121oC for about 15 min.
b. Sabouraud Agar (Sabouraud Glucose Agar)
D- Glucose monohydrate 4.0 g
Peptones (meat and casein) 10.0 g
Agar 15.0 g
Water 1000 mL
Dissolve the ingredients in the water, adjust to give an expected pH of 5.4 to 5.8 and sterilize by heating in an autoclave at 121oC for 15 min.
All the above media should be incubated for 24 h at 37oC before use. Any contaminated media should be discarded.
Instead of preparing media, one can also use dehydrated media of Hi media/Difco and rehydrate the required quantify as per instructions on the bottle label, dispense in required quantities and sterilize at 15 psi at 121oC for 15 min.
Preparation of Inoculum
1. Preparatory to the test, inoculate the surface of a Tryptone soya agar plate for bacteria or sabouraud agar plate for fungi, with the recently grown stock culture of each of the specified micro-organisms.
2. Incubate the bacterial cultures at 30oC - 35oC for 18 - 24 h, the culture of Candida albicans at 20oC to 25oC for 48 h, and culture of Aspergillus niger at 20oC to 25oC for 7 days or until good sporulation is obtained.
3. Sub-cultures may be needed after revival before the organism is in its optimal state, but it is recommended that the number of subcultures be kept to a minimum.
4. To harvest the bacterial and Candida albicans cultures, use a sterile suspending fluid containing 0.9% w/v of Sodium Chloride and 0.1%w/v of Peptone for dispersal and transfer of the surface growth into a suitable vessel.
5. Add sufficient suspending fluid to reduce microbial count to about 108 micro-organisms per mL.
6. To harvest the Aspergillus niger culture, use a sterile suspending fluid containing 0.9% w/v of Sodium Chloride and 0.05% w/v of Plysorbate 80 and adjust the spore count to abut 108 per mL by adding the same solution.
7. Remove immediately a suitable sample from each suspension and determine the number of colony-forming units per mL in each suspension by plate count or by membrane filtration.
8. The value serves to determine the inoculum and the base line to use in the test.
9. The suspensions shall be used immediately.
As per IP
Procedure
1. Inoculate each original product container or product tube (when original container is not suitable for inoculation with sterile syringe fitted with needle, transfer 20 mL per capped bacteriological tube) with one of the standardized microbial suspension using a ration equivalent to 0.1 mL of inoculum suspension to 20 mL of product, and mix.
2. Final concentration should be between 1 x 105 and 1 x 106 micro-organisms per mL of product.
3. Determine, the number of viable micro-organisms by plate count method in each inoculum suspension and from there calculate the initial concentration of micro-organisms per mL of product under test.
4. Incubate the inoculated containers or tubes at 20o C to 25o C.
5. Determine the viable count (by plate count method at 7,14,21 and 28 days subsequent to inoculation).
6. Record any change, if observed in appearance.
As per BP
Procedure
1. To count the viable micro-organisms in the inoculated products, use the Agar medium
2. Inoculate a series of containers of the product to be examined, each with a suspension of one of the test organisms to give an inoculum of 105 to 106 micro-organisms per g or per mL of the preparation.
3. The volume of the suspension of inoculum does not exceed 1 % of the volume of the product.
4. Mix thoroughly to ensure homogeneous distribution.
5. Maintain the inoculated containers at 20 oC to 25 oC, protected from light.
6. Remove a suitable sample from each container, typically 1 mL of 1 g quantities, at zero hour and at appropriate intervals according to the type of the product and determine the number of viable microorganisms by plate count or membrane filtration.
7. Ensure that any residual anti-microbial activity of the product is eliminated by dilution, by filtration or by the use of a specific incubator.
8. When dilution procedures are used, due allowance is made for the reduced sensitivity in the recovery of small numbers of viable micro-organisms.
9. When the specific inactivator is used, the ability of the system to support the growth of the test organisms in confirmed by the use of appropriate controls. The procedure is validated to verify its ability to demonstrate the required reduction in count of viable micro-organisms.
As per USP
Procedure
1. Where the product container can be entered aseptically, such as with a needle and syringe through a rubber stopper, conduct the test in 5 original product containers.
2. If the product container is such that it can not be entered aseptically, transfer 20 mL samples of the product to each of five sterile, capped bacteriological tubes of suitable size.
3. Inoculate each tube or product container with one of the standardized microbial suspensions, using a ratio equivalent to 0.10 mL of INOCULUM to 20 mL of product, and mix.
4. A suitable concentration of test micro-organisms should be added so that the concentration in the test preparation immediately after inoculation is between 100,000 and 1,000,000 micro-organisms per mL.
5. Determine the number of viable micro-organisms in each inoculum suspension, and calculate the initial concentration of micro-organisms per mL of product under test by the plate-count method.
6. Incubate the inoculated containers or tubes at 20 oC to 25 oC.
7. Examine the containers or tubes at 7, 14, 21, and 28 days subsequent to inoculation.
8. Record any changes observed in appearance, and determine by the plate-count procedure the number of viable micro-organisms present at each of these time intervals.
9. Using the theoretical concentrations of micro-organisms present at the start of the test, calculate the percentage change in the concentration of each micro-organisms during the test.
INTERPRETATION:
The preservative is effective in the product examined if:
1. The concentration of viable bacteria are not more than 0.1% of the initial concentrations by the fourteenth day.
2. The concentrations of viable yeasts and molds remain at or below the initial concentrations during the first 14 days.
3. The concentration of each test micro-organism remains at or below these designated levels during the remainder of 28 day test period.
The criteria for evaluation of anti-microbial activity are given in the appropriate table below in terms of the log reduction in the number of viable micro-organisms using as baseline the value obtained for the inoculum.





