Adduct Formation Reactions Research Articles

Identifying the Transients and Transformation Products in Hydroxyl Radical-Methimazole Reactions Using DFT and UPLC-Q-TOF MS/MS Approaches.

Oxidative reactions of the hydroxyl radical (·OH) with methimazole (MMI), an antithyroid drug, are crucial for understanding its fate in oxidizing environments. By synergistically integrating density functional theory and ultraperformance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF MS/MS) techniques, we elucidated the transients and transformation products (TPs) arising from the ·OH-MMI reactions. We probed two hydrogen-atom abstraction (HA) reactions, three radical adduct formation reactions, and single electron transfer (SET) at the M06-2X/6-311++G(d,p)/SMD(water) level. All proposed reaction channels, except for HA from the methyl group and SET, were found to be barrier-free. SET is the dominant oxidation pathway, accounting for 44% of oxidations, as determined by branching ratio analysis. The selenium analogue, MSeI, exhibited minor reactivity differences compared to MMI, yet its overall patterns resembled those of ·OH-MMI reactions. TPs were generated experimentally by reacting MMI with ·OH produced by UV-photolysis of H2O2. Eight TPs were identified from an approximately 24% degradation of MMI using UPLC-Q-TOF MS/MS analysis, and an additional two TPs were identified from the approximately 52% degraded MMI sample. The exact identities of all of the TPs were established through their corresponding fragmentation patterns. This study elucidates the drug's susceptibility to free radical species under physiologically relevant conditions.

Effect of pH on UV/H2O2-mediated removal of single, mixed and halogenated parabens from water

Adjusting pH values in aqueous environments can significantly improve the efficiency by which parabens and halo-parabens are removed. In this study, 20 neutral and deprotonated species were selected as models to investigate their pH-dependent removal mechanisms and kinetics by a UV/H2O2 process using density functional theory (DFT). Compared to neutral species, deprotonated species exhibit higher reactivity to HO• due to their high electron cloud density. H atom abstraction (HAA) reactions on the substitution groups are the most favorable pathways for neutral species, while radical adduct formation (RAF) reactions are the most favorable for deprotonated species. Single electron transfer (SET) reactions can be neglected for neutral species, while these reactions become a viable route for deprotonated molecules. The total reaction rate constants range from 1.63 × 109 to 3.74 × 1010 M− 1 s− 1 at pH 7.0, confirming the experimental results. Neutral and weakly alkaline conditions are favorable for the degradation of MeP and halo-parabens in the UV/H2O2 process. The order of removal efficiency at optimum pH is dihalo-parabens > mono-halo-parabens ≈ F, F-MeP > MeP. Furthermore, the transformation products must undergo oxidative degradation due to their high toxicity. Our findings provide new insights into the removal of parabens and their halogenated derivatives from wastewater.

Theoretical investigation on the degradation of sulfadiazine in water environments: Oxidation of •OH, SO4•−and CO3•− and reactivity of (TiO2)n clusters (n = 1–6)

This study reported the degradation of micro pollutants through reactive radicals like •OH, SO4•−and CO3•−. Sulfadiazine (SDZ) is widely used antibiotic and emerging water pollutant. The synergistic degradation of sulfadiazine by •OH, SO4•−and CO3•− was investigated through quantum chemistry methods. The degradation of SDZ by •OH radicals was dependent on radical adduct formation (RAF) reactions while degradation of SDZ by SO4•− radical was caused by RAF, hydrogen atom transfer (HAT) and also single electron transfer (SET). The degradation of SDZ by CO3•− radical mainly caused by RAF and HAT. The degradation rate constants of SDZ initiated through •OH, SO4•−and CO3•− radicals were calculated separately: kSDZ-SO4•− (5.29 × 1010 M−1 s−1) > kSDZ-•OH (1.38 × 1010 M−1 s−1) > kSDZ-CO3•− (1.28 × 108 M−1 s−1). The degradation mechanism of SDZ by (TiO2)n (n = 1–6) cluster was investigated. The conformation between benzene ring and heterocyclic ring of SDZ has a lower adsorption energy and the (TiO2)n cluster (n = 5) is the most stable adsorption conformation with SDZ. The toxicity assessment showed that most of the degradation products were less toxic and eco-friendly. However, O-P1, P4 and P9 exhibited more severe aquatic toxicity.

