Vitamin D Metabolite Extraction Using Displacement Agents
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Solution Overview
Problem
Current methods for determining vitamin D metabolites in biological fluids, such as blood and serum, face challenges including laborious chemical processes, unreliable immunological and LC-MS methods, and interference from hemolysis and protein binding, leading to inaccurate measurements and potential wrong medication.
Innovation Solution
A method involving the use of a solid sorption material that adsorbs body fluids, protecting them from moisture and light, followed by a controlled release of vitamin D metabolites using a solvent buffer and elution solution, allowing for stable storage and analysis without interference from hemolysis or protein binding, using a kit that includes a capillary tube and protective device for easy sampling and transport.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If immunological determination methods are used for vitamin D metabolites, then measurement can be performed, but more than 85% of vitamin D metabolites are bound to VDBP and proteins requiring complex sample preparation
Solution Approach 1:
The invention extracts vitamin D metabolites from their protein-bound state by adding displacement agents (salicylic acid, warfarin, or aniline sulfonic acid) that compete for binding sites on VDBP and albumin, releasing the metabolites into the supernatant for direct measurement without complex preparation
Solution Approach 2:
The invention uses displacement agents as intermediaries that mediate the release of vitamin D metabolites from protein binding. These agents temporarily occupy binding sites on VDBP and albumin, facilitating metabolite release while being easily removable in subsequent processing steps
2Measurement precision
If protein precipitation methods (ethanol or periodate) are used, then vitamin D metabolites can be analyzed, but additional processing steps and reagents are required
Solution Approach 1:
The invention employs disposable displacement agents that are added in excess, perform their function of releasing metabolites, and are then discarded in the waste supernatant, eliminating the need for complex cleanup or recovery steps
Solution Approach 2:
The invention changes the chemical environment by adding displacement agents that alter the binding equilibrium, allowing metabolites to be released and measured directly without physical separation steps required by precipitation methods
3Reliability
If deproteinization and delipidation using acetonitrile are used, then serum samples can be processed, but the procedure becomes time-consuming and complex
Solution Approach 1:
The invention applies partial deproteinization by adding displacement agents that specifically release metabolites from proteins without requiring complete protein precipitation or removal, achieving sufficient sample preparation in a fraction of the time
Solution Approach 2:
The invention performs preliminary release of metabolites from protein binding before analysis, allowing direct measurement without subsequent time-consuming extraction or purification steps
4Measurement precision
If fluorine-containing surfactants are used for metabolite release, then vitamin D metabolites can be extracted, but the method requires additional reagents and processing steps
Solution Approach 1:
The invention uses displacement agents that serve multiple functions: they release metabolites from VDBP and albumin, facilitate metabolite solubility in the supernatant, and can be easily removed in subsequent processing, eliminating the need for specialized surfactants
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 provides stable and accurate determination of vitamin D metabolites, reducing interference from hemolysis and protein binding, and allows for long-term storage and transport of samples, improving measurement reliability and reproducibility.
Implementation Method 1
a solid sorption material that adsorbs body fluids
Implementation Method 2
a controlled release of vitamin D metabolites using a solvent buffer and elution solution
Data Source
AI summary
Method and test kit for quantitative determination of Vitamin D metabolites in blood, wherein a predetermined amount of blood is pre-analytically immobilized on a solid sorption material. Thereby, hemolysis of the blood has no effect on the analysis of vitamin D metabolites. For quantitative analysis the dried blood spot on the sorption material is dissolved with an aqueous solvent buffer containing detergent, pH 7.0 to 10.0, and the vitamin D metabolites are eluted with a protic organic solution having a permittivity of less than 35. The eluate is analyzed for vitamin D metabolites using conventional methods.