Modified Nucleoside LC/MS Gradient Elution for Faster Separation
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Solution Overview
Problem
The increased hydrophobicity of modified nucleosides due to chemical modification leads to prolonged separation times in LC/MS analysis, delaying the diagnosis and severity prediction of conditions like COVID-19.
Innovation Solution
A method involving gradient elution with a specific solvent mixing ratio change rate in LC/MS to separate and elute modified nucleosides and reference components efficiently, incorporating a third period with a higher solvent mixing ratio change rate between first and second periods.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If gradient elution with conventional solvent mixing ratio change is used to separate modified nucleosides and reference components, then separation accuracy is maintained, but analysis time is prolonged due to increased hydrophobicity of modified nucleosides
Solution Approach 1:
The patent applies dynamics by making the solvent mixing ratio change rate variable rather than constant. The gradient elution process is divided into multiple stages with different change rates: a first stage with a slower change rate for initial separation, a second stage with an accelerated change rate for rapid elution of hydrophobic modified nucleosides, and a third stage with reduced change rate for final separation. This dynamic adjustment of the gradient profile optimizes both separation quality and analysis time.
Solution Approach 2:
The patent changes the parameter of solvent mixing ratio change rate during the gradient elution process. By increasing the change rate in the second stage specifically, the system accelerates the elution of hydrophobic modified nucleosides that would otherwise require prolonged analysis time, while maintaining sufficient separation through the multi-stage approach.
2Reliability
If conventional gradient elution is used for LC/MS analysis of modified nucleosides, then reliable separation is achieved, but diagnostic efficiency is reduced due to prolonged measurement time
Solution Approach 1:
The dynamic multi-stage gradient elution approach maintains reliable separation by preserving the essential gradient progression while adding controlled acceleration phases. The second stage's accelerated solvent mixing ratio change rapidly elutes the target modified nucleosides without compromising separation quality, thereby improving diagnostic efficiency.
Solution Approach 2:
The patent applies the skipping principle by rushing through the elution of hydrophobic modified nucleosides in the second stage with an accelerated solvent mixing ratio change rate. This allows the system to quickly pass through the time-consuming separation phase for these compounds while still achieving reliable separation, thus enhancing overall diagnostic efficiency.
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 significantly shortens the measurement time for modified nucleosides, enabling accurate and early diagnosis and severity prediction of conditions like COVID-19.
Implementation Method 1
separating the target component and the reference component from each other by gradient elution in which a mixing ratio of a plurality of solvents constituting a mobile phase is changed with time to elute from the column
Implementation Method 2
detecting each of the target component and the reference component by mass spectrometry
Data Source
AI summary
An analyzing method includes: an elution step of introducing a sample containing a target component as a modified nucleoside having hydrophobicity increased by modification and a reference component as a component different from the target component into a column, and separating the target and reference components by gradient elution; a step of detecting the target and reference components by mass spectrometry; and a step of calculating a ratio between detection values of the target component and reference components. In the elution step, a mixing ratio of the solvents is changed such that between a first period in which the target component is eluted and a second period in which the reference component is eluted, a third period is provided in which a change rate of a mixing ratio at a column outlet is larger than that of a mixing ratio in each of the first and second periods.