TEST FOR PYROGENS

The test for Pyrogens is provided to determine compliance with the requirements given in individual monograph / specifications.
PRINCIPLE
Pyrogen test is the measurement of the rise in body temperature of rabbits following the intravenous injection of a sterile solution of a substance being examined. It is designed for products that can be tolerated by the test rabbits in a dose not to exceed 10.0 mL per kg. Weight injected intravenously within a period of to more than 10 min.
SELECTION OF ANIMALS
1. Use healthy, adult rabbits of either sex, weighing not less than 1.5 kg, fed a complete and balanced diet not containing antibiotics and showing no loss of body weight during the week preceding the test.
2. The rabbits must not have been used in a similar test :
a. during the preceding 3 days (as per BP) or 2 days (as per USP and IP)
b. during the preceding 3 weeks (as per BP) or 2 weeks (as per USP and IP) unless the material being examined passed the test Rabbits used in a test for Pyrogens where the mean rise in the rabbits temperature has exceeded 1.2 OC are permanently excluded (as per BP) and (as per USP and IP)
ANIMALS QUARTERS
1. Keep the rabbits individually in a quiet area with an appropriate uniform temperature with a variation of ± 2 OC and uniform humidity and free from disturbance.
2. Carry out the test in a quiet room where there is no risk of disturbance exciting the animals and in which the room temperature is within 3 OC of that of the rabbits living quarters or in which the rabbits have been kept for at least 18 h before the test.
3. Withhold food from the rabbits overnight and until the test is completed; withhold water only during the test.
EQUIPMENT
Thermometer
1. The thermometer or electrical device used indicates the temperature with a precision of 0.1 OC and is inserted in the rectum of the rabbit to a depth of about 5 cm.
2. The depth of inserted is constant for any one rabbit in any one test.
3. When an electrical device is used it should be inserted in the rectum of the rabbit 90 min. before the injection of the solution being examined and left in position throughout the test.
Glassware, Syringes and Needles
All glassware, syringes and needles must be thoroughly washed with water for injections and heated in a hot air oven at 250 OC for 30 min. or at 200 OC for 1 h.
Retaining Boxes
1. The retaining boxes for rabbits in which the temperature is being measured by an electrical device are made in such a way that the animals are retained only by loosely-fitting neck stocks; the rest of the body remains relatively free so that the rabbits may sit in a normal position.
2. The rabbits are not retrained by the use of straps or other similar methods that may harm the animal.
3. The animals must be put into boxes less than 1 h before the test and remain in them throughout the test.
Diluents
1. Treat all diluents and solutions for washing and rinsing of devices or parenteral injections assemblies in a manner that all assure that they are sterile and pyrogen-free.
2. Periodically perform control pyrogen tests on the representative portions of the diluents and solutions for washing or rinsing of the apparatus.
3. Where Sodium Chloride injection is specified as a diligent, use injection containing 0.9 % of Sodium Chloride.
Recording of Temperature
1. Use an accurate temperature-sensing device such as clinical thermometer or thermistor or other suitable probes that have been calibrated t assure an accuracy of ± 0.1 OC and have been tested to determine that a maximum reading is reached in less than 5 min.
2. Insert the thermometer or temperature-sensing probe into the rectum of the test rabbit to a depth of not less than 7.5 cm, and, after a period of time not less than that previously determined as sufficient, record the rabbit’s body temperature, do not use any rabbit having a temperature exceeding 39.8 OC.
3. Not more than 30 min. prior to the injection of the test determine the control temperature of each rabbit. This is the base for the determination of any temperature increase resulting from the injection of test solution.
4. In any one group of test rabbits, use only those rabbits whose control temperatures do not vary by more than 1 OC from each other.
As per IP
Preliminary Test (Sham Test)
1. If animals are used for the first time in a pyrogen test or have not been used during the two previous weeks, condition them one to three days before testing the substance being examined, by injection intravenously into the 10 mL per Kg of body weight of a pyrogen-free saline solution.
2. Record the temperature of the animals, beginning at least 90 min. before injection and continuing for 3 h after injection of the solution being examined. Any animal showing a temperature variation of 0.6 OC or more must not be used in the main test.
Main Test
Carry out the test using a group of three orbits.
Preparation of the Sample
1. Dissolve the substance being examined in, or dilute with, a pyrogen-free saline solution.
2. Warm the liquids being examined to approx. 38 OC before injection.
3. The amount of the sample to be injected varies according to the preparation being examined and is prescribed in the individual monograph.
4. The volume of injection is not less than 0.5 mL/Kg and not more than 10 mL/Kg of body weight.
Procedure
1. Record the temperature of each animal at intervals of not more than 30 min. beginning at least 90 min. before the injection of the solution being examined and continuing for 3 h after the injection.
2. Not more than 40 min. immediately preceding the injection of the test does, record the ‘initial temperature’ of each rabbit, which is the mean of two temperature readings recorded for the rabbit at an interval of 30 min. in the 40 min. period.
3. Rabbits showing a temperature variation greater than 0.2 OC between two successive readings in the determination of ‘initial temperature’ should not be used for the test.
4. Inject slowly the solution being examined into the marginal vein the ear of each rabbit over a period not exceeding 10 min., unless otherwise prescribed in the monograph.
5. Record the temperature of each animal at half-hourly intervals for 3 h after the injection
6. The difference between the ‘initial temperature’ and the ‘maximum temperature’ which is the highest temperature recorded for a rabbit is taken to be its response.
7. When this difference is negative, the result is counted as a zero response.
INTERPRETATION
1. If the sum of the responses of the group of three rabbits does not exceed 1.4 OC and if the response of any individual rabbit is less than 0.6 OC, the preparation being examined passes the test.
2. If the response of any rabbit exceeds 1.4 OC, continue the test using 5 other rabbits.
3. If not more than 3 of the 8 rabbits show individual responses of 0.6 OC or more, and if the sum of the responses of the group of 8 rabbits does not exceed 3.7 OC, the preparation being examined passes the test.
As per BP
Preliminary Test
1. One to three days before testing the product, inject intravenously into animals, selected as prescribed above but that have not been used during the 2 previous weeks. 10 mL/Kg of body weight of a pyrogen-free 0.9 % w/v solution of Sodium Chloride warmed to about 38.5 OC .
2. Record the temperature of the animals, beginning at least 90 min. before injection and continuing for 3 h after injection of the solution being examined.
3. Any animal showing a temperature variation greater than 0.6 OC, must not be used in the main test.
Main Test
Carry out the test using a group of three rabbits.
Preparation and Injection of the Sample
1. The preparation being examined may be dissolved in, or diluted with, a pyrogen-free 0.9 % w/v solution prescribed in the monograph.
2. Warm the liquid being examined to approx. 38.5 OC, before the injection.
3. Inject the solution slowly into the marginal vein of the ear of each rabbit over a period not exceeding 4 min. unless other wise prescribed in the monograph.
4. The amount of the sample to be injected varies according to the preparation being examined and is prescribed in the monograph.
5. The volume of the injections 0.5 to 10 mL/kg of body weight.
Determination of the Initial and Maximum Temperature
1. The ‘initial temperature’ of each rabbit is that mean of two temperature reading recording for that rabbit at an interval of 30 min. in the 40 min. immediately preceding the injection of the material being examined.
2. The ‘maximum temperature’ of each of rabbit is the highest temperature recorded for that rabbit in the 3 h. after the injection.
3. Record the temperature of each animal at intervals of not more than 30 min. beginning at least 90 min. before the injection of the solution being examined and continuing for 3 h after the injection.
4. The difference between the initial temperature and the maximum temperature of each rabbit is taken to be its response.
5. When this difference is negative, the result is counted as a zero response.
6. Rabbits showing a temperature difference greater than 0.2 OC, between any two successive reading taken during the 90 min. before the injection are withdrawn from the test.
7. In any one test, only rabbits having initial temperature that do not differ from one another be more than 1 OC may be used.
8. All rabbits having an initial temperature higher than 39.8 OC pr ;lower than 38.0 OC are excluded from the test.
INTERPRETATION
1. Having carried out the test first on a group of three rabbits, repeat if necessary on further groups of three rabbits to a total of four groups, depending on the results obtained.
2. If the summed response of the first group does not exceed the figure given in second column of the following table the preparation being examined passes the test.
3. If the summed response exceeds the figure of the second column but does not exceed the figure in the third column of the table repeat the test as indicated above.
4. If the summed response is greater than the figure given in the third column of the table the preparation being examined fails the test.


