Extraction-Free Fatty Acid Quantitation for Polysorbate Hydrolysis

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Solution Overview

Problem

Current methods for quantifying free fatty acids released from polysorbate hydrolysis in pharmaceutical formulations require an extraction step, which is time-consuming and limits high-throughput analysis.

Innovation Solution

A method for quantifying free fatty acids in pharmaceutical formulations without an extraction step, using liquid chromatography and mass spectrometry, with the inclusion of internal standards and calibration curves to ensure accuracy.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If an extraction step is used to quantify free fatty acids, then measurement precision is improved, but productivity deteriorates due to time-consuming extra processing

Engineering Contradiction:

Improvequantification accuracy of free fatty acidsVSAvoidanalysis throughput

Core Design Contradiction:

Measurement precisionVSProductivity

Solution Approach 1:

The patent eliminates the extraction step entirely by using direct injection of the pharmaceutical formulation into the liquid chromatography system. The mass spectrometer with selected ion monitoring capability provides sufficient sensitivity to detect free fatty acids without prior extraction, thereby resolving the contradiction by removing the problematic extraction step while maintaining detection capability through advanced instrumentation.

Inventive Principle:

Principle #2Taking out (Extraction)

2Measurement precision

If an extraction step is used to quantify free fatty acids, then measurement precision is improved, but loss of time increases due to extra processing steps

Engineering Contradiction:

Improvequantification accuracy of free fatty acidsVSAvoidsample processing time

Core Design Contradiction:

Measurement precisionVSLoss of time

Solution Approach 1:

The patent incorporates internal standards into the formulation before incubation, allowing for accurate quantification without subsequent extraction steps. This preliminary action of spiking with internal standards enables direct analysis, eliminating time-consuming extraction and processing steps while maintaining measurement precision through the internal standard correction method.

Inventive Principle:

Principle #10Preliminary action

3Productivity

If direct quantification without extraction is used, then productivity is improved, but measurement precision deteriorates due to matrix interference

Engineering Contradiction:

Improveanalysis throughputVSAvoidquantification accuracy of free fatty acids

Core Design Contradiction:

ProductivityVSMeasurement precision

Solution Approach 1:

The patent uses internal standards as intermediaries to bridge the gap between direct injection and accurate quantification. The internal standards co-elute with the free fatty acids and experience the same matrix effects, allowing for correction of any interference. This intermediary approach enables direct injection (high productivity) while compensating for matrix effects (maintaining precision).

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent implements a feedback mechanism by comparing the signal of free fatty acids to that of internal standards. The internal standards provide a reference that accounts for matrix effects and instrument variability, allowing the system to self-correct and provide accurate quantification without extraction, thus resolving the contradiction between productivity and precision.

Inventive Principle:

Principle #23Feedback

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

Enables sensitive and efficient quantitation of free fatty acids, allowing for the detection of low levels of polysorbate degradation within a short time frame and prediction of long-term degradation.

Implementation Method 1

subjecting said formulation to liquid chromatography to separate said free fatty acids

Methodology Applied

Scientific EffectChromatography: Chromatography

Implementation Method 2

using a mass spectrometer to quantify said free fatty acids

Methodology Applied

Scientific EffectMass spectrometry:

Implementation Method 3

Enzymatic hydrolysis of polysorbate has been recognized as the primary route of polysorbate degradation

Methodology Applied

Scientific EffectHydrolysis: Hydrolysis

Data Source

PatentUS12517098B2Monitoring polysorbate hydrolysis in pharmaceutical formulations using an ultrasensitive extraction-free fatty acid quantitation method

Publication Date: 2026.01.06 REGENERON PHARMACEUTICALS INC

AI summary

The present invention generally pertains to methods of quantifying free fatty acids in a pharmaceutical formulation. In particular, the present invention pertains to a method of quantifying free fatty acids released from polysorbate hydrolysis in a pharmaceutical formulation comprising a pharmaceutical product, polysorbate and free fatty acids, using liquid chromatography-mass spectrometry, without the use of an extraction step.