Model Compounds Research Articles

Exploring the Photocatalytic Cleavage Pathway of the β-5 Linkage Lignin Model Compound on Carbon Nitride.

As a globally abundant source of biomass, lignocellulosic biomass has been the centre of attention as a potential resource for green energy generation and value-added chemical production. A key component of lignocellulosic biomass, lignin, which is comprised of aromatic monomers, is a potential feedstock for value added chemical production. The cleavage processes of the linkages between monomers to obtain high value products, however, requires significant investigation as it is a complex, non-facile process. This study focuses on the photocatalytic valorization of a β-5 lignin model compound, a key linkage in the lignin structure. It was found that greater yields of aromatic products were obtained from the photocatalytic conversion of β-5 lignin model compound using carbon nitride (CN) when compared to Evonik P25 titanium dioxide (TiO2). Products of the β-5 model compound photocatalytic conversion were determined and C-C bond cleavage was observed. It was also determined that the solvent participated in the reactions with the introduction of a cyano group to one of the products. Radical quenching experiments revealed that superoxide radicals participated in the CN photocatalytic conversion. These results reveal for the first time the products and possible mechanism of the photocatalytic transformation of β-5 model compounds using CN photocatalysis.

Novel Piperazine Based Compounds Target Alzheimer's Disease Relevant Amyloid β42 and Tau Derived Peptide AcPHF6, and the Lead Molecule Increases Viability in the Flies Expressing Human Tau Protein.

Alzheimer's disease (AD) is the leading form of dementia in the United States and the world. The pathophysiology of AD is complex and multifaceted. Accumulation of senile plaques and neurofibrillary tangles (NFTs) are hallmarks of AD. The aggregation of amyloid β (senile plaques) and tau tangles (NFTs) results in the death of neurons in the cortex and hippocampus, which manifests itself in cognitive decline and memory loss. Current therapies rely on conventional approaches that have only treated the underlying symptoms without disease modification. Data from clinical studies point to a complex role of amyloid β (Aβ) in a way that enhances the tau phenotype throughout the disease process. To address the co-pathogenic role of Aβ and tau, we undertook development of multitarget compounds aiming at both tau and Aβ to slow or stop disease progression and provide neuroprotection. Here, we demonstrate a dose-dependent effect of the novel test compounds that inhibit aggregation of AcPHF6 (a shorter version of tau protein) and Aβ1-42 peptides in thioflavin T fluorescent assays. The compounds were also shown to disaggregate preformed aggregates dose dependently. To further validate these findings, circular dichroism experiments were carried out to examine the nature of inhibition. Additionally, transmission electron microscopy experiments were carried out to gain insights into the morphologies of aggregates obtained from dose-dependent inhibition of AcPHF6 and Aβ1-42 as well as dissociation of preformed aggregates from these peptides. Compounds D-687 and D-688 reversed Aβ1-42 induced toxicity in SH-SH5Y cells, significantly demonstrating neuroprotective properties. Finally, in a study with Drosophila melanogaster expressing human tau protein isoform (2N4R) in all the neurons, compound D-688 significantly increased the survival of flies compared to vehicle treated controls. Future studies will further examine the neuroprotective properties of these lead compounds in various animal models.

Fluorescence-guided tumor visualization of colorectal cancer using tumor-initiating probe yellow in preclinical models.

Fluorescence-guided surgery has emerged as an innovative technique with promising applications in the treatment of various tumors, including colon cancer. Tumor-initiating probe yellow (TiY) has been discovered for identifying tumorigenic cells by unbiased phenotypic screening with thousands of diversity-oriented fluorescence library (DOFL) compounds in a patient-derived lung cancer cell model. This study demonstrated the clinical feasibility of TiY for tumor-specific fluorescence imaging in the tissues of patients with colorectal cancer (CRC). To evaluate the efficacy of TiY in tumor imaging, surgical specimens were obtained, consisting of 36 tissues from 18 patients with CRC, for ex vivo molecular fluorescence imaging, histology, and immunohistochemistry. Orthotopic and chemically induced CRC mice models were administered TiY topically, and distinct tumor lesions were observed in 10min by real-time fluorescence colonoscopy and ex vivo imaging. In a hepatic metastasis mouse model using splenic injection, TiY accumulation was detected in metastatic liver lesions through fluorescence imaging. Correlation analysis between TiY intensity and protein expression, assessed via immunohistochemistry and Western blotting, revealed a positive correlation between TiY and vimentin and Zeb1, which are known as epithelial-mesenchymal transition (EMT) markers of cancers. A comparative analysis of TiY with other FDA-approved fluorescence probes such as ICG revealed greater quantitative differences in TiY fluorescence intensity between tumor and normal tissues than those observed with ICG. Altogether, these results demonstrated that TiY has a strong potential for visualizing CRC by fluorescence imaging in various preclinical models, which can be further translated for clinical use such as fluorescence-guided surgery. Furthermore, our data indicate that TiY is preferentially uptaken by cells with EMT induction and progression, and overexpressing vimentin and Zeb1 in patients with CRC.