As per USP Procedure
1. Unless otherwise specified in the individual monograph, inject to an ear vein of each of three rabbits 10 mL of the test solution per kg of body weight, completing each injection within 10 min. after start of administration.
2. The test solution is either the product, constituted if necessary as directed in the labeling or the material under test treated as directed in the individual monograph and injected in the dose specified therein.
3. For Pyrogen testing of devices or injection assemblies, use washing or rinsing of the surfaces that come in contact with the parenterally administered material or with the injection site or internal tissues of the patient.
4. Assure that all test solutions are protected from contamination.
5. Perform the injection after warming the test solution to a temperature of 37 ± 2 OC.
6. Record the temperature at 30 min. intervals between 1 and 3 h subsequent to the injection.
Test interpretation and continuation
1. Consider any temperature decreases as zero rise.
2. If no rabbit shows an individual rise in temperature of 0.5 OC or more above its respective control temperature, the product meets the requirements for the absence of the pyrogens.
3. If any rabbits shows individual temperature rise of 0.5 OC or more, continue the test using five other rabbits.
4. If not more than three of the eight rabbits show individual rises in temperature 0.5 OC or more and if the sum of the eight individual maximum temperature rises does not exceed 3.3 OC the material under examination meets the requirements for the absence of Pyrogens.

WASHING AND STERILIZATION OF APPARATUS

1.0 OBJECTIVE
To lay down a procedure for washing and sterilization of glass apparatus.
2.0 SCOPE
This SOP covers the procedure for washing and sterilization of glass apparatus is applicable to Quality Control Department Formulation Units
3.0 RESPONSIBILITY
Microbiologist.
4.0 ACCOUNTABILITY
Department Head.
5.0 PROCEDURE
5.1 Wash the glassware thoroughly using soap solution (Extran 1.0%) followed by potable water rinses to remove all the traces of the residues. Finally rinse with purified water.
5.2 Dry the washed glassware at 105°C in oven.
5.3 Cover the open ends of glass apparatus with a caps / cotton plugs and wrap with aluminum foil.
5.4 Sterilize in the autoclave at 15 psi, 121 OC for 20 min.
5.5 Keep the sterilized material in separate and labeled area to avoid mix up with unsterilized material.
5.6 Record the details.
5.7 For new glassware follow the following procedure
5.7.1 Treat the glassware’s with 5%w/v sodium carbonate solution and then soak overnight in 1% v/v solution of Hydrochloric acid.
5.7.2 Rinse the glassware’s with sufficient quantity of potable water and then by purified water.
5.7.3 Further clean the glassware’s as per the routine procedure as above.

Consumption & Accounting of Excess/Shortage material received from vendor

1.0 OBJECTIVE
To lay down a procedure for Consumption and accounting of Excess/Shortage material received from the vendor.
2.0 SCOPE
This SOP is applicable to Raw and Packing Material Warehouse.
3.0 RESPONSIBILITY
Officer - Warehouse.
4.0 ACCOUNTABILITY
Asst.Manager – Warehouse
5.0 PROCEDURE
5.1 Weigh and label (Loose container label) for all materials after each issue.
5.2 Check the remaining available physical quantity with the recorded SAP R/3 stock.
5.3 At the completion of a particular consignment, check the physical stock with SAP R/3 stock.
5.4 Prepare an Excess/Shortage certificate(as per Annexure enclosed ) for the quantity in excess/shortage.
5.6 Update the stocks in the SAP R/3 system as & when the Excess/Shortage certificate is Approved and Authorized.
5.6 The posting of issues in SAP R/3 system related toExcess quantity than the specified norms as below, is done only after the Authorization of the certificate.
5.7 An Investigation report is enclosed along with the Excess/Shortage certificate in case the quantity is More than 0.5 % in case of excess More than 0.2 % in case shortage
for Active Ingredients More than 1.0 % in case of excess More than 0.5 % in case of shortage
for Excepients, Primary & Secondary Packing materials. More than 5.0 % for Solvents

Handling of loose container

1.0 OBJECTIVE
To lay down a procedure for Handling of loose containers, of raw materials.
2.0 SCOPE
2.1.0 This SOP is applicable to dispensing area
3.0 RESPONSIBILITY
Officer Warehouse
4.0 ACCOUNTABILITY
Assistant Manager-Warehouse
5.0 PROCEDURE:
5.1.0 After dispensing of material,container & inner polybag of the loose material to be closed properly.
5.1.1 Weigh the loose container and record the Gross weight on the label.
5.1.2 Based on the original tare weight calculate the net weight of the loose material and record the net weight on the label as per annexure attached.
5.1.3 Sign the label and affix the loose container label on the container over the earlier loose container label after verifying the details.
5.1.4 Before affixing the loose container label strike off the earlier loose container label.
5.1.5 Send back the loose container to warehouse through the material exit box after dedusting and cleaning
5.1.6 The same loose container to be taken first for next dispensing.

OPERATION OF HI-REACH TRUCK - MAKE : MACNEILL, MODEL: 115-EN(T/L)

