Negative Electron Transfer Collision-Induced Dissociation of G-Quadruplexes: Uncovering the Guanine Radical Anion Loss Pathway.

G-quadruplex (G4) DNA can form highly stable secondary structures in the presence of metal cations, and research has shown its potential as a transcriptional regulator for oncogenes in the human genome. In order to explore the interactions of DNA with metal cations using mass spectrometry, employing complementary fragmentation methods can enhance structural information. This study explores the use of ion-ion reactions for sequential negative electron transfer collision-induced dissociation (nET-CID) as a complement to traditional ion-trap CID (IT-CID). The resulting nET-CID data for G4 anions with and without metal cations show an increase in fragment ion type diversity and yield of structurally informative ions relative to IT-CID. The nET-CID yields greater sequence coverage by virtue of fragmentation at the 3'-side of thymine residues, which is lacking with IT-CID. Potassium adductions to backbone fragments in IT-CID and nET-CID spectra were nearly identical. Of note is a prominent fragment resulting from a loss of a 149 Da anion seen in nET-CID of large, G-rich sequences, proposed to be radical anion guanine loss. Neutral loss of neutral guanine (151 Da) and deprotonated nucleobase loss (150 Da) have been previously reported, but this is the first report of radical anion guanine loss (149 Da). Confirmation of the identity of the 149 Da anion results from the examination of the homonucleobase sequence 5'-GGGGGGGG-3'. Loss of a charged adenine radical anion at much lower relative abundance was also noted for the sequence 5'-AAAAAAAA-3'. DFT modeling indicates that the loss of a nucleobase as a radical anion from odd-electron nucleic acid anions is a thermodynamically favorable fragmentation pathway for G.

Nitrate and Nitrite Reductions at Copper(II) Sites: Role of Noncovalent Interactions from Second-Coordination-Sphere.

Reductions of nitrate and nitrite (NOx-) are of prime importance in combatting water pollution arising from the excessive use of N-rich fertilizers. While examples of NOx- reductions are known, this report illustrates hydrazine (N2H4)-mediated transformations of NOx- to nitric oxide (NO)/nitrous oxide (N2O). For nitrate reduction to NO, initial coordination of the weakly coordinating NO3- anion at [(mC)CuII]2+ cryptate has been demonstrated to play a crucial role. A set of complementary analyses (X-ray diffraction and Fourier-transform infrared spectroscopy (FTIR), UV-vis, and NMR spectroscopies) on NO3--bound metal-cryptates [(mC)MII(NO3)](ClO4) (1-M, M = Cu/Zn) demonstrates the binding of NO3- through noncovalent (NH···O, CH···O, and anion···π) and metal-ligand coordinate interactions. Subsequently, reactions of [(mC)CuII(14/15NO3)](ClO4) (1-Cu or 1-Cu/15N) with N2H4·H2O have been illustrated to reduce 14/15NO3- to 14/15NO. Intriguingly, in the absence of the second-coordination-sphere interactions, a closely related coordination motif [(Bz3Tren)CuII]2+ (in 3-Cu) does not bind NO3- and is unable to assist in N2H4·H2O-mediated NO3- reduction. In contrast, nitrite coordinates at the tripodal CuII sites in both [(mC)CuII]2+ and [(Bz3Tren)CuII]2+ irrespective of the additional noncovalent interactions, and hence, the N2H4 reactions of the copper(II)-nitrite complexes [(mC)CuII(O14/15NO)]+ and [(Bz3Tren)CuII(O14/15NO)]+ (in 2-Cu/4-Cu) result in a mixture of 14/15NO and N14/15NO.

High-fidelity Recognition of Organotrifluoroborate Anions (R-BF3 - ) as Designer Guest Molecules.

The recognition of boron compounds is well developed as boronic acids but untapped as organotrifluoroborate anions (R-BF3 - ). We are exploring the development of these and other designer anions as anion-recognition motifs by considering them as substituted versions of the parent inorganic ion. To this end, we demonstrate strong and reliable binding of organic trifluoroborates, R-BF3 - , by cyanostar macrocycles that are size-complementary to the inorganic BF4 - progenitors. We find that recognition is modulated by the substituent's sterics and that the affinities are retained using the common K+ salts of R-BF3 - anions.

Hydrazine Radiolysis by Gamma-Ray in the N2H4-Cu+-HNO3 System.

