MPK | STANDARD OPERATING PROCEDURE OF THE MICROBIAL LIMIT TEST OF NON STERILE SAMPLE | MPK

 

1.0.      Purpose:

To describe the procedure of the Microbial limit Test of Non Sterile samples.

(Raw Materials and Finished Products)

                                         

RAW MATERIAL

2.0.      Scope:

This procedure is applicable to Microbiology lab in (Pvt) ltd.

 

3.0.      Responsibilities:

Action	Responsibility
Follow and implement this SOP	Microbiologist
Compliance of SOP	Laboratory Incharge

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4.0.      General Requirements:

4.1. Media / Diluent / Reagents:

4.2  Soybean-Casein Digest broth (SCDB) / Tryptone Soy Broth (TSB)

4.3  Polysorbate 80

4.4  Sabouraud Dextrose Agar. (SDA)

4.5  Sabouraud Dextrose Broth (SDB)/ Sabouraud Liquid Medium

4.6  Peptone Water

4.7  MacConkey Broth. (Mac Br)

4.8  MacConkey Agar. (Mac Ag)

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4.9  Rappaport Vassiliadis Broth. (RVB)

4.0.1  Xylose Lysine Deoxycholate Agar. (XLD)

4.0.2  Cetramide Agar. (CA)

4.0.3  Mannitol Salt Agar. (MSA)

4.0.4  Mossel Broth. (MB)

4.0.5  Violet Red Bile Glucose Agar. (VRBGA)

4.2. Glasswares:

4.2.1  Depyrogenated Sterile Petri Plates

4.2.2  Sterile Diluent Bottles / Test Tubes.

 

4.3. Others:

4.3.1  Micro Pipette with Sterile tips (0.1, 1.0 and 5.0 ml).

4.3.2  Sterile Colony Spreader.

4.3.3  Wire Loop

4.3.4  Isopropyl Alcohol 70%

 

4.4. Equipments:

4.4.1  Incubator                                 (QC – 00)

4.4.2  Incubator (for E coli)               (QC – 00)

4.4.3  Cool Incubator                        (QC – 00)

4.4.4  Depyrogenator                        (QC – 00)

4.4.5  Autoclave                                (QC – 00)

4.4.6  Water Bath                              (QC – 00)

4.4.7  Laminar Air Flow                     (QC – 00)

4.4.8  Bio Safety Cabinet                  (QC – 00)

4.4.9  Analytical Balance                  (QC – 00)

 

4.5. Frequency of Test:

Ø  Every New Non Sterile Product.

Ø  Every Batch of Non Sterile Products / As per Frequency (In house)

5.0.      Procedure

5.1.   Sampling Criteria:

         5.1.1. Steps for Raw Material Sampling:

S. No	Action	Responsibility
5.1.1.1	After receiving intimation Slip of raw material, sampling will be done from the concerned area.	Microbiologist
5.1.1.2	Prior to sampling of Raw material, observe the relevant information. Ø  Material Description / Name of Raw material Ø  Manufacture’s Batch / lot number Ø  I.S number Ø  Quantity Ø  Number of container Ø  Physical condition of containers and Raw material	Microbiologist
5.1.1.3	Raw material sampling is done as per respective sampling plan. Ø  Active: All containers of active material must be sampled as per sampling plan. Ø  Excepient: If the numbers of containers are less than 5 all containers should be sampled. If more than 5 containers Ön+ 1 container should be sampled. Make a composite by mixing equal quantity of all samples and perform test from composite samples.	Microbiologist
5.1.1.4	Raw materials to be sampled are placed under the Laminar Flow in the sampling room.	Microbiologist
5.1.1.5	Sampling will be done in Sterile container with the help of sterile spatula.	Microbiologist

 

5.1.2. Precautions while sampling of Raw Materials:

S. No	Action	Responsibility
5.1.2.1	Sampling of Raw material should be done Under Laminar Flow Hood.	Microbiologist
5.1.2.2	While sampling all excess packaging is to be removed except the primary pack.	Microbiologist
5.1.2.3	Immediately prior to sampling all container to be sampled are to be wiped with a damp cloth or vacuumed, depending on the nature of pack. Bags sampled with a spear (or cut open) must have the sampling area swabbed with 70% IPA.	Microbiologist
5.1.2.4	Always use appropriate sampling tools.	Microbiologist
5.1.2.5	Any Raw Material requiring microbiological check is to be sampled with sterilized tools and into sterilized jars.	Microbiologist
5.1.2.6	Mask, lab coat, shoe covers, and disposable gloves are to be worn for all microbiological raw material sampling; always immediately wash off any spillage of material on your eyes.	Microbiologist
5.1.2.7	Before sampling any raw material carefully read the sampling instructions given on the Raw Material Sampling Specification.	Microbiologist