1.0 OBJECTIVE
To laydown a procedure for Operation of Hi-Reach Truck.
2.0 SCOPE
This SOP is applicable to Raw Material ,Secondary packing Material and Finished Goods Warehouse Hi-Reach Trucks, Capacity: 1500 Kg
3.0 RESPONSIBILITY
Operator
4.0 ACCOUNTABILITY
Executive Warehouse / Executive Services
5.0 PROCEDURE
5.1 Precautions
5.1.1 Check for Hydraulic Oil leakage and arrest.
5.1.2 Ensure that charger plug is removed from batteries.
5.2 Pre Start Up
5.2.1 Check for distilled water level in the cells of the battery unit and top up if necessary.
5.2.2 Check for specific gravity of the pilot cells that to be 1240 mm of Hydrometer.
5.2.3 Connect truck plug to the battery plugs.
5.2.4 Select mode of truck direction with direction lever.
5.3 Set Up
5.3.1 Set the rear wheel steering in straight.
5.3.2 Set forks clear from the ground
5.4 Operation
5.4.1 Sit firmly in the driving seat.
5.4.2 Insert the key in the key switch and turn on.
5.4.3 Release parking brake by depressing the foot brake.
5.4.4 Select direction with the help of direction switch.
5.4.5 Slowly depress the roller type accelerator pedal for movement of truck.
5.4.6 For Hydraulic Operations:
5.4.6.1Lift Lever:
Pull the lever back to lift and push the lever forward to lower.
5.4.6.2 Reach Lever:
Push the lever forward to reach OUT and pull the lever back to reach IN.
5.4.6.3 Tilt Lever:
Pull the lever back for backward tilt and push the lever forward for forward tilt.
5.4.7 While lifting the load drive the truck as close to the load as possible and turn the vehicles so that the forks are in line with the load.
5.4.8 Ensure that the forks are correctly spaced and conform with the width of the pallet entry.
5.4.9 Raise the forks to the required height and push the tilt lever forward until the masts are vertical.
5.4.10Drive slowly forward until the reach legs almost touch the pallet.
5.4.11Apply the parking brake and push the reach slowly forward until the forks are fully entered into the pallet.
5.4.12 Raise the fork carriage until the weight of the load is taken up.
5.4.13 Tilt the masts back and pull the reach lever back until the masts are fully retracted.
5.4.14 Lower the forks to within 6” (152 mm) of the reach legs and move the direction switch lever into the reverse position and release the parking brake.
5.4.15 Ensure that the way is clear and reverse slowly away from the stack.
5.4.16 Before reaching the truck’s destination fully release the accelerator pedal and allow the electric brake to slow down the truck..
5.5 Shut Down
5.5.1 For stopping the truck, fully release the accelerator pedal is one option for allow the electric brake slowly.
5.5.2 By pressing the foot brake pedal for quick stopping the Hi-Reach Truck.
5.5.3 Ensure that the forks are fully lowered when parking or leaving the truck.
5.5.4 Ensure that masts reached back to original position.
5.5.5 Ensure that parking brake applied.
5.5.6 Check for key switch ‘OFF’ position.
5.5.7 Check for disconnections of battery plug from the truck wiring.
5.5.8 Connect the charger leads to the truck wiring for battery charger.

Control on materials sent for job works

1.0 OBJECTIVE
To lay down a procedure for material sent for job works and its stock accounting.
2.0 SCOPE
2.1.0 Raw Materials & Primary packaging materials.
3.0 RESPONSIBILITY
Officer - Warehouse and Executive-Supply Chain Management Department.
4.0 ACCOUNTABILITY
Asst.Manager – Stores & Excise and Executive -Supply Chain Management Department.
5.0 PROCEDURE
5.1 Supply Chain Management Department will coordinate with warehouse department on job works.
5.2 Material to be sent out for job work only on the advice of Supply Chain Management Department.
5.3 The details of the material to be sent for job work such as Item description,quantity received,date of receipt,invoice number against which received & its date,quantity being sent,nature of job work,place or unit where job work is to be done,expected date of return of finished good are to be given to Excise Department (at Factory) one day in advance.
5.4 Assemble the required quantities and get cross checked by another Officer – Warehouse
5.5 Load the goods in the presence of Security Guard.
5.6 Prepare the Delivery Challan cum RGP for the material loaded in the vehicle and pass the same to excise department for preparation of Excise Invoice/Challan.
5.7 Post the entries in SAP against the job order raised by Supply Chain Management Department.
5.8 Send the truck along with Delivery Challan cum RGP,C.O.As for raw materials,Excise Invoice/Challan & Way bill to the destiny after due checking by security at material gate.
5.9 On receipt of processed goods, prepare Goods Received Note in SAP for the actual quantity received.
5.10 Prepare Goods Received Note for unprocessed returned quantity in SAP separately.
5.11 Supply Chain Management Department to raise Write-Off authorization form for wastage and scrap generated during job work.
5.12 On receipt of approved Write-Off note,post the same in SAP.
5.13 Keep track of identity, traceability and link of the total transactions till it completely gets squared up.

OPERATION OF BATTERY OPERATED PALLET TRUCK, MAKE: MACNEILL

1.0 OBJECTIVE
To laydown a procedure for Operation of Battery Operated Pallet Truck.
2.0 SCOPE
This SOP is applicable to Finished Goods Packing Material Warehouse Battery Operated Pallet Truck, Model: Commuter /1115, Capacity: 1500Kg
3.0 RESPONSIBILITY
Technical Assistant.
4.0 ACCOUNTABILITY
Executive-Warehouse and Executive-Services
5.0 PROCEDURE
5.1 Precautions
5.1.1. Check and arrest any oil leakages .
5.1.2. Ensure that the plug is removed from the battery after charger is switched off.
5.2 Pre Start Up
5.2.1. Check for distilled water level in the cells of the battery unit and top up if necessary.
5.2.2. Check for specific gravity of the pilot cells to be 1240mm of Hydrometer.
5.2.3. Ensure that battery cable connections are tight and that cell caps are dry and clean.
5.2.4. Check for load carriage and forks are free from visible defects i.e deformation, cracks or excessive wear
5.2.5. Ensure that brakes are functioning correctly.
5.3 Set Up
5.3.1 Set steering handle in straight position.
5.3.2 Set forks clear from the ground.
5.4 Operation
5.4.1 Insert Key in the key switch and turn ON with the handle in its vertical position.
5.4.2 Turn hydraulic screw control in clock wise direction and depress the push button lift switch thus raising the forks.
5.4.3 Lower the control handle from its normal upright position to an angle of 450 approximately thus release the mechanical brake.
5.4.4 Stand behind the truck and slowly depress one of the buttons. The buttons should be governed by the direction required. i.e. “Away - Toward”.
5.4.5 Depress the selected button slowly for normal speed.
5.4.6 Depress the button fully for obtaining more speed (i.e.,) 5.0 Km/hr approximately.
5.4.7 Push the control handle and it will move upper range by spring action and stops the vehicle for emergency stopping.
5.5 Shut Down
5.5.1 For braking the vehicle release pole slowly for its upward position.
5.5.2 Ensure that lifting equipment must be completely lowered.
5.5.3 Ensure that key switch is turned OFF.
5.5.4 Ensure disconnection of battery plug from the truck wiring.
5.5.5 Connect the battery charger leads to the truck wiring for battery charging

