Cell Pretreatment With Acid Heating for Microbial Mass Spectrometry
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Solution Overview
Problem
Existing methods for pretreating cells for mass spectrometry, such as using formic acid on a sample plate or in a tube, often result in low peak intensity of cytoplasmic components, particularly for microorganisms with strong cell walls, leading to inadequate identification and discrimination.
Innovation Solution
A pretreatment method involving contacting cells with a first acidic solution followed by heating improves cytoplasmic component extraction efficiency, using organic acids like formic acid, trifluoroacetic acid, or acetic acid, and heating at specific temperatures to enhance peak intensity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If formic acid treatment is performed on a sample plate, then the treatment is simple and quick, but the extraction efficiency of cytoplasmic components is low due to insufficient cell destruction
Solution Approach 1:
The patent applies parameter changes by introducing heating as a thermal parameter to enhance the formic acid treatment. By controlling temperature (e.g., 95°C for 5 minutes), the extraction efficiency of cytoplasmic components is significantly improved while maintaining the simplicity of the overall method. This resolves the contradiction by modifying physical parameters rather than complicating the procedural steps.
Solution Approach 2:
The patent employs preliminary action by performing formic acid treatment and heating before mass spectrometry analysis. This pre-treatment step ensures complete cell lysis and extraction of cytoplasmic components (including ribosomal proteins) prior to analysis, thereby improving extraction efficiency without adding complexity to the main analytical process.
2Productivity
If formic acid treatment is performed in a container, then cell contact with formic acid is improved, but extraction efficiency remains insufficient for microorganisms with strong cell walls
Solution Approach 1:
The patent applies parameter changes by introducing heating as a thermal parameter to enhance the formic acid treatment. By controlling temperature (e.g., 95°C for 5 minutes), the extraction efficiency of cytoplasmic components is significantly improved while maintaining the simplicity of the overall method. This resolves the contradiction by modifying physical parameters rather than complicating the procedural steps.
3Productivity
If bead grinding is used to improve cell destruction, then extraction efficiency improves, but device failure and operational difficulties occur
Solution Approach 1:
The patent replaces the mechanical bead grinding system with a thermal-chemical system combining formic acid treatment and heating. This substitution eliminates the operational difficulties and device failure risks associated with bead grinding while achieving comparable or superior extraction efficiency through chemical lysis and thermal denaturation.
Solution Approach 2:
The patent introduces formic acid as a chemical intermediary that facilitates cell lysis without requiring mechanical force. The acid acts as a mediator between the sample and the extraction process, enabling efficient release of cytoplasmic components through chemical action rather than mechanical disruption.
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
The method significantly enhances the peak intensity of cytoplasmic components, enabling more accurate identification and discrimination of microorganisms, including those with strong cell walls, without the drawbacks of bead grinding or device failure.
Implementation Method 1
contacting the cell with a first acidic solution containing an organic acid
Implementation Method 2
extracting a cytoplasmic component of the cell by heating the cell in contact with the first acidic solution
Data Source
AI summary
A method of pretreatment of a sample containing a cell for mass spectrometry, including contacting the cell with a first acidic solution containing an organic acid; and extracting a cytoplasmic component of the cell by heating the cell in contact with the first acidic solution.