Radiolysis of chemical agents occurs during the decontamination of nuclear power plants. The γ-ray irradiation tests of the N2H4-Cu+-HNO3 solution, a decontamination agent, were performed to investigate the effect of Cu+ ion and HNO3 on N2H4 decomposition using a Co-60 high-dose irradiator. After the irradiation, the residues of N2H4 decomposition were analyzed by Ultraviolet-visible (UV) spectroscopy. NH4+ ions generated from N2H4 radiolysis were analyzed by ion chromatography. Based on the results, the decomposition mechanism of N2H4 in the N2H4-Cu+-HNO3 solution under γ-ray irradiation condition was derived. Cu+ ions form Cu+N2H4 complexes with N2H4, and then N2H4 is decomposed into intermediates. H+ ions and H● radicals generated from the reaction between H+ ion and eaq- increased the N2H4 decomposition reaction. NO3- ions promoted the N2H4 decomposition by providing additional reaction paths: (1) the reaction between NO3- ions and N2H4●+, and (2) the reaction between NO● radical, which is the radiolysis product of NO3- ion, and N2H5+. Finally, the radiolytic decomposition mechanism of N2H4 obtained in the N2H4-Cu+-HNO3 was schematically suggested.

Analyses of Inositol Phosphates and Phosphoinositides by Strong Anion Exchange (SAX)-HPLC.

The phosphate esters of myo-inositol (Ins) occur ubiquitously in biology. These molecules exist as soluble or membrane-resident derivatives and regulate a plethora of cellular functions including phosphate homeostasis, DNA repair, vesicle trafficking, metabolism, cell polarity, tip-directed growth, and membrane morphogenesis. Phosphorylation of all inositol hydroxyl groups generates phytic acid (InsP6), the most abundant inositol phosphate present in eukaryotic cells. However, phytic acid is not the most highly phosphorylated naturally occurring inositol phosphate. Specialized small molecule kinases catalyze the formation of the so-called myo-inositol pyrophosphates (PP-InsPs), such as InsP7 and InsP8. These molecules are characterized by one or several "high-energy" diphosphate moieties and are ubiquitous in eukaryotic cells. In plants, PP-InsPs play critical roles in immune responses and nutrient sensing. The detection of inositol derivatives in plants is challenging. This is particularly the case for inositol pyrophosphates because diphospho bonds are labile in plant cell extracts due to high amounts of acid phosphatase activity. We present two steady-state inositol labeling-based techniques coupled with strong anion exchange (SAX)-HPLC analyses that allow robust detection and quantification of soluble and membrane-resident inositol polyphosphates in plant extracts. These techniques will be instrumental to uncover the cellular and physiological processes controlled by these intriguing regulatory molecules in plants.

A GdW10@PDA-CAT Sensitizer with High-Z Effect and Self-Supplied Oxygen for Hypoxic-Tumor Radiotherapy.

Anticancer treatment is largely affected by the hypoxic tumor microenvironment (TME), which causes the resistance of the tumor to radiotherapy. Combining radiosensitizer compounds and O2 self-enriched moieties is an emerging strategy in hypoxic-tumor treatments. Herein, we engineered GdW10@PDA-CAT (K3Na4H2GdW10O36·2H2O, GdW10, polydopamine, PDA, catalase, CAT) composites as a radiosensitizer for the TME-manipulated enhancement of radiotherapy. In the composites, Gd (Z = 64) and W (Z = 74), as the high Z elements, make X-ray gather in tumor cells, thereby enhancing DNA damage induced by radiation. CAT can convert H2O2 to O2 and H2O to enhance the X-ray effect under hypoxic TME. CAT and PDA modification enhances the biocompatibility of the composites. Our results showed that GdW10@PDA-CAT composites increased the efficiency of radiotherapy in HT29 cells in culture. This polyoxometalates and O2 self-supplement composites provide a promising radiosensitizer for the radiotherapy field.

Role of salt ions and molecular weights on the formation of Mesona chinensis polysaccharide-chitosan polyelectrolyte complex hydrogel.