Theoretical and Experimental Investigation of the Antioxidation Mechanism of Loureirin C by Radical Scavenging for Treatment of Stroke.

Recent pharmacological studies have shown that dragon's blood has an anti-cerebral ischemia effect. Loureirin C (LC), a kind of dihydrochalcone compound in dragon's blood, is believed to be play an important role in the treatment of ischemia stroke, but fewer studies for LC have been done. In this paper, we report the first experimental and theoretical studies on the antioxidation mechanism of LC by radical scavenging. The experimental studies show that LC has almost no effect on cell viability under 15 μM for the SH-SY5Y cells without any treatments. For the SH-SY5Y cells with oxygen and glucose deprivation-reperfusion (OGD/R) treatment, LC increased the viability of SH-SY5Y cells. The results of 2',7'-Dichlorodihydrofluorescein diacetate (DCFH-DA) and MitoSox Red experiments indicate that LC is very efficient in inhibiting the generation of the intracellular/mitochondrial reactive oxygen species (ROS) or removing these two kinds of generated ROS. The density functional theory (DFT) calculations allowed us to elucidate the antioxidation mechanisms of LC. Fukui function analysis reveals the radical scavenging of LC by hydrogen abstraction mechanism, the complex formation by e-transfer, and radical adduct formation (RAF) mechanism. Among the H-abstraction, the complex formation by e-transfer, and radical adduct formation (RAF) reactions on LC, the H-abstraction at O-H35 position by OH• is favorable with the smallest energy difference between the product and two reactants of the attack of OH• to LC of -0.0748 Ha. The bond dissociation enthalpies (BDE), proton affinities (PA), ionization potential (IP), proton dissociation enthalpy (PDE), and electron transfer enthalpy (ETE) were calculated to determine thermodynamically preferred reaction pathway for hydrogen abstraction mechanism. In water, IP and the lowest PDE value at O3-H35 position are lower than the lowest BDE value at O3-H35 position; 41.8986 and 34.221 kcal/mol, respectively, indicating that SEPT mechanism is a preferred one in water in comparison with the HAT mechanism. The PA value of O3-H35 of LC in water is -17.8594 kcal/mol, thus the first step of SPLET would occur spontaneously. The minimum value of ETE is higher than the minimum value of PDE at O3-H35 position and IP value, 14.7332 and 22.4108 kcal/mol, respectively, which suggests that the SEPT mechanism is a preferred one in water in comparison with the SPLET mechanism. Thus, we can draw a conclusion that the SEPT mechanism of is the most favorite hydrogen abstraction mechanism in water, and O-H35 hydroxyl group has the greatest ability to donate H-atoms.

Unravelling the effects of complexation of transition metal ions on the hydroxylation of catechol over the whole pH region

Catechol pollutants (CATPs) serving as chelating agents could coordinate with many metal ions to form various CATPs-metal complexes. Little information is available on the effects of complexation of metal ions on CATPs degradation. This work presents a systematical study of •OH-mediated degradation of catechol and catechol-metal complexes over the whole pH range in advanced oxidation processes (AOPs). Results show that the pH-dependent complexation of metal ions (Zn2+, Cu2+, Ti4+ and Fe3+) promotes the deprotonation of catechol under neutral and even acidic conditions. The radical adduct formation (RAF) reactions are both thermodynamically and kinetically favorable for all dissociation and complexation species, and OH/O− group-containing C positions are more vulnerable to •OH attack. The kinetic results show that the complexation of the four metal ions offers a wide pH range of effectiveness for catechol degradation. At pH 7, the apparent rate constant (kapp) values for different systems follow the order of catechol+Ti4+ ≈ catechol+Zn2+ > catechol+Cu2+ > catechol+Fe3+ > catechol. The mechanistic and kinetic results would greatly improve our understanding of the degradation of CATPs-metal and other organics-metal complexes in AOPs. The toxicity assessment indicates that the •OH-based AOPs have the ability for decreasing the toxicity and increasing the biodegradability during the processes of catechol degradation.

