Unexpectedly regioselective Diels-Alder reactions of new unsymmetrical benzoquinones: A convenient synthetic entry to uniquely substituted decalins.

We report flexible synthesis of new unsymmetrically 2,6-disubstituted benzoquinones (33 examples) and a systematic study of their reactivity in the Diels-Alder reaction. The Diels-Alder reactions of selected unsymmetrical benzoquinones with seemingly similar substituents were found to proceed with high regioselectivity and the formation of selected experimentally observed main products was rationalized by theoretical (DFT) calculations. The findings can be exploited in the convenient preparation of densely substituted and stereochemically defined decalins with unique angular substituents at ring fusion. We also demonstrate the usefulness of this methodology in complex molecule synthesis through the total synthesis of a novel forskolin analog possessing an ethyl group at the fusion of the rings B and C.

Development of Visible Spectrophotometric Methods for the Determination of Tricyclic Antidepressants Based on Formation of Molecular Complexes with p-Benzoquinones.

Tricyclic antidepressants are commonly employed in the management of major depressive disorders. The present work describes two visible (VIS) spectrophotometric techniques that utilize the formation of charge transfer complexes between four antidepressant compounds, namely, amitriptyline hydrochloride (AMI), imipramine hydrochloride (IMI), clomipramine hydrochloride (CLO), and trimipramine maleate (TRI) acting as electron donors and two p-benzoquinones, namely, p-chloranilic acid (pCA) and 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ), serving as electron acceptors. The stoichiometry of the compounds produced exhibited a consistent 1:1 ratio in all instances, as established by Job's method. Molar absorptivities, equilibrium association constants, and several other spectroscopic properties were determined for all complexes. The developed spectrophotometric techniques were validated intra-laboratory and successfully applied for quantitative assessment of the four antidepressant active ingredients in several commercial pharmaceutical formulations. The methods are relatively simple, fast, and use readily available laboratory instrumentation, making them easily applicable by most quality control laboratories worldwide.

Potential and mechanism of disinfection by-products removal in drinking water by bubbling corona discharge.

Disinfection by-products (DBPs) with significant teratogenic and carcinogenic properties have become a growing concern among the public. As an efficient and environmentally friendly technology, non-thermal plasma offers potential for removing emerging micro-pollutants. In this study, the degradation performance of bubbling corona discharge was evaluated on 24 halogenated alicyclic and aliphatic DBPs present in drinking water at concentrations ranging from ng/L to µg/L. The degradation of DBPs followed pseudo-first-order kinetics with rate constants (kobs) in the descending order of halonitromethanes (HNMs), halogenated benzoquinones (HBQs), haloacetonitriles, trihalomethanes (THMs), haloketones, halogenated aldehydes, and haloacetic acids (HAAs). THMs, HNMs, and HBQs were effectively removed within 5 min under a discharge power of 28 W. Degradation rates achieved by plasma treatment surpass those of other conventional treatment technologies. The required energy consumption was in the range of 5-30 kW·h/m3/order. Furthermore, the study investigated the effects of discharge power, initial concentration, and economic analysis on the degradation of four selected DBPs as representatives of mono-, di- and multi-carbon-containing DBPs, namely chloroform (TCM) and bromoform (TBM), tribromoacetic acid (TBAA), and 2,3,5,6-tetrachloro-1,4-benzoquinone (TetraC-BQ). Reactive radicals in the plasma system were investigated using electron paramagnetic resonance, optical emission spectroscopy, fluorimetry, and radical scavengers. Hydrated electrons and hydroxyl radicals played an important role in the removal of DBPs. The intermediates generated during the degradation of TCM, TBM, TBAA, and TetraC-BQ were identified, and the possible degradation pathways for mono- and binary C-DBPs and HBQs were deduced. The breakdown of HBQs did not produce secondary contamination with aliphatic DBPs. The carbon in DBPs was primarily converted to formic acid, acetic acid, and oxalic acid, and the halogens were mainly converted to halogen ions. Additionally, luminescent bacteria toxicity testing confirmed that plasma treatment could reduce the acute toxicity of water samples. These findings demonstrate the potential of plasma treatment as a post-treatment device at the household level.

