Dehydration-fragmentation mechanism of cathinones and their metabolites in ESI-CID.

Various cathinone-derived designer drugs (CATs) have recently appeared on the drug market. This study examined the mechanism for the generation of dehydrated ions for CATs during electrospray ionization collision-induced dissociation (ESI-CID). The generation mechanism of dehydrated ions is dependent on the amine classification in the cathinone skeleton, which is used in the identification of CATs. The two hydrogen atoms eliminated during the dehydration of cathinone (primary amine) and methcathinone (secondary amine) were determined, and the reaction mechanism was elucidated through the deuterium labeling experiments. The hydrogen atom bonded to the amine nitrogen was eliminated with the proton added during ESI, in both of the tested compounds. This provided evidence that CATs with tertiary amine structures (such as dimethylcathinone and α-pyrrolidinophenones [α-PPs]) do not undergo dehydration. However, it was shown that the two major tertiary amine metabolites (1-OH and 2″-oxo) of CATs generate dehydrated ions in ESI-CID. The dehydration mechanisms of the metabolites of α-pyrrolidinobutiophenone (α-PBP) belongs to α-PPs were also investigated. Stable-isotope labeling showed the dehydration of the 1-OH metabolite following a simple mechanism where the hydroxy group was eliminated together with the proton added during ESI. In contrast, the dehydration mechanism of the 2″-oxo metabolite involved hydrogen atoms in three or more locations along with the carbonyl group oxygen, indicating that dehydration occurred via multiple mechanisms likely including the rearrangement reaction of hydrogen atoms. These findings presented herein indicate that the dehydrated ions in ESI-CID can be used for the structural identification of CATs.

Synergistic Ion-pair Extraction and Separation of Trivalent Lanthanoid Ions with 4-Isopropyltropolone and 1,10-Phenanthroline into o-Dichlorobenzene.

The synergistic extraction of trivalent lanthanoid (Ln(III)) ions with 4-isopropyltropolone (Hipt) and 1,10-phenanthroline (phen) in o-dichrolobenzene (DCB) was investigated. The synergistic effect in DCB is more significant than that in toluene, and the polynuclear complexes found in toluene are not formed in DCB. Based on the 3-dimensional equilibrium analysis, the extracted species for La(III), Eu(III), and Lu(III) are found to be ion-pairs, such as Ln(ipt)2phen·ClO4 and Ln(ipt)2(phen)·ClO4 in the presence of NaClO4 as a salt, and the extraction constants of the respective species were determined. The simultaneous extraction of different lanthanoids in the present extraction system was demonstrated. The separation factors between lighter lanthanoids were larger than those with 2-ethylhexyl phosphonic acid mono-2-ethylhexyl ester, an excellent extractant for the separation of lanthanoid ions, in decane.

Extended internal standard method for quantitative 1H NMR assisted by chromatography (EIC) for analyte overlapping impurity on 1H NMR spectra.

We devised a novel extended internal standard method of quantitative 1H NMR (qNMR) assisted by chromatography (EIC) that accurately quantifies 1H signal areas of analytes, even when the chemical shifts of the impurity and analyte signals overlap completely. When impurity and analyte signals overlap in the 1H NMR spectrum but can be separated in a chromatogram, the response ratio of the impurity and an internal standard (IS) can be obtained from the chromatogram. If the response ratio can be converted into the 1H signal area ratio of the impurity and the IS, the 1H signal area of the analyte can be evaluated accurately by mathematically correcting the contributions of the 1H signal area of the impurity overlapping the analyte in the 1H NMR spectrum. In this study, gas chromatography and liquid chromatography were used. We used 2-chlorophenol and 4-chlorophenol containing phenol as an impurity as examples in which impurity and analyte signals overlap to validate and demonstrate the EIC, respectively. Because the 1H signals of 2-chlorophenol and phenol can be separated in specific alkaline solutions, 2-chlorophenol is suitable to validate the EIC by comparing analytical value obtained by the EIC with that by only qNMR under the alkaline condition. By the EIC, the purity of 2-chlorophenol was obtained with a relative expanded uncertainty (k = 2) of 0.24%. The purity matched that obtained under the alkaline condition. Furthermore, the EIC was also validated by evaluating the phenol content with the absolute calibration curve method by gas chromatography. Finally, we demonstrated that the EIC was possible to evaluate the purity of 4-chlorophenol, with a relative expanded uncertainty (k = 2) of 0.22%, which was not able to be separated from the 1H signal of phenol under any condition.