OPERATION OF ELECTRIC PEDESTRIAN OPERATED PALLET TRUCK (BOPT) MAKE:JOSTS

1.0 OBJECTIVE
To lay down a procedure for Operation of Electric Pedestrian Operated Pallet Truck.
2.0 SCOPE
This SOP is applicable to Raw Material Stores Electric Pedestrian Operated Pallet Truck Type: EJE20, Capacity: 1500 Kg
3.0 RESPONSIBILITY
Technical Assistant
4.0 ACCOUNTABILITY
Electrical Executive
5.0 PROCEDURE
5.1 Precautions
5.1.1 Check for any Hydraulic Oil leakages and arrest.
5.1.2 Ensure that plug is removed from the battery after charger is switched off.
5.2 Pre Start Up
5.2.1 Check for distilled water level in the cells of the battery unit and top up if necessary.
5.2.2 Check for specific gravity of the pilot cells and to be at 1240 mm of Hydrometer.
5.2.3 Ensure that battery cable connections are tight and that cell caps are dry and clean.
5.2.4 Ensure that battery plug is securely connected.
5.2.5 Check for play in the steering system.
5.2.6 Check for load carriage and fork tins are free from visible defects i.e. deformation, cracks or excessive wear.
5.2.7 Ensure that service brake and parking brake are functioning correctly.
5.3 Set Up
5.3.1 Set steering handle in straight position.
5.3.2 Set fork tins clear from the ground.
5.4 Operation
5.4.1 Insert key in the key switch and turn ‘ON’
5.4.2 Release parking brake by moving the pole handle in to the driving range.
5.4.3 Select direction with help of wing-type levers mounted on head of the steering control handle.
5.4.4 Slowly press the wing-type lever for movement of the pallet truck.
5.4.5 For lifting of fork tins press the push button which is located at the pole head for required height of fork tins
5.4.6 For lowering push the lever of the lowering valve in front direction. The lowering speed is controlled by the amount of movement applied to the valve lever.
5.4.7 Ensure that collision safe guard button should be depressed in case of emergency only.
5.4.8 For stopping the truck move the pole upwards or downwards into the braking range. when the pole is released, it will automatically move in to the upper range by spring action and stops the vehicle.
5.5 Shut Down
5.5.1 For breaking the vehicle, release pole slowly for it’s upward position.
5.5.2 Ensure that lifting equipment must be completely lowered.
5.5.3 Check for operating controls to be normal.
5.5.4 Ensure that key switch turned to the OFF.
5.5.5 Ensure disconnection of battery plug from the truck wiring.
5.5.6 Connect the battery charger leads to the truck wiring for battery charging.

Wednesday, December 24, 2008

Receipt of excess Raw/ Packing materials from production

1.0 OBJECTIVE
To lay down a procedure for Receipt of excess Raw/ Packing Material from Production.
2.0 SCOPE
Raw Material, Packing Material Warehouse and Production hold area.
3.0 RESPONSIBILITY
Assistant / Officer - Warehouse Assistant / Officer - Production.
4.0 ACCOUNTABILITY
Assistant Manager - Production and Assistant Manager - warehouse.
5.0 PROCEDURE
5.1 Excess packing material to be received with proper document through Stock Return Note (SRN) only.
5.2 Stock Return Note (SRN) contains the following details.
A) S.No
B) Date, Product and Batch No.
C) S.No of materials.
D) Code
E) Item
F) Quantity returned
G) Analytical Report No
H) Quantity Verified by signature and date.
I) Reasons for return / remarks.
5.3 Stock Return Note (SRN) to contain the signatures of Officer – Production,Quality Assurance Personnel and Assistant Manager - Production.
5.4 Material being returned has to be labeled with details of Batch No/ Analytical Report No. / Item Code / Item description.
5.5 Receive unused production returns in properly labeled and packed condition and store at specified area.
5.6 In case of Raw-material, returns to be sent to the Dispensing hold area by the Officer - Production.
5.7 In case of Raw-material, weighment should be done inside the Dispensing room and signed by the Assistant / Officer - Warehouse and countersigned and checked by the Dispensing Officer - Production.
5.8 In case of primary packing material verify the quantity by Weight and No. of Rolls.
5.9 In case of secondary packing material verify the quantity by counting.
5.10 Sign the Stock Return Note (SRN) for quantity received.
5.11 Enter the transaction in SAP.
5.12 Stock Return Note (SRN) material to be issued during the next immediate requirement from Production.

Receipt of On-line Rejections

1.0 OBJECTIVE
To lay down a procedure for Receipt of “On-line Rejected Materials”.
2.0 SCOPE
Production, Packing hold area and dispensing area.
3.0 RESPONSIBILITY
Officer- Production / Officer - Warehouse
4.0 ACCOUNTABILITY
Assistant Manager - Production and Asst.Manager -Warehouse.
5.0 PROCEDURE
5.1 On-line rejected materials to be received with On-line rejection note duly signed by the Assistant Manager - Production./Quality Assurance.
5.2 On-line rejected material to be labelled as “On-line rejects”.
5.3 On-line rejected labels are to be signed by the Officer Production /Officer - Quality Assurance Department.
5.4 Stock return note to be raised by the Officer - Production for returning the On-line rejects and counter signed by Quality Assurance.
5.5 The stock return note to contain the S.No. and Date of On line Rejection with reasons.
5.6 The On-line rejected material to be sent to Dispensing room properly labelled along with a Online rejection note.
5.7 The On-line rejected material to be weighed inside the Dispensing room by Officer - Warehouse in presence of Officer - Production.
5.8 The stock return note to be signed by Officer - Warehouse and countersigned by the Officer - Production.
5.9 The On-line rejected material to be taken to Raw-material/Packing material Warehouse through the pass box and stored in rejected area.
5.10 On line rejections should be informed to Purchase Department with in a week for their further action.
5.11 Post the online rejected quantities with details in SAP.

Receipt of Labels

1.0 OBJECTIVE
To lay down a Procedure for receipt of labels.
2.0 SCOPE
Packing Material Warehouse.
3.0 RESPONSIBILITY
Officer - Warehouse.
4.0 ACCOUNTABILITY
Asst.Manager -- Warehouse
5.0 PROCEDURE
5.1 Labels received from the suppliers are cross-checked with Delivery Challan for quantity by Warehouse Assistant/officer.
5.2 Quantity is checked by counting 50/100 Nos. and taking the weight of the same, the remaining total consignment is weighed.
5.3 Goods Received Note (GRN) has to be prepared in SAP