Redox-directed identification of toxic transformation products during ozonation of aromatics.

The toxicity assessment of transformation products (TPs) formed in oxidative water treatment is crucial but challenging because of their low concentration, structural diversity, and mixture complexity. Here, this study developed a novel redox-directed approach for identification of toxic TPs without the individual toxicity and concentration information. This approach based on sodium borohydride reduction comprised an integrated process of toxicological evaluation, fluorescence excitation-emission matrix characterization, high-resolution mass spectrometry detection, followed by ecological toxicity assessment of identified TPs. The redox-directed identification of primary causative toxicants was experimentally tested for the increased nonspecific toxicity observations in the ozonated effluents of model aromatics. Reduction reaction caused a remarkable decrease in toxicity and increase in fluorescence intensity, obtaining a good linear relation between them. More than ten monomeric or dimeric p-benzoquinone (p-BQ) TPs were identified in the ozonated effluents. The occurrence of the p-BQ TPs was further verified through parallel sodium sulfite reduction and actual wastewater ozonation experiments. In vitro bioassays of luminescent bacteria, as well as in silico genotoxicity and cytotoxicity predictions, indicate that the toxicity of p-BQ TPs is significantly higher than that of their precursors and other TPs. These together demonstrated that the identified p-BQ TPs are primary toxicity contributors. The redox-directed approach facilitated the revelation of primary toxicity contribution, illustrating emerging p-BQs are a concern for aquatic ecosystem safety in the oxidative treatment of aromatics-contaminated wastewater.

Immune Stimulation via Wounding Alters Chemical Profiles of Adult Tribolium castaneum.

Group-living individuals experience immense risk of disease transmission and parasite infection. In social and in some non-social insects, disease control with immunomodulation arises not only via individual immune defenses, but also via infochemicals such as contact cues and (defensive) volatiles to mount a group-level immunity. However, little is known about whether activation of the immune system elicits changes in chemical phenotypes, which may mediate these responses. We here asked whether individual immune experience resulting from wounding or injection of heat-killed Bacillus thuringiensis (priming) leads to changes in the chemical profiles of female and male adult red flour beetles, Tribolium castaneum, which are non-social but gregarious. We analyzed insect extracts using GC-FID to study the chemical composition of (1) cuticular hydrocarbons (CHCs) as candidates for the transfer of immunity-related information between individuals via contact, and (2) stink gland secretions, with analysis of benzoquinones as main active compounds regulating 'external immunity'. Despite a pronounced sexual dimorphism in CHC profiles, wounding stimulation led to similar profile changes in males and females with increases in the proportion of methyl-branched alkanes compared to naïve beetles. While changes in the overall secretion profiles were less pronounced, absolute amounts of benzoquinones were transiently elevated in wounded compared to naïve females. Responses to priming were insignificant in CHCs and secretions. We suggest that changes in different infochemicals after wounding may mediate immune status signaling in the context of both internal and external immune responses in groups of this non-social insect, thus showing parallels to social immunity.

Antifungal Activity of Benzoquinones Produced by Tribolium castaneum in Maize-Associated Fungi.

