Electrochemical simulation of psychotropic drug metabolism compared to in vivo processes using liquid chromatography and mass spectrometry
At a Glance
Section titled âAt a Glanceâ| Metadata | Details |
|---|---|
| Publication Date | 2025-08-28 |
| Journal | Frontiers in Pharmacology |
| Authors | Paulina JerszyĆska, MaĆgorzata SzultkaâMĆyĆska |
| Institutions | Nicolaus Copernicus University |
| Citations | 1 |
| Analysis | Full AI Review Included |
Technical Documentation & Analysis: MPCVD Diamond for Electrochemical Drug Metabolism Simulation
Section titled âTechnical Documentation & Analysis: MPCVD Diamond for Electrochemical Drug Metabolism Simulationâ6CCVD specializes in providing high-quality, custom MPCVD diamond materials (SCD, PCD, BDD) essential for advanced electrochemical and analytical applications, such as the EC-MS/MS methodology detailed in this research.
Executive Summary
Section titled âExecutive SummaryâThis research successfully validates Electrochemical (EC) simulation coupled with LC-MS/MS as a powerful, complementary tool for predicting psychotropic drug metabolism, directly leveraging the unique properties of Boron-Doped Diamond (BDD) electrodes.
- Core Achievement: EC-LC-MS/MS successfully mimicked oxidative Phase I and Phase II (GSH conjugation) metabolism for 11 psychotropic drugs (e.g., Quetiapine, Clozapine, Venlafaxine).
- Material Superiority: The Boron-Doped Diamond (BDD) working electrode was identified as the optimal material, providing the highest signal intensity and enabling electrochemical reactions across a wide potential range (up to 3,000 mV).
- Validation: Electrochemical transformation products (TPs) showed strong agreement with metabolites identified through traditional Human Liver Microsome (HLM) incubations and real patient plasma samples.
- Key Reactions Simulated: The BDD electrode effectively simulated critical Phase I reactions, including N-dealkylation, hydroxylation, and S-oxidation.
- Technical Advantage: The EC-MS approach offers a rapid, ethical, and purely instrumental method for generating oxidative metabolic fingerprints, circumventing time-consuming biological sample preparation.
- 6CCVD Value: 6CCVD is the expert supplier of custom BDD electrodes and substrates required to replicate and scale this high-efficiency analytical technique.
Technical Specifications
Section titled âTechnical SpecificationsâThe following hard data points relate to the critical electrochemical and analytical parameters used in the EC-LC-MS/MS setup:
| Parameter | Value | Unit | Context |
|---|---|---|---|
| Working Electrode Material | Boron-Doped Diamond (BDD) | N/A | Selected for highest signal intensity and widest potential range |
| BDD Potential Range (Oxidation) | 0 to 3,000 | mV | Applied linearly at 10 mV/s |
| Optimal EC Temperature | 36.7 | °C | Controlled oven compartment temperature |
| Optimal Buffer pH (EC) | 5 and 7 | N/A | Ammonium acetate buffer (10 mM concentration) yielded highest signals |
| Electrochemical Cell Type | Thin-layer cell (ReactorCellâą) | N/A | Used for online/offline transformation |
| Drug Solution Concentration | 5 and 10 | ”M | Prepared in selected mobile phase |
| Mobile Phase Flow Rate (EC) | 10 | ”L/min | Syringe pump flow rate |
| ESI Ionization Mode | Positive | N/A | Mass Spectrometry operation |
| Spray Voltage | 4000 | V | LC-MS/MS parameter |
| Drying Gas (Nitrogen) Flow | 6.0 | L/min | LC-MS/MS parameter |
| LC Column | ACE C18 (150 mm x 4.6 mm) | N/A | Used for separation of TPs |
Key Methodologies
Section titled âKey MethodologiesâThe electrochemical simulation of drug metabolism relied on precise control of the BDD working electrode within a specialized thin-layer cell (ReactorCellâą).
- Electrode Selection and Optimization: Four electrode types (Pt, Au, Glassy Carbon (GC), and BDD) were tested. The BDD electrode, consisting of an ultra-thin crystalline diamond layer deposited on a silicon substrate, was chosen for its ability to conduct electrochemical processes in a wide potential range (0-3,000 mV).
- Phase I (Oxidation) Simulation: Drug solutions (5 or 10 ”M) were infused into the electrochemical cell (heated to 36.7 °C) via a syringe pump (10 ”L/min). The BDD potential was linearly increased from 0 to 3,000 mV to induce oxidative transformation products (TPs).
- Phase II (Conjugation) Simulation: For Phase II mimicry, the effluent from the electrochemical cell was mixed online with a solution containing the conjugation agent, Glutathione (GSH), in a reaction coil.
- LC-MS/MS Detection: TPs were separated using LC (ACE C18 column) and analyzed online/offline using a triple quadrupole mass spectrometer operating in positive ion mode (ESI(+)).
- Metabolite Identification: Structures of TPs were proposed based on potential-dependent signal intensities and MS/MS fragmentation spectra, confirming metabolic reactions such as N-dealkylation, hydroxylation, and S-oxidation.
