Dilution Solution for Free Cortisol Measurement
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Solution Overview
Problem
Current methods for measuring free cortisol in blood are inaccurate and time-consuming, as they require complex sample pretreatment and measure both free and protein-bound cortisol, making real-time monitoring and diagnosis challenging.
Innovation Solution
A blood sample dilution solution comprising zinc sulfate and an alkaline neutralizing solution is used to precipitate proteins, allowing for the selective measurement of free cortisol without additional pretreatment, using enzyme immunoreaction or mass spectrometry.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If complex sample pretreatment methods (high-efficiency filtration or equilibrium dialysis) are used to measure free cortisol, then measurement precision is improved, but loss of time and device complexity increase significantly
Solution Approach 1:
The invention changes the chemical parameters of the sample by adjusting pH to alkaline conditions (pH 8.0-10.0) and adding zinc ions, which causes protein precipitation and releases bound cortisol. This parameter change simplifies the measurement process while maintaining accuracy, eliminating the need for complex filtration or dialysis procedures.
Solution Approach 2:
The invention extracts free cortisol from the complex blood sample by precipitating proteins that bind cortisol. By adding zinc sulfate or zinc acetate to precipitate proteins, the bound cortisol is released and can be measured separately, effectively extracting the analyte of interest from the complex matrix.
2Measurement precision
If complex sample pretreatment methods are used to measure free cortisol, then measurement precision is improved, but device complexity increases
Solution Approach 1:
The invention uses simple parameter changes (pH adjustment and zinc ion addition) to achieve protein precipitation and cortisol release, replacing complex mechanical or procedural systems with straightforward chemical modifications that can be performed in a single test tube.
Solution Approach 2:
The sample pretreatment method is self-service in nature, where the addition of reagents automatically triggers protein precipitation and cortisol release without requiring external equipment or complex operational steps. The system uses the chemical properties of the sample itself to facilitate the separation process.
3Ease of operation
If antibody-based immunoassays are used to measure cortisol, then ease of operation is improved, but measurement precision deteriorates because total cortisol (free + bound) is measured instead of free cortisol
Solution Approach 1:
The invention extracts free cortisol from the protein-bound form by precipitating the binding proteins (CBG and albumin) with zinc ions at alkaline pH. This separation allows the subsequent immunoassay to measure only free cortisol without interference from bound cortisol, maintaining operational simplicity while improving measurement precision.
Solution Approach 2:
The invention performs preliminary protein precipitation and cortisol release before the immunoassay measurement. This preliminary action ensures that only free cortisol is available for detection by the antibody, preventing the measurement of total cortisol and ensuring accurate free cortisol quantification.
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 enables accurate and rapid evaluation of physiologically active free cortisol, with high recovery rates of free cortisol and other steroids, suitable for clinical diagnostics and emergency situations.
Implementation Method 1
precipitating proteins in serum separated from blood using a protein precipitation solution
Implementation Method 2
adding an alkaline neutralizing solution to inactivate the remaining precipitation solution
Data Source
AI summary
The present invention relates to a diluted solution for measuring free steroids in blood and a method for measuring free steroids using the same, and more particularly, to a method for preparing a diluted blood sample for measuring free steroids in blood and a composition for blood sample dilution solution.In the method for preparing a diluted blood sample for measuring free steroids in blood of the present invention, it was confirmed that physiologically active free cortisol could be evaluated with the cortisol enzyme immunoreaction detection kit without additional sample pretreatment by removing cortisol which is present by binding to excess albumin and cortisol-specific corticosteroid-binding globulin (CBG) contained in serum using a protein precipitation and neutralizing agent. In addition, by selectively removing the protein in the blood sample, it is possible to measure low molecular weight compounds, including steroids, which are strongly bound to proteins in the blood, such as cortisol, in free form.Therefore, the method of the present invention can be applied simply and effectively to medical sites requiring blood tests for disease diagnosis, monitoring of free cortisol before and after surgery, and emergency patients.