Tribolium castaneum (Herbst) adults produce 1,4-benzoquinone (BQ), methyl-1,4-benzoquinone (MBQ), and ethyl-1,4-benzoquinone (EBQ). These components are chemical defenses used as repellents and irritants, and BQ has a negative impact on the growth of some fungal species. In this work, the inhibitory and/or lethal effects of these benzoquinones on the development of six fungi identified in maize, namely Aspergillus flavus, A. fumigatus, A. niger, Fusarium sp., Penicillium sp., and Trichoderma sp., were evaluated. Ten-day-long disk diffusion trials were performed using benzoquinones. The experiments simulated the activity of BQ (B1) or "EBQ + MBQ" (B2) released by 40-day-old insect adults (n = 200), considering a total average release of 45 µg per adult. Inhibition halos imposed by benzoquinones on fungal growth showed a significant effect when compared with the controls (water and solvent). Mycelial growth was decreased for all fungi, with the level of response depending on the fungal species. B1 and B2 displayed an inhibitory effect against all fungi, but Trichoderma sp. and A. niger showed rapid recoveries. B2 showed a lethal effect on Penicillium sp. The inhibitory and lethal activities of benzoquinones released by T. castaneum adults may contribute to regulate fungal growth, and understanding their interaction is important to develop innovative control strategies.

Secretion and Detection of Defensive Compounds by the Red Flour Beetle Tribolium castaneum Interacting with the Insect Pathogenic Fungus Beauveria bassiana.

Entomopathogenic fungi such as Beauveria bassiana are extensively used for the control of insect pests worldwide. They infect mostly by adhesion to the insect surface and penetration through the cuticle. However, some insects, such as the red flour beetle Tribolium castaneum (Herbst), have evolved resistance by embedding their cuticle with antifungal compounds. Thus, they avoid fungal germination on the cuticle, which result in low susceptibility to entomopathogenic fungi. In adult T. castaneum, these antifungals are the well-known defensive compounds methyl-1,4- and ethyl-1,4-benzoquinone. In this study, we added B. bassiana conidia on the diet of adult beetles to study the effect of the entomopathogen on the secretion and detection of the beetle volatile blend containing both benzoquinones. The compounds were analyzed by solid phase microextraction coupled to gas chromatography-flame ionization detection, and were detected by electroantennography. In addition, we measured the expression level of four genes encoding for two odorant-binding proteins (OBP), one chemosensory protein (CSP), and one odorant receptor (OR) in both healthy and fungus-treated insects. Significant alterations in the secretion of both benzoquinones, as well as in the perception of methyl-1,4-benzoquinone, were found in fungus-treated insects. TcOBP7D, TcOBP0A and TcCSP3A genes were down-regulated in insects fed conidia for 12 and 48 h, and the latter gene was up-regulated in 72 h samples. TcOR1 expression was not altered at the feeding times studied. We conclude that fungus-treated insects alter both secretion and perception of benzoquinones, but additional functional and genetic studies are needed to fully understand the effects of fungal infection on the insect chemical ecology.

Quantitative identification of halo-methyl-benzoquinones as disinfection byproducts in drinking water using a pseudo-targeted LC-MS/MS method.

Halobenzoquinones (HBQs) as disinfection byproducts (DBPs) in drinking water is prioritized for research due to their prevalent occurrence and high toxicity. However, only fifteen HBQs can be identified among a high diversity using targeted LC-MS/MS analysis in previous studies due to the lack of chemical standards. In this study, we developed a pseudo-targeted LC-MS/MS method for detecting and quantifying diverse HBQs. Distinct fragment characteristics of HBQs was observed according to the halogen substituent effects, and extended to the development of a multiple-reaction-monitoring (MRM) method for the quantification of the 46 HBQs that were observed in simulated drinking water using non-targeted analysis. The fragmentation mechanism was supported by the changes of Gibbs free energy (ΔG), and a linear relationship between the ΔG and the ionization efficiency of analytes was developed accordingly for quantification of these 46 HBQs, 30 of which were lack of chemical standards. It is noted that 29 of the 30 newly-identified HBQs were halo-methyl-benzoquinones (HMBQs), which were predicted to be carcinogens related with drinking-water bladder cancer risk and be more toxic than non-methyl HBQs. Using the new method, twelve HMBQs were detected in actual drinking water samples with concentrations up to 100.4 ng/L, 3 times higher than that reported previously. The cytotoxicity in CHO cells of HMBQs was over 1-fold higher than that of non-methyl-HBQs. Therefore, HMBQs are an essential, highly toxic group of HBQs in drinking water, which deserve particular monitoring and control.

