ABSTRACT
In this study, we developed a multi-site acylation strategy to improve the lipophilicity and cellular uptake of troxerutin, a natural flavonoid with many health-promoting bioactivities. By clarifying the acylation properties of troxerutin catalyzed by lipases from different sources, a series of troxerutin ester derivatives acylated at different sites was synthesized, including troxerutin dipropyl (TDP), tripropyl (TTP), tetrapropyl (TEP), dibutyl (TDB), monohexyl (TMH), monooctyl (TMO) and monodecyl (TMD) esters. Interestingly, the troxerutin esters acylated at multiple sites with shorter fatty chains (TDP, TTP and TEP) had similar lipophilicity to the mono-acylated esters bearing longer fatty chains (TMH, TMO and TMD, respectively) and meanwhile demonstrated surprisingly lower cytotoxicity than that of the long fatty-chain mono-esters. In particular, the multi-acylated esters with shorter fatty chains showed remarkably higher cellular uptake than the mono-esters with long fatty chains. In vitro gastrointestinal digestion suggested that the multi-acylated esters of troxerutin were more resistant to gastrointestinal degradation than the mono-esters. These results indicated that multi-site acylation with short fatty chains could be an effective alternative to introducing one-site mono-acylation for the modification of troxerutin and other flavonoid compounds.
Subject(s)
Hydroxyethylrutoside , Lipase , Acylation , Humans , Hydroxyethylrutoside/analogs & derivatives , Hydroxyethylrutoside/pharmacology , Hydroxyethylrutoside/chemistry , Hydroxyethylrutoside/metabolism , Lipase/metabolism , Lipase/chemistry , AnimalsABSTRACT
Troxerutin (TRX) is a mixture of semisynthetic hydroxyethylrutosides (Hers) arising from hydroxyethylation of rutin, a natural occurring flavonoid. TRX is commonly used for its anti-oxidant and anti-inflammatory properties in chronic venous insufficiency and other vascular disorders. In recent studies, the protective effects of TRX in Alzheimer's disease, colon carcinogenesis and hepatocellular carcinoma are emerged. However, the chemical stability of TRX has never been studied. Hence, the aims of the work were to study the TRX chemical stability through a forced degradation study and to develop and validate a new stability indicating LC-UV method for determination of TRX. In order to perform the study, TRX stability was tested in various stress conditions analysing the degradation samples by LC-MS. Three degradation products (DPs; D1, D2 and D3, 3',4',7-Tri-O-(ß-hydroxyethyl)quercetin, 3',4',5,7-Tetra-O-(ß-hydroxyethyl)quercetin and 3',4'-Di-O-(ß-hydroxyethyl)quercetin respectively) arising from degradation in acidic conditions were identified and synthesized: among them, D1 resulted the stability indicator for hydrolytic degradation. Furthermore, a stability-indicating LC-UV method for simultaneous determination of triHer (3',4',7-Tri-O-(ß-hydroxyethyl)rutin, the principal component of the mixture) and D1 was developed and validated. The LC-UV method consisted in a gradient elution on a Phenomenex Kinetex EVO C18 (150 × 3â¯mm, 5⯵m) with acetonitrile and ammonium bicarbonate buffer (10â¯mM, pH 9.2). The method was linear for triHer (20-60⯵gâ¯mL-1) and D1 (5.1-35⯵gâ¯mL-1). The intraday and interday precision were determined and expressed as RSDs: all the values wereâ¯≤â¯2% for both triHer and D1. The method demonstrated also to be accurate and robust and the average recoveries were 98.8 and 97.9% for triHer and D1, respectively. Moreover, the method resulted selective and specific for all of the components present in the degradation pattern of TRX (diHer (3',4'-Di-O-(ß-hydroxyethyl)rutin), triHer, tetraHer (3',4',5,7-Tetra-O-(ß-hydroxyethyl)rutin), D3, D1 and D2) and it was successfully applied for the stability studies of both drug substances and drug products.
