Formation and stability of peptide enolates in aqueous solution

Second-order rate constants k(DO) (M(-1) s(-1)) were determined in D(2)O for deprotonation of the N-terminal alpha-amino carbon of glycylglycine and glycylglycylglycine zwitterions, the internal alpha-amino carbon of the glycylglycylglycine anion, and the acetyl methyl group and the alpha-amino carbon of the N-acetylglycine anion and N-acetylglycinamide by deuterioxide ion. The data were used to estimate values of k(HO) (M(-1) s(-1)) for proton transfer from these carbon acids to hydroxide ion in H(2)O. Values of the pK(a) for these carbon acids ranging from 23.9 to 30.8 were obtained by interpolation or extrapolation of good linear correlations between log k(HO) and carbon acid pK(a) established in earlier work for deprotonation of related neutral and cationic alpha-carbonyl carbon acids. The alpha-amino carbon at a N-protonated N-terminus of a peptide or protein is estimated to undergo deprotonation about 130-fold faster than the alpha-amino carbon at the corresponding internal amino acid residue. The value of k(HO) for deprotonation of the N-terminal alpha-amino carbon of the glycylglycylglycine zwitterion (pK(a) = 25.1) is similar to that for deprotonation of the more acidic ketone acetone (pK(a) = 19.3), as a result of a lower Marcus intrinsic barrier to deprotonation of cationic alpha-carbonyl carbon acids. The cationic NH(3)(+) group is generally more strongly electron-withdrawing than the neutral NHAc group, but the alpha-NH(3)(+) and the alpha-NHAc substituents result in very similar decreases in the pK(a) of several alpha-carbonyl carbon acids.     

Determination of the pKa of cyclobutanone: Brønsted correlation of the general base-catalyzed enolization in aqueous solution and the effect of ring strain

The induction of strain in carbocycles, thereby increasing the amount of s-character in the C-H bonds and the acidity of these protons, has been probed with regard to its effect on the rate constants for the enolization of cyclobutanone. The second-order rate constants for the general base-catalyzed enolization of cyclobutanone have been determined for a series of 3-substituted quinuclidine buffers in D(2)O at 25 °C, I = 1.0 M (KCl). The rate constants for enolization were determined by following the extent of deuterium incorporation (up to ～30% of the first α-proton) into the α-position, as a function of time. The observed pseudo-first-order rate constants correlated to the [basic form] of the buffer and yielded the second-order rate constants for the general base-catalyzed enolization of cyclobutanone for four tertiary amine buffers. A Br?nsted β-value of 0.59 was determined from the second-order rate constants determined. Comparison of the results for cyclobutanone to those previously reported for acetone and a 1-phenylacetone derivative, under similar conditions, indicated that the ring strain of the carbocycle appeared to have only a small effect on the general base-catalyzed rate constants for enolization. The similarity of the rate constants for the general base-catalyzed enolization of cyclobutanone to those determined for acetone allowed for an estimation of the limits of the rate constant for protonation of the enolate intermediate of cyclobutanone by the conjugate acid of 3-quinuclidinone (k(BH) = 5 × 10(8) - 2 × 10(9) M(-1) s(-1)). Combining the rate constants for deprotonation of cyclobutanone (k(B)) and protonation of the enolate of cyclobutanone (k(BH)) by 3-quinuclidinone and its conjugate acid, the pK(a) of the α-protons of cyclobutanone has been estimated to be pK(a) = 19.7-20.2.     

Covalent catalysis by pyridoxal: evaluation of the effect of the cofactor on the carbon acidity of glycine

First-order rate constants for deprotonation of the alpha-imino carbon of the adduct between 5'-deoxypyridoxal (1) and glycine were determined as the rate constants for Claisen-type addition of glycine to 1 where deprotonation is rate determining for product formation. There is no significant deprotonation at pH 7.1 of the form of the 1-glycine iminium ion with the pyridine nitrogen in the basic form. The value of kHO for hydroxide ion-catalyzed deprotonation of the alpha-imino carbon increases from 7.5 x 10(2) to 3.8 x 10(5) to 3.0 x 10(7) M(-1) s(-1), respectively, with protonation of the pyridine nitrogen, the phenoxide oxyanion, and the carboxylate anion of the 1-glycine iminium ion. There is a corresponding decrease in the pKas for deprotonation of the alpha-imino carbon from 17 to 11 to 6. It is proposed that enzymes selectively bind and catalyze the reaction of the iminium ion with pKa = 17. A comparison of kB = 1.7 x 10(-3) s(-1) for deprotonation of the alpha-imino carbon of this cofactor-glycine adduct (pKa = 17 by HPO4(2-) with k(cat)/K(m) = 4 x 10(5) M(-1) s(-1) for catalysis of amino-acid racemization by alanine racemase shows that the enzyme causes a ca 2 x 10(8)-fold acceleration of the rate of deprotonation the alpha-imino carbon. This corresponds to about one-half of the burden borne by alanine racemase in catalysis of deprotonation of alanine.     