The effects of the addition of salt ions and molecular weights (Mw) of CH on Mesona chinensis polysaccharide (MCP)-chitosan (CH) hydrogel were investigated. Result indicated both low concentration of monovalent salt ions (Na+ and K+), divalent cations (Ca2+) and oxoanions (SO42-) could promote the gel properties of MCP-CH hydrogel. The Mw of CH has huge impact on the formation and properties of hydrogel. Combining the relationship between rheology and structural, monovalent salt ions such as Na+ and K+ affect gel formation and its properties by influencing electrostatic interaction and chain conformation. Both divalent cations (Ca2+) and oxoanions (SO42-) facilitated the formation of gel networks via electrostatic interaction, coordination bonds and hydrogen bonds. Moreover, Mw of CH influenced formation and texture of MCP-CH hydrogel via affecting the conformation of CH molecular chain. These findings will provide a few theoretical bases to understand the formation mechanism of MCP-CH hydrogel.

Effect of anion type on enzymatic hydrolysis of starch-(thermostable α-amylase)-calcium system in a low-moisture solid microenvironment of bioextrusion.

The effect of six anions (Cl-, OH-, NO3-, SO42-, C6H10O62- and PO43-) on a starch (St)-enzyme (thermostable α-amylase, TαA)-calcium (Ca) system was investigated in a low-moisture solid state. Two levels of Ca salts (1 and 10 mmol/100 g St) added to potato starch with and without TαA were analyzed by FT-IR, DSC and SEM. The surface morphologies of the St-Ca complexes were different in the presence of various anions, and the residual Ca salts around the St granules might decrease the enzymatic action. For bioextrusion, TαA (0.5‰ and 1.5‰) were introduced for a relatively low Ca content (1 mmol/100 g). Significant differences in enzyme activity were observed, increasing the activity of TαA by SO42- (146.54 %) > C6H10O62- > Cl- > control > NO3- > OH- ≈ PO43- and C6H10O62- (123.20 %) ≈ Cl- ≈ SO42- > control > PO43 > OH- > NO3- for the low and high enzyme levels, respectively.

Pyridinium-substituted tetraphenylethylene salt-based photosensitizers by varying counter anions: a highly efficient photodynamic therapy for cancer cell ablation and bacterial inactivation.

Cancer and bacterial infection seriously threaten the health of human beings. The development of an image-guided photosensitizer with a "Two-in-One" function that can be simultaneously used for both efficient cancer cell ablation and rapid bacterial inactivation is highly in demand. In this project, we designed and prepared two aggregation-induced emission luminogens (AIEgens) (called TPEPy-I and TPEPy-PF6) with a strong electron push-pull effect. They have a near-infrared (NIR) emission, a high 1O2 quantum yield up to 0.93 and a fluorescence turn-on effect in mitochondria. Upon white light irradiation, the two mitochondria-targeting AIEgens exhibit a highly efficient photodynamic ablation of HeLa cells as well as excellent photodynamic inactivation of both Gram-positive S. aureus and Gram-negative E. coli. The time-dependent density functional theory (TD-DFT) results indicate that compared to TPEPy-PF6, TPEPy-I can easily produce the triplet state that is a prerequisite for 1O2 formation. Moreover, the positive effect of iodide anions gives TPEPy-I a higher photodynamic efficacy in cancer cell ablation and bacterial inactivation as compared with TPEPy-PF6.

Anion Recognition by Neutral Chalcogen Bonding Receptors: Experimental and Theoretical Investigations.

The utilization of neutral receptors for the molecular recognition of anions based on chalcogen bonding (ChB) is an undeveloped area of host-guest chemistry. In this manuscript, the synthesis of two new families of sulfur, selenium, and tellurium-based ChB binding motifs are reported. The stability of the thiophene, selenophene, and tellurophene binding motifs has enabled the determination of the association constants for ChB halide anion binding in the polar aprotic solvent THF by 1 H, 77 Se, and 125 Te NMR experiments. Two different aromatic cores are used and one or two Ch-binding motifs are incorporated with the purpose of encapsulating the anion, offering up to two concurrent chalcogen bonds. Theoretical calculations and NMR experiments reveal that, for S and Se receptors, hydrogen-bonding interactions involving the acidic H atom adjacent to the chalcogen atom are energetically favored over the ChB interaction. However, for the tellurophene binding motif, the σ-hole interaction is competitive and more favored than the hydrogen bond.

Co-immunomodulatory Activities of Anionic Macromolecules Extracted from Codium fragile with Red Ginseng Extract on Peritoneal Macrophage of Immune-Suppressed Mice.