Opening and Closing of Ware house

1.0 OBJECTIVE
To lay down a procedure for Opening & closing of Raw/Packing /Finished Goods/and Engineering material Warehouses.
2.0 SCOPE
2.1. Raw Material / Packing Material / Finished Goods Engineering material Warehouses.
3.0 RESPONSIBILITY
Assistant / Officer - Warehouse.
4.0 ACCOUNTABILITY
Asst.Manager - Warehouse
5.0 PROCEDURE :
5.1.1 At the beginning of every working day the Warehouse Key to be taken by Assistant / Officer - Warehouse from the security by making necessary entries in Key’s Register (Maintained by -Security).
5.1.2 Security to hand over the Key to the person Authorised to draw the Raw Material / Packing Material Finished goods/ & Engineering material Warehouse keys.
5.1.3 Security to ensure the drawee signature and counter sign in the Key register.
5.1.4 In the absence of authorised person Security Officer to handover the keys to the person named by Assistant Manager - Warehouse on written request.
5.1.5 Immediately on opening the Warehouse check the log books for any transactions made during non-working hours and holidays.
5.1.6 On opening of Warehouse, Officer / Assistant - Warehouse to go round inside the Warehouse for checking any abnormalities and report to the Assistant Manager - Warehouse.
5.1.7. Before closing the Warehouse ensure all the electrical appliances are switched off such as lighting, fans, air handling units and computer. Ensure closure of all valves of purified water lines.
5.1.8 At the close of each working day the keys after locking to be handed over by Authorised person to the Security by making necessary entries in the Key register.
5.1.9 Security to counter sign the Key Register on receipt of the keys.
5.1.10 Security to keep the keys in the key rack under lock and key.
5.2 Issue of key in Case of Emergency
5.2.1 Warehouse key can be issued in emergencies by Security only when the request is made on an Internal communication mentioning the reasons for opening the Ware House. The Internal Communication to be signed by person not below the Rank of Assistant Manager. Security to inform over phone to the Assistant Manager - Warehouse /Manager – Warehouse/Plant - Manager before giving the key.
5.2.2 Opening and closing of the Warehouse in case of emergency to be done in the presence of a Security person and a third person ( other than the one opening the Warehouse) who should not be below the rank of an Officer.
5.2.3 Security to ensure that entries are made in the log book maintained at Warehouse indicating for the Warehouse was opened during non -working hours, any material drawn to be recorded for suitable action by Warehouse personnel next working day.
5.2.4 Security to inform in writing to Asst.Manager – Warehouse about the opening of Warehouse, immediately the next day morning

Monday, December 22, 2008

Handling & storage of flammable Materials

1.0 OBJECTIVE
To lay down a procedure for Handling and Storage of Flammable and Hazardous Materials.
2.0 SCOPE
Raw Material Warehouse and Solvent Yard.
3.0 RESPONSIBILITY
Assistant / Officer / Executive - Warehouse.
4.0 ACCOUNTABILITY
Asst.Manager - Warehouse.
5.0 PROCEDURE
5.1 All the flammable items to be identified and a specific area to be allotted.
5.2 The following instructions to be strictly followed in monitoring the area.
5.2.1 The place of storage to be safe guarded by fencing to restrict the movement of people.
5.2.2 Materials to be unloaded in tankers with pipe line connections.
5.2.3 While unloading the material, they are to be handled properly . for example pressurized drums to be unloaded with the help of tyres to avoid blasting due to sudden impact.
5.2.4 No live wires to be passed through the area where inflammable materials can be exposed to the sparks.
5.2.5 No smoking in the area is allowed.
5.2.6 The material is to be completely tightened with lids and the drums to be covered with tarpaulin.
5.2.7 Pipe line valves to be closed whenever they are not in use.
5.2.8 At high temperatures to reduce heat the tarpaulin covers to be made wet with water twice daily.
5.2.9 To fight against fire uncertainities, suitable fire extinguishers and sand buckets near the area to be provided.
5.2.10 Do not store Hydrochloric Acid inside the Warehouse.

Friday, December 19, 2008

Stock Verification and Reconciliation

1.0 OBJECTIVE
To lay down a procedure for “Stock Verification and Reconciliation” of raw material and packing material.
2.0 SCOPE
Raw Material, Packing Material Warehouses, and Solvent Yard.
3.0 RESPONSIBILITY
Officer - Warehouse.
4.0 ACCOUNTABILITY
Assistant Manager - Warehouse.
5.0 PROCEDURE
5.1 Stock verification and reconciliation of all the material in the warehouse is done as below.
5.1.1 Create a Physical Inventory document in SAP with reference to inhouse batch number in all working days & take printout of the same.
5.1.2 Note the physical stock against the SAP stock in the Physical Inventory document.
5.1.3 Post the physical stocks in SAP.
5.1.4 Assess the variance & prepare Excess/Shortage certificate after completion of the stock against that particular in-house batch number as per SOP No:XXXXX.
5.2 The above verification and reconciliation should be done in addition to the perpetual stock verification done during dispensing.
5.3 Stocks of solvents to be verified at the end of every month and for the differences such as evaporation losses and handling losses “Excess /Shortage Certificate” has to be raised as per SOP No.XXXXX.
5.4 Update the stocks in SAP R/3 system as when the Excess /Shortage certificate is authorized & approved.

Disposal of Obsolete Materials

1.0 OBJECTIVE
To lay down a Procedure for Disposal of Obsolete Materials.
2.0 SCOPE
This SOP is applicable to Raw and packing material Warehouses of Formulations Units
3.0 RESPONSIBILITY
Officer - Warehouse.
4.0 ACCOUNTABILITY
Assistant.Manager - Warehouse
5.0 PROCEDURE
5.1 Materials which are no more in use and lying in the Warehouse idle for more than six months has to be informed to the Supply Chain Management Department in writing.
5.2 Supply Chain Management Department to explore any alternative exits with Formulations Research and Development.
5.3 Obsolete material to be stacked on top slot of the rack duly labelled.
5.4 Write-off note to be generated by Supply Chain Management Department based on the decision taken on the disposal.
5.5 Destroy write-off materials within 10 days from the date of receipt of duly approved “Write-off “ authorization form.
5.6 Provide proof of destruction copy to the concerned person at Central Excise Department of Factory and Accounts.
5.7 Write-off material has to be destroyed suitably in the presence of the Quality Assurance person and ensure signature and date.
5.8 Enter the transaction in SAP System wherever required.
5.9 Preserve action completed write-off forms separately.

Thursday, December 18, 2008

Handling and Disposal of On-line Rejected Material

1.0 OBJECTIVE
To lay down a Procedure for Disposal of on line rejections
2.0 SCOPE
2.1.0 Area
2.1.1 Raw & Packing Material Warehouse.
3.0 RESPONSIBILITY
Officer - Warehouse
4.0 ACCOUNTABILITY
Asst.Manager - Warehouse
5.0 PROCEDURE
5.1 Online rejections should be received from Production on authorized documents
“ON-LINE REJECTION NOTE” only as per the Annexure-1
5.2 “ON LINE REJECTION NOTE” to be signed by Executive -Production and Executive -
Quality Assurance Departments.
5.3 Online rejected materials to be received with clear Online Rejected labels signed by the Officer - Production and Officer - Quality Assurance Departments.
5.4 The action plan regarding the remaining qty. of the particular consignment which is Online rejected ,should be clearly mentioned in the On-Line Rejection Note.
5.5 Warehouse to forward the On-Line Rejection Note to Purchase Department for suitable action within 3 days.
5.6 Less than 0.5 Kg. rolls of Blister Foil, less than 1.0 Kg. of Aluminium Foil, Strips Foil and PVC Film are not to be transferred to Warehouse as on line rejections.This should be destroyed by production it self.
5.7 On line rejected material which is returned to party as per Purchase advise is to be informed to Accounts with a copy to Purchase Department to recover the amount as they are approved and paid goods.
5.8 On receipt of instructions from Purchase, action to be completed within a month. Statement indicating the stocks to be prepared fortnightly and copies to be sent to Supply Chain Management Department.
5.9 If Quality Assurance re-approves any on line rejections for any reason Warehouse to re-issue the same on First Priority to Production with re-approval status label.
5.10 For the rejections which cannot be recovered / replaced “WRITE OFF NOTE” has to be raised by the Supply Chain Management Department.
5.11 Write-Off material to be destroyed in the presence of the Quality Assurance persons.
5.12 Any deviation in the above procedure is to be informed to the Asst.Manager – Warehouse.

