Improving and Comparing Probiotic Plate Count Methods by Analytical Procedure Lifecycle Management.

Area of Science:

• Microbiology
• Analytical Chemistry
• Food Science

Background:

• Probiotics must contain adequate viable microorganisms for efficacy, with colony-forming units (CFU) as the standard measure.
• Classical plating methods for CFU enumeration exhibit high variability, complicating comparisons and quality control.
• Measurement uncertainties in current methods hinder reliable assessment of probiotic potency.

Purpose of the Study:

• To introduce Analytical Procedure Lifecycle Management (APLM) as a tool to reduce uncertainty in probiotic enumeration.
• To demonstrate how APLM enhances the reliability and comparability of CFU counting methods.
• To provide practical guidance for implementing APLM in the probiotics industry.

Main Methods:

• Application of APLM principles to define and manage the lifecycle of CFU enumeration procedures.
• Utilizing a step-by-step process to establish procedure performance based on fitness for intended use.
• Analysis of simulated and real-world data, including manufacturer development data, to illustrate APLM application.

Main Results:

• APLM implementation identified key factors contributing to variability, such as the dilution step.
• Procedural modifications guided by APLM led to process improvements and reduced variability.
• Enhanced understanding and control over analytical procedures for CFU enumeration were achieved.

Conclusions:

• APLM provides a robust framework for improving the reliability of probiotic viable cell count determinations.
• Implementing APLM enables confident decision-making in probiotic formulation and process control, including overage requirements.
• This approach leads to cost savings through reduced overage and ensures products deliver the intended beneficial doses.