Subject(s)
Chromatography, High Pressure Liquid , Hydroxyethylrutoside/analogs & derivatives , Spectrophotometry, Ultraviolet , Technology, Pharmaceutical/methods , Calibration , Chromatography, High Pressure Liquid/standards , Drug Contamination , Drug Stability , Hydroxyethylrutoside/chemical synthesis , Hydroxyethylrutoside/chemistry , Limit of Detection , Linear Models , Molecular Structure , Reference Standards , Reproducibility of Results , Spectrometry, Mass, Electrospray Ionization , Spectrophotometry, Ultraviolet/standards , Technology, Pharmaceutical/standardsABSTRACT
Troxerutin is a bioflavonoid, which can be used to treat venous disorders, thrombosis and cerebrovascular diseases. Recent studies have demonstrated that it may also be used to prevent edemas. However, it is not known whether troxerutin protects against the cardiomyopathic complications of diabetes. In the present study, a rat model of type 2 diabetes was used to investigate the potential for troxerutin to protect against diabetic cardiomyopathy, through changes to nuclear factorκB (NFκB) expression. Troxerutin administration significantly reduced heart rate, blood pressure, blood glucose and plasma triglyceride levels across all measured time points. Furthermore, troxerutin significantly reduced reactive oxygen species levels, NFκB protein expression, and suppressed the phosphorylated forms of AKT, insulin receptor substrate 1 (IRS1) and cJun Nterminal kinase (JNK). These results suggested that troxerutin protects against cardiomyopathy via alterations in NFκB, AKT and IRS1 signaling, in a rat model of type 2 diabetes.
Subject(s)
Diabetes Mellitus, Type 2/complications , Diabetic Cardiomyopathies/metabolism , Hydroxyethylrutoside/analogs & derivatives , Insulin Receptor Substrate Proteins/metabolism , NF-kappa B/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Signal Transduction/drug effects , Animals , Biomarkers , Blood Glucose/drug effects , Body Weight/drug effects , Diabetes Mellitus, Experimental , Diabetic Cardiomyopathies/drug therapy , Diabetic Cardiomyopathies/pathology , Disease Models, Animal , Hydroxyethylrutoside/chemistry , Hydroxyethylrutoside/pharmacology , JNK Mitogen-Activated Protein Kinases/metabolism , Male , Rats , Reactive Oxygen Species/metabolismSubject(s)
Albumins/chemistry , Glycation End Products, Advanced/chemistry , Hydroxyethylrutoside/analogs & derivatives , Protein Aggregation, Pathological/drug therapy , Glycation End Products, Advanced/antagonists & inhibitors , Glycosylation/drug effects , Humans , Hydroxyethylrutoside/chemistry , Hydroxyethylrutoside/pharmacology , Maillard Reaction/drug effectsABSTRACT
Troxerutin, a flavonoid best known for its radioprotective and antioxidant properties is of considerable interest of study due to its broad pharmacological activities. The present study on troxerutin highlights its abilities to bind DNA and enhance cancer cell killing in response to radiation. Troxerutin showed strong binding with calf thymus DNA in vitro. Troxerutin-DNA interaction was confirmed by CD spectropolarimetry. The mode of binding of troxerutin to DNA was assessed by competing troxerutin with EtBr or DAPI, known DNA intercalator and a minor groove binder, respectively. DAPI fluorescence was drastically reduced with linear increase in troxerutin concentration suggesting possible binding of troxerutin to DNA minor groove. Further, computational studies of docking of troxerutin molecule on mammalian DNA also indicated possible troxerutin-DNA interaction at minor groove of DNA. Troxerutin was found to mainly localize in the nucleus of prostate cancer cells. It induced cytotoxicity in radioresistant (DU145) and sensitive (PC3) prostate cancer cells. When troxerutin pre-treated DU145 and PC3 cells were exposed to γ-radiation, cytotoxicity as estimated by MTT assay, was found to be further enhanced. In addition, the % subG1 population detected by propidium iodide staining also showed similar response when combined with radiation. A similar trend was observed in terms of ROS generation and DNA damage in DU145 cells when troxerutin and radiation were combined. DNA binding at minor groove by troxerutin may have contributed to strand breaks leading to increased radiation induced cell death.