6CCVD Solutions & Capabilities
Section titled â6CCVD Solutions & CapabilitiesâThe successful application of EC-MS/MS for drug metabolism simulation hinges on the quality and performance of the Boron-Doped Diamond (BDD) electrode. 6CCVD is uniquely positioned to supply the high-specification diamond materials necessary to replicate and advance this critical research.
| Research Requirement | 6CCVD Solution & Value Proposition |
|---|---|
| High-Performance BDD Electrodes | Applicable Materials: Heavy Boron-Doped PCD or BDD-Coated Silicon Substrates. The research confirms BDDâs superior performance (0-3,000 mV window). 6CCVD provides custom BDD films with optimized doping levels for maximum electrochemical efficiency and stability in complex buffer systems. |
| Custom Dimensions and Thickness | Custom Dimensions: The experiment used a thin-layer cell. 6CCVD offers custom diamond plates/wafers up to 125mm (PCD) and SCD/BDD films from 0.1”m to 500”m thick, ideal for both miniaturized microfluidic reactors and large-scale analytical systems. |
| Electrode Surface Finish | Polishing: While BDD is used for electrochemistry, 6CCVD offers ultra-smooth polishing (Ra < 1nm for SCD, Ra < 5nm for PCD) for integration into high-precision analytical systems or for use as high-purity thermal management substrates within the EC oven. |
| Metal Contact Layers (Pt, Au, Ti) | Metalization: The paper tested Pt and Au electrodes. 6CCVD offers internal, custom metalization capabilities (Au, Pt, Pd, Ti, W, Cu) for creating robust electrical contacts or integrated electrode arrays on diamond substrates, streamlining reactor fabrication. |
| Replication of EC-MS/MS Methodology | Engineering Support: 6CCVDâs in-house PhD material scientists can assist researchers in selecting the precise BDD specifications (doping concentration, film thickness, and substrate type) required to optimize the simulation of specific oxidative metabolism pathways for psychotropic drugs and similar compounds. |
| Global Research Supply | Shipping: We provide reliable global shipping (DDU default, DDP available) to ensure that specialized BDD materials reach research facilities and pharmaceutical labs worldwide without delay. |
For custom specifications or material consultation, visit 6ccvd.com or contact our engineering team directly.
View Original Abstract
Introduction Psychotropic drugs strongly affect the human psyche through their ability to modulate the neurotransmitter activity and to treat mental disorders and diseases. Monitoring of psychotropic drugs in clinical studies is significant. Thus. establishing methodologies for analyzing these drugs and their pharmacologically active metabolites in biological matrices is essential for patientsâ safety. Therefore, therapeutic drug monitoring (TDM) of these drugs in patients receiving pharmacotherapy in psychiatric hospitals is necessary to avoid medical complications, psychiatric adverse effects, or poisoning. In addition to TDM, the main factor in pharmacokinetics that should be monitored along with the drug is its metabolic pathway. The literature on transformation products (TPs) resulting from the psychotropic drug degradation is limited. Hence, to investigate the potential TPs of target compounds, electrochemistry (EC) and liver microsome assays were used to generate TPs, which were further characterized using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The results obtained by EC-(LC)-MS and liver microsome assays were compared with conventional in vivo studies by analyzing biological samples (human plasma) from patients. Methods The electrochemical mimicry of the oxidative phase I and II metabolism was achieved in a thin-layer cell equipped with a boron-doped diamond (BDD) working electrode under controlled potential conditions. Structures were proposed for the electrochemically generated products based on the MS/MS experiments. Moreover, in order to examine the proposed metabolic pathways of target compounds, the incubation with human liver microsomes was applied. Additionally, a sensitive, specific, and rapid LC-MS/MS method was developed and validated to quantify selected drugs and their metabolites in biological samples. The preparation of biological samples was accomplished through microextraction by a packed sorbent (MEPS). Finally, the results from LC-MS/MS analysis of biological samples, liver microsomes and electrochemical TPs were compared to evaluate the quality of electrochemical metabolism mimicry. Results and discussion Data from in vivo experiments agreed with the data from electrochemical oxidation, which predicted some of the potential metabolites found in the human liver microsomes. EC-(LC)-MS is well-suited for the simulation of the oxidative metabolism of selected psychotropic drugs and acts as the orthogonal source of information about drug metabolites compared to liver microsomes and biological matrices. EC-(LC)-MS enables the direct identification of reactive TPs, circumvents time-consuming sample preparation and is ethically advantageous because it reduces the need for animal experiments.
Tech Support
Section titled âTech SupportâOriginal Source
Section titled âOriginal SourceâReferences
Section titled âReferencesâ- 2012 - Metabolism of the active metabolite of quetiapine, N-desalkylquetiapine in vitro [Crossref]
- 2009 - Metabolic studies of tetrazepam based on electrochemical simulation in comparison to in vivo and in vitro methods [Crossref]
- 2000 - Citalopram-a review of pharmacological and clinical effects
- 2015 - Electrochemistry-mass spectrometry to study reactive drug metabolites and CYP450 simulations [Crossref]
- 2017 - Identification and characterization of vilazodone metabolites in rats and microsomes by ultrahigh-performance liquid chromatography/quadrupole time-of-flight tandem mass spectrometry [Crossref]
- 2016 - Electrochemistry-coupled to mass spectrometry in simulation of metabolic oxidation of methimazole: identification and characterization of metabolites [Crossref]
- 2021 - GLORYx: prediction of the metabolites Resulting from Phase 1 and Phase 2 Biotransformations of Xenobiotics [Crossref]
- 2019 - BioTransformer: a comprehensive computational tool for small molecule metabolism prediction and metabolite identification [Crossref]
- 1999 - O- and N-demethylation of venlafaxine in vitro by human liver microsomes and by microsomes from cDNA-transfected cells: effect of metabolic inhibitors and SSRI antidepressants [Crossref]
- 2009 - Overview of therapeutic drug monitoring [Crossref]