In this study we investigated the immune effects of oral administration of anionic macromolecules extracted from Codium fragile (CFAM) and red ginseng extract mixture on the peritoneal macrophage cells in immune-suppressed mice. Cyclophosphamide (CY) induces the immune-suppressed condition. CY-treated mice were orally fed with different concentrations of CFAM supplemented with red ginseng extract and the peritoneal macrophages collected. CY treatment significantly decreased the immune activities of peritoneal macrophages, compared to the normal mice. The administration of CFAM mixed with red ginseng extract significantly boosted the viability of macrophage cells and nitric oxide production of peritoneal macrophages. Further, the oral administration of CFAM mixed with red ginseng extract up-regulated the expression of iNOS, COX-2, and TLR-4 as well as cytokines such as IL-1ß, IL-6, TNF-α, and IFN-γ more than the red ginseng-treated group. This study showed that CFAM enhanced the immune activity of red ginseng extract in the peritoneal macrophage cells of immune-suppressed mice. Furthermore, CFAM might be used as a co-stimulant of red ginseng extract through the regulation of macrophage cells for the enhancement of human health and immunity.

Adenine nucleobase directed supramolecular architectures based on ferrimagnetic heptanuclear copper(II) entities and benzenecarboxylate anions.

Two planar organic anions, benzoate and benzene-1,4-dicarboxylate (terephthalate), have been selected as potential π-stacking intercalators among ferrimagnetic [Cu7(µ-adeninato)6(µ3-OH)6(µ-H2O)6]2+ heptameric discrete entities. The resulting supramolecular architecture is highly dependent on the negative charge density distribution, mainly located in the carboxylate groups of the organic anions. In this sense, the benzoate anion, with just one carboxylate group, does not allow its intercalation between the adeninato ligands as it would imply a high steric hindrance among the heptameric entities. As a consequence, these benzoate anions are located inside the voids of the crystal structure reducing the accessible volume of compound [Cu7(µ-adeninato)6(µ3-OH)6(µ-H2O)6](benzoate)2·~17H2O (1). On the contrary, the terephthalate anion, containing two carboxylate groups at opposite sites, adopts a π-stacking sandwich arrangement between two adeninato ligands that affords the porous open structure of formula [Cu7(µ-adeninato)6(µ3-OH)6(µ-H2O)6](terephthalate)·nH2O (2a, 2b; n: 12 and 24, respectively). In addition to that, the less directional nature of the π-stacking interactions in comparison to the complementary hydrogen bonding based supramolecular metal-organic frameworks (SMOFs), suits them with a flexible architecture able to reversibly adsorb/desorb water (up to a 25-30% at 20 °C) altogether with the expansion/shrinkage of the crystal structure. The bridging adeninato and hydroxido ligands are effective magnetic exchange mediators to provide a ST = 5/2 ferrimagnetic state for the heptanuclear entity.

Macrocycle-Directed Construction of Tetrahedral Anion-π Receptors for Nesting Anions with Complementary Geometry.

Manipulation of the emerging anion-π interactions in a highly cooperative manner through sophisticated host design represents a very challenging task. In this work, unprecedented tetrahedral anion-π receptors have been successfully constructed for complementary accommodation of tetrahedral and relevant anions. The synthesis was achieved by a macrocycle-directed approach by using large macrocycle precursors bearing four reactive sites, which enabled a kinetic-favored pathway and afforded the otherwise inaccessible tetrahedral cages in considerable yields. Crystal structure suggested that the tetrahedral cages have an enclosed three-dimensional cavity surrounded by four electron-deficient triazine faces in a tetrahedral array. The complementary accommodation of a series of tetrahedral and relevant anions including BF4 - , ClO4 - , H2 PO4 - , HSO4 - , SO4 2- and PF6 - was revealed by ESI-MS and DFT calculations. Crystal structures of ClO4 - and PF6 - complexes showed that the anion was nicely encapsulated within the tetrahedral cavity with up to quadruple cooperative anion-π interactions by an excellent shape and size match. The strong anion-π binding was further confirmed by negative ion photoelectron spectroscopy measurements.

Phytotoxicity of ionic liquids.

