New Methodology for the Identification of Metabolites of Saccharides and Cyclitols by Off-Line EC-MALDI-TOF-MS

At a Glance

Metadata	Details
Publication Date	2020-07-24
Journal	International Journal of Molecular Sciences
Authors	Gulyaim Sagandykova, Justyna Walczak‐Skierska, Fernanda Monedeiro, Paweł Pomastowski, Bogusław Buszewski
Institutions	Nicolaus Copernicus University
Citations	7
Analysis	Full AI Review Included

Technical Documentation & Analysis: Off-Line EC-MALDI-TOF-MS using BDD Electrodes

This document analyzes the research paper “New Methodology for the Identification of Metabolites of Saccharides and Cyclitols by Off-Line EC-MALDI-TOF-MS” to highlight the critical role of high-quality Boron-Doped Diamond (BDD) electrodes and to propose specific material solutions available through 6ccvd.com.

Executive Summary

The research successfully demonstrates a novel off-line electrochemical (EC) system coupled with MALDI-TOF-MS for simulating Phase I oxidative cell metabolism.

Core Technology: The methodology relies on the stability and wide potential window of the Boron-Doped Diamond (BDD) electrode as the working surface for electrochemical conversion.
Application: Simulation and identification of candidate metabolites (via hydrogenation, dehydrogenation, and hydroxylation) of biologically active saccharides and cyclitols (e.g., D-glucose, D-pinitol).
Material Performance: The BDD electrode maintained stability across a wide potential range (0 to 3000 mV) necessary for driving complex redox reactions.
Novelty: This is one of the first applications of an off-line EC system for generating electrochemical products from these low-molecular-weight compounds for subsequent MS analysis.
Analytical Power: The combination allows for the production, detection, and identification of reactive intermediate metabolites, crucial for drug activation studies and metabolomics.
6CCVD Value Proposition: 6CCVD specializes in providing the high-purity, custom-dimensioned BDD substrates required to replicate and scale this advanced electrochemical research.

Technical Specifications

The following hard data points define the operational parameters of the BDD-based electrochemical system and the subsequent MALDI-TOF-MS analysis:

Parameter	Value	Unit	Context
Working Electrode Material	Boron-Doped Diamond (BDD)	N/A	Ultra-thin layer deposited on a silicon substrate
Electrochemical Potential Range	0 to 3000	mV	Required for analyte conversion (EC simulation)
EC System Flow Rate	10	µL/min	Infusion pump operation
EC System Oven Temperature	37	°C	Used during EC conversion
Working Solution pH	7.4	N/A	10 mM Ammonium acetate buffer
MALDI Laser Wavelength	355	nm	Modified Nd:YAG (Smart beam II^TM)
MALDI Laser Frequency	2	kHz	Used for all measurements
MALDI m/z Range	60-1600	m/z	Reflective positive and negative ionization modes
MALDI Laser Power	80	%	Global attenuator at 50%

Key Methodologies

The off-line EC-MALDI-TOF-MS methodology relies on precise control of the BDD working electrode environment:

Electrode System: A three-electrode arrangement was utilized, featuring a Boron-Doped Diamond (BDD) working electrode, a Pd counter electrode, and a HyREF (Pd/H₂) reference electrode.
BDD Substrate: The BDD layer was an ultra-thin film deposited on a silicon substrate, capable of operating up to 3000 mV.
Solution Preparation: Cyclitols and saccharides were prepared at 10 µg/mL concentration in 10 mM ammonium acetate (pH 7.4) mixed with acetonitrile.
EC Conversion: Solutions were injected into the ReactorCell™ at a controlled flow rate of 10 µL/min and maintained at 37 °C.
Fraction Collection: Fractions containing the electrochemically generated products were collected manually, evaporated, and re-dissolved in methanol-water (1:1).
MS Analysis: Samples were analyzed using MALDI-TOF-MS with 2,5-dihydroxybenzoic acid (DHB) and $\alpha$-cyano-4-hydroxycinnamic acid (HCCA) matrices to identify electrochemical products and fragmentation patterns.

