Method Optimization — API

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Method Optimization — API

The Challenge

Optimization of Commercial API Assay and Related Substance Method within USP<621>

The Barriers

  • 7 weeks to optimize method within confines of USP<621> and conduct pre-validation
  • Method operates in a narrow pH window (± 0.05) near pKa of the API
  • Method utilizes ion pairing agent
  • Multiple peaks are evaluated for resolution and PDA spectral evaluation during system suitability
  • Tight control of peak tailing and resolution
  • Long analysis time (72 minutes)

Analytical Development Services

  • First 5 weeks: Optimize method conditions
    • Screen columns, mobile phases, column temperature, etc. within bounds of USP<621>
    • Perform forced degradation and evaluate specificity on lead condition
  • Last 2 weeks: Execute full pre-validation on lead condition
    • Evaluate API and impurity accuracy, linearity, LOQ, etc. on primary condition
    • Evaluate specificity with forced degradation / mass balance across three additional conditions

Scientific Data / Explanation

  • 2 Chemists, bi-weekly correspondence, 4 Waters ARC UHPLCs utilized in parallel throughout study
  • 7 weeks of continuous acquisition time from project start to finish (185 sample sequences, 1,083 injections)
  • Two final conditions optimized with either no reversal in elution order or reversal of elution order
    • Best resolution condition reversed elution order but only one column manufacturer could be used
    • Next best condition maintained elution order with expanded flexibility of column selection but required existing conditions
    • Generated reports on Method Development, Pre-validation and Forced Degradation

The Outcome

  • Program initiated on time
  • Efficiently completed project on schedule, using 2 FTEs fewer than the client thought would be required
  • Provided a comprehensive summary of available options for improving method
  • Demonstrated that multiple improved conditions are suitable for validation
  • Performed a full pre-validation with specificity and forced degradation across multiple columns

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