Glyphosate Detection Without Derivatization via LC/MS/MS

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Solution Overview

Problem

Current methods for detecting glyphosate and its metabolites in various test matrices are limited by the need for analyte derivatization and lack specificity, particularly in analyzing new metabolites like N-acetylglyphosate and N-acetyl AMPA, which are not efficiently detected without causing matrix effects.

Innovation Solution

The development of methods and compositions that allow for the detection and quantitation of glyphosate, N-acetylglyphosate, N-acetyl AMPA, and AMPA using LC/MS/MS technology without derivatization, employing phosphoric acid as a weak ion-pairing reagent and stable isotopes as internal standards to normalize matrix effects, enabling robust quantitative analysis across different samples.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If derivatization is used to detect glyphosate and metabolites, then detection capability is improved, but method complexity and loss of information increase

Engineering Contradiction:
Improvedetection capabilityVSAvoidmethod complexity
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The patent extracts and detects glyphosate and its metabolites (N-acetylglyphosate, N-acetyl AMPA, AMPA) directly from test matrices without requiring derivatization steps. The method uses liquid-liquid extraction followed by LC/MS/MS analysis to identify and quantify these compounds in their native forms, eliminating the need for additional derivatization reagents and steps while maintaining detection precision.

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The patent employs phosphoric acid as a weak ion-pairing reagent that acts as an intermediary to enhance detection response and linearity in the LC/MS/MS analysis. This intermediary substance facilitates the detection process without requiring derivatization, improving measurement precision while reducing method complexity.

Inventive Principle:
Principle #24Intermediary (Mediator)

2Device complexity

If conventional detection methods are used for new metabolites, then analysis is simplified, but specificity and detection response deteriorate

Engineering Contradiction:
Improveanalysis simplicityVSAvoidspecificity
Core Design Contradiction:
Device complexityVSMeasurement precision

Solution Approach 1:

The patent changes the detection parameters by using LC/MS/MS technology with specific mass-to-charge ratio filtering to detect glyphosate and its metabolites. The method monitors specific m/z ratios (e.g., m/z 63 for glyphosate, m/z 147 for N-acetylglyphosate) to achieve high specificity. This parameter change enables the detection of new metabolites like N-acetyl AMPA and AMPA with high specificity without increasing method complexity.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent replaces conventional detection mechanisms with LC/MS/MS technology that uses electrical and magnetic fields to separate and detect ions based on their mass-to-charge ratios. This substitution of detection mechanism provides enhanced specificity for new metabolites while maintaining analytical simplicity through automated sample processing.

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

3Reliability

If matrix effects are present in analysis, then detection accuracy is improved, but measurement precision and linearity worsen

Engineering Contradiction:
Improvedetection accuracyVSAvoidlinearity
Core Design Contradiction:
ReliabilityVSMeasurement precision

Solution Approach 1:

The patent uses phosphoric acid as a weak ion-pairing reagent that acts as an intermediary to mitigate matrix effects during LC/MS/MS analysis. This intermediary substance improves detection accuracy while maintaining measurement precision and linearity by reducing ion suppression and enhancement effects caused by the test matrix components.

Inventive Principle:
Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent employs stable isotope-labeled internal standards as feedback mechanisms to normalize and correct for matrix effects. The internal standards undergo the same extraction and ionization processes as the analytes, allowing for real-time compensation of matrix-induced variations in detection response, thereby maintaining both accuracy and precision across different test matrices.

Inventive Principle:
Principle #23Feedback

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

These methods provide a highly specific and sensitive analysis of glyphosate and its metabolites, improving detection response and linearity, and allowing for accurate monitoring in various crop and animal matrices without matrix interference, with a target limit of quantitation of 0.050 mg/kg.

Implementation Method 1

employing phosphoric acid as a weak ion-pairing reagent

Methodology Applied
Scientific EffectIon-pairing:

Implementation Method 2

using LC/MS/MS technology without derivatization

Methodology Applied
Scientific EffectMass spectrometry:

Implementation Method 3

employing phosphoric acid as a weak ion-pairing reagent and stable isotopes as internal standards to normalize matrix effects

Methodology Applied
Scientific EffectIsotope labeling:

Implementation Method 4

extracting a composition comprising the N-acetylglyphosate and/or N-acetyl AMPA from the test sample

Methodology Applied
Scientific EffectLiquid-liquid extraction: Liquid-Liquid Extraction

Data Source

PatentEP2124570B1Methods for detecting glyphosate and metabolites thereof
Publication Date: 2016.11.16 EI DU PONT DE NEMOURS & CO
  • EP2124570B1 patent drawingFigure 1
  • EP2124570B1 patent drawingFigure 2
  • EP2124570B1 patent drawingFigure 3

AI summary

The present invention provides various methods and compositions which allow for determining the presence or amount of glyphosate, N-acetylglyphosate, N-acetyl AMPA or aminomethyl phosphoric acid (AMPA) and its various metabolites in a variety of test matrices. In one method, determining the presence or amount of N-acetylglyphosate and/or N-acetyl AMPA in a test sample comprises providing the test sample suspected of containingN-acetylglyphosate and/or N-acetyl AMPA; extracting the N-acetylglyphosate and/or N-acetyl AMPA from the test sample; and, detecting the N-acetylglyphosate and/or N-acetyl AMPA in the extract. In other methods, the presence or amount of at least one of glyphosate, N-acetylglyphosate, N-acetyl AMPA or aminomethyl phosphonic acid (AMPA) or a metabolite thereof in a test sample is determined. The method comprises providing the test sample suspected of containing at least one of glyphosate, N-acetylglyphosate, N-acetyl AMPA or AMPA or a metabolite thereof; extracting from the test sample at least one of the glyphosate, N-acetyl glyphosate, N-acetyl AMPA or AMPA; and, detecting at least one of the glyphosate, the N-acetylglyphosate, the N-acetyl AMPA and the AMPA from the test sample; wherein detection of the glyphosate, N-acetylglyphosate, N-acetyl AMPA or AMPA occurs without derivatization of the glyphosate, the N-acetylglyphosate, N- acetyl AMPA or the AMPA.