Neo-Glycolipid Oximes as Intestinal Permeation Enhancers for Peptide Hormone PYY3-36.

Herein, we describe the design and synthesis of 16 neo-glycolipids that are potential permeation enhancers for oral drug delivery of peptide therapeutics. These amphiphilic neo-glycolipids are composed of fatty acids and various carbohydrates (D-glucose, lactose, cellobiose, maltose) via an oxime linker. The ability of the synthesized neo-glycolipids to enhance permeation of fluorescein-labelled dextran (4 kDa) or 3H-mannitol across intestinal epithelium was investigated in vitro using monolayers of human epithelial Caco-2 cells. Their effects were compared with (pre-)clinically known enhancers as reference compounds; sodium salts of octanoic, decanoic, and dodecanoic acid, and sodium salcaprozate (SNAC). Most neo-glycolipids increased the permeation of the model compounds, proving that neo-glycolipids, which possess vastly different properties from the reference compounds, e.g., in terms of clogD and polar surface area, are effective permeation enhancers. The neo-glycolipid based on decanoic acid and glucose was more potent than related compounds based on disaccharides. Significant differences in solubility and cellular compatibility were found for neo-glyolipids based on different carbohydrates. Finally, neo-glycolipids were evaluated as permeation enhancers for the peptide hormone PYY3-36. Glucose- and maltose-derived neo-glycolipids based on decanoic and dodecanoic acid showed promising enhancements in PYY3-36 permeation while maintaining good cellular compatibility, relevant for oral delivery of obesity treatments.

Impact of Potassium Addition on the Performance of Ni/MgAl2O4 Catalysts in Steam Reforming of Bio-Oil Model Compounds

Impact of Potassium Addition on the Performance of Ni/MgAl₂O₄ Catalysts in Steam Reforming of Bio-Oil Model Compounds

Hydrodeoxygenation of Isoeugenol-Derived Model Compound over Carbon-Supported Pt and Pt-SnS Catalysts for the Production of Sustainable Jet Fuel.

This study focuses on the sustainable production of bio-jet fuel through the catalytic hydrodeoxygenation (HDO) of isoeugenol (IE). Sucrose-based activated carbon supported bimetallic Platinum-Tin metal sulphides (PtO-SnS/AC) catalyst was prepared for HDO process. Physicochemical properties of catalysts with different spraying synthesis methods (in situ and ex situ metal doping) and Pt loading (0.1-1.0 %) were further investigated. The PtO-SnS/AC catalysts were characterised using various techniques such as X-ray absorption spectroscopy (XAS), X-ray photoelectron spectroscopy (XPS), X-ray diffraction (XRD), high-resolution transmission electron microscopy (HRTEM), field-emission scanning electron microscopy (FESEM) and thermogravimetric analysis (TGA). Both HRTEM and FESEM results show the successful preparation of a spherical nanoparticles doped over activated carbon, and Pt was dispersed on the outer shell of the particles. The catalytic HDO of IE was evaluated in a batch system and showed a high yield and conversion as follows: IE conversion of 100 %, liquid-phase mass balance of 95.92 %, dihydroeugenol (DH) conversion of 99.32 %, propylcyclohexane (PCH) yield of 88.94 % and 2-methoxy-4-propylcyclohexanol (HYD) yield of 76.19 %. Moreover, the PtO-SnS/AC catalyst exhibited high reusability with low metal leaching and high coke resistance for 10 cycles. The catalyst was evaluated in a continuous flow reactor for 100 h at different reaction temperatures, and interestingly, the catalyst showed low deactivation with a high half-time.