The roles of HO•, ClO• and BrO• radicals in caffeine degradation: A theoretical study

ClO• and BrO• are newly discovered reactive radicals that contribute to the degradation of micropollutants. However, the research on pollutant degradation by ClO• and BrO• is still lacking. Thus, the mechanism, kinetics, and toxicity of caffeine degradation by HO•, ClO•, and BrO• were computationally studied and compared. Results showed that radical adduct formation (RAF) reaction was dominant for HO•, ClO•, and BrO• initiated reactions of caffeine. The main reaction sites were C5 and C8 of caffeine for HO•, while only the RAF reaction on C8 was prominent for ClO• and BrO•. The initiated reaction rate constants of caffeine by HO•, ClO•, and BrO• were in the order of HO• (5.29 × 109 M−1 s−1) > ClO• (1.40 × 109 M−1 s−1) > BrO• (2.17 × 108 M−1 s−1). The kinetic simulation verified that ClO• played a crucial role in the degradation of caffeine by the UV/chlorine process. In addition to HO-adducts, the subsequent reaction mechanisms of ClO- and BrO-adducts have also been investigated. The formation mechanisms of several important products, namely dimethylparabanic acid (P2), di(N-hydroxymethyl) parabanic acid (P5), 1,3,7-trimethyluric acid (P6), and 8-oxocaffeine (P11), were elucidated. Remarkably, stable chlorinated and brominated intermediates or products were not generated in ClO•- and BrO•-mediated subsequent degradations of caffeine. The assessment of aquatic toxicity and health effects showed that caffeine could penetrate the blood-brain barrier (human), and caffeine and its degradation products were potentially harmful to the aquatic environment.

Theoretical investigation on the contribution of HO[rad], SO4[rad]- and CO3[rad]- radicals to the degradation of phenacetin in water: Mechanisms, kinetics, and toxicity evaluation

Indirect oxidation induced by reactive free radicals, such as hydroxyl radical (HO), sulfate radical (SO4-) and carbonate radical (CO3-), plays an important or even crucial role in the degradation of micropollutants. Thus, the coadjutant degradation of phenacetin (PNT) by HO, SO4- and CO3-, as well as the synergistic effect of O2 on HO and HO2 were studied through mechanism, kinetics and toxicity evaluation. The results showed that the degradation of PNT was mainly caused by radical adduct formation (RAF) reaction (69% for Г, the same as below) and H atom transfer (HAT) reaction (31%) of HO. For the two inorganic anionic radicals, SO4- initiated PNT degradation by sequential radical addition-elimination (SRAE; 55%), HAT (28%) and single electron transfer (SET; 17%) reactions, while only by HAT reaction for CO3-. The total initial reaction rate constants of PNT by three radicals were in the order: SO4- > HO > CO3-. The kinetics of PNT degradation simulated by Kintecus program showed that UV/persulfate could degrade target compound more effectively than UV/H2O2 in ultrapure water. In the subsequent reaction of PNT with O2, HO and HO2, the formation of mono/di/tri-hydroxyl substitutions and unsaturated aldehydes/ketones/alcohols were confirmed. The results of toxicity assessment showed that the acute and chronic toxicity of most products to fish increased and to daphnia decreased, and acute toxicity to green algae decreased while chronic toxicity increased.