Mechanistic Analysis of Ion Association between Dendrigraft Poly-l-lysine and 8-Anilino-1-naphthalenesulfonate at Liquid|Liquid Interfaces.

Molecular association between biocompatible dendritic polymers, dendrigraft poly-l-lysines (DGLs), and an anionic fluorescent probe, 8-anilino-1-naphthalenesulfonate (ANS-), was studied at the polarized water|1,2-dichloroethane (DCE) interface. The fluorescence intensity of ANS measured in aqueous solution was enhanced by the coexistence of DGLs over a wide pH range (2 < pH < 10), where ANS and DGL exist as a monoanionic form and a polycation, respectively. The voltammetric responses indicated that the positively charged DGLs were adsorbed at the water|DCE interface, whereas ANS- was transferred across the interface accompanied by the adsorption process. The interfacial behavior of the DGL-ANS associates was analyzed by potential-modulated fluorescence (PMF) spectroscopy. The PMF results demonstrated that the ion association between DGLs and ANS at the water|DCE interface is strongly affected by the applied potential and the dendritic generation of DGL. By applying appropriate potentials, the ANS anion was dissociated from its ion associate with DGLs at the interface and transferred into the organic phase, whereas DGLs remained in the aqueous phase. The Gibbs free energy of ion association (Δ GD···ANS) was estimated for the second-fourth generation DGLs (DGL-G2-G4) and the G4 polyamidoamine (PAMAM) dendrimer as a control. The highest stability of the DGL-G4-ANS associate manifested itself through Δ GD···ANS: DGL-G4-ANS (>G4 PAMAM dendrimer-ANS) > DGL-G3-ANS > DGL-G2-ANS. The results elucidated the efficient anion-binding ability of higher generation DGLs and its potential dependence at the liquid|liquid interface.

Potential-Induced Aggregation of Anionic Porphyrins at Liquid|Liquid Interfaces.

The adsorption and self-aggregation of anionic porphyrins were studied at the polarized water|1,2-dichloroethane (DCE) interface by polarization-modulation total internal reflection fluorescence (PM-TIRF) spectroscopy. 5,10,15,20-Tetrakis(4-sulfonatophenyl)porphyrin diacid (H4TPPS2-) and protoporphyrin IX (H2PP2-) exhibited high surface activities at the interface. The selective excitation of interfacial species in PM-TIRF measurements elucidated the potential-induced aggregation mechanism of the porphyrins. The J-aggregates of H4TPPS2- were reversibly formed only at the water|DCE interface by applying appropriate potentials even when the porphyrins exist as monomers in the aqueous and organic solutions. In the H2PP2- system, the slow aggregation process was found in the negative potential region. The spectral characteristics and the signal phase of PM-TIRF indicated that the H2PP2- monomers were adsorbed with relatively standing orientation and that the long axis of the J-aggregates was nearly in plane of the interface. H2PP2- was also investigated at the biomimetic phospholipid-adsorbed water|DCE interface. The competitive adsorption of neutral glycerophospholipids effectively inhibited the potential-dependent adsorption and interfacial aggregation processes of H2PP2-. The results demonstrated that the aggregation state of the charged species can reversibly be controlled at liquid|liquid interfaces as a function of externally applied potential.

Vitamin B6 Prevents IL-1ß Protein Production by Inhibiting NLRP3 Inflammasome Activation.