Degradation of sulfamethoxazole in a falling film dielectric barrier discharge system: Performance, mechanism and toxicity evaluation

The ubiquitous presence of sulfonamides (SAs) in wastewater poses serious risks to human health and ecosystem safety. This study evaluated the performance of a falling film dielectric barrier discharge (DBD) system on the removal of five SAs, namely sulfamethoxazole (SMX), sulfisoxazole (SIZ), sulfathiazole (STZ), sulfadiazine (SDZ) and sulfamerazine (SMR). Removal efficiencies >99 % were observed for all target SAs within 30 min of treatment, with pseudo-first order rate constants varying between 0.17 and 0.27 min−1. Superior removal efficiencies were achieved under acidic conditions compared to neutral and alkaline conditions. Using SMX as a model compound, mechanistic investigations revealed that the synergy of reactive oxygen species (ROS) and reactive nitrogen species (RNS) led to its efficient degradation, with peroxynitrites (ONOO−/ONOOH) and hydroxyl radical (OH) playing pivotal roles. SMX degradation pathways encompassing nitration/nitrosation, hydroxylation, deamination, CS and SN bond cleavage were proposed. The toxicity evaluation results demonstrated that the solution toxicity diminished following the plasma treatment under specific conditions. In particular, the solution treated with air or oxygen discharge enhanced the growth of wheat seedlings, suggesting the potential for reusing plasma-treated wastewater in agriculture. This study enhances our understanding of the underlying mechanisms involved in the plasma degradation of SAs and reveals the significant potential of plasma technology as a sustainable approach for treating wastewater.

Steam reforming of toluene as model compounds of biomass tar by Fe–K-based biochar nano-catalysts at mild temperatures

Development of efficient catalysts for steam reforming of biomass tar is crucial for the widespread application of biomass gasification. In this work, a novel biochar nano-catalyst (BN) is conveniently prepared via one-step catalytic pyrolysis, using K and Fe to adjust the surface structure of the biochar, thereby enhancing the activity for toluene reforming. Fe–K/BN is tested in a fixed-bed reactor for steam reforming of toluene, a model compound of biomass tar, achieving a 100% toluene conversion rate at 650 °C. Fe–K/BN is activated in situ with steam to increase their specific surface area and enhance the O-containing functional groups on the surface, improving toluene adsorption and reforming. Finally, the stability of Fe–K/BN is tested in the simulated syngas, maintaining a 100% toluene conversion rate during a 26-h continuous catalytic reforming experiment. This work provides a promising strategy for the design of an active and stable catalyst for toluene reforming, which has potential for application of tar steam reforming during biomass gasification.

Insights into the oxygen vacancies in titanium-aluminum composite oxides for Ru-catalyzed bio-oil upgrading using guaiacol as a model compound

Insights into the oxygen vacancies in titanium-aluminum composite oxides for Ru-catalyzed bio-oil upgrading using guaiacol as a model compound

Hydrodeoxygenation of guaiacol over modified coconut carbon supported Ni nanoparticles catalysts under alkaline condition

The efficient utilization of lignin from the papermaking black liquor has attracted interest due to its carbon-neutral value and industrial application. This work illustrates a simple support-phosphate-pretreatment strategy to develop superdispersed Ni species in activated carbon (AC) to enhance the hydrodeoxygenation (HDO) reactions of the lignin model compound guaiacol into biohydrocarbons under alkaline conditions. For the first time, sodium polyphosphate was applied as a pretreatment agent for coconut carbon in the synthesis of carbon-supported Ni catalysts. Superdispersed Ni nanoparticles were achieved on Ni/NaPnOAC with a particle size of 2.5 nm, which was much smaller than that on unmodified Ni/C (13.1 nm). The characterization results and reactions revealed that the P−O that formed served as anchoring sites for the Ni species and strengthened the interaction between the Ni species and support (SMSI), which resulted in significantly improved dispersion of the Ni metal sites and, thus, a nearly 6-fold greater yield of hydrocarbons was obtained on Ni/NaPnOAC (58.1%) than on Ni/C (10.1%). In addition, compared with Ni2P/NaPnOAC and NiS/NaPnOAC, Ni/NaPnOAC exhibited higher HDO activity, especially higher direct deoxygenation (DDO) activity and higher benzene yield, reducing hydrogen consumption during the HDO reaction.

1,4-Benzodioxane Substituted Aza-BODIPY: Towards Photostable yet Efficient Triplet Photosensitizer.