Insights into the Mechanism of Hydroxyl Radical Mediated Oxidations of 2-Aminopurine: A Computational and Sonochemical Product Analysis Study.

Mechanistic details of hydroxyl radical (•OH) mediated oxidations of 2-aminopurine (2AP) in the aqueous phase have been established in this study via a combination of DFT calculations (at the M05-2X/6-311+G(d,p) level with SMD solvation) and sonochemical end product analyses by the LC-Q-TOF-MS/MS method. Rate constants and branching ratios for single electron transfer (SET), two H-abstractions (HA), and seven radical adduct formation (RAF) reactions of •OH with 2AP were evaluated using transition state theory (TST). The RAF at the C8-position of 2AP is noted as the dominant process, which constitutes almost 46.1% of overall reaction routes. The SET mechanism accounts for the second major pathway (39.6%) followed by RAF at the C6-position (14.3%). Formations of 14 transformation products (TPs, i.e., the nonradical end products) in the sonochemical reactions of •OH with 2AP have been identified by means of the LC-Q-TOF-MS/MS technique. Among the 14 TPs (designated as TP1 to TP14), the lowest and highest mass to charge ratio (m/z) were respectively observed at 129 and 269 in ESI-MS positive ionization mode. The identities of all TPs have been proposed on the basis of elemental composition of [M + H]+ ions and their respective MS-MS fragmentation pattern. Four TPs (including guanine) are considered as obtained directly from primary transients by radical elimination, radical-radical combination/disproportionation reactions. The remaining 10 TPs are postulated as a result of successive self- and/or cross-reactions of primary transients/four first generation TPs with reagents such as •OH, O2, and solvent H2O molecules.

Model for Metal Extraction from Chloride Media with Basic Extractants: A Coordination Chemistry Approach

The metal extraction mechanism of basic extractants is typically described as an anion exchange process, but this mechanism does not correctly explain all observations. This paper introduces a novel model for the extraction of metals by basic extractants from chloride media supported by experimental data on methyltrioctylammonium chloride and Aliquat 336 chloride systems. This model relies on the hypothesis that the metal species least stabilized in the aqueous phase by hydration (i.e., the metal species with the lowest charge density) is extracted more efficiently than the more water stabilized species (i.e., species with higher charge densities). Once it is transferred to the organic phase, the extracted species can undergo further Lewis acid-base adduct formation reactions with the chloride anions available in the organic phase to form negatively charged chloro complexes, which than associate with the organic cations. Salting-out agents influence the extraction, most likely by decreasing the concentration of free water molecules, which destabilizes the metal complex in the aqueous phase. The evidence provided includes (1) the link between extraction and transition-metal speciation, (2) the trend in extraction efficiency as a function of the concentration of different salting-out agents, and (3) the behavior of HCl in the extraction system. The proposed extraction model better explains the experimental observations in comparison to the anion exchange model and allows the prediction of optimal conditions for metal extractions and separations a priori, by selecting the most suitable salting-out agent and its concentration.

Synthesis, Characterization, Cytotoxicity, and Time‐Dependent NMR Spectroscopic Studies of (SP‐4‐3)‐Oxalato[(1R,2R,4R/1S,2S,4S)‐(4‐trifluoromethyl‐cyclohexane‐1,2‐diamine)]platinum(II)

The oxaliplatin derivative (SP‐4‐3)‐oxalato[(1R,2R,4R/1S,2S,4S)‐(4‐trifluoromethyl‐cyclohexane‐1,2‐diamine)]platinum(II) hosting a trifluoromethyl group at position 4 of the cyclohexane ring is presented. The ligand was synthesized as an 1R,2R,4R/1S,2S,4S racemic mixture starting from 4‐trifluoromethyl‐cyclohexanol over five steps and coordinated to platinum to yield first the dichlorido and subsequently the oxalato complex. This novel compound and its analogue featuring a (13C2)oxalate ligand were characterized by multinuclear (1H, 13C, 15N, 19F, 195Pt) one‐ and two‐dimensional NMR spectroscopy. Ligand exchange and adduct formation reactions with DMSO, NaCl, CaCl2, l‐methionine, and 5′‐GMP were investigated via time‐dependent measurements by 19F and 13C NMR spectroscopy. The cytotoxicity of the title complex was investigated in three human cancer cell lines; the IC50 values were found in the low micromolar range, attesting moderate to high antiproliferative activity, depending on the cell line.