Ring strain and its effect on the rate of the general-base catalyzed enolization of cyclobutanone

[reaction: see text] The 3-quinuclidinone-catalyzed (pK(BH) = 7.5) enolization of cyclobutanone (1) in D(2)O at 25 degrees C, I = 1.0 (KCl) was followed by deuterium incorporation, which was determined by (1)H NMR. The second-order rate constant for the buffer-catalyzed deprotonation of 1 was found to be k(B) = 3.3 x 10(-4) M(-1) s(-1), which is compared to rates for acetone and 2-(2'-oxopropyl)benzaldehyde under similar conditions. The data shows that ring strain has very little effect on the energy barrier to deprotonation of 1 vs the unstrained systems.     

Substituent effects on electrophilic catalysis by the carbonyl group: anatomy of the rate acceleration for PLP-catalyzed deprotonation of glycine

First-order rate constants, determined by (1)H NMR, are reported for deuterium exchange between solvent D(2)O and the α-amino carbon of glycine in the presence of increasing concentrations of carbonyl compounds (acetone, benzaldehyde, and salicylaldehyde) and at different pD and buffer concentrations. These rate data were combined with (1)H NMR data that define the position of the equilibrium for formation of imines/iminium ions from addition of glycine to the respective carbonyl compounds, to give second-order rate constants k(DO) for deprotonation of α-imino carbon by DO(-). The assumption that these second-order rate constants lie on linear structure-reactivity correlations between log k(OL) and pK(a) was made in estimating the following pK(a)'s for deprotonation of α-imino carbon: pK(a) = 22, glycine-acetone iminium ion; pK(a) = 27, glycine-benzaldehyde imine; pK(a) ≈ 23, glycine-benzaldehyde iminium ion; and, pK(a) = 25, glycine-salicylaldehyde iminium ion. The much lower pK(a) of 17 [Toth, K.; Richard, J. P. J. Am. Chem. Soc. 2007, 129, 3013-3021] for carbon deprotonation of the adduct between 5'-deoxypyridoxal (DPL) and glycine shows that the strongly electron-withdrawing pyridinium ion is unique in driving the extended delocalization of negative charge from the α-iminium to the α-pyridinium carbon. This favors carbanion protonation at the α-pyridinium carbon, and catalysis of the 1,3-aza-allylic isomerization reaction that is a step in enzyme-catalyzed transamination reactions. An analysis of the effect of incremental changes in structure on the activity of benzaldehyde in catalysis of deprotonation of glycine shows the carbonyl group electrophile, the 2-O(-) ring substituent and the cation pyridinium nitrogen of DPL each make a significant contribution to the catalytic activity of this cofactor analogue. The extraordinary activity of DPL in catalysis of deprotonation of α-amino carbon results from the summation of these three smaller effects.     

Flash photolytic generation of ortho-quinone methide in aqueous solution and study of its chemistry in that medium

