Transport Validation of Microbial Test Samples 

Purpose

The Purpose of the study was to establish the evidence that the Microbial sample does not get contaminated during transport after sampling from sampling location to site of analysis (Microbiology Laboratory).

Details of Study

Three different Batches of Raw Material (Microcrystalline Cellulose PH 101) were selected for the Study. The materials were selected based on the basis of scientific judgement that the material serves as excellent growth medium for growth and multiplication of Microorganism if contamination occurs

Sampling System

Sampling for microbial test sample was done in duplicate from two different container of each Control Number using aseptic technique. Sampling was done under RLAF. Sample quantity of approximately 10 gram was done. Sampling was done from three different control number (A, B and C) of Microcrystalline Cellulose PH101 and details of sampling were recorded.

Sampling Preparation, Sampling procedure and Transport Method

a.      Sampling preparation

Sampling vials were cleaned with 1% Labolene solution and rinsed with purified water. After cleaning, the vials were sterilized at 160°C for 2 hours in Hot Air Oven. Spatulas were wrapped in aluminum foil and sterilized at 160°C for 2 hours in Hot Air Oven.

After sterilization, sampling vials and spatulas were kept in clean and dried sampling box (Prior cleaned with 70% Iso propyl alcohol).The sampling box were packed in plastic bag and transported to warehouse by carrying the sampling box by Sampling Personnel.

Sampling Procedure

Ø  Sampling were done under RLAF of Sampling Booth.

Ø  Sampling vials and spatulas were taken out from sampling box under RLAF after sanitizing hands with 70% IPA and wearing sterile gloves by sampling Personnel.

Ø  Sample containers were opened under RLAF.

Ø  Sampling vials were opened and sampling was done with sterile spatula and vials were closed carefully to prevent from Contamination.

Ø  The vials containing sample were kept in sampling box and the lid of sampling box were closed.

Ø  The sampling box were kept in Plastic Bag.

a.      Transport Method

Ø  The plastic bag containing sampling box were carried from warehouse to Microbiology Lab by Sampling Personnel.

Ø  The samples were stored at 2-8°C in refrigerator until tested.

Analysis Method

1 gm of sample in was dissolved in 10 ml sterile Soyabean casein digest medium and 0.01g polylobate 80 was added to sample containing tube. Using sterile pipette tips, 1ml sample was added in 90 mm diameter of sterile Petri dishes in Triplicate. 15-20 ml sterile and previously cooled at about 45°C Soyabean Casein Digest Agar (SCDA) was added in Petri dishes containing the sample for total aerobic microbial count and swirled to mix the sample. Positive Control was performed using Bacillus Subtilis ATCC 6633 and Negative control without microorganism with test simultaneously. The Petri dishes were allowed to solidify and SCDA Petri dishes were incubated at 35°C for 3 days.

Analytical Result

S.No.	Sample	Control No.	Remarks
			Total Aerobic Microbial Count (Cfu/g)
1	Microcrystalline Cellulose PH 101	A	10
2	Microcrystalline Cellulose PH 101	B	20
3	Microcrystalline Cellulose PH 101	C	10
Limit			NMT 1000 Cfu/g

 

Conclusion

After analyzing, the Sample of three different control number ((A, B and C) of Microcrystalline Cellulose 101, the result was found to be in defined limit. The data obtained from the test indicates that the transport method employed for transport of microbial test sample after sampling is accurate and it has no any significant effect of contamination during transport of sample. Hence the transport method is verified and can be used to transport of microbial test samples.

 

References

1. United States Pharmacopoeial Convention. (2025). <61> Microbiological examination of

Non-sterile products: Microbial enumeration tests. In USP–NF 2025. Rockville, MD.

2. Annexure 4, WHO guidelines for sampling of pharmaceutical products and related materials.

WHO Technical Report Series, No. 929, 2005.

3. Appendix 7, TRS 992. Non-sterile process validation .Annex 3, 2015 (3).