Quantum–classical approach to the reaction dynamics in a superfluid helium nanodroplet. The Ne2 dimer and Ne–Ne adduct formation reaction Ne + Ne-doped nanodroplet

The dynamics of the Ne2 dimer and Ne-Ne adduct formation in a superfluid helium nanodroplet [(4He)N; T = 0.37 K], Ne + Ne@(4He)N→ Ne2@(4He)N'/Ne-Ne@(4He)N' + (N-N')4He with N = 500, has been investigated using a hybrid approach (quantum and classical mechanics (QM-CM) descriptions for helium and the Ne atoms, respectively) and taking into account the angular momentum of the attacking Ne atom, Ne(1). Comparison with zero angular momentum QM results of our own shows that the present results are similar to the quantum ones for the initial Ne(1) velocities (v0) of 500 and 800 m s-1 (the former one being the most probable velocity of Ne at 300 K), in all cases leading to the Ne2 dimer (re = 3.09 Å). However, significant differences appear below v0 = 500 m s-1, because in the QM-CM dynamics, instead of the dimer, a Ne-Ne adduct is formed (r0 = 5.45 Å). The formation of this adduct will probably dominate as the contribution to reactivity of angular momenta larger than zero is the leading one and angular momentum strongly acts against the Ne2 production. Angular momentum adds further difficulties in producing the dimer, since it makes it more difficult to remove the helium density between both Ne atoms to lead, subsequently, to the Ne2 molecule. Hence, the formation of the neon-neon adduct, Ne-Ne@(4He)N', clearly dominates the reactivity of the system, which results in the formation of a "quantum gel"/"quantum foam", because the two Ne atoms essentially maintain their identity inside the nanodroplet. Large enough Ne(1) initial angular momentum values can induce the formation of vortex lines by the collapse of superficial excitations (ripplons), but they occur with greater difficulty than in the case of the capture of the Ne atom by a non doped helium nanodroplet, due to the wave interferences induced by the Ne induced by the solvation layers of the Ne atom originally placed inside the nanodroplet. We hope that this work will encourage other researchers to investigate the reaction dynamics in helium nanodroplets, an interesting topic on which there are few studies available.

A computational foray into the mechanism and catalysis of the adduct formation reaction of guanine with crotonaldehyde.

Crotonaldehyde, a common environmental pollutant and product of endogenous lipid peroxidation, reacts with guanine to form DNA adducts with pronounced genotoxicity and mutagenicity. Here, we explore the molecular mechanism of this adduct formation using double-hybrid density functional theory methods. The reaction can be envisaged to occur in a two-step fashion via an aza-Michael addition leading to an intermediate ring-open adduct followed by a cyclization reaction giving the mutagenic ring-closed adduct. We find that (i) a 1,2-type addition is favored over a 1,4-type addition for the aza-Michael addition, and (ii) an initial tautomerization of the guanine moiety in the resulting ring-open adduct significantly reduces the barrier toward cyclization compared to the direct cyclization of the ring-open adduct in its keto-form. Overall, the aza-Michael addition is found to be rate-determining. We further find that participation of a catalytic water molecule significantly reduces the energy barriers of both the addition and cyclization reaction. © 2018 Wiley Periodicals, Inc.