Vitamin B6 includes six water-soluble vitamers: pyridoxal (PL), pyridoxamine (PM), pyridoxine (PN), and their phosphorylated forms. Pyridoxal 5'-phosphate (PLP) is an important cofactor for many metabolic enzymes. Several lines of evidence demonstrate that blood levels of PLP are significantly lower in patients with inflammation than in control subjects and that vitamin B6 has anti-inflammatory effects, with therapeutic potential for a variety of inflammatory diseases. Although one of our group previously demonstrated that PL inhibits the NF-κB pathway, the molecular mechanism by which vitamin B6 suppresses inflammation is not well understood. Here, we showed that both PL and PLP suppressed the expression of cytokine genes in macrophages by inhibiting Toll-like receptor (TLR)-mediated TAK1 phosphorylation and the subsequent NF-κB and JNK activation. Furthermore, PL and PLP abolished NLRP3-dependent caspase-1 processing and the subsequent secretion of mature IL-1ß and IL-18 in LPS-primed macrophages. In contrast, PM and PN had little effect on IL-1ß production. PLP, but not PL, markedly reduced the production of mitochondrial reactive oxygen species (ROS) in peritoneal macrophages. Importantly, PL and PLP reduced IL-1ß production induced by LPS and ATP, or by LPS alone, in mice. Moreover, PL and PLP protected mice from lethal endotoxic shock. Collectively, these findings reveal novel anti-inflammatory activities for vitamin B6 and suggest its potential for preventing inflammatory diseases driven by the NLRP3 inflammasome.

Photoluminescent Detection of Nitrite with Carbon Nanodots Prepared by Microwave-assisted Synthesis.

A photoluminescent detection method for nitrite with high selectivity and sensitivity using carbon nanodots (CNDs) is demonstrated. The selectivity of nitrite is accomplished by a highly specific diazotization reaction between nitrite and p-phenylenediamine (p-PDA). In the presence of nitrite, p-PDA easily reacts to form the diazonium cation in the acidic aqueous solution. By alkalization of the reaction mixture, diazonium cation of p-PDA was converted to an aryl radical to form aggregated CNDs, which causes the change in the photoluminescent intensity of CNDs. In the present method, nitrite can be selectively detected down to 1 µM over several anions, such as nitrate, perchlorate, sulfate, fluoride, chloride, and bromide at mM levels.

Photoinduced Electron Transfer of PAMAM Dendrimer-Zinc(II) Porphyrin Associates at Polarized Liquid|Liquid Interfaces.

The heterogeneous photoinduced electron-transfer reaction of the ion associates between NH2-terminated polyamidoamine (PAMAM) dendrimers and 5,10,15,20-tetrakis(4-sulfonatophenyl)porphyrinato zinc(II) (ZnTPPS(4-)) was studied at the polarized water|1,2-dichloroethane (DCE) interface. The positive photocurrent arising from the photoreduction of ZnTPPS(4-) by a lipophilic quencher, decamethylferrocene, in the interfacial region was significantly enhanced by the ion association with the PAMAM dendrimers. The photocurrent response of the dendrimer-ZnTPPS(4-) associates was dependent on the pH condition and on the generation of dendrimer. A few cationic additives such as polyallylamine and n-octyltrimethyammonium were also examined as alternatives to the PAMAM dendrimer, but the magnitude of the photocurrent enhancement was rather small. The high photoreactivity of the dendrimer-ZnTPPS(4-) associates was interpreted mainly as a result of the high interfacial concentration of photoreactive porphyrin units associated stably with the dendrimer which was preferably adsorbed at the polarized water|DCE interface. The photochemical data observed in the second and fourth generation PAMAM dendrimer systems demonstrated that the higher generation dendrimer which can incorporate a porphyrin molecule more completely in the interior is less efficient for the photocurrent enhancement at the interface. These results indicated that the photoreactivity of ionic reactant at a polarized liquid|liquid interface can readily be modified via ion association with the charged dendrimer.

Highly selective synergism for the extraction of lanthanoid(III) ions with ß-diketones and trioctylphosphine oxide in an ionic liquid.