We report herein the synthesis of aza-BODIPY substituted with 1,4-benzodioxane-6-yl substituents at 3,5 positions of the chromophore system. Both pyrrole rings of the aza-BODIPY in question were substituted with bromine atoms in order to induce highly desirable photophysical properties, such as highly populated excited triplet state (T1) and long excited triplet-state lifetime (τT) of 21 μs. The photosensitized oxygenation of a model compounds, viz. DPBF, points to a high singlet oxygen and/or other ROS formation quantum yield of 0.42. The photosensitizer studied exhibited an absorption band within the so-called "therapeutic window", with λabs 678 nm. As estimated by CV/DPV measurements the 1,4-benzodioxane-6-yl substituted aza-BODIPYs studied exhibited a multi-electron oxidations at a relatively low potentials (Eox), pointing to the very good electron-donating properties of these molecules. High photostability and thermal stability was observed for all compounds studied. The good singlet oxygen quantum yield measured combined with an exceptional photostability makes this aza-BODIPY a promising candidate for applications such as photocatalysis and photodynamic therapy (PDT).

Gastrointestinal fate of proteins from commercial plant-based meat analogs: Silent passage through the stomach, oxidative stress in intestine, and gut dysbiosis in Wistar rats.

Plant-based meat analogs (PBMAs) are common ultra-processed foods (UPFs) included in the vegan/vegetarian diets as presumed healthy alternatives to meat and meat products. However, such health claims need to be supported by scientific evidence. To gain further insight into this topic, two commercial UPFs typically sold as meat analogs, namely, seitan (S) and tofu (T), were included in a cereal-based chow and provided to Wistar rats for 10weeks. A group of animals had, simultaneously, an isocaloric and isoprotein experimental diet formulated with cooked beef (B). In all cases, experimental chows (∼4kcal/g feed) had their basal protein concentration increased from 14% to 30% using proteins from S, T, or B. Upon slaughter, in vivo protein digestibility was assessed, and the entire gastrointestinal tract (digests and tissues) was analyzed for markers of oxidative stress and untargeted metabolomics. Metagenomics was also applied to assess the variation of microbiota composition as affected by dietary protein. Diets based on PBMAs showed lower protein digestibility than those containing meat and promoted an intense luminal glycoxidative stress and an inflammatory intestinal response. The fermentation of undigested oxidized proteins from T in the colon of Wistar rats likely led to formation of mutagenic metabolites such as p-cresol. The presence of these compounds in the animal models raises concerns about the potential effects of full replacement of meat by certain PBMAs in the diet. Therefore, future research might target on translational human studies to shed light on these findings.

Electroextraction of Large Volume Samples Using Paper Points Coupled With Hollow Fiber Membranes: Study of Parameters and Strategies to Enhance Analytical Performance.

Electroextraction (EE) encompasses a range of sample preparation methods whose effectiveness, selectivity, and efficiency are significantly influenced by the physical-chemical characteristics of analytes, samples, and instrumental conditions. This article explores, for the first time, various strategies aimed at enhancing the extraction efficiency of a recent approach of EE utilizing a paper point (PP) combined with a hollow fiber (HF) (abbreviated as PP-HF-EE) to extract various cationic and anionic model compounds from water samples. The study also explores, experimentally, the impact of agitation, organic filter composition, PP diameter, and PP brand on extraction performance, and proves that all these factors are quite important, especially when digital image analysis is utilized for determination. Furthermore, this work demonstrates the ease and feasibility of simultaneously extracting cations and anions using PP-HF-EE and proposes a straightforward method to enhance analyte concentration on the vertex of the PP through a base-to-vertex focusing. Lastly, it is demonstrated, using capillary electrophoresis coupled to a UV-Vis detector, that for PP-HF-EE, the extraction efficiency and pre-concentration factor are less dependent on other parameters when multiple PPs per sample are utilized, with signal enhancement values of up to 111 and 339 for nortriptyline and haloperidol, respectively. All the findings and strategies presented herein constitute significant contributions that can facilitate future research in method development, particularly in the utilization of PP-HF-EE and similar EE approaches.

Glaser-Hay-Coupled Random Copolymers Containing Boron Difluoride Formazanate Dyes.

𝜋-Conjugated polymers, including those based on acetylenic repeating units, are an exciting class of materials that offer narrow optical band gaps and tunable frontier orbital energies that lead to their use in organic electronics. This work expands the knowledge of structure-property relationships of acetylenic polymers through the synthesis and characterization of a series of Glaser-Hay-coupled model compounds and random copolymers comprised of BF2 formazanate, fluorene, and/or bis(alkoxy)benzene units. The model compounds and copolymers synthesized exhibit redox activity associated with the reversible reduction of the BF2 formazanate units and the irreversible reduction of the fluorene and bis(alkoxy)benzene units. The copolymers exhibit absorption profiles characteristic or intermediate of their respective models and homopolymers, leading to broad absorption of UV-vis light. The alkyne linkages of the model compounds and copolymers are reacted with [Co2(CO)8] to convert the alkyne functional groups into cobalt carbonyl clusters. This transformation leads to blue-shifted absorption profiles due to a decrease in π-conjugation, demonstrating the ability to tune the properties of these materials through post-polymerization functionalization. The redox activity and broad absorption bands of the polymers reported make them excellent candidates for use in photovoltaics and other light-harvesting applications.