Flash photolysis of o-hydroxybenzyl alcohol, o-hydroxybenzyl p-cyanophenyl ether, and (o-hydroxybenzyl)trimethylammonium iodide in aqueous perchloric acid and sodium hydroxide solutions, and in acetic acid and biphosphate ion buffers, produced o-quinone methide as a short-lived transient species that underwent hydration back to benzyl alcohol in hydrogen-ion catalyzed (k(H+) = 8.4 x 10(5) M(-1) s(-1)) and hydroxide-ion catalyzed (k(HO)- = 3.0 x 10(4) M(-1) s(-1)) reactions as well as an uncatalyzed (k(UC) = 2.6 x 10(2) s(-1)) process. The hydrogen-ion catalyzed reaction gave the solvent isotope effect k(H+)/k(D)+ = 0.42, whose inverse nature indicates that this process occurs by rapid and reversible equilibrium protonation of the carbonyl oxygen atom of the quinone methide, followed by rate-determining capture of the carbocation so produced by water. The magnitude of the rate constant of the uncatalyzed reaction, on the other hand, indicates that this process occurs by simple nucleophilic addition of water to the methylene group of the quinone methide. Decay of the quinone methide is also accelerated by acetic acid buffers through both acid- and base-catalyzed pathways, and quantitative analysis of the reaction products formed in these solutions shows that this acceleration is caused by nucleophilic reactions of acetate ion rather than by acetate ion assisted hydration. Bromide and thiocyanate ions also accelerate decay of the quinone methide through both hydrogen-ion catalyzed and uncatalyzed pathways, and the inverse nature of solvent isotope effects on the hydrogen-ion catalyzed reactions shows that these reactions also occur by rapid equilibrium protonation of the quinone methide carbonyl oxygen followed by rate-determining nucleophilic capture of the ensuing carbocation. Assignment of an encounter-controlled value to the rate constant for the rate-determining step of the thiocyanate reaction leads to pK(a) = -1.7 for the acidity constant of the carbonyl-protonated quinone methide.     

Acid-base equilibria and dynamics in sodium dodecyl sulfate micelles: geminate recombination and effect of charge stabilization

The synthetic flavylium salt 4-carboxy-7-hydroxy-4'-methoxyflavylium chloride (CHMF) exhibits two acid-base equilibria in the range of pH 1-8 in both aqueous and micellar sodium dodecyl sulfate (SDS) solutions. The values of pK(a1) and pK(a2) for the cation-zwitterion (AH(2)(+) <--> Z + H(+)) and the zwitterion-base (Z <--> A(-) + H(+)) equilibria increase from 0.73 and 4.84 in water to 2.77 and 5.64 in SDS micelles, respectively. The kinetic study of the Z <--> A(-) + H(+) ground-state reactions in SDS points to the diffusion-controlled protonation of A(-) in the aqueous phase (k(p2w) = 4.2 x 10(10) M(-)(1) s(-)(1)) and in the micelle (k(p2m) = 2.3 x 10(11) M(-)(1) s(-)(1)). The deprotonation rate of Z did not significantly change upon going from water (k(d2) = 6.3 x 10(5) s(-)(1)) to SDS (k(d2) = 5.2 x 10(5) s(-)(1)), in contrast with the behavior of ordinary cationic flavylium salts, for which k(d2) strongly decreases in SDS micelles. These results suggest that deprotonation of the zwitterionic acid is not substantially perturbed by the micellar charge. Electronic excitation of the Z form of CHMF induces fast adiabatic deprotonation of the hydroxyl group of Z() (2.9 x 10(10) s(-)(1) in water and 8.4 x 10(9) s(-)(1) in 0.1 M SDS), followed by geminate recombination on the picosecond time scale. Interestingly, while recombination in water (k(rec) = 1.7 x 10(9) s(-)(1)) occurs preferentially at the carboxylate group, at the SDS micelle surface, recombination (k(rec) = 9.2 x 10(9) s(-)(1)) occurs at the hydroxyl group. The important conclusion is that proton mobility at the SDS micelle surface is substantially reduced with respect to the mobility in water, which implies that geminate recombination should be a general phenomenon in SDS micelles.     

Protonated benzofuran,anthracene, naphthalene,benzene, ethene,and ethyne: measurements and estimates of pK(a) and pK(R)