Selective synergism for the extraction of lanthanoids(III) (Ln) with ß-diketones such as 2-thenoyltrifluoroacetone, 2-naphthoyltrifluoroacetone, and benzoylacetone has been investigated in the presence of trioctylphosphine oxide as a hydrophobic neutral ligand in 1-butyl-3-methylimidazolium bis(trifluoromethanesulfonyl)imide as an ionic liquid. The extractability of heavier Ln was remarkably enhanced, resulting in a significant improvement in the separation of Ln. It was found that the present synergism is ascribed to the formation of cationic ternary complexes, followed by ion exchange into the ionic liquid.

Communication: Coordination structure of bromide ions associated with hexyltrimethylammonium cations at liquid/liquid interfaces under potentiostatic control as studied by total-reflection X-ray absorption fine structure.

Total-reflection X-ray absorption fine structure (TR-XAFS) technique was applied for the first time to an interface between two immiscible electrolyte solutions under potentiostatic control. The hydration structure of bromide ions was investigated at polarized 2-octanone/water interfaces. TR-XAFS spectra at Br K-edge measured in the presence of hexyltrimethylammonium bromide (C6TAB) were slightly modified depending on the Galvani potential difference (Δ(o)(w)φ). The extended X-ray absorption fine structure analysis exposed hydration structure changes of bromide ions at the polarized interface. The coordination structure of bromide ions at the interface could be analyzed as compared with bromide ions dissolved in aqueous solution and Br(-)-exchanged resin having quaternary ammonium groups. The results indicated that bromide ions were associated with C6TA(+) at the polarized interface. The relative contribution of ion association form of bromide ions with quaternary ammonium groups was enhanced at a potential close to the ion transfer of C6TA(+), where the interfacial concentration of C6TA(+) is increased as a function of Δ(o)(w)φ.

Spectroelectrochemical characterization of dendrimer-porphyrin associates at polarized liquid|liquid interfaces.

Molecular encapsulation of anionic porphyrins in NH2-terminated polyamidoamine (PAMAM) dendrimers and the interfacial behavior of the dendrimer-porphyrin associates were studied at the polarized water|1,2-dichloroethane (DCE) interface. Formation of the ion associates was significantly dependent on the pH condition and on generation of dendrimers. 5,10,15,20-Tetrakis(4-sulfonatophenyl)porphyrin (ZnTPPS(4-)) associated with the positively charged fourth-generation (G4) PAMAM dendrimer was highly stabilized in acidic aqueous solution without protolytic demetalation in a wide range of pH values (pH > 2). In contrast to the zinc(II) complex, the free base porphyrin (H2TPPS(4-)) was readily protonated under acidic conditions even in the presence of the dendrimers. In addition, the J-aggregates of diprotonated species, (H4TPPS(2-))n, were preferably formed on the dendrimer. The interfacial mechanism of the dendrimer-porphyrin associates was analyzed in detail by potential-modulated fluorescence (PMF) spectroscopy. PMF results indicated that the dendrimers incorporating porphyrin molecules were transferred across the positively polarized water|DCE interface via adsorption step, whereas the transfer responses of the porphyrin ions released from the dendrimers were observed at negatively polarized conditions. A negative shift of the transfer potential of porphyrin ions compared to the intrinsic transfer potential was apparently observed for each ion association system. The ion association stability between the dendrimer and the porphyrin molecules could be estimated from a negative shift of the transfer potential. ZnTPPS(4-) exhibited relatively strong interaction with the higher generation dendrimer, whereas H2TPPS(4-) was less effectively associated with the dendrimers.

Valence discriminative detection of metal cations by a chromotropic acid-grafted glassy carbon electrode.

Covalent grafting of chromotropic acid (CA) moiety to a glassy carbon electrode was performed by electrochemical reduction of the corresponding diazonium cation generated in situ. Grafted electrodes were subjected to the electrochemical detection of multivalent cations using Fe(CN)(6)(3-/4-) as an electrochemical probe molecule. In the absence of metal cations, redox reaction of the anionic probe was depressed by electrostatic repulsion between the probe and negatively charged CA moieties on the electrode surface. Binding of tetravalent cations such as Zr(IV) and Hf(IV) and trivalent cations such as Al(III) and lanthanide(III) to the CA moiety on the electrode surface resulted in an increase in the current intensity of the probe due to a decrease in electrostatic repulsion. Highly selective detection of the tetravalent cations was achieved in the micromolar concentration range at pH 1.0.