Shortage and damage intimation/Disposal procedure for Insurance claim

1.0 OBJECTIVE
To lay down a procedure for intimation of shortages and damages of Incoming Goods.
2.0 SCOPE
This SOP is applicable to all Warehouses of Formulations Units.
3.0 RESPONSIBILITY
Assistant /Officer - Warehouse.
4.0 ACCOUNTABILITY
Asst.Manager - Warehouse
5.0 PROCEDURE
5.1 If any shortage or damage in incoming Goods found, the same has to be mentioned on the Delivery Challan before acknowledging the receipt.
5.2 The damages should be checked in presence of Quality Assurance personnel / the supplier / transporter.
5.3 If the damage is on the outer container / packing the same should be changed in presence of Quality Assurance
5.4 Shortages has to be indicated in Goods Received Note and inward actual quantity received and inform the same with in 24 hrs to the Purchase Department.
5.5 If the consignment is extensively damaged and Quality Assurance confirms not fit for receiving, do not accept and act as per Purchase Department’s advice for returning the consignment.
5.6 Partially damaged / leaking material to be stored separately in under test area.
5.7 Check material containers/packages for any leakage’s and suitable action to be taken if any leakage is observed.
5.8 Show the damaged items / short received item to the insurance surveyors as and when they come for assessment.
5.9 Hold the damaged goods till insurance company directs the action to be taken on settlement of claims.
5.10 Arrange destruction or despatch within 10 days time from the receipt of instructions from Purchase / Insurance Company.
5.11. Arrange destructions in the presence of Quality Assurance personnel if the instructions are for destructions
5.12 Provide destruction certificate to Purchase and Accounts.
5.13 In case of outside despatch provide mode of despatch details to Purchase Department.

Cleaning of Dispensing Tools

1.0 OBJECTIVE
To laydown a procedure for Cleaning of Dispensing Tools
2.0 SCOPE
This SOP is applicable for Raw Material Warehouse at Formulations Units.
2.1 Area
2.1.1 All the dispensing areas - Raw Material Warehouse
2.1.2 Scoops, Spoons and servers.
3.0 RESPONSIBILITY
Assistant / Officer -Warehouse and Dispensing Chemist -Production.
4.0 ACCOUNTABILITY
Assistsnt Manager - Warehouse and Assistant Manager - Production.
5.0 PROCEDURE
5.1 Keep the once used (i.e., one material) dispensing tools such as scoops, spoons and big size server spoons in the box containing polybag as specified “accessories to be cleaned”
5.2 On completion of each batch dispensing, arrange for cleaning of used dispensing tools in the wash room.
5.3 Ensure cleaning person to wash the tools with purified water and final rinsing with purified water.
5.4 After wash, tools to be wiped with a fresh lint-free cloth.
5.5 Ensure the drying and cover the same with shrink wrap film.
5.6 Keep the covered tools in the box provided for use.
5.7 Keep the wash area clean.

Wednesday, December 17, 2008

Storage of Raw and Primary Packing Materials

1.0 OBJECTIVE
To lay down a procedure for Storage of Raw and Primary Packing Materials.
2.0 SCOPE
This SOP is applicable to Raw Material Warehouse.
3.0 RESPONSIBILITY
Assistant / Officer - Warehouse.
4.0 ACCOUNTABILITY
Asst.Manager – Warehouse.
5.0 PROCEDURE
5.1 All material to be stored at temperature and relative humidity as specified in
Annexure-1.
5.2 Record the temperature & R.H of Warehouse
5.3 Status label has to be affixed neatly to all material container possibly adjacent to the original Manufacturers label.
5.4 Materials should be neatly stacked on HDPE pallets and should be convenient for issue.
5.5 Clear PVDC films are to be stored in cool rooms.
5.6 Load on each pallet should be moderate and not exceed 500 kgs. Stack the material properly and uniformly on pallets.
5.7 Materials should not be protruding outside the pallets.
5.8 Arrangement of materials on pallets should be convenient for physical checking and counting.
5.9 Do not stack or block the gangways.
5.10 Materials such as Starch bags / purified Talc / Magnesium stearate should be covered with polythene bags and stored, to avoid contamination.
5.11 Undertest materials to be stacked in undertest area only.
5.12 Approved material to be stacked in approved area only.
5.13 Rejected material to be stacked in rejected area only.
5.14 On approval of undertest material,after affixing the approved label shift the material to approved area .
5.15 On Rejection of undertest material,after affixing the rejected label
shift the material to rejected area.
5.16 Follow the storage conditions of materials as recommended by Formulations Research and Development Department.

Tuesday, December 16, 2008

Goods Received Note -Preparation

1.0 OBJECTIVE
To lay down a procedure for Preparation of Goods Received Note for Raw and Packing materials.
2.0 SCOPE
2.1 Raw Material and Packing Material Warehouse.
3.0 RESPONSIBILITY
Officer - Warehouse.
4.0 ACCOUNTABILITY
Executive - Warehouse / Asst. Manager – Stores & excise.
5.0 PROCEDURE
5.1 For all the material received, prepare a Goods Received Note (GRN).
5.2 Prepare GRN immediately upon receipt and send the Certificate ofAnalysis (COA ),mentioning the GRN No.,to Quality control Dept..


5.3 Note the GRN No. and Date on the Delivery challan/Invoice and file the same.

5.4 Raise fresh GRN for materials in case of subsequent reapproval after receiving the duly approved Non Conformance Report.