Aqueous solvolyses of acyl derivatives of hydrates (water adducts) of anthracene and benzofuran yield carbocations which undergo competitive deprotonation to form the aromatic molecules and nucleophilic reaction with water to give the aromatic hydrates. Trapping experiments with azide ions yield rate constants k(p) for the deprotonation and k(H2O) for the nucleophilic reaction based on the "azide clock". Combining these with rate constants for (a) the H(+)-catalyzed reaction of the hydrate to form the carbocation and (b) hydrogen isotope exchange of the aromatic molecule (from the literature) yields pK(R) = -6.0 and -9.4 and pK(a) = -13.5 and -16.3 for the protonated anthracene and protonated benzofuran, respectively. These pK values may be compared with pK(R) = -6.7 for naphthalene hydrate (1-hydroxy-1,2-dihydronaphthalene), extrapolated to water from measurements by Pirinccioglu and Thibblin for acetonitrile-water mixtures, and pK(a) = -20.4 for the 2-protonated naphthalene from combining k(p) with an exchange rate constant. The differences between pK(R) and pK(a) correspond to pK(H2O), the equilibrium constant for hydration of the aromatic molecule (pK(H2O) = pK(R) - pK(a)). For naphthalene and anthracene values of pK(H2O) = +13.7 and +7.5 compare with independent estimates of +14.2 and +7.4. For benzene, pK(a) = -24.3 is derived from an exchange rate constant and an assigned value for the reverse rate constant close to the limit for solvent relaxation. Combining this pK(a) with calculated values of pK(H2O) gives pK(R) = -2.4 and -2.1 for protonated benzenes forming 1,2- and 1,4-hydrates, respectively. Coincidentally, the rate constant for protonation of benzene is similar to those for protonation of ethylene and acetylene (Lucchini, V.; Modena, G. J. Am. Chem Soc. 1990, 112, 6291). Values of pK(a) for the ethyl and vinyl cations (-24.8) may thus be derived in the same way as that for the benzenonium ion. Combining these with appropriate values of pK(H2O) then yields pK(R) = -39.8 and -29.6 for the vinyl and ethyl cations, respectively.     

Protolytic reactions of 3,3'-dimethylnaphthidine and 3,3'-dimethoxybenzidine

The dissociation constants of diprotonated 3,3'-dimethylnaphthidine (DMN) and 3,3'-dimethoxybenzidine (DMB) have been determined spectrophotometrically. They are: pK(a1) = 2.62 +/- 0.03, pK(a2) = 3.33 +/- 0.09 for DMN: pK(a1) = 2.83 +/- 0.07, pK(a2) = 4.05 +/- 0.12 for DMB. The molar absorptivities (l.mole(-1).cm(-1)) of all forms of the indicators have been also determined: epsilon(B) = 1.68 x 10(4), epsilon(BH(+)) = 9.34 x 10(3), epsilon(BH(2+)(2)) = 1.80 x 10(3) at 300 nm for DMB; epsilon(B) = 7.33 x 10(3), epsilon(BH(+)) = 3.73 x 10(3), epsilon(BH(2+)(2)) = 0 at 330 nm for DMN.     

Novel ground- and excited-state prototropic reactivity of a hydroxycarboxyflavylium salt

Synthetic and natural hydroxyflavylium salts are super-photoacids, exhibiting values of the rate constant for proton transfer to water in the excited state as high as 1.5 x 10(11) s(-1). The synthetic flavylium salt 4-carboxy-7-hydroxy-4'-methoxyflavylium chloride (CHMF) has an additional carboxyl group at the 4-position of the flavylium cation that deprotonates in the ground state at a lower pH (pK(a1) = 0.73; AH2+ --> Z) than the 7-hydroxy group (pK(a2) = 4.84; Z --> A-). Ground-state deprotonation of the carboxyl group of the acid (AH2+) to form the zwitterion (Z) is too fast to be detected by nanosecond laser flash perturbation of the ground-state equilibrium, while deprotonation of the hydroxyl group of Z to form the anionic base (A-) occurs in the microsecond time range (k(d2) = 0.6 x 10(6) s(-1) and k(p2) = 4.2 x 10(10) M(-1) x s(-1)). In the excited state, the cationic form (AH2+) deprotonates in approximately 9 ps, resulting in the excited neutral base form (AH), which is unstable in the ground state. Deprotonation of Z occurs in 30 ps (k(d2) = 2.9 x 10(10) s(-1)), to form excited A-, which either reprotonates (k(p3)* = 3.7 x 10(10) M(-1) x s(-1)) or decays in 149 ps, and shows an important contribution from geminate recombination to give the excited neutral base (AH). Predominant reprotonation of A- at the carboxylate group reflects both the presence of the negative charge on the carboxylate and the increase in the excited-state pK(a) of the carboxyl group. Thus, while the hydroxyl pK(a) decreases by approximately 5 units upon going from the ground state (pK(a) = 4.84) to the excited state (pK(a) = -0.2), that of the carboxyl group increases by at least this much. Consequently, the excited state of the Z form of CHMF acts as a molecular proton transporter in the picosecond time range.     