6CCVD Solutions & Capabilities

The successful replication and advancement of this EC-MALDI-TOF-MS methodology depend entirely on the quality and customization of the Boron-Doped Diamond electrode. 6CCVD is uniquely positioned to supply the necessary high-performance diamond materials and fabrication services.

Research Requirement	6CCVD Solution & Capability	Technical Advantage
Applicable Materials	Heavy Boron-Doped Polycrystalline Diamond (PCD-BDD)	Provides the high conductivity and wide electrochemical window (up to 3000 mV) essential for robust redox studies and metabolite simulation.
Electrode Substrate & Dimensions	Custom Dimensions & Substrates	We supply BDD plates/wafers up to 125mm (PCD) and custom substrates up to 10mm thick, enabling precise integration into specialized flow cells like the ReactorCell™.
BDD Layer Thickness	Precision Thickness Control	SCD and PCD films are available from 0.1 µm up to 500 µm, ensuring optimal doping uniformity and long-term stability under high potential stress.
Integrated Electrode Systems	Advanced Metalization Services	The experiment required Pd counter and reference electrodes. 6CCVD offers in-house deposition of metals (Au, Pt, Pd, Ti, W, Cu) for creating integrated, robust three-electrode systems directly on the diamond substrate, simplifying assembly and improving contact reliability.
Surface Quality	Ultra-Low Roughness Polishing	While the paper used an ultra-thin layer, 6CCVD offers polishing down to Ra < 1nm (SCD) and < 5nm (PCD), crucial for minimizing non-specific adsorption and ensuring reproducible surface chemistry in sensitive EC/MS applications.
Global Research Support	Engineering Support & Global Shipping	Our in-house PhD team can assist with material selection and design optimization for similar metabolomics and redox simulation projects. Global shipping (DDU default, DDP available) ensures timely delivery worldwide.

Customization Potential

The paper utilized a BDD layer on a silicon substrate. 6CCVD offers the flexibility to deposit BDD films on various substrates (including Si) and provides advanced laser cutting services to achieve the exact geometric requirements for microfluidic or flow-cell EC systems.

Call to Action

To replicate this cutting-edge metabolomics research or to design custom BDD electrodes for your next electrochemical project, 6CCVD provides the highest quality MPCVD diamond materials and expert fabrication services.

For custom specifications or material consultation, visit 6ccvd.com or contact our engineering team directly.

View Original Abstract

A combination of electrochemistry (EC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (off-line EC-MALDI-TOF-MS) was applied for determination of the studied biologically active compounds (D-glucose, D-fructose, D-galactose, D-pinitol, L-chiro-inositol, and myo-inositol) and their possible electrochemical metabolites. In this work, boron-doped diamond electrode (BDD) was used as a working electrode. MALDI-TOF-MS experiments were carried out (both in positive and negative ion modes and using two matrices) to identify the structures of electrochemical products. This was one of the first applications of the EC system for the generation of electrochemical products produced from saccharides and cyclitols. Moreover, exploratory data analysis approaches (correlation networks, hierarchical cluster analysis, weighted plots) were used in order to present differences/similarities between the obtained spectra, regarding the class of analyzed compounds, ionization modes, and used matrices. This work presents the investigation and comparison of fragmentation patterns of sugars, cyclitols, and their respective products generated through the electrochemistry (EC) process.

Tech Support

Original Source

DOI: https://doi.org/10.3390/ijms21155265

References

2019 - Simultaneous Determination of Cyclitols and Sugars Following a Comprehensive Investigation of 40 Plants [Crossref]
2017 - Sugars and health: A review of current evidence and future policy [Crossref]
2019 - Pressurized liquid extraction of cyclitols and sugars: Optimization of extraction parameters and selective separation [Crossref]
2017 - A window on cyclitols: Characterization and analytics of inositols [Crossref]
2018 - Determination of sugars and cyclitols isolated from various morphological parts of Medicago sativa L. [Crossref]
2015 - Discovery and resupply of pharmacologically active plant-derived natural products: A review [Crossref]
2012 - Chemical and Biochemical Applications of MALDI TOF-MS Based on Analyzing the Small Organic Compounds [Crossref]
2019 - The emerging interface of mass spectrometry with materials [Crossref]
2007 - A novel software tool for copolymer characterization by coupling of liquid chromatography with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry [Crossref]