Subject(s)
DNA/metabolism , Flavonoids/metabolism , Flavonoids/pharmacology , Hydroxyethylrutoside/analogs & derivatives , Neoplasms/radiotherapy , Antineoplastic Agents/chemistry , Antineoplastic Agents/metabolism , Antineoplastic Agents/pharmacology , Antineoplastic Agents/therapeutic use , DNA/chemistry , DNA Damage , Flavonoids/therapeutic use , Humans , Hydroxyethylrutoside/chemistry , Hydroxyethylrutoside/metabolism , Hydroxyethylrutoside/pharmacology , Hydroxyethylrutoside/therapeutic use , Male , Models, Molecular , Molecular Docking Simulation , Neoplasms/drug therapy , Prostatic Neoplasms/drug therapy , Reactive Oxygen SpeciesABSTRACT
Troxerutin (TRX) is a flavonoid present in tea, coffee, cereal grains, various fruits and vegetables have been reported to exhibit radioprotective, antithrombotic, nephro and hepato-protective effects. A systematic study was undertaken to evaluate its free radical scavenging ability and anti-apoptotic activity in cell-free and cellular systems. TRX scavenged superoxide, nitric oxide and also other model stable radicals like 1,1-diphenyl-2-picryl-hydazyl, and 2,2'-azinobis3-ethylbenzothiazoline-6-sulfonic acid. It reacted with hydroxyl radicals, carbonate and thiocyanate anions, as evaluated by pulse radiolysis and stopped flow techniques. TRX protected different cell types (epithelial cells, fibroblasts and lymphocytes) against peroxyl radical-induced apoptosis, necrosis and mitotic death. It scavenged intracellular basal and inducible ROS levels and also restored depletion of intracellular GSH levels, suggesting that free radical scavenging ability may be responsible for the observed cytoprotection of different cell types. TRX may find application as an adjuvant in treating various diseases attributed to oxidative stress.
Subject(s)
Antioxidants/chemistry , Free Radicals/chemistry , Hydroxyethylrutoside/analogs & derivatives , Plant Extracts/chemistry , Reactive Oxygen Species/chemistry , Cell Death , Flavonoids , Hydroxyethylrutoside/chemistry , Oxidative StressABSTRACT
The studies on the interaction between tRNA (transfer RNA) and small molecules are an area of remarkable recent attention. For this notion a fundamental knowledge of the molecular features involving the interaction of small molecules with tRNA is crucial. Hence, in the present study we have investigated the interaction of TXER (troxerutin), natural bioflavonoid rutin derivative with yeast tRNA by using various spectroscopic techniques and molecular docking studies. The UV absorption and fluorescence emission studies demonstrated external binding of TXER on tRNA with low binding constant values as compared to strong binders. Circular dichroism (CD) spectroscopy study revealed that TXER did not show any significant modification on native conformation of tRNA. Furthermore in electrochemical study, the complex of TXER-tRNA did not expose any noticeable positive potential peak shift which indicated an interaction of TXER with tRNA by electrostatic or external binding mode. The docking study showed that the hydrogen and hydrophobic interactions were involved in binding of TXER-tRNA with docking score -7.0 kcal/mol. These findings led us to confirm the interaction of TXER on tRNA through external binding with low binding affinity, indicating its potential bioapplication in the future.