Mechanisms of acid-base catalysis of beta-elimination reactions in systems activated by a pyridine ring

Beta-elimination reactions from 1 (in quinuclidine/quinuclidinium chloride, imidazole/imidazolium, and acetate/acetic acid buffers) and from 2 (in imidazole/imidazolium and acetate/acetic acid buffers) with formation of 4-vinylpyridine and 2-vinylpyridine, respectively, were studied. The results of a kinetic study of acid-base catalysis and H/D exchange are consistent with NH(+), the protonated substrate, as the species that undergoes carbon deprotonation with an E1cb mechanism. The comparison with previously studied reactions in acetohydroxamate/acetohydroxamic acid buffer confirms this assignment. The high proton activating factor, PAF, value observed (PAF = 1.2 x 10 (6) with isomer 1 in quinuclidine/quinuclidinium buffer) can be explained with the high stability by the resonance of the intermediate carbanion.     

Kinetics and mechanism of chromate reduction with hydrogen peroxide in base

[Cr(VI)O(4)](2)(-) is reduced to [Cr(V)(O(2))(4)](3)(-) by hydrogen peroxide in strongly basic media where the acid dissociation of H(2)O(2) (pK(a) = 11.65) is appreciable. The reaction is first order in chromium(VI) and inhibited by hydroxide. The hydrogen peroxide dependence is defined by the form of the effective pseudo-first-order rate constant: k(eff) = [H(2)O(2)](3)/(K(1) + K(2)[H(2)O(2)] + K(3)[HO(2)(-)]) with K(1) = 175(43) s x M(3), K(2) = 403(18) s x M(2), and K(3) = 1422(34) s x M(2). Hydrogen peroxide anion initially attacks chromate, and subsequent equilibrium steps that exchange oxo groups for three peroxo groups precede a rate-determining, one-electron, intramolecular reduction step.     

Spectral properties and reactivity of diarylmethanol radical cations in aqueous solution. Evidence for intramolecular charge resonance

Spectral properties and reactivities of ring-methoxylated diarylmethane and diarylmethanol radical cations, generated in aqueous solution by pulse and gamma-radiolysis and by the one-electron chemical oxidant potassium 12-tungstocobalt(III)ate, have been studied. The radical cations display three bands in the UV, visible, and vis-NIR regions of the spectrum. The vis-NIR band is assigned to an intramolecular charge resonance interaction (CR) between the neutral donor and charged acceptor rings, as indicated by the observation that the relative intensity of the vis-NIR band compared to that of the UV and visible bands does not increase with increasing substrate concentration and that the position and intensity of this band is influenced by the ring-substitution pattern. In acidic solution (pH = 4), monomethoxylated diarylmethanol radical cations 1a.(+ -)1e.(+) decay by C(alpha)-H deprotonation [k = (1.7-1.9) x 10(4)s(-1)] through the intermediacy of a ketyl radical, which is further oxidized in the reaction medium to give the corresponding benzophenones, as evidenced by both time-resolved spectroscopic and product studies. With the dimethoxylated radical cation 2.(+), C(alpha)-H deprotonation is instead significantly slower (k = 6.7 x 10(2)s(-1)). In basic solution, 1a.(+)-1e.(+) undergo (-)OH-induced deprotonation from the alpha-OH group with k(OH.)approximately equal to 1.4 x 10(10)M(-1)s(-1), leading to a ketyl radical anion, which is oxidized in the reaction medium to the corresponding benzophenone.     

