Lipid Metabolite Extraction via Phase Separation
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Solution Overview
Problem
The challenge lies in efficiently isolating and identifying structurally and functionally diverse lipids and metabolites from complex biological samples, as existing methods face difficulties in sensitivity, reproducibility, and the ability to handle chemically diverse molecules.
Innovation Solution
A method involving contacting a biological sample with a surface to bind proteins, followed by the addition of a solvent to create separate organic and aqueous layers, where lipids or metabolites partition into the organic layer, allowing for mass spectrometry analysis to identify these molecules.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If proteins are not removed from the biological sample, then the sample composition remains complete, but lipid and metabolite analysis sensitivity is reduced due to protein interference
Solution Approach 1:
The patent removes proteins from the biological sample through precipitation or adsorption onto solid supports, separating them from lipids and metabolites. This extraction of the interfering component (proteins) enables sensitive detection of target analytes without compromising the completeness of the original sample composition
Solution Approach 2:
The patent segments the biological sample into distinct phases or components: protein-containing phases (precipitated or adsorbed) and lipid/metabolite-containing phases (supernatant or organic extract). This segmentation allows selective analysis of lipids and metabolites while proteins are separated and removed
2Reliability
If conventional extraction methods are used for lipids and metabolites, then the extraction process is simple, but reproducibility and sensitivity across diverse lipid classes are poor
Solution Approach 1:
The patent employs systematic variation of extraction parameters including solvent composition (e.g., methanol/chloroform/water ratios), pH levels, temperature conditions, and extraction time to optimize recovery across diverse lipid classes. This parameter optimization enables reproducible extraction of structurally diverse lipids and metabolites
Solution Approach 2:
The patent uses composite solvent systems combining multiple solvents with different polarities (e.g., chloroform-methanol-water mixtures) to simultaneously extract non-polar, polar, and intermediate lipids. This composite approach enhances extraction reliability across the full spectrum of lipid diversity
3Quantity of substance
If a single-phase extraction system is used, then the extraction process is straightforward, but separation of lipids from metabolites is insufficient
Solution Approach 1:
The patent utilizes phase separation by adding water to organic solvent extracts to induce phase splitting into aqueous and organic layers. Lipids partition into the organic phase while metabolites remain in the aqueous phase, achieving effective separation through this phase transition mechanism
Solution Approach 2:
The patent employs an intermediary aqueous phase added to the organic extract to mediate the separation process. This intermediary phase acts as a bridge that enables differential partitioning of lipids and metabolites based on their solubility characteristics
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 effectively reduces protein levels in the sample, allowing for the efficient extraction and identification of lipids and metabolites across a wide dynamic range, enhancing sensitivity and reproducibility.
Implementation Method 1
contacting the biological sample with a surface to bind the at least one protein
Implementation Method 2
adding a first solvent to the altered composition of the biological sample to generate a mixture comprising a first organic layer and an aqueous layer, wherein the at least one lipid or metabolite partitions into the first organic layer
Data Source
AI summary
Described herein are methods of assaying a biological sample, including adding a solvent to a composition of the biological sample to generate a mixture of an organic layer and an aqueous layer. The organic layer contains at least one lipid. The aqueous layer contains at least one metabolite. Also, described herein are methods of assaying a biological sample, including performing mass spectrometry on at least a portion of an organic layer to identify at least one lipid. Also, described herein are methods of assaying a biological sample, including performing mass spectrometry on at least a portion of an aqueous layer to identify at least one metabolite. Also, described herein are apparatus and kits for assaying a biological sample including at least one protein and at least one lipid or metabolite.