Ionic liquids (ILs) have been one of the most interesting chemical entities over the last two decades and have been investigated by numerous scientists all over the world. However, during IL research, it has been shown that these compounds present toxicity to both terrestrial and aquatic plants, among others. The phytotoxicity of ILs depends on the type of cation, the length of the alkyl chain in the substituent or enantioselectivity, on the concentration used, and, it appears that the type of anion may also have an impact on toxicity. The toxic effects of ILs on plants also depend on the conditions under which such tests are conducted. The results may help facilitate the development of protective environmental measures against IL-induced negative effects, but they may also be used in various landscape-related areas, such as herbology, to design new substances with weed killing properties.

Preferential Binding of Cytochrome c to Anionic Ligand-Coated Gold Nanoparticles: A Complementary Computational and Experimental Approach.

Membrane-bound proteins can play a role in the binding of anionic gold nanoparticles (AuNPs) to model bilayers; however, the mechanism for this binding remains unresolved. In this work, we determine the relative orientation of the peripheral membrane protein cytochrome c in binding to a mercaptopropionic acid-functionalized AuNP (MPA-AuNP). As this is nonrigid binding, traditional methods involving crystallographic or rigid molecular docking techniques are ineffective at resolving the question. Instead, we have implemented a computational assay technique using a cross-correlation of a small ensemble of 200 ns long molecular dynamics trajectories to identify a preferred nonrigid binding orientation or pose of cytochrome c on MPA-AuNPs. We have also employed a mass spectrometry-based footprinting method that enables the characterization of the stable protein corona that forms at long time-scales in solution but remains in a dynamic state. Through the combination of these computational and experimental primary results, we have established a consensus result establishing the identity of the exposed regions of cytochrome c in proximity to MPA-AuNPs and its complementary pose(s) with amino-acid specificity. Moreover, the tandem use of the two methods can be applied broadly to determine the accessibility of membrane-binding sites for peripheral membrane proteins upon adsorption to AuNPs or to determine the exposed amino-acid residues of the hard corona that drive the acquisition of dynamic soft coronas. We anticipate that the combined use of simulation and experimental methods to characterize biomolecule-nanoparticle interactions, as demonstrated here, will become increasingly necessary as the complexity of such target systems grows.

C60 Mediated Ion Pair Interaction for Label-Free Electrochemical Immunosensing with Nanoporous Anodic Alumina Nanochannels.

Label-free biosensing based on the nanoporous anodic alumina (NAA) membrane emerged as a versatile biosensing platform in the recent decade. In the present work, we developed a new immunosensing strategy based on the nanochannels of NAA and the ion pair interaction mediated by electrochemistry of C60. The NAA served as the matrix for the immobilization of the capture antibodies. The incubation of target antigens resulted in the formation of the immunocomplexes and thus an increase of the steric hindrance of the nanochannels. Therefore, the concentration of the redox probe transported through the nanochannels decreases, which can be detected at the working electrode modified with C60. Herein, we initially found that the cathodic peak ascribed to the reduction of C60 to C60- was obviously enhanced by the presence of the redox probe K3[Fe(CN)6] and which was contributed to the formation of a ternary ion association complex among C60, tetraoctylammonium bromide, and K3[Fe(CN)6]. Therefore, the transportation of K3[Fe(CN)6] though the NAA-based bionanochannels can be detected by a C60 modified electrode with an amplified signal. Choosing human epididymis protein 4 (HE4) as the model target, a linear range of 1.0 ng mL-1 to 100 ng mL-1 can be established between the peak current obtained from the differential pulse voltammetric response of the platform and the concentration of HE4. The detection limit was 0.2 ng mL-1. This study not only provides a new avenue to develop the other nanochannel-based biosensing platform for a variety of other disease biomarkers but also contributes to the electrochemistry of fullerene.

Exploring the Effect of Choline-Based Ionic Liquids on the Stability and Activity of Stem Bromelain.

Enzymes are very important components which are vital for the existence of every cellular life. There is significant interest in the use of structurally stable and catalytically active enzymes in pharmaceutical, food, fine chemicals industries, and in various industrial processes as catalysts. Stem bromelain (BM) is a proteolytic enzyme which is widely used in chemical, medical, and pharmaceutical fields. However, harsh process conditions are the main barriers to the effective use of this enzyme in different applications. To overcome these drawbacks, biocompatible bio-based ionic liquids (ILs), composed of the choline cation (an essential nutrient) and different anions are used. The ILs namely choline chloride [Ch]+[Cl]-, choline acetate [Ch]+[Ac]-, choline dihydrogen phosphate [Ch]+[Dhp]-, choline bitartrate [Ch]+[Bit]-, choline iodide [Ch]+[I]-, and choline hydroxide [Ch]+[OH]- are chosen for the current work. Therefore, in the present study, structural stability and activity of BM have been evaluated in the presence of choline-based ILs using various biophysical techniques at different concentrations. The present work demonstrated that [Ch]+[OH]- is the strongest destabilizer, whereas [Ch]+[Cl]- is the best stabilizer for the native structure of BM among all studied ILs. This work revealed the suitability of some choline-based ILs as potential media for sustained stability and activity of BM.