Reactivity of chlorogenic acid toward hydroxyl and methyl peroxy radicals relative to trolox in nonpolar media

The quantum mechanics-based test for overall free-radical scavenging activity was applied for the investigation of antioxidative capacity of chlorogenic acid (5-O-caffeoylquinic acid, 5CQA) relative to trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid, Tx) as a reference compound. Hydrogen atom transfer (HAT), radical adduct formation (RAF), electron transfer (ET), and proton loss (PL) reactions of 5CQA and Tx with HO• and CH3OO• radicals in benzene and pentyl ethanoate were examined. For this purpose, two theoretical models, M06-2X/6-311++G(d,p) in combination with the CPCM solvation model, and M05-2X/6-311++G(d,p) in combination with the SMD solvation model, were employed. It was found that M05-2X/6-311++G(d,p)—SMD failed to evaluate the influence of pentyl ethanoate, whereas M06-2X—CPCM in benzene and pentyl ethanoate and M05-2X—SMD in benzene proved to be operative and showed similar trends. Both compounds can react with HO• via HAT and RAF mechanisms, whereas HAT is the only reaction pathway with CH3OO•. 5CQA is more efficient scavenger of HO• than Tx, but less efficient scavenger of CH3OO•. Less reactive free radicals are more suitable for the determination of antioxidative activity of a compound relative to Tx. Highly reactive free radicals need to be included in the investigation of all potential reaction pathways of the compound, in which case the results from different approaches can be inconsistent.

Free radicals interacting with Cu, Ag and Au clusters

We investigated free radical scavenger capacity of small neutral Cu, Ag and Au clusters (Cu, Ag and Au up to ten atoms, and Au20), using three molecules (OH, OOH, OCH3) and two reaction mechanisms (Electron Transfer and Radical Adduct Formation). Principally this indicates that metal clusters are good free radical scavengers. They are good electron donors and the radical adduct formation reaction is thermodynamically feasible. Here, we show how Molecular Electrostatic Potential is very useful for determining the active site of the cluster in terms of interaction with the free radical. This investigation affirms that free radicals interact with the balanced electron density zone of the cluster. This may be very important for further studies that contemplate more than one free radical bonded to the metal clusters. These findings should serve as a significant guide for experimentalists when attempting to analyze the effectiveness of small Cu, Ag and Au clusters, as antioxidant materials.

Is Vitamin A an Antioxidant or a Pro-oxidant?

Antioxidant efficiency of all-trans-retinol has been studied on the basis of characteristic thermochemical properties using density functional theory. The influence of the solvent polarity has also been evaluated. It is found that retinol may act in parallel as an effective antioxidant via H atom donating as well as a pro-oxidant in yielding reactive hydroxyl radical. In fact, the lowest values of bond dissociation enthalpy were found at the C18-H and C18-OH positions. Retinol was also determined to be a good electron donor but bad acceptor in the single electron transfer (ET) reaction with hydroperoxyl (HOO•) radical. In addition, potential energy surfaces of H atom transfer (HAT) and radical adduct formation (RAF) reactions between retinol and HOO• radical were also investigated in the gas phase and in the solvent. The results demonstrated that the RAF mechanism was generally more predominant than ET and HAT ones. The most favored radical addition position was found at the C2═C3 double bond in the cyclohexenyl ring. Moreover, the radical scavenging reactivity via RAF reactions was strongly exergonic and thermodynamically feasible while the ET one was endergonic. Natural bond orbital analysis showed that the lone pairs of electrons on the oxygen atom of the HOO• radical were donated to the unoccupied antibonding orbital of transferred H atom in HAT reactions. In contrast, in the case of RAF reactions, strong interactions between 2p orbitals on oxygen atoms of the radical and the π orbital of the double bond on the retinol molecule were recognized. The results obtained in this work were in agreement with previous experimental observations.

