Zwitterionic Stationary Phases for Nucleic Acid Gradient Separation
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Solution Overview
Problem
Existing methods are inadequate for efficiently separating nucleic acid component compounds, such as RNA and DNA, in sample fluids, which are crucial for disease monitoring, metabolomic pathway analysis, and nutrient monitoring.
Innovation Solution
The use of a chromatographic column with a zwitterionic stationary phase, combined with a mobile phase comprising a polar aprotic solvent, a protic solvent, and a volatile buffer salt, allows for the separation of nucleic acid compounds by varying the ratio of solvents and ionic strength over time, followed by mass spectrometry analysis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional chromatographic methods are used for separating nucleic acid component compounds, then the separation process becomes complex and time-consuming, but the separation efficiency and precision are insufficient
Solution Approach 1:
The patent employs parameter changes by systematically varying mobile phase composition (polar aprotic solvent to protic solvent ratio), ionic strength (buffer salt concentration), and pH to optimize the separation of nucleic acid component compounds on zwitterionic stationary phases, achieving high precision separation through controlled parameter adjustments rather than complex methodological changes
Solution Approach 2:
The patent utilizes composite material principles by combining zwitterionic stationary phases with specific mobile phase compositions (polar aprotic solvents like acetonitrile combined with protic solvents like water or alcohol and volatile buffer salts), creating a synergistic system that enhances separation precision while maintaining method simplicity
2Productivity
If conventional chromatographic methods are used for separating nucleic acid component compounds, then the analysis time is extended, but the separation efficiency remains inadequate
Solution Approach 1:
The patent achieves improved separation efficiency by optimizing parameter changes including increasing the proportion of polar aprotic solvent in the mobile phase, adjusting ionic strength to enhance peak capacity, and modifying pH to improve selectivity, thereby reducing analysis time while maintaining high separation efficiency
Solution Approach 2:
The patent applies dynamics by implementing gradient elution where the mobile phase composition changes over time (increasing polar aprotic solvent proportion and adjusting ionic strength dynamically), allowing for faster elution of later-retained compounds and reducing overall analysis time while preserving separation efficiency
3Measurement precision
If zwitterionic stationary phases are used with optimized mobile phases, then separation precision is improved, but the method complexity increases
Solution Approach 1:
The patent simplifies operation by establishing standardized parameter ranges for mobile phase composition (specific ratios of polar aprotic to protic solvents), ionic strength (buffer salt concentrations), and pH values that can be directly applied without requiring complex optimization procedures, making the method easy to implement while maintaining high separation precision
Solution Approach 2:
The patent employs volatile buffer salts that can be easily removed by evaporation, allowing for simple sample preparation and analysis without requiring complex cleanup procedures, thereby maintaining operational simplicity while achieving high separation precision through the zwitterionic stationary phase
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 method effectively separates and identifies nucleic acid components, enabling precise analysis and quantification, enhancing the understanding of metabolic pathways and disease monitoring.
Implementation Method 1
separation of nucleic acid component compounds in sample fluids from one another on zwitterionic stationary phases
Implementation Method 2
chromatographic column comprising a zwitterionic stationary phase contained inside the column
Data Source
AI summary
The present disclosure pertains to methods of separating nucleic acid component compounds from one another. In some embodiments, the methods comprise: (a) loading a sample fluid comprising a plurality of nucleic acid component compounds onto a chromatographic column comprising a zwitterionic stationary phase contained inside the column; (b) flowing a mobile phase through the chromatographic column over a time period thereby forming an eluent in which at least some of the plurality of the nucleic acid component compounds are separated from each other, the mobile phase comprising a polar aprotic solvent, a protic solvent, and a volatile buffer salt, wherein flowing the mobile phase comprises varying a ratio of the protic solvent to the polar aprotic solvent over at least a portion of the time period and varying an ionic strength of the volatile buffer salt over at least a portion of the time period.