Scope

This SOP applies to all non-sterile dosage forms (oral, topical, nasal, and ophthalmic) analyzed in the Analytical Method Development (AMD) department for microbial quality assessment under pharmacopeial and regulatory guidelines.

3. Responsibilities

• Microbiologist: Performs testing for microbial enumeration and specified organism detection, and conducts identification using biochemical and molecular techniques.
• Analytical Scientist: Assesses risk based on dosage form, target population, and route of administration.
• QA Executive: Reviews identification reports and determines objectionability based on regulatory criteria.

4. Accountability

The Head of Analytical Method Development is accountable for ensuring objectionable organism testing and risk assessments are performed for all relevant products and batches as part of routine microbiological quality control.

5. Procedure

5.1 Initial Detection

1. Perform microbial enumeration as per USP <61>.
2. If unknown colonies are observed during TAMC/TYMC testing or during specified organism screening (USP <62>), proceed to isolation.

5.2 Isolation and Purification

1. Streak representative colonies onto selective and differential media (e.g., MacConkey Agar, Mannitol Salt Agar, Cetrimide Agar, SDA).
2. Incubate plates at appropriate temperatures (30–35°C for bacteria, 20–25°C for fungi) for 24–72 hours.

5.3 Identification Methods

1. Perform Gram staining and microscopic examination.
2. Use biochemical identification kits (e.g., API®, HiBac™, VITEK®) or automated systems for genus/species identification.
3. If needed, confirm identity using 16S rRNA or ITS sequencing for bacteria and fungi respectively.

5.4 Determination of Objectionability

1. Assess the organism’s objectionability based on:
   • Pathogenicity (e.g., E. coli, S. aureus, P. aeruginosa)
   • Route of administration (e.g., ophthalmic, nasal, topical)
   • Patient population (e.g., pediatric, immunocompromised)
   • Product type (e.g., water-based formulations)
2. Refer to FDA Guidance, USP <1111>, and EMA microbial limits guidelines.

5.5 Action Steps for Objectionable Findings

1. Inform QA and initiate deviation/CAPA procedures.
2. Retest retained sample and confirm result.
3. Assess batch impact and recommend disposition (e.g., rejection, recall).

6. Abbreviations

• SOP: Standard Operating Procedure
• CFU: Colony Forming Unit
• API: Active Pharmaceutical Ingredient
• QA: Quality Assurance

7. Documents

1. Isolation and Identification Record – Annexure-1
2. Objectionable Organism Risk Assessment – Annexure-2
3. Microbial Detection Report – Annexure-3

8. References

• USP <61>, <62>, <1111>
• FDA Guidance for Industry: Microbiological Quality Considerations in Non-Sterile Drugs
• Ph. Eur. 5.1.4
• ICH Q6A Specifications

9. SOP Version

Version: 2.0

10. Approval Section

	Prepared By	Checked By	Approved By
Signature
Date
Name	Rohit Verma	Nikita Mehra	Sunita Reddy
Designation	Microbiologist	QA Reviewer	Head – AMD
Department	Analytical Method Development	QA	Analytical Method Development

11. Annexures

Annexure-1: Isolation and Identification Record

Sample ID	Colony Morphology	Media Used	Organism Identified
NSP-056	Pink, circular, dry	MacConkey Agar	E. coli

Annexure-2: Objectionable Organism Risk Assessment

Organism	Product Type	Route	Risk Level	Conclusion
E. coli	Syrup	Oral	High	Objectionable

Annexure-3: Microbial Detection Report

Date	Sample ID	Detected Organism	Reported To	Remarks
20/05/2025	NSP-056	E. coli	QA & Regulatory	Confirmed, deviation raised

Revision History:

Revision Date	Revision No.	Details	Reason	Approved By
21/05/2025	2.0	Updated procedure with risk assessment table and 16S rRNA confirmation option	Annual Review and Compliance Update	Sunita Reddy
01/05/2022	1.0	Initial SOP Release	New SOP	QA Head