Degradation mechanisms of Microcystin-LR during UV-B photolysis and UV/H2O2 processes: Byproducts and pathways

The removal and degradation pathways of microcystin-LR (MC-LR, [M+H]+ = 995.6) in UV-B photolysis and UV-B/H2O2 processes were examined using liquid chromatography-tandem mass spectrometry. The UV/H2O2 process was more efficient than UV-B photolysis for MC-LR removal. Eight by-products were newly identified in the UV-B photolysis ([M+H]+ = 414.3, 417.3, 709.6, 428.9, 608.6, 847.5, 807.4, and 823.6), and eleven by-products were identified in the UV-B/H2O2 process ([M+H]+ = 707.4, 414.7, 429.3, 445.3, 608.6, 1052.0, 313.4, 823.6, 357.3, 245.2, and 805.7). Most of the MC-LR by-products had lower [M+H]+ values than the MC-LR itself during both processes, except for the [M+H]+ value of 1052.0 during UV-B photolysis. Based on identified by-products and peak area patterns, we proposed potential degradation pathways during the two processes. Bond cleavage and intramolecular electron rearrangement by electron pair in the nitrogen atom were the major reactions during UV-B photolysis and UV-B/H2O2 processes, and hydroxylation by OH radical and the adduct formation reaction between the produced by-products were identified as additional pathways during the UV-B/H2O2 process. Meanwhile, the degradation by-products identified from MC-LR during UV-B/H2O2 process can be further degraded by increasing H2O2 dose.

Molybdenum Dithiolene Complexes: Trinuclear Mixed-Valence Complexes and Geometrical Isomers of Dinuclear Complexes

Abstract The reaction of an oxygen- and sulfur-bridged incomplete cubane-type molybdenum complex [Mo3(µ3-S)(µ-S)2(µ-O)(dtp)3(µ-dtp)(CH3CN)] (3) (dtp, diethyl dithiophosphate) with methyl propiolate (HC≡CCOOCH3, MP) and phenyl acetylene (HC≡CC6H5, PA) yields trinuclear mixed-valence Mo3(IV, V, V) complexes [Mo3(O)2(µ3-S)(µ3-SCH=C(COOCH3)S)(dtp)3(µ-OAc)] (3MP) and [Mo3(O)2(µ3-S)(µ3-SCH=C(C6H5)S)(dtp)3(µ-OAc)] (3PA), respectively, and also dinuclear Mo2(V, V) complexes [Mo2(O)2(µ-S)(µ-SCH=C(COOCH3)S)(dtp)2] (2aMP and 2bMP (a geometrical isomer of 2aMP)) and [Mo2(O)2(µ-S)(µ-SCH=C(C6H5)S)(dtp)2] (2aPA and 2bPA (a geometrical isomer of 2aPA)), respectively. The origin of the geometrical isomerism of the dimers comes from the unsymmetric nature of the acetylene derivatives. All of the structures were determined by X-ray crystallography, which revealed that each complex has two carbon-sulfur (µ-S) bonds arising from the adduct formation reaction between complex 3 and an acetylene derivative MP or PA. 1H NMR spectroscopy has proven that the dinuclear complexes are formed via the trinuclear complexes. The XPS spectra of 3MP show that the peaks of Mo 3d3/2 and Mo 3d5/2 are clearly split into two in the ratio of two to one, respectively, while those of 3 show no splitting, which indicates that 3MP is a mixed-valence complex with Mo3(IV, V, V) oxidation states. Cyclic voltammograms of 3MP and 3PA show chemically reversible one-electron oxidation processes at E1/2 = 0.21 V and E1/2 = 0.15 V vs. Ag/Ag+, respectively, and irreversible reduction peaks at −0.92 V and −1.00 V vs. Ag/Ag+, respectively. The dinuclear complexes, especially 2aPA and 2bPA having phenyl groups, are photosensitive to give their isomeric counter parts.

Mass Spectrometry Based Mechanistic Insights into Formation of Tris Conjugates: Implications on Protein Biopharmaceutics.