Subject(s)
Hydroxyethylrutoside/analogs & derivatives , Molecular Docking Simulation , RNA, Transfer/chemistry , Binding Sites , Circular Dichroism , Electrochemical Techniques , Hydrophobic and Hydrophilic Interactions , Hydroxyethylrutoside/chemistry , Hydroxyethylrutoside/metabolism , Nucleic Acid Conformation , RNA, Transfer/metabolism , Spectrometry, Fluorescence , Static ElectricityABSTRACT
Troxerutin (TXER) is a derivative of naturally occurring bioflavonoid rutin. It possesses different biological activities in rising clinical world. The biological activity possessed by most of the drugs mainly targets on macromolecules. Hence, in the current study we have examined the interaction mechanism of TXER with calf thymus DNA (CT-DNA) by using various spectroscopic methods, isothermal titration calorimetry (ITC) and molecular docking studies. Further, DNA cleavage study was carried out to find the DNA protection activity of TXER. UV-absorption and emission spectroscopy showed low binding constant values via groove binding. Circular dichroism study indicates that TXER does not modify native B-form of DNA, and it retains the native B-conformation. Furthermore, no effective positive potential peak shift was observed in TXER-DNA complex during electrochemical analysis by which it represents an interaction of TXER with DNA through groove binding. Molecular docking study showed thymine guanine based interaction with docking score -7.09 kcal/mol. This result was compared to experimental ITC value. The DNA cleavage study illustrates that TXER does not cause any DNA damage as well as TXER showed DNA protection against hydroxyl radical induced DNA damage. From this study, we conclude that TXER interacts with DNA by fashion of groove binding.
Subject(s)
DNA/chemistry , Hydroxyethylrutoside/analogs & derivatives , Molecular Docking Simulation , Spectrum Analysis , Animals , Cattle , DNA/metabolism , Hydroxyethylrutoside/chemistry , Hydroxyethylrutoside/metabolism , Models, Molecular , Molecular Conformation , Spectrum Analysis/methodsABSTRACT
Four simple, accurate, sensitive and precise spectrophotometric methods were developed and validated for simultaneous determination of Troxerutin (TXN) and Carbazochrome (CZM) in their bulk powders, laboratory prepared mixtures and pharmaceutical dosage forms. Method A is first derivative spectrophotometry (D(1)) where TXN and CZM were determined at 294 and 483.5 nm, respectively. Method B is first derivative of ratio spectra (DD(1)) where the peak amplitude at 248 for TXN and 439 nm for CZM were used for their determination. Method C is ratio subtraction (RS); in which TXN was determined at its λmax (352 nm) in the presence of CZM which was determined by D(1) at 483.5 nm. While, method D is mean centering of the ratio spectra (MCR) in which the mean centered values at 300 nm and 340.0 nm were used for the two drugs in a respective order. The two compounds were simultaneously determined in the concentration ranges of 5.00-50.00 µg mL(-1) and 0.5-10.0 µg mL(-1) for TXN and CZM, respectively. The methods were validated according to the ICH guidelines and the results were statistically compared to the manufacturer's method.