 

                                                     STERILIZATION 

         5.1.3. Steps for Finished Products Sampling:

S. No	Action	Responsibility
5.1.3.1.	For every New Product first three batches will be sample in three portion (start, mid and end) for validation purpose.	Microbiologist
5.1.3.2.	For routine Sampling of Syrups/Suspensions, 1 pack of bottle will collect randomly from the whole batch.	Microbiologist
5.1.3.3.	For routine Sampling of Tablets and Capsules, sample will be collected after blistering. Quantity of Blisters for sampling depends upon the weight of the tablet and capsule. (minimum 10 grams)	Microbiologist
	Transdermal patches: Remove the protective cover sheets (“release liner”) of the transdermal patches and place them, adhesive side upwards, on sterile glass or plastic trays. Cover the adhesive surface with sterile porous material, for example sterile gauze, to prevent the patches from sticking together, and transfer the patches to a suitable volume of the chosen diluent containing inactivators such as Polysorbate 80 and/or lecithin. Shake the preparation vigorously for at least 30 min.	Microbiologist

 

5.3. General Method for Dilution Preparation:

S. No	Action	Responsibility
5.3.1.	Using the appropriate dilution factor, usually 1:10 dilution by suspending 10 gm or ml of test sample in the 100 ml or 90 ml of diluent, preferably TSB with Polysorbate 80 (1%).	Microbiologist
5.3.2.	Mix the suspensions to disperse the test sample. Note: Care must be taken so that excessive mixing and heating of the sample does not occur during this step.	Microbiologist

 

5.4. General Testing Method for Non Sterile Product (Raw Materials / Finished Goods):

S. No	Action	Responsibility
5.4.1.	Perform the test by using Pour Plate Method. Perform the test at least in duplicate for each Medium and note down the results by using the mean count of the cfu’s.	Microbiologist
5.4.2.	Pour Plate Method: Prepare at least two Petri plates for each medium and each level of dilution. Shake the sample preparation to mix well and pipette 1 ml in duplicate to sterile labeled Petri dishes. Add 15 – 20 ml of tempered appropriate agar in particular plates. Allow liquid sample agar to mix by rotating plates gently clockwise and anticlockwise. Allow agar sample to solidify in Petri plates. Note: Whole test will perform under Laminar Flow Hood.	Microbiologist
5.4.3.	Negative Controls: Diluent Negative Control: Prepare Diluent negative controls by adding 1 ml of each Diluent into two sets of Petri plates. Add 15-20 ml of melted TSA into one set of Petri plates and SDA into another set of Petri plates and incubate appropriately with the product plates. Media Negative Control: Prepare media negative controls by adding 15 – 20 ml of melted TSA and SDA into two Petri Plates respectively. Incubate appropriately with the product plates. Following incubation these plates will serve as negative control for the Total count procedure.	Microbiologist

 

 6.0.      Definitions:

Term/word/statement	Meaning/Description
MLT	Microbial Limit Test
Inoculum	Cells used in an inoculation, such as cells added to start a culture
Pathogens	A pathogen an infectious agent, or more commonly germ, is a biological agent that causes disease to its host.

 

8.0.      List of Records Generated:

S. No	Description
1.	Report of Microbial Limit Test for Raw Materials.
2.	Report of Microbial Limit Test for Finished Products.
3.	Report of Microbial Limit Test for Johnson & Johnson.

 

 

10.0.   Annexure:

S. No	Description
1.	Annexure I - Acceptance Criteria for Microbiological Quality
2.	Annexure II - Morphological Characteristic of Specified Microorganisms on Selective Media
3.	Annexure III - Report of Microbial Limit Test for Raw Materials
4.	Annexure IV - Report of Microbial Limit Test for Finished Product
5.	Annexure V - Report of Microbial Limit