Thermodynamics of Anion Binding by Chalcogen Bonding Receptors.

The application of chalcogen bonding (ChB) to anion recognition is an underdeveloped area of host-guest supramolecular chemistry. The chemical instability of heavier chalcogen derivatives may in part be responsible for the lack of progress. Herein, the synthesis of a new structurally simple, tellurium-based ChB binding motif is reported, the robust stability of which has enabled the thermodynamic properties for ChB halide anion binding in polar aprotic and wet protic organic solvent media to be elucidated. The thermodynamic data reveals how the subtle interplay between ChB host, anion guest and solvent dictates halide binding selectivity and affinity trends. These findings help to provide a deeper insight into the nature of the ChB-anion interaction.

Interaction between a Nonsteroidal Anti-inflammatory Drug (Ibuprofen) and an Anionic Surfactant (AOT) and Effects of Salt (NaI) and Hydrotrope (4-4-4).

Ibuprofen (IBF), 2-(4-isobutylphenyl) propionic acid, is a surface-active, common nonsteroidal anti-inflammatory drug (NSAID), and it possesses a high critical micelle concentration (cmc) compared to that of conventional surfactants. The interactions of this common NSAID with an anionic surfactant, sodium octyl sulfosuccinate, were studied by tensiometric, fluorimetric, and calorimetric measurements to investigate this system as a possible model drug-delivery system for an NSAID like IBF, particularly in a high-dose regime for IBF. The interactions between the drug and the surfactant were modeled using a regular solution theory approach in the presence and absence of a model electrolyte (sodium iodide) and a novel nonaromatic, gemini hydrotrope, tetramethylene-1,4-bis( N, N-dimethyl- N-butylammonium)bromide (4-4-4). Both the simple and the hydrotropic electrolyte were shown to have an effect on the solution properties (aggregation parameters, interfacial properties, and thermodynamics of aggregate formation) of the drug-surfactant mixtures and on the interaction between the drug and the surfactant. Surface charges of all self-assembled systems were estimated from ζ-potential measurements, whereas density functional theory calculations showed the interaction energy comparison among all of the binary and ternary combinations. All of these results were interpreted in terms of how altering the subtle balance of hydrophobic and electrostatic forces can significantly improve the ability of these self-assembled systems to transport drug molecules.

Simultaneous determination of rutin and ascorbic acid in a sequential injection lab-at-valve system.

A green, simple, accurate and highly sensitive sequential injection lab-at-valve procedure has been developed for the simultaneous determination of ascorbic acid (Asc) and rutin using 18-molybdo-2-phosphate Wells-Dawson heteropoly anion (18-MPA). The method is based on the dependence of the reaction rate between 18-MPA and reducing agents on the solution pH. Only Asc is capable of interacting with 18-MPA at pH 4.7, while at pH 7.4 the reaction with both Asc and rutin proceeds simultaneously. In order to improve the precision and sensitivity of the analysis, to minimize reagent consumption and to remove the Schlieren effect, the manifold for the sequential injection analysis was supplemented with external reaction chamber, and the reaction mixture was segmented. By the reduction of 18-MPA with reducing agents one- and two-electron heteropoly blues are formed. The fraction of one-electron heteropoly blue increases at low concentrations of the reducer. Measurement of the absorbance at a wavelength corresponding to the isobestic point allows strictly linear calibration graphs to be obtained. The calibration curves were linear in the concentration ranges of 0.3-24mgL-1 and 0.2-14mgL-1 with detection limits of 0.13mgL-1 and 0.09mgL-1 for rutin and Asc, respectively. The determination of rutin was possible in the presence of up to a 20-fold molar excess of Asc. The method was applied to the determination of Asc and rutin in ascorutin tablets with acceptable accuracy and precision (1-2%).