Specific cooperative effect of a macrocyclic receptor for metal ion transfer into an ionic liquid.

An intramolecular cooperative extraction system for the removal of strontium cations (Sr(2+)) from water by use of a novel macrocyclic receptor (H(2)ßDA18C6) composed of diaza-18-crown-6 and two ß-diketone fragments in ionic liquid (IL) is reported, together with X-ray spectroscopic characterization of the resulting extracted complexes in the IL and chloroform phases. The covalent attachment of two ß-diketone fragments to a diazacrown ether resulted in a cooperative interaction within the receptor for Sr(2+) transfer, which remarkably enhanced the efficiency of Sr(2+) transfer relative to a mixed ß-diketone and diazacrown system. The intramolecular cooperative effect was observed only in the IL extraction system, providing a 500-fold increase in extraction performance for Sr(2+) over chloroform. Slope analysis and potentiometric titration confirmed that identical extraction mechanisms operated in both the IL and chloroform systems. Extended X-ray absorption fine structure spectroscopy revealed that the average distance between Sr(2+) and O atoms in the Sr(2+) complex was shorter in IL than in chloroform. Consequently, Sr(2+) was held by H(2)ßDA18C6 more rigidly in IL than in chloroform, representing an important factor dominating the magnitude of the intramolecular cooperative effect of H(2)ßDA18C6 for Sr(2+). Furthermore, competitive extraction studies with alkaline earth metal ions revealed that the magnitude of the intramolecular cooperative effect depended on the suitability between metal ion size and the cavity size of H(2)ßDA18C6. Sr(2+) was successfully recovered from IL by controlling the pH in the receiving phase, and the extraction performance of H(2)ßDA18C6 in IL was maintained after five repeated uses.

Evaluation of a hydrophilic ionic liquid as a salting-out phase separation agent to a water-tetrahydrofuran homogeneous system for aqueous biphasic extraction separation.

The use of a hydrophilic ionic liquid (IL), 1-butyl-3-methylimidazolium chloride (C(4)mimCl), as a salting-out phase separation agent to a water-tetrahydrofuran homogeneous system was studied for possible applications to novel aqueous biphasic extraction separation. The IL showed a salting-out phase-separation ability. Also, differences in the polarity between the formed two phases were smaller than that when using NaCl as a salting-out agent. This result suggested that C(4)mimCl remaining in water-rich phase acts not only as a salting-out agent, but also a component of a mixed-solvent. Possible uses of C(4)mimCl/NaCl mixed salting-out agent system were also discussed.

Substoichiometric isotope dilution mass spectrometry of boron by the ion-pair extraction with halogenated salicyl alcohol derivatives and a quaternary ammonium salt.

Novel salicyl alcohol derivatives (H(2)X(n)sal), 5-bromo-, 3,5-dibromo-, and 3,5-diiodosalicyl alcohol which were abbreviated to H(2)Brsal, H(2)Br(2)sal, and H(2)I(2)sal, respectively, were synthesized and used for the selective extraction of boric acid. Boric acid was extracted with each H(2)X(n)sal into chlorobenzene containing trioctylmethylammonium chloride (TOMACl) as an ion-pair complex, TOMA·B(X(n)sal)(2), at a different pH range. The extraction constant (K(ex)) of boric acid was determined by the equilibrium analyses including the formation of hydrogen-bonded complex of each H(2)X(n)sal with TOMACl in the organic phase. The K(ex) values obtained by salicyl alcohol (H(2)sal) and its derivatives were decreased in the order of H(2)I(2)sal ≥ H(2)Br(2)sal > H(2)Brsal > H(2)sal. The most powerful extractant, H(2)I(2)sal, was employed for the substoichiometric extraction of boric acid, which was extracted at pH 5 - 9 with a substoichiometric amount of TOMACl in the presence of an excess of H(2)I(2)sal. The present substoichiometric separation method combined with the stable isotope dilution analysis using inductively coupled plasma mass spectrometry (ICP-MS) could be successfully applied to the determination of boron in a reference material of high-analysis compound fertilizer (FAMIC-A-08) without any correction as to the isotopic abundance.

