Microcrystalline MALDI Matrix Spot for Hepcidin Detection
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Solution Overview
Problem
Current methods for analyzing hepcidin and small molecules in complex biological samples, such as urine and blood, face challenges due to high contaminant levels and the need for extensive sample preparation, which reduces sensitivity and increases costs, limiting throughput and accuracy in diagnostic applications.
Innovation Solution
A mass spectrometry method using a microcrystalline MALDI matrix spot with a hydrophobic region allows direct analysis of complex samples without prior purification, enabling the detection and quantification of hepcidin and small molecules by applying a complex sample to a pre-applied microcrystalline MALDI matrix spot, washing, and subsequent analysis via mass spectrometry.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If standard dried droplet preparation or SELDI technique is used for analyzing hepcidin in complex biological samples, then the analysis can be performed, but the sensitivity is insufficient to detect decreased hepcidin levels and the method is time-consuming, limiting high throughput analysis
Solution Approach 1:
The patent applies preliminary action by pre-applying microcrystalline MALDI matrix spots to the sample carrier before sample application. This pre-prepared matrix enables direct analysis of complex biological samples without time-consuming purification steps, thereby improving both detection sensitivity and analysis throughput simultaneously
Solution Approach 2:
The patent uses microcrystalline MALDI matrix with specific crystal size distribution (predominantly 5-20 μm) created by controlled sublimation. This local optimization of crystal size and distribution in the matrix spot enhances ionization efficiency and detection sensitivity while maintaining rapid analysis capability
2Reliability
If extensive sample preparation including dilution or desalting is performed prior to MALDI-MS analysis, then the sample quality is improved, but the detection is limited and overall throughput decreases due to additional steps
Solution Approach 1:
The patent extracts only the essential function of sample preparation by using the hydrophobic region to automatically retain hydrophobic analytes while allowing hydrophilic contaminants to be washed away. This eliminates the need for manual dilution or desalting steps, maintaining sample quality while preserving high throughput
Solution Approach 2:
The sample carrier structure performs self-service by using its built-in hydrophobic region to automatically separate and retain analytes during the washing step. The system cleans itself without requiring external purification equipment or complex sample preparation protocols
3Measurement precision
If LC-MS/MS assay is used for determining serum hepcidin with purification steps, then the target molecules are purified, but the overall throughput decreases and large quantities of sample volume (at least 50 ml) are required
Solution Approach 1:
The patent applies local quality by creating a localized hydrophobic region on the sample carrier that selectively interacts with hydrophobic analytes. This localized functional area enables efficient analyte retention from small sample volumes while allowing rapid washing of contaminants, achieving purification accuracy without requiring large sample volumes
Solution Approach 2:
The patent changes the physical-chemical parameters of the sample carrier by incorporating a hydrophobic region with specific contact angle properties. This parameter change enables selective analyte retention based on hydrophobicity, allowing effective purification from minimal sample volumes and eliminating the need for extensive sample pre-treatment
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 robust, reproducible, and sensitive detection of hepcidin and small molecules, reducing the need for extensive sample preparation, increasing throughput, and improving diagnostic accuracy by allowing direct analysis from unpurified body fluids, even at low concentrations, with high specificity and linearity across a wide range of concentrations.
Implementation Method 1
matrix-assisted laser desorption/ionisation mass spectrometry (MALDI-MS)
Implementation Method 2
analysing said sample for the presence or absence of at least one target analyte via mass spectrometry
Implementation Method 3
a pre-applied microcrystalline MALDI matrix spot which is at least partially encompassed by a hydrophobic region
Data Source
Figure 1a~1g
Figure 2a~2b
Figure 3a
AI summary
The present invention pertains to a mass spectrometry method for analysing the presence or absence of at least one target analyte in a complex sample, comprising at least the following steps: a) Using a sample carrier suitable for mass spectrometry comprising a pre-applied microcrystalline MALDI matrix spot which is at least partially encompassed by a hydrophobic region; b) Applying a complex sample such that it becomes located on said microcrystalline MALDI matrix spot; c) washing said sample on said microcrystalline MALDI matrix spot; d) analysing said sample for the presence or absence of at least one target analyte via mass spectrometry. The method is in particular suitable for quantitatively analysing target analytes such as hepcidin and other peptides, drug compounds and metabolites in body fluids.