Pigmentación violácea dolorosa, reto clínico y enfoque diagnóstico en un país tropical / Painful violaceous pigmentation, a clinical challenge and diagnostic approach in a tropical country

RESUMEN Contrario a lo que se cree, los milpiés no se consideran animales venenosos, su mecanismo de defensa puede resultar perjudicial para el ser humano, manifestándose como lesiones de carácter eritematoso, purpúrico y cianótico, concomitante dolor local y parestesia; Según la exposición, el cuadro clínico puede evolucionar a lesiones moderadas y severas, ampollas y úlceras. Describimos el caso de un paciente pediátrico de sexo masculino de la región andina de Colombia, con temperaturas diarias que oscilan entre 15° y 30°C y una humedad promedio de 72 %, quien accidentalmente pisó un milpiés, lo cual fue un desafío diagnóstico entre el personal de salud que generó su atención.

ABSTRACT Millipedes are not considered poisonous animals, although in some cases their chemical defense mechanisms may be deleterious for human beings, manifesting as erythematous, purpuric and/ or cyanotic lesions alongside local pain and paresthesia. According to the exposure, the clinical condition may progress to moderate and severe lesions such as ulcers and blisters. We describe a case of a male child from the Andean region in Colombia, an area with daily temperatures between 15° and 30° C and 72% average humidity. The boy accidentally stepped on a millipede, and this occurrence became a challenge for its diagnosis amongst staff who provided emergency health care.

Alkenylbenzoquinones and other compounds from the fruit of Maesa lanceolata exhibited potent cytotoxic, antibacterial, and antiradical scavenging activities§.

A phytochemical study of the methanol extract of the fruit of Maesa lanceolata resulted in the isolation of a new alkenylbenzoquinone (1), alongside the known compounds (Z)-2,5-dihydroxy-6-methyl-3-(pentadec-10'-enyl)-1,4-benzoquinone (2), 2,5-dihydroxy-6-methyl-3-(nonadec-14'-enyl)-1,4-benzoquinone (3), 2,5-dihydroxy-6-methyl-3-(tridecyl)-1,4-benzoquinone (4), (2S,3S,4R,2'R,9E)-[2'-hydroxytetraeicosanoyl]-2-aminooctadec-9-ene-1,3,4-triol (5), monopalmitin (glyceryl palmitate) (6), lupeol (7), and 3-O-(ß-D-glucopyranoside)-ß-sitosterol (8). The structures of the compounds were established by the means of spectroscopic (1 D- and 2 D-NMR) and spectrometric techniques (MS). The isolated compounds were assessed for their antibacterial, cytotoxic, and antiradical activities. Compound 2 showed moderate activity against Staphylococcus warneri (DSMZ 20036), while the other compounds were inactive. The two quinones 1 and 2 were significantly cytotoxic, with IC50 values of 0.005 µM and 12.5 µM respectively, and were weakly active towards DPPH radical (IC50 >250 µg/mL).

Fungal quinones: diversity, producers, and applications of quinones from Aspergillus, Penicillium, Talaromyces, Fusarium, and Arthrinium.

Quinones represent an important group of highly structurally diverse, mainly polyketide-derived secondary metabolites widely distributed among filamentous fungi. Many quinones have been reported to have important biological functions such as inhibition of bacteria or repression of the immune response in insects. Other quinones, such as ubiquinones are known to be essential molecules in cellular respiration, and many quinones are known to protect their producing organisms from exposure to sunlight. Most recently, quinones have also attracted a lot of industrial interest since their electron-donating and -accepting properties make them good candidates as electrolytes in redox flow batteries, like their often highly conjugated double bond systems make them attractive as pigments. On an industrial level, quinones are mainly synthesized from raw components in coal tar. However, the possibility of producing quinones by fungal cultivation has great prospects since fungi can often be grown in industrially scaled bioreactors, producing valuable metabolites on cheap substrates. In order to give a better overview of the secondary metabolite quinones produced by and shared between various fungi, mainly belonging to the genera Aspergillus, Penicillium, Talaromyces, Fusarium, and Arthrinium, this review categorizes quinones into families such as emodins, fumigatins, sorbicillinoids, yanuthones, and xanthomegnins, depending on structural similarities and information about the biosynthetic pathway from which they are derived, whenever applicable. The production of these quinone families is compared between the different genera, based on recently revised taxonomy. KEY POINTS: • Quinones represent an important group of secondary metabolites widely distributed in important fungal genera such as Aspergillus, Penicillium, Talaromyces, Fusarium, and Arthrinium. • Quinones are of industrial interest and can be used in pharmacology, as colorants and pigments, and as electrolytes in redox flow batteries. • Quinones are grouped into families and compared between genera according to the revised taxonomy.