A kinetic and product study of the Cl + HO2 reaction

Absolute rate data and product branching ratios for the reactions Cl + HO2 --> HCl + O2 (k1a) and Cl + HO2 --> OH + ClO (k1b) have been measured from 226 to 336 K at a total pressure of 1 Torr of helium using the discharge flow resonance fluorescence technique coupled with infrared diode laser spectroscopy. For kinetic measurements, pseudo-first-order conditions were used with both reagents in excess in separate experiments. HO2 was produced by two methods: through the termolecular reaction of H atoms with O2 and also by the reaction of F atoms with H2O2. Cl atoms were produced by a microwave discharge of Cl2 in He. HO2 radicals were converted to OH radicals prior to detection by resonance fluorescence at 308 nm. Cl atoms were detected directly at 138 nm also by resonance fluorescence. Measurement of the consumption of HO2 in excess Cl yielded k1a and measurement of the consumption of Cl in excess HO2 yielded the total rate coefficient, k1. Values of k1a and k1 derived from kinetic experiments expressed in Arrhenius form are (1.6 +/- 0.2) x 10(-11) exp[(249 +/- 34)/T] and (2.8 +/- 0.1) x 10(-11) exp[(123 +/- 15)/T] cm3 molecule(-1) s(-1), respectively. As the expression for k1 is only weakly temperature dependent, we report a temperature-independent value of k1 = (4.5 +/- 0.4) x 10(-11) cm3 molecule(-1) s(-1). Additionally, an Arrhenius expression for k1b can also be derived: k1b = (7.7 +/- 0.8) x 10(-11) exp[-(708 +/- 29)/T] cm3 molecule(-1) s(-1). These expressions for k1a and k1b are valid for 226 K < or = T < or = 336 and 256 K < or = T < or = 296 K, respectively. The cited errors are at the level of a single standard deviation. For the product measurements, an excess of Cl was added to known concentrations of HO2 and the reaction was allowed to reach completion. HCl product concentrations were determined by IR absorption yielding the ratio k1a/k1 over the temperature range 236 K < or = T < or = 296 K. OH product concentrations were determined by resonance fluorescence giving rise to the ratio k1b/k1 over the temperature range 226 K < or = T < or = 336 K. Both of these ratios were subsequently converted to absolute numbers. Values of k1a and k1b from the product experiments expressed in Arrhenius form are (1.5 +/- 0.1) x 10(-11) exp[(222 +/- 17)/T] and (10.6 +/- 1.5) x 10(-11) exp[-(733 +/- 41)/T] cm3 molecule(-1) s(-1), respectively. These expressions for k1a and k1b are valid for 256 K < or = T < or = 296 and 226 K < or = T < or = 336 K, respectively. A combination of the kinetic and product data results in the following Arrhenius expressions for k1a and k1b of (1.4 +/- 0.3) x 10(-11) exp[(269 +/- 58)/T] and (12.7 +/- 4.1) x 10(-11) exp[-(801 +/- 94)/T] cm3 molecule(-1) s(-1), respectively. Numerical simulations were used to check for interferences from secondary chemistry in both the kinetic and product experiments and also to quantify the losses incurred during the conversion process HO2 --> OH for detection purposes.     

Hydroxy double salt anion exchange kinetics: effects of precursor structure and anion size

(1)H NMR spectroscopy and powder X-ray diffraction have been used to explore the details of anion exchange reactions of two layered hydroxy double salts (HDSs), zinc copper hydroxy acetate (ZCA), nickel zinc hydroxy acetate (NZA), and a related layered material, zinc hydroxy acetate (ZHA), at room temperature (21-22 degrees C). Reactions that followed Avrami-Erofe'ev kinetics with respect to temporal profiles for acetate release, ZCA with butyrate (k = 1.7 x 10(-3) s(-1)), and octanoate (k = 0.79 x 10(-3) s(-1)) anions, as well as ZHA with octanoate (k = 2.6 x 10(-3) s(-1)), demonstrate that rate constants for acetate release are influenced by the exchange anion relative size as well as by the solid precursor structure/composition. The reaction of NZA with octanoate deviated from expected Avrami-Erofe'ev behavior, with evidence for an intermediate species in the solid phase that may influence the rate of acetate release into solution. The reaction of ZCA with formate anions exhibited a unique zeroth-order kinetics release of acetate, providing the possibility of developing tunable nanostructured anion release sources by use of variations in the size of the exchange species.     

