HDL3 Cholesterol Quantification Without Ultracentrifugation
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Solution Overview
Problem
Existing methods for quantifying high-density lipoprotein 3 (HDL3) cholesterol are laborious, requiring skilled operations, special equipment, and prolonged time, and often suffer from interference from other lipoproteins, making them impractical for routine use.
Innovation Solution
A method involving the use of phospholipase and sphingomyelinase to transfer cholesterol from HDL3 to the outside of the reaction system, followed by quantification of remaining cholesterol, allowing for specific measurement of HDL3 cholesterol without laborious operations.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If ultracentrifugation is used to measure HDL3 cholesterol, then measurement precision is improved, but device complexity and operation difficulty increase
Solution Approach 1:
The patent replaces the mechanical ultracentrifugation system with a chemical reaction system using phospholipase and sphingomyelinase enzymes. The enzymatic reaction selectively hydrolyzes phospholipids and sphingomyelin in HDL3, releasing cholesterol that can be measured by standard colorimetric methods, thereby eliminating the need for complex ultracentrifugation equipment while maintaining measurement precision.
Solution Approach 2:
The patent introduces phospholipase and sphingomyelinase as intermediary substances that selectively act on HDL3 cholesterol. These enzymes serve as mediators between the target analyte (HDL3 cholesterol) and the detection system, enabling specific measurement through enzymatic hydrolysis followed by standard cholesterol quantification methods.
2Measurement precision
If HPLC is used to separate HDL2 and HDL3, then measurement precision is improved, but operation time and device complexity increase
Solution Approach 1:
The patent replaces the HPLC mechanical separation system with a biochemical approach using phospholipase and sphingomyelinase. The enzymatic reaction selectively processes HDL3 in the presence of HDL2, allowing direct measurement without time-consuming chromatographic separation. This reduces operation time from hours to minutes while maintaining measurement precision through enzyme specificity.
3Measurement precision
If HDL3 precipitation method is used, then measurement precision is improved, but operation complexity and time increase
Solution Approach 1:
The patent uses phospholipase and sphingomyelinase as intermediary enzymes that selectively bind to and hydrolyze components of HDL3. This enzymatic mediation simplifies the operation by replacing manual precipitation steps with automated enzymatic reactions that can be performed in standard laboratory equipment, reducing both operation complexity and time while maintaining precision.
4Measurement precision
If NMR is used to measure lipoprotein particles, then measurement precision is improved, but device complexity and cost increase
Solution Approach 1:
The patent replaces the complex NMR physical measurement system with a biochemical assay using phospholipase and sphingomyelinase. The enzymatic hydrolysis of HDL3 components releases cholesterol that can be measured by simple colorimetric methods, eliminating the need for expensive and complex NMR equipment while achieving comparable measurement precision through biochemical specificity.
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
Enables accurate and efficient quantification of HDL3 cholesterol using an automatic analyzer, with the option to calculate HDL2 cholesterol by subtraction, eliminating the need for ultracentrifugation or pretreatment.
Implementation Method 1
phospholipase and/or sphingomyelinase is/are allowed to act on a test sample to transfer cholesterol to the outside of the reaction system
Data Source
AI summary
A method that enables quantification of cholesterol in high-density lipoprotein 3 (HDL3) in a test sample without requiring a laborious operation is disclosed. The method for quantifying cholesterol in HDL3 comprises: Step 1 wherein phospholipase and/or sphingomyelinase is/are allowed to act on a test sample to transfer cholesterol to the outside of the reaction system; and Step 2 wherein cholesterol remaining in the reaction system is quantified. The method enables specific quantification of HDL3 cholesterol in a test sample using an automatic analyzer without requirement of a laborious operation such as ultracentrifugation or pretreatment. Further, quantification of the HDL2 cholesterol level can also be carried out by subtracting the HDL3 cholesterol level from the total HDL cholesterol level obtained by a conventional method for quantifying the total HDL cholesterol in a test sample.