Anticancer activity and toxicity of new quaternary ammonium geldanamycin derivative salts and their mixtures with potentiators.

Geldanamycin (GDM) has been modified by different type neutral/acidic/basic substituents (1-7) and by quinuclidine motif (8), transformed into ammonium salts (9-13) at C(17). These compounds have been characterised by spectroscopic and x-ray methods. Derivative 8 shows better potency than GDM in MCF-7, MDA-MB-231, A549 and HeLa (IC50s = 0.09-1.06 µM). Transformation of 8 into salts 9-13 reduces toxicity (by 11-fold) at attractive potency, e.g. MCF-7 cell line (IC50∼2 µM). Our studies show that higher water solubility contributes to lower toxicity of salts than GDM in healthy CCD39Lu and HDF cells. The use of 13 mixtures with potentiators PEI and DOX enhanced anticancer effects from IC50∼2 µM to IC50∼0.5 µM in SKBR-3, SKOV-3, and PC-3 cancer cells, relative to 13. Docking studies showed that complexes between quinuclidine-bearing 8-13 and Hsp90 are stabilised by extra hydrophobic interactions between the C(17)-arms and K58 or Y61 of Hsp90.

Application of the Meerwein reaction of 1,4-benzoquinone to a metal-free synthesis of benzofuropyridine analogues.

Several new heterocyclic systems based on a hydroxybenzofuro[2,3-b]pyridine building block were prepared. This benzofuropyridine is easily available from the Meerwein reaction of benzoquinone and a heterocyclic diazonium salt, followed by reduction and cyclization. Electrophilic substitution and further condensations give polycyclic systems, including oxazolo- and chromeno-fused analogues.

3-O-glycosylation of kaempferol restricts the supply of the benzenoid precursor of ubiquinone (Coenzyme Q) in Arabidopsis thaliana.

Ubiquinone (Coenzyme Q) is a vital respiratory cofactor and antioxidant in eukaryotes. The recent discovery that kaempferol serves as a precursor for ubiquinone's benzenoid moiety both challenges the conventional view of flavonoids as specialized metabolites, and offers new prospects for engineering ubiquinone in plants. Here, we present evidence that Arabidopsis thaliana mutants lacking kaempferol 3-O-rhamnosyltransferase (ugt78d1) and kaempferol 3-O-glucosyltransferase (ugt78d2) activities display increased de novo biosynthesis of ubiquinone and increased ubiquinone content. These data are congruent with the proposed model that unprotected C-3 hydroxyl of kaempferol triggers the oxidative release of its B-ring as 4-hydroxybenzoate, which in turn is incorporated into ubiquinone. Ubiquinone content in the ugt78d1/ugt78d2 double knockout represented 160% of wild-type level, matching that achieved via exogenous feeding of 4-hydroxybenzoate to wild-type plants. This suggests that 4-hydroxybenzoate is no longer limiting ubiquinone biosynthesis in the ugt78d1/ugt78d2 plants. Evidence is also shown that the glucosylation of 4-hydroxybenzoate as well as the conversion of the immediate precursor of kaempferol, dihydrokaempferol, into dihydroquercetin do not compete with ubiquinone biosynthesis in A. thaliana.

