HDL Cholesterol Measurement Using Selective Aggregation Reagents
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Solution Overview
Problem
Conventional methods for measuring HDL cholesterol are inefficient, time-consuming, and prone to inaccuracy due to complications in separation processes and potential for nonspecific reactions or incomplete cholesterol elimination.
Innovation Solution
A method involving cholesterol esterase and cholesterol oxidase or dehydrogenase in the presence of specific nitrogen-containing surfactants and polyanions, such as dextran sulfate, to specifically target HDL cholesterol, avoiding aggregation or elimination of other lipoproteins, thereby enabling accurate measurement.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional fractionation methods (ultracentrifugation, immunochemical method, electrophoresis, precipitation) are used to separate HDL from other lipoproteins, then measurement specificity is improved, but operation complexity and time consumption increase significantly
Solution Approach 1:
The invention extracts and eliminates only the interfering component (non-HDL cholesterol) from the sample using selective aggregation reagents, rather than performing complex separation to isolate HDL. By removing VLDL and LDL through aggregation with dextran sulfate and other reagents, the method directly obtains measurable HDL cholesterol without requiring ultracentrifugation, electrophoresis, or other complex fractionation procedures
Solution Approach 2:
The invention introduces intermediary substances (aggregation reagents including dextran sulfate, divalent metal salts, and specific surfactants) that mediate the selective aggregation of non-HDL lipoproteins. These intermediaries facilitate the separation process by causing VLDL and LDL to aggregate and precipitate, allowing HDL to remain in solution for direct enzymatic measurement without complex instrumentation
2Measurement precision
If conventional fractionation methods are used, then measurement accuracy is improved, but measurement time increases
Solution Approach 1:
The invention performs preliminary selective aggregation of non-HDL lipoproteins in a single reaction step before measurement, eliminating the need for time-consuming sequential separation procedures. The aggregation reagents are added directly to the sample and allowed to act for a brief period, pre-concentrating and removing interfering substances before enzymatic measurement begins
Solution Approach 2:
The invention merges multiple functions into a single reaction system: selective aggregation of non-HDL lipoproteins, clarification of supernatant, and enzymatic measurement of HDL cholesterol all occur in one integrated process. This combines what would traditionally require separate ultracentrifugation, decanting, and measurement steps into a single test tube reaction
3Ease of operation
If aggregation reagents are used to eliminate non-HDL cholesterol, then measurement simplicity is improved, but turbidity and deposition problems occur
Solution Approach 1:
The invention optimizes reaction parameters including pH, temperature, and reagent concentrations to control the aggregation process. By adjusting these parameters, the method achieves complete aggregation of non-HDL lipoproteins while minimizing turbidity interference and preventing excessive deposition that could clog measurement systems or interfere with optical detection
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 simple and accurate measurement of HDL cholesterol without the need for complex separation steps, reducing measurement time and improving specificity, thus enhancing diagnostic capabilities for arteriosclerosis prediction.
Implementation Method 1
reacting the sample with i) cholesterol esterase and cholesterol oxidase or ii) cholesterol esterase, oxidized coenzyme and cholesterol dehydrogenase
Implementation Method 2
cholesterol oxidase or dehydrogenase in the presence of specific nitrogen-containing surfactants and polyanions
Implementation Method 3
specific nitrogen-containing surfactants and polyanions, such as dextran sulfate
Implementation Method 4
forming a complex with lipoproteins other than HDL
Data Source
AI summary
The present invention provides a reagent, a reagent and a kit for the simple and accurate measurement of HDL cholesterol. The method for the measurement of cholesterol in high-density lipoprotein in a sample comprises reacting the sample with i) cholesterol esterase and cholesterol oxidase or ii) cholesterol esterase, oxidized coenzyme and cholesterol dehydrogenase in an aqueous medium comprising a specific nitrogen-containing surfactant having the structure of amine or ammonium salt and a polyanion, and measuring the formed hydrogen peroxide or reduced coenzyme.