Copper Incorporated Covalent Organic Framework as a Heterogeneous Catalyst for CuAAC Reaction

AbstractCovalent organic frameworks (COFs) incorporating metal are attractive alternatives for metal‐catalyzed organic transformations. For effective metal incorporation in COF a favorable ligand environment is required. Pyridine and hydrazone units can provide effective binding sites for transition metals. The major challenge in synthesizing hydrazone‐linked COFs is the inherent flexibility of the linker, causing differences in lengths and orientations during solvothermal synthesis. We demonstrate that incorporation of enol form in the framework facilitates non‐covalent interactions such as hydrogen bonding, reduces degrees of freedom and enhances rigidity. Here, we synthesized TFP‐PyHz COF utilizing 2,4,6‐trihydroxybenzene‐1,3,5‐tricarbaldehyde (TFP) and pyridine‐2,6‐dicarbohydrazide. Enol form in the framework was confirmed by comparing the IR and 13C solid‐state NMR spectra of TFP‐PyHz with its model compound. The presence of this enol form also facilitates the incorporation of Cu2+ through post‐modification as confirmed by IR and XPS analysis of postmodified Cu‐TFP‐PyHz. The copper‐incorporated material Cu‐TFP‐PyHz is utilized as a heterogeneous catalyst for copper‐catalyzed click reactions, enabling the synthesis of 1,4‐triazoles.

Regulation of Adsorption Properties of Whey Protein Isolate Fibril-Calcium Alginate Aerogels by Controlling Protein Fibrillation Time.

Amyloid-like fibrils have a relatively high specific surface area, which makes them suitable as absorbents. In this study, we prepared absorbent composite aerogels from whey protein isolate fibril (WPIF) and calcium alginate (CA). The impact of protein fibrillation time (4-24 h) on the adsorption properties of these aerogels was assessed, as this influences the nature of the WPIFs formed. Fourier transform infrared spectroscopy analysis indicated that hydrogen bonding, hydrophobic interactions, and electrostatic interactions were the main molecular interactions in the composite aerogels. Then, the adsorption properties of the aerogels were investigated using crystal violet as a model compound. The adsorption properties of all composite aerogels were significantly improved compared to CA aerogels, which was mainly attributed to the numerous functional groups of the surfaces of the WPIFs, as well as the rougher surface topology of the composite aerogels. The adsorption capacity of the composite aerogels first increased and then decreased with increasing fibrillation time, with 8 h fibrillation leading to the largest adsorption capacity (1886.11 mg/g). Thioflavin T fluorescence, atomic force microscopy, light extinction, average particle size, and zeta potential analysis indicated that the high fibril content, mature fibril morphology, large number of available functional groups on the surface, and relatively low zeta potential of WPIF8 might be the main reasons. Adsorption kinetics and isothermal adsorption profile analysis suggested that monolayer adsorption occurred through chemical adsorption processes. The edible aerogel with high adsorption properties developed in this work has potential application in food and biomedical fields.

HZSM-5 supported with N-doped Fe for microwave catalytic pyrolysis of lignin

Microwave catalytic pyrolysis of lignin is one of emerging technologies for efficiently converting lignin into fuels and chemicals. Molecular sieves (HZSM-5) are widely used as carriers to load metallic components for biomass pyrolysis but often encounter challenges such as high diffusion resistance and insufficient acidity of acidic sites. Traditionally, HZSM-5 is directly modified to enhance the activity of the acidic sites, but this modification is ineffective in improving the metallic components’ activity. Instead, we made nitrogen doping on Fe and then supported on HZSM-5 to construct Fe-N25/HZSM-5 catalysts for microwave catalytic pyrolysis of lignin. The N-doped Fe has the effect of “killing two birds with one stone” on enhancing the catalytic activity. The characterization results indicated that N replaced C of the graphitic carbon network to form carbon nitride structure, and further combined with Fe to generate Fe-N bonds, improving the electronegativity of the metallic component. Besides, the loading of N-doped Fe increased the acidity of the catalyst. The utilization of 1.00Fe-N25/HZSM-5 obtained the greatest bio-gas yield (35.2 wt%), a hydrogen yield of 27.8 mL/min, bio-oil yield (36.4 wt%) and phenol selectivity of 46.2 %. The likely pathway of microwave catalytic pyrolysis on Fe-N25/HZSM-5 was proposed through DFT calculation, using guaiacol as a model compound. The results suggested that N-doped Fe on HZSM-5 8 T tended to break the side-chain methoxy during pyrolysis of guaiacol, with energy barrier of 0.14 eV. This paper provides a promising strategy for modification of microwave pyrolysis catalysts.