Wild Blonde Capuchins (Sapajus flavius) Perform Anointing Behaviour Using Toxic Secretions of a Millipede (Spirobolida: Rhinocricidae).

Defensive secretions of millipedes are remarkable for containing toxic quinones known to efficiently repell hematophagous arthropods. Here we show that Endangered blonde capuchin monkeys make use of such secretions. We (i) describe the anointing behavior performed by the monkeys (ii) identify the millipede species used in the process (iii) describe the volatile chemical composition of its secretion. The blonde capuchin monkeys selectively searched for millipedes hidden under the ground. We observed three bouts of anointing behavior, performed by 13 individuals of all age classes (from adults to independent infants), both solitarily (1 event) and socially (10 events). The only millipede species used by the monkeys is an undescribed species of the genus Poecilocricus (Spirobolida, Rhinocricidae). The volatile chemical composition of the secretions was predominantly comprised of a mixture of benzoquinones and hydroquinones. The social nature of the behavior and time of the observations (mosquito season), suggest that social bonding and mosquito avoidance is linked to the anointing behavior of the monkeys.

Antimicrobial and antibiofilm activity of the benzoquinone oncocalyxone A.

Resistance to antimicrobials is a challenging issue that complicates the treatment of infections caused by bacteria and fungi, thus requiring new therapeutic options. Oncocalyxone A, a benzoquinone obtained from Auxemma oncocalyx (Allem) Taub has several biological effects; however, there is no data on its antimicrobial action. In this study, its antimicrobial and antibiofilm activities were evaluated against bacteria and fungi of clinical interest. Strains of Gram-positive and Gram-negative bacteria, and filamentous fungi and yeasts were selected to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of oncocalyxone A. The antibacterial effect of oncocalyxone A was studied using survival curves, atomic force microscopy (AFM), and the involvement of oxidative stress. We examined the inhibitory action of the molecule on biofilm formation and its hemolytic activity against human erythrocytes. Our results showed that among the strains tested, Staphylococcus epidermidis was highly sensitive to the action of oncocalyxone A, with an MIC of 9.43 µg/mL. In most bacterial strains analyzed, a bacteriostatic effect was observed, though the molecule showed no antifungal activity. Antibiofilm activity was observed against the methicillin-resistant S. aureus bacteria. Additionally, results from atomic force microscopy imaging showed that oncocalyxone A significantly altered bacterial morphology. Further, oncocalyxone A showed no hemolytic activity at concentrations ≥151 µg/mL. Together, our results demonstrate the antibacterial and antibiofilm potential of oncocalyxone A, indicating its therapeutic potential against bacterial resistance.

Synthesis, structure and anticancer activity of new geldanamycin amine analogs containing C(17)- or C(20)- flexible and rigid arms as well as closed or open ansa-bridges.

The nucleophilic attack of amines at C(17) or C(17)/C(20) positions of geldanamycin's (GDM) benzoquinone, via initial 1,4-Michael conjugate addition mechanism, yield new analogs with closed or open ansa-bridges (1-31), respectively. X-ray structures of analogs 22 and 24 reveals an unexpected arrangement of the ansa-bridge in solid (conformer B), that is located between those of conformers A, prevailing in solution (trans-lactam), and C, crucial at binding to Hsp90 (cis-lactam). The structure of a new-type conformer B allows to better understand the molecular recognition mechanism between the GDM analogs and the target Hsp90. Combined analysis of: anticancer test results (SKBR-3, SKOV-3, PC-3, U-87, A-549) and those performed in normal cells (HDF), KD values and docking modes at Hsp90 as well as clogP parameters, reveals that the rigid C(17)-arm (piperidyl, cyclohexyl) with a H-bond acceptor as carbonyl group together with a lipophilicity clogP∼3 favor high potency of analogs, even up to IC50 ∼0.08 µM, at improved selectivity (SIHDF > 30), when compared to GDM. The most active 25 show higher anticancer potency than 17-AAG (in SKOV-3 and A-549) as well as reblastatin (in SKBR-3 and SKOV-3). Opening of the ansa-bridge within GDM analogs, at the best case, decreases activity (IC50∼2 µM) and toxicity in HDF cells (SIHDF∼2-3), relative to GDM.

