Poloxamer Quantification in Protein Samples via pH-Adjusted SEC
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Solution Overview
Problem
Current methods are unable to quantitatively determine poloxamers in protein samples with molecular masses between 5-70 kDa, which is crucial for assessing the stability and shelf life of liquid pharmaceutical formulations.
Innovation Solution
A method involving size exclusion chromatography with a mobile phase pH adjusted below 3, followed by refractive index detection, is used to separate and quantify poloxamers in liquid protein samples, effectively distinguishing them from proteins and other ingredients.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If size exclusion chromatography is used to separate poloxamers from proteins, then separation is achieved, but poloxamers and proteins with molecular masses between 5-70 kDa cannot be effectively distinguished
Solution Approach 1:
The patent changes the pH parameter of the mobile phase to below 3 (highly acidic conditions). This parameter change causes proteins to denature and aggregate, increasing their apparent molecular size so they are retained by the size exclusion column, while poloxamers remain soluble and pass through, achieving effective separation and accurate quantification
2Quantity of substance
If conventional SEC methods are used, then separation of small molecules from poloxamers is achieved, but quantitative determination of poloxamers in protein samples is not possible
Solution Approach 1:
The method adapts SEC for protein samples by changing the mobile phase pH to below 3. This makes the method versatile for quantifying poloxamers in protein-containing formulations where the protein molecular mass is comparable to the poloxamer, enabling quantitative determination that was previously impossible with conventional SEC methods
Solution Approach 2:
The highly acidic mobile phase acts as an intermediary that differentially affects proteins and poloxamers. It causes protein denaturation and aggregation while maintaining poloxamer solubility, thereby enabling the separation and quantitative determination of poloxamers in complex protein samples
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
This approach allows for precise and accurate measurement of poloxamer concentration in protein samples, ensuring the stability and potency of pharmaceutical formulations over their shelf life, particularly in formulations containing proteins like FSH, LH, and hCG.
Implementation Method 1
a separation step using a size exclusion chromatography column
Implementation Method 2
The poloxamer is detected in the context of a further step by analysing the eluted phase, for example using a RI (refractory index) detection system
Data Source
Figure 1
AI summary
The invention relates to the analytical determination of poloxamers in a liquid protein sample.