Exploring the efficacy of Re-Ni embedded KIT-6/H-Mor catalysts for renewable energy – A comprehensive experimental and theoretical (DFT) exploration

The global market for n-propyl benzene value is projected to reach $30.47 billion by 2030, with a CAGR of 4.3% from 2023 to 2030. This market analysis identifies diverse applications for propyl benzene, spanning the petroleum industry, industrial chemicals, automotive sector, and more. The research introduces eugenol, a compound derived from lignin, as a model compound for bio-oil to n-propyl benzene (bio-fuel analogs) production. A novel Re-Ni/KIT-6-H-Mordenite catalyst is developed for efficient hydrodeoxygenation of eugenol, showcasing high conversion rates and selectivity under moderate temperatures. Comprehensive characterization techniques and theoretical calculations highlight the synergistic interaction between Re and Ni metals. The catalyst’s reusability and successful application for bio-fuel production under atmospheric conditions further validate its potential. This study pioneers the exploration of Re-Ni bimetallic catalysts for HDO of lignin-derived compounds, offering valuable insights into sustainable bio-fuel development.

Rational screening of metal single-atom-doped ZSM-5 to promote ring-opening cracking of cycloalkanes to light olefins

The ring-opening cracking of cycloalkanes to light olefins poses a formidable challenge due to the sluggish activation of the C–C bond. In this study, we realize the rational screening of various single metal doped M−ZSM−5 catalysts (M = Mn, Co, Ni, Cu, Zn, Zr, Ru, Rh, Pd, Ag, Pt) to promote the ring-opening cracking of cycloalkanes to light olefins. Our calculation results show that methyl cyclopentane, a model compound of cycloalkanes, tends to adsorb in the form of a “half-chair” configuration in the intersecting channels of the M−ZSM−5, which is caused by the drastic distortion of the five-membered ring, accompanied by large changes in the bond angles. Based on the host–guest interaction between the active site and hydrocarbon in ring-opening cleavage, we reveal that the binding energies of metals to H and C can respectively quantify the metal’s ability to activate C–H and C–C bonds in cycloalkanes. By solving the microkinetic model, we plot activity-selectivity TOF volcano diagrams, where Mn, Ru, Rh-ZSM-5 within the EH range of −2.5 to −3.5 eV and the EC range of −6 to −8 eV can efficiently induce the ring-opening reaction of cycloalkanes without excessive dehydrogenation leading to aromatic hydrocarbon formation. The calculation shows good agreement with catalytic experimental tests and in situ DRIFT spectra results, providing a series of potential candidate catalysts for the ring-opening cracking of cycloalkane-rich diesel fuel.

Detection and Quantification of Drug-Protein Adducts in Human Liver.

Covalent protein adducts formed by drugs or their reactive metabolites are risk factors for adverse reactions, and inactivation of cytochrome P450 (CYP) enzymes. Characterization of drug-protein adducts is limited due to lack of methods identifying and quantifying covalent adducts in complex matrices. This study presents a workflow that combines data-dependent and data-independent acquisition (DDA and DIA) based liquid chromatography with tandem mass spectrometry (LC-MS/MS) to detect very low abundance adducts resulting from CYP mediated drug metabolism in human liver microsomes (HLMs). HLMs were incubated with raloxifene as a model compound and adducts were detected in 78 proteins, including CYP3A and CYP2C family enzymes. Experiments with recombinant CYP3A and CYP2C enzymes confirmed adduct formation in all CYPs tested, including CYPs not subject to time-dependent inhibition by raloxifene. These data suggest adducts can be benign. DIA analysis showed variable adduct abundance in many peptides between livers, but no concomitant decrease of unadducted peptides. This study sets a new standard for adduct detection in complex samples, offering insights into the human adductome resulting from reactive metabolite exposure. The methodology presented will aid mechanistic studies to identify, quantify and differentiate between adducts that result in adverse drug reactions and those that are benign.