Quantifying the efficiency of o-benzoquinones reaction with amino acids and related nucleophiles by cyclic voltammetry.

The reaction efficiency of o-benzoquinones with amines (L-lysine, Nα-acetyl-L-lysine, glycine, L-methionine and L-arginine), thiols (L-cysteine and Nα-acetyl-L-cysteine) and protein (bovine serum albumin) were determined at pH 5.0, 7.0 and 8.0 and scan rate of 10, 50 and 100 mV/s by cyclic voltammetry. Nucleophiles containing multiple nucleophilic groups and nucleophilic group possessing low pKa value would enhance the reactivity of nucleophiles towards o-benzoquinones. The reactivity of different o-benzoquinones with L-lysine/L-cysteine followed the order: protocatechuic acid quinone ≈ catechol quinone > 4-methylbenzoquinone ≈ caffeic acid quinone > rosmarinic acid quinone > chlorogenic acid quinone. The reactivity of quinones would be decreased by the steric hindrance of substituents on quinone ring, and it would also be weakened by enhancing electron cloud density of quinone ring. Adducts generated by the interaction of 4-methylbenzoquinone with amines and thiols were tentatively identified as amine-quinone adduct and thiol-phenol adduct respectively by UPLC-QTOF-MS/MS and cyclic voltammetry.

Study of reactions of Nε-(carboxymethyl) lysine with o-benzoquinones by cyclic voltammetry.

The reaction of Nε-(carboxymethyl) lysine (CML) with eight kinds of non-flavonoid o-benzoquinones and five kinds of flavonoid o-benzoquinones were investigated by cyclic voltammetry at pH 5.0, 7.0 and 8.0 and scan rate of 10, 50 and 100 mV/s. The reactivity of o-benzoquinones towards CML is weakened by the electron-donating substituent and strengthened by the electron-withdrawing substituent on the o-benzoquinone rings. The steric hindrance of the substituents on o-benzoquinone rings also weakens the quinone reactivity. Reaction of 4-methylbenzoquinone with CML (38.0 ±â€¯1.3%) was found to be faster than that with l-lysine (31.3 ±â€¯1.5%) and Nα-acetyl-l-lysine (14.5 ±â€¯0.1%) but slower than that with l-cysteine (≥100.0%) and Nα-acetyl-l-cysteine (≥100.0%) at pH 7.0 and scan rate of 10 mV/s. Products obtained by the reaction of CML with o-benzoquinones were found to include a CML-quinone adduct according to the cyclic voltammetry and UPLC-QTOF-MS/MS analysis.

Rapid analysis of chemical constituents of Huoxue Zhitong Capsules based on UPLC-Q-TOF/MS / 中草药

Objective: To rapidly analyze the chemical constituents of Huoxue Zhitong Capsules performed by ultra-high performance liquid chromatography quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF/MS). Methods: A Waters Acquity UPLC BEH C18 column (100 mm × 2.1 mm, 1.7 μm) was used with 0.1% formic acid aqueous solution and 0.1% formic acid acetonitrile as a mobile phase. The flow rate was 0.4 mL/min; The samples were collected in positive and negative ion modes, respectively. The processed data was analyzed by Masslynx 4.1TM software. Results: A total of 113 compounds were identified in positive and negative ion mode, including terpenoids, saponins, organic acids, benzoquinones and others, and the medicinal sources of the compounds were assigned. Conclusion: The established method can systematically, quickly and accurately identify various chemical components of Huoxue Zhitong Capsule. This study provides a reference for the selection of quality evaluation indicators and the basis of pharmacodynamics of Huoxue Zhitong Capsule.