Absorption and emission study of 2',7'-difluorofluorescein and its excited-state buffer-mediated proton exchange reactions

2',7'-Difluorofluorescein (Oregon Green 488) is a new fluorescein-based dye, which has found many applications, above all in biochemistry and neurosciences, and its use has become very popular in the last years. In recent years, we have been investigating the excited-state proton exchange reactions of fluorescein and the effect of suitable proton acceptors and donors which promote these reactions. The excited-state proton transfer reactions may appreciably influence the fluorescence results when using these dyes. We present steady-state emission evidence that acetate buffer species promote an excited-state proton transfer between neutral, monoanionic, and dianionic forms of 2',7'-difluorofluorescein. The time course of the excited species in this reaction was characterized through time-resolved fluorescence measurements, and the kinetics of the reaction was solved by using the global compartmental analysis. A previous identifiability study on the compartmental system set the conditions to design the fluorescence decay surface. This is the first experimental system, studied within this kinetic model, solved under identifiability conditions through global compartmental analysis. The recovered rate constant values for deactivation were 2.94 x 10(8) s(-1) for the monoanion and 2.47 x 10(8) s(-1) for the dianion, whereas the rate constant values of the buffer-mediated excited-state reaction were 9.70 x 10(8) and 1.79 x 10(8) M(-1) s(-1) for the deprotonation and protonation, respectively. With these values, a pK(a) = 4.02 was obtained. In this work, we additionally provide an absorption study, including acid-base equilibria, determination of ground-state pK(a) values (1.02, 3.61, and 4.69), and recovery of molar absorption coefficients of every prototropic species, including absorption and NMR evidence for the existence of three tautomers in neutral species. Steady-state emission spectra of 2',7'-difluorofluorescein in aqueous solution are also described, where the strong photoacid behavior of the cation is noteworthy.     

Kinetics and mechanism of bromate-bromide reaction catalyzed by acetate

The initial rate of the bromate-bromide reaction, BrO3- + 5Br- + 6H+ --> 3Br2 + 3H2O, has been measured at constant ionic strength, I = 3.0 mol L(-1), and at several initial concentrations of acetate, bromate, bromide, and perchloric acid. The reaction was followed at the Br2/Br3- isosbestic point (lambda = 446 nm) by the stopped-flow technique. A very complex behavior was found such that the results could be fitted only by a six term rate law, nu = k1[BrO3-][Br-][H+]2 + k2[BrO3-][Br-]2[H+]2 + k3[BrO3-][H+]2[acetate]2 + k4[BrO3-][Br-]2[H+]2[acetate] + k5[BrO3-][Br-][H+]3[acetate]2 + k6[BrO3-][Br-][H+]2[acetate], where k1 = 4.12 L3 mol(-3) s(-1), k2 = 0.810 L4 mol(-4) s(-1), k3 = 2.80 x 10(3) L4 mol(-4) s(-1), k4 = 278 L5 mol(-5) s(-1), k5 = 5.45 x 10(7) L6 mol(-6) s(-1), and k6 = 850 L4 mol(-4) s(-1). A mechanism, based on elementary steps, is proposed to explain each term of the rate law. This mechanism considers that when acetate binds to bromate it facilitates its second protonation.     

Thermodynamics, kinetics, and mechanism of the stepwise dissociation and formation of Tris(L-lysinehydroxamato)iron(III) in aqueous acid

pK(a) values for the hydroxamic acid, alpha-NH(3)(+), and epsilon-NH(3)(+) groups of L-lysinehydroxamic acid (LyHA, H(3)L(2+)) were found to be 6.87, 8.89, and 10.76, respectively, in aqueous solution (I = 0.1 M, NaClO(4)) at 25 degrees C. O,O coordination to Fe(III) by LyHA is supported by H(+) stoichiometry, UV-vis spectral shifts, and a shift in nu(CO) from 1648 to 1592 cm(-1) upon formation of mono(L-lysinehydroxamato)tetra(aquo)iron(III) (Fe(H(2)L)(H(2)O)(4)(4+)). The stepwise formation of tris(L-lysinehydroxamato)iron(III) from Fe(H(2)O)(6)(3+) and H(3)L(2+) was characterized by spectrophotometric titration, and the values for log beta(1), log beta(2), and log beta(3) are 6.80(9), 12.4(2), and 16.1(2), respectively, at 25 degrees C and I = 2.0 M (NaClO(4)). Stopped-flow spectrophotometry was used to study the proton-driven stepwise ligand dissociation kinetics of tris(L-lysinehydroxamato)iron(III) at 25 degrees C and I = 2.0 M (HClO(4)/NaClO(4)). Defining k(n) and k(-n) as the stepwise ligand dissociation and association rate constants and n as the number of bound LyHA ligands, k(3), k(-3), k(2), k(-2), k(1), and k(-1) are 3.0 x 10(4), 2.4 x 10(1), 3.9 x 10(2), 1.9 x 10(1), 1.4 x 10(-1), and 1.2 x 10(-1) M(-1) s(-1), respectively. These rate and equilibrium constants are compared with corresponding constants for Fe(III) complexes of acetohydroxamic acid (AHA) and N-methylacetohydroxamic acid (NMAHA) in the form of a linear free energy relationship. The role of electrostatics in these complexation reactions to form the highly charged Fe(LyHA)(3)(6+) species is discussed, and an interchange mechanism mediated by charge repulsion is presented. The reduction potential for tris(L-lysinehydroxamato)iron(III) is -214 mV (vs. NHE), and a comparison to other hydroxamic acid complexes of Fe(III) is made through a correlation between E(1/2) and pFe.     