We present here extensive mass spectrometric studies on the formation of a Tris conjugate with a therapeutic monoclonal antibody. The results not only demonstrate the reactive nature of the Tris molecule but also the sequence and reaction conditions that trigger this reactivity. The results corroborate the fact that proteins are, in general, prone to conjugation and/or adduct formation reactions and any modification due to this essentially leads to formation of impurities in a protein sample. Further, the results demonstrate that the conjugation reaction happens via a succinimide intermediate and has sequence specificity. Additionally, the data presented in this study also shows that the Tris formation is produced in-solution and is not an in-source phenomenon. We believe that the facts given here will open further avenues on exploration of Tris as a conjugating agent as well as ensure that the use of Tris or any ionic buffer in the process of producing a biopharmaceutical drug is monitored closely for the presence of such conjugate formation. Graphical Abstract ᅟ.

Competitive Deprotonation and Superoxide [O₂⁻•)] Radical-Anion Adduct Formation Reactions of Carboxamides under Negative-Ion Atmospheric-Pressure Helium-Plasma Ionization (HePI) Conditions.

Carboxamides bearing an N-H functionality are known to undergo deprotonation under negative-ion-generating mass spectrometric conditions. Herein, we report that N-H bearing carboxamides with acidities lower than that of the hydroperoxyl radical (HO-O(•)) preferentially form superoxide radical-anion (O2(-•)) adducts, rather than deprotonate, when they are exposed to the glow discharge of a helium-plasma ionization source. For example, the spectra of N-alkylacetamides show peaks for superoxide radical-anion (O2(-•)) adducts. Conversely, more acidic amides, such as N-alkyltrifluoroacetamides, preferentially undergo deprotonation under similar experimental conditions. Upon collisional activation, the O2(-•) adducts of N-alkylacetamides either lose the neutral amide or the hydroperoxyl radical (HO-O(•)) to generate the superoxide radical-anion (m/z 32) or the deprotonated amide [m/z (M - H)(-)], respectively. For somewhat acidic carboxamides, the association between the two entities is weak. Thus, upon mildest collisional activation, the adduct dissociates to eject the superoxide anion. Superoxide-adduct formation results are useful for structure determination purposes because carboxamides devoid of a N-H functionality undergo neither deprotonation nor adduct formation under HePI conditions.

Optimized sample preparation strategy for the analysis of low molecular mass adducts of a fluorescent cisplatin analogue in cancer cell lines by CE-dual-LIF.

Pt-based anticancer drugs, such as cisplatin, are known to undergo several (bio-)chemical transformation steps after administration. Hydrolysis and adduct formation with small nucleophiles and larger proteins are their most relevant reactions on the way to the final reaction site (DNA), but there are still many open questions regarding the identity and pharmacological relevance of various proposed adducts and intermediates. Furthermore, the role of buffer components or additives, which are inevitably added to samples during any type of analytical measurement, has been frequently neglected in previous studies. Here, we report on adduct formation reactions of the fluorescent cisplatin analogue carboxyfluorescein diacetate platinum (CFDA-Pt) in commonly used buffers and cell culture medium. Our results indicate that chelation reactions with noninnocent buffers (e.g., Tris) and components of the cell culture/cell lysis medium must be taken into account when interpreting results. Adduct formation kinetics was followed up to 60h at nanomolar concentrations of CFDA-Pt by using CE-LIF. CE-MS enabled the online identification of such unexpected adducts down to the nanomolar concentration range. By using an optimized sample preparation strategy, unwanted adducts can be avoided and several fluorescent adducts of CFDA-Pt are detectable in sensitive and cisplatin-resistant cancer cell lines. By processing samples rapidly after incubation, we could even identify the initial, but transient, Pt species in the cells as deacetylated CFDA-Pt with unaltered complexing environment at Pt. Overall, the proposed procedure enables a very sensitive and accurate analysis of low molecular mass Pt species in cancer cells, involving a fast CE-LIF detection within 5min.