Solvent effect of ionic liquids on the distribution constant of 2-thenoyltrifluoroacetone and its nickel(II) and copper(II) chelates and the evaluation of the solvent properties based on the regular solution theory.

Distribution constants of 2-thenoyltrifluoroacetone (Htta) and its Ni(II)and Cu(II) chelates between 1-alkyl-3-methylimidazolium bis(trifluoromethanesulfonyl)imide (C(n)mimTf(2)N; n=4, 6, and 8) as ionic liquid (IL) and an aqueous phase were determined. The enol fraction of Htta in ILs was spectrophotometrically measured to calculate the distribution constant of the enol form (K(D,HE)) of Htta. The K(D,HE) values in ILs were evaluated by comparing those in various molecular solvents such as alkanes, aromatic hydrocarbons, chlorohydrocarbons, ethers, ketones, and esters previously reported on the basis of the regular solution theory (RST). It was elucidated that the IL solutions of Htta (enol) can be taken as apparently regular solutions as expected in the organic solvents. On the other hand, the effect of ILs on the distribution constant of metal(II) chelates (K(D,M)) was fairly complicated. The Cu(tta)(2)-IL solutions behaved like the alkane and aromatic hydrocarbon solutions but the Ni(tta)(2)-IL (C(4)mimTf(2)N) like ether and ketone solutions. In the Ni(II) case, some specific interactions between the Ni(II) chelate and IL was suggested. Finally, the solubility parameters of ILs were calculated using K(D,HE) by RST and were in good agreement with the literature values obtained by the enthalpy of vaporization.

An 8-sulfonamidoquinoline derivative with imidazolium unit as an extraction reagent for use in ionic liquid chelate extraction systems.

To evaluate the superiority of using a task-specific onium salt (TSOS), a novel TSOS reagent 1-methyl-3-[2-(8-quinolinylaminosulfonyl)ethyl]imidazolium chloride (HmimesqCl) was synthesized, and its performance in extraction of different divalent metal cations was investigated. This TSOS reagent has both a cationic imidazolium unit and an anionic (proton-dissociable) complexation unit for the extraction of metal cations into an ionic liquid. In the use of HmimesqCl, relatively high extraction of metals resulted in comparison with the use of its non-imidazolium analogs, without changing the extraction selectivity.

Synergistic effect of 18-crown-6 derivatives on chelate extraction of lanthanoids(III) into an ionic liquid with 2-thenoyltrifluoroacetone.

The synergistic effect of 18-crown-6 derivatives, such as 18-crown-6 (18C6), cis-dicyclohexano-18-crown-6 (DC18C6) and dibenzo-18-crown-6 (DB18C6), on the extraction of trivalent lanthanoids (Ln(3+)) into an ionic liquid, 1-butyl-3-methylimidazolium bis(trifluoromethanesulfonyl)imide, with 2-thenoyltrifluoroacetone (Htta) was investigated. The extractability of lighter Ln(3+) was enhanced by adding 18C6 or DC18C6, whereas no enhancement of the extractability was observed by adding DB18C6. Moreover, the synergistic effect by the crown ether (CE) was increased along with the decrease in the atomic number of Ln. In the synergistic extraction system, Ln(3+) was extracted as cationic ternary complexes Ln(tta)(2)(CE)(+) and Ln(tta)(CE)(2+), and it was suggested that the formation of the Ln(tta)(CE)(2+) complex as an extracted species results in the large synergistic effect. This synergistic effect originated in a size-fitting effect of CE on complexation to Ln(3+).

Cooperative intramolecular interaction of diazacrown ether bearing beta-diketone fragments on an ionic liquid extraction system.

A novel extractant beta-diketone-substituted diaza-18-crown-6 demonstrated very efficient extraction of Sr(2+) due to an intramolecular synergistic effect on the ionic liquid extraction system and recovery of Sr(2+) from the ionic liquid was successfully achieved under acidic conditions.