Direct evidence for base-mediated decomposition of alkyl hydroperoxides (ROOH) in the gas phase

The reaction of F(-) with CH(3)OOH has been studied in the gas phase using a tandem flowing afterglow-selected ion flow tube apparatus. The reaction is rapid (k = 1.23 x 10(-9) cm(3) s(-1), 49% efficiency), and formation of HO(-) + CH(2)O + HF is the major reaction channel observed (85%). Isotopic labeling, reactions of F(-) with larger alkyl hydroperoxides, and computational studies demonstrate that the major product ion, HO(-), is formed via a concerted elimination mechanism that appears to be general to all alkyl hydroperoxides possessing an alpha-hydrogen. This mechanism represents a base-mediated decomposition of alkyl hydroperoxides in the gas phase that may have important implications for solution and biochemical reactions. The reverse reaction, CH(3)OO(-) + HF is also efficient (k = 2.43 x 10(-9) cm(3) s(-1)). The major product ensemble HO(-) + CH(2)O + HF (81%) is identical to that of the forward reaction, and represents a novel neutral-catalyzed decomposition of the anion.     

Investigation of the mechanism of formation of a thiolate-ligated Fe(III)-OOH

Kinetic studies aimed at determining the most probable mechanism for the proton-dependent [Fe(II)(S(Me2)N(4)(tren))](+) (1) promoted reduction of superoxide via a thiolate-ligated hydroperoxo intermediate [Fe(III)(S(Me2)N(4)(tren))(OOH)](+) (2) are described. Rate laws are derived for three proposed mechanisms, and it is shown that they should conceivably be distinguishable by kinetics. For weak proton donors with pK(a(HA)) > pK(a(HO(2))) rates are shown to correlate with proton donor pK(a), and display first-order dependence on iron, and half-order dependence on superoxide and proton donor HA. Proton donors acidic enough to convert O(2)(-) to HO(2) (in tetrahydrofuran, THF), that is, those with pK(a(HA)) < pK(a(HO(2))), are shown to display first-order dependence on both superoxide and iron, and rates which are independent of proton donor concentration. Relative pK(a) values were determined in THF by measuring equilibrium ion pair acidity constants using established methods. Rates of hydroperoxo 2 formation displays no apparent deuterium isotope effect, and bases, such as methoxide, are shown to inhibit the formation of 2. Rate constants for p-substituted phenols are shown to correlate linearly with the Hammett substituent constants σ(-). Activation parameters ((ΔH(++) = 2.8 kcal/mol, ΔS(++) = -31 eu) are shown to be consistent with a low-barrier associative mechanism that does not involve extensive bond cleavage. Together, these data are shown to be most consistent with a mechanism involving the addition of HO(2) to 1 with concomitant oxidation of the metal ion, and reduction of superoxide (an "oxidative addition" of sorts), in the rate-determining step. Activation parameters for MeOH- (ΔH(++) = 13.2 kcal/mol and ΔS(++) = -24.3 eu), and acetic acid- (ΔH(++) = 8.3 kcal/mol and ΔS(++) = -34 eu) promoted release of H(2)O(2) to afford solvent-bound [Fe(III)(S(Me2)N(4)(tren))(OMe)](+) (3) and [Fe(III)(S(Me2)N(4)(tren))(O(H)Me)](+) (4), respectively, are shown to be more consistent with a reaction involving rate-limiting protonation of an Fe(III)-OOH, than with one involving rate-limiting O-O bond cleavage. The observed deuterium isotope effect (k(H)/k(D) = 3.1) is also consistent with this mechanism.