6.0 REVISION

6.1. Due to SOP review date expiry.

Operation of Decontamination Booth

1.0 OBJECTIVE
To lay down a procedure for operation of de-contamination booth.
2.0 SCOPE
Raw material and Primary Packing material Warehouse.
3.0 RESPONSIBILITY
Executive - Warehouse.
4.0 ACCOUNTABILITY
Asst. Manager - Warehouse.
5.0 PROCEDURE
5.1 PRECAUTIONS
5.1.1 Ensure that the chain guards must be closed.
5.1.2 Containers should be placed on the conveyor, ensure that it should not touch the frame.
5.2 PRE START UP
5.2.1 Ensure for cleanliness of conveyor rollers.
5.2.2 Check the cleaning of suction filter, in case of any dust, get it cleaned.
5.3 SET UP
5.3.1 Set the selector switch in required mode i.e., Forward or Reverse.
5.4 OPERATION
5.4.1 Inform the warehouse personnel inside warehouse to organize pallets for unloading the material and to open the inside shutter.
5.4.2 Switch on the mains.
5.4.3 Start the side air blower by operating push button.
5.4.4 Start the top suction blower by operating push button.
5.4.5 Ensure both the blowers are running
5.4.6 Start the roller conveyor by operating push button.
5.4.7 Ensure that the shutter opened fully.
5.4.8 Place the container on the roller conveyor.
5.4.9 Container is collected at the end of the conveyor by Warehouse.
5.5 SHUT DOWN
5.5.1 Ensure that all the containers have been taken inside.
5.5.2 Inform Warehouse personnel to close the inside shutter.
5.5.3 Stop the conveyor by operating “ RED push button”.
5.5.4 Stop the top suction blower by operating “RED push button”.
5.5.5 Stop the side air blower by operating “RED push button”.
5.5.6 Keep the forward / Reverse Selector switch in “IDLE” condition.

Sunday, December 14, 2008

HPLC COLUMN RECEIPT AND ITS USAGE

1.0 OBJECTIVE
To lay down a procedure for receiving HPLC columns registering and its usage.
2.0 SCOPE
This SOP is applicable to Quality Control Department.
3.0 RESPONSIBILITY
Quality Control Executive / Officer.
4.0 ACCOUNTABILITY
Department Head.
5.0 PROCEDURE
5.1 On receipt of a new column, enter the following details in the HPLC column register.
A. Name of the column.
B. Make
C. Dimension and its particle size
D. Date of receipt
E. Mfr. Certificate : Received / Not received
F. Application
G. Manufacturers column number
H. Column number allotted
I. Usage with effective from (w.e.f.) [date]
5.3 All new columns in the lab should be numbered at the time of receipt.
5.4 Q.C Executive/Officer will be responsible to allot the number on receipt of the column.
5.5 All new columns / spare columns should be kept properly under lock & key and columns under usage shall be kept in allotted places.
5.6 Five digit alpha numeric is designed to number the column, for example F3HC000,
Where F3 represents Formulations 3 Quality control, HC represents HPLC Column and 000 is a serial number,Which represents the column serial number 001 to 999
5.7 Depending up on the Specificity of the column, its application is decided immediately
or prior to use. The same has to recorded in the HPLC columns register.
5.8 If the column Performance is found unsatisfactory as per analytical method, the same
Should be discarded from the routine usage, and The same column should be labeled and kept separately for its regeneration or destruction.
5.9 After regeneration, if a column performance is found satisfactory , the column has to be kept for usage again.
5.10 If the column performance is not satisfactory, the column should be labeled FOR DESTRUCTION and record of such columns should be maintained
5.11 New column will be issued to the laboratory after proper authorization.

Thursday, December 11, 2008

Preparation of Volumetric solutions/ Reagents/ Mobile phase and Standardization of volumetric solutions

1.0 OBJECTIVE
To lay down a procedure for preparation of volumetric solutions / reagents / mobile phase & standardization of volumetric solution used for chemical analysis
2.0 SCOPE
This SOP is applicable to Quality Control Department
3.0 RESPONSIBILITY
Quality Control Chemist.
4.0 ACCOUNTABILITY
Department Head.
5.0 GENERAL INSTRUCTIONS
5.1 All chemicals / solvents used for preparation of volumetric solutions and reagents shall be LR Grade / AR Grade / GR Grade.
5.2 All volumetric solutions / reagent shall be prepared in purified water unless otherwise the use of carbon-dioxide-free water is specified.
5.3 Volumetric Solutions :
5.3.1 All volumetric solutions / reagent shall be freshly prepared once in every three months or whenever the solution shows any fungal growth or sedimentation or deviation of the strength [Molarity / Normality] from the desired value by more than ± 10%.
5.3.2 The restandardization date shall be one month for all volumetric solutions
5.3.3 Records will be maintained in the Volumetric Solution Standardization Record.
5.3.4 Standardization shall be determined in duplicate either by manually or potentiometrically (wherever applicable) as per STP. Strength of volumetric solution, shall be within ± 10..0 % of the specified strength.
5.3.5 Store all volumetric solutions / reagents in borosilicate stoppered glass bottle, protected from heat, air & moisture. Wherever specified in STP, volumetric solutions / reagents shall be stored in amber colored stoppered glass bottles.
5.3.6 Label the volumetric solutions with details :
1.Name of the Solution
2.Strength:- Exact molarity or normality (wherever applicable),
3.Prepared by,
4.Date of Preparation,
5.Standardized by,
6.Date of Standardization,
7.Use before.
5.3.7 Label the reagents with details
1. Name of the solution
2. Strength:- Molarity / Normality or percentage w/w or w/v (wherever applicable)
3. Prepared by,
4. Date of preparation,
5. Use before,
Record the details for volumetric solutions :-
1. Name of the volumetric solution,
2. Date of preparation,
3. Date of standardization,
4. Weight of primary standard taken or volume of equimolar / equinormal standard solution (wherever applicable ),
5. Volume of titrant consumed,
6. Normality / Molarity (wherever applicable),
7. Average Normality / Molarity (wherever applicable),
8. Signature of Analyst,
5.3.8 Calculate the strength of the volumetric solution using one of the following formula (wherever applicable)
1. Using volumetric solution of known strength.
S2 = V1 x S1
V2
Where,
V1 = Volume of the Standard solution in ml.
S1 = Strength of Standard Solution (Normality or Molarity)
V2 = Volume of test solution in ml.
S2 = Strength of test solution (Normality or Molarity)
2. Using primary standard
Normality / Molarity = W _
V x F
Where,
W = Weight taken of primary standard (g/mg)
V = Volume of titrant consumed (ml.)
F = Equivalent factor for corresponding solution.
5.3.9 Shelf life of reagents shall be 3 months, the same shall be prepared freshly wherever it is mentioned in the pharmacopoeia / STP
5.3.10 The volumetric solutions / reagents shall be destroyed after expiry and fresh solution shall be prepared. The expiry of Volumetric solutions is mentioned in the STP.
5.3.11 The volumetric solutions / reagents shall never be used if any fungal growth, sedimentation or precipitation is observed.
5.40 Mobile phase :
5.4.1 Mobile phase should be prepared only with 0.45 m membrane filtered water only. Mobile phase should not be stored beyond 24 hours.
5.4.2 Label of mobile phase should consists of the following details :
1. Product / Material for which the mobile phase is being used
2. Prepared by3.Date of preparation.