Subject(s)
Adrenochrome/analogs & derivatives , Dosage Forms , Hydroxyethylrutoside/analogs & derivatives , Spectrophotometry/methods , Adrenochrome/analysis , Adrenochrome/chemistry , Hydroxyethylrutoside/analysis , Hydroxyethylrutoside/chemistry , Regression Analysis , Reproducibility of ResultsABSTRACT
Troxerutin is a trihydroxyethylated derivative of the flavonoid, rutin. It has been reported to possess the hepatoprotective, nephroprotective, antioxidant, anti-inflammatory, and antihyperlipidemic activities. Troxerutin treatment reduced the blood glucose and glycosylated hemoglobin levels in high-cholesterol-induced insulin-resistant mice and in type-2 diabetic patients. However, the mechanism by which it exhibits antidiabetic property was unknown. Therefore, the present study was designed to evaluate the effect of troxerutin on insulin signaling molecules in gastrocnemius muscle of high fat and sucrose-induced type-2 diabetic rats. Wistar male albino rats were selected and divided into five groups. Group I: Control. Group II: High fat and sucrose-induced type-2 diabetic rats. Group III: Type-2 diabetic rats treated with troxerutin (150 mg/kg body weight/day orally). Group IV: Type-2 diabetic rats treated with metformin (50 mg/kg body weight/day orally). Group V: Normal rats treated with troxerutin (150 mg/kg body weight/day orally). After 30 days of treatment, fasting blood glucose, oral glucose tolerance, serum lipid profile, and the levels of insulin signaling molecules, glycogen, glucose uptake, and oxidation in gastrocnemius muscle were assessed. Diabetic rats showed impairment in insulin signaling molecules (IR, p-IRS-1(Tyr632), p-Akt(Ser473), ß-arrestin-2, c-Src, p-AS160(Thr642), and GLUT4 proteins), glycogen concentration, glucose uptake, and oxidation. Oral administration of troxerutin showed near normal levels of blood glucose, serum insulin, lipid profile, and insulin signaling molecules as well as GLUT4 proteins in type-2 diabetic rats. It is concluded from the present study that troxerutin may play a significant role in the management of type-2 diabetes mellitus, by improving the insulin signaling molecules and glucose utilization in the skeletal muscle.
Subject(s)
Blood Glucose/metabolism , Diabetes Mellitus, Experimental/drug therapy , Glucose Transporter Type 4/metabolism , Hydroxyethylrutoside/analogs & derivatives , Hypoglycemic Agents/administration & dosage , Muscle, Skeletal/drug effects , Administration, Oral , Animals , Diabetes Mellitus, Experimental/metabolism , Diabetes Mellitus, Experimental/pathology , Diet, High-Fat/adverse effects , Hydroxyethylrutoside/administration & dosage , Hydroxyethylrutoside/chemistry , Hydroxyethylrutoside/pharmacology , Hypoglycemic Agents/chemistry , Hypoglycemic Agents/pharmacology , Insulin/blood , Male , Muscle, Skeletal/metabolism , Rats , Rats, Wistar , Signal Transduction/drug effectsABSTRACT
During the scavenging of free radicals flavonoids are oxidized to electrophilic quinones. Glutathione (GSH) can trap these quinones, thereby forming GSH-flavonoid adducts. The aim of this study was to compare the stability of the GSH-flavonoid adduct of 7-mono-O-(ß-hydroxyethyl)rutoside (monoHER) with that of quercetin. It was found that GSH-quercetin reacts with the thiol N-acetyl-L-cysteine (NAC) to form NAC-quercetin, whereas GSH-monoHER does not react with NAC. In addition, the adduct of the monoHER quinone with the dithiol dithiothreitol (DTT) is relatively stable, whereas the DTT-quercetin adduct is readily converted into quercetin and DTT disulfide. These differences in reactivity of the thiol-flavonoid adducts demonstrate that GSH-monoHER is much more stable than GSH-quercetin. This difference in reactivity was corroborated by molecular quantum chemical calculations. Thus, although both flavonoid quinones are rapidly scavenged by GSH, the advantage of monoHER is that it forms a stable conjugate with GSH, thereby preventing a possible spread of toxicity. These findings demonstrate that even structurally comparable flavonoids behave differently, which will be reflected in the biological effects of these flavonoids.
Subject(s)
Glutathione/chemistry , Hydroxyethylrutoside/analogs & derivatives , Quercetin/chemistry , Hydroxyethylrutoside/chemistry , Molecular Structure , Sulfhydryl Compounds/chemistryABSTRACT
The clinical use of the anticancer drug doxorubicin is limited by severe cardiotoxicity. In mice, the semisynthetic antioxidant flavonoid 7-mono-O-(ß-hydroxyethyl)-rutoside (monoHER) has been successfully used as a protector against doxorubicin-induced cardiotoxicity. However, most monoHER has already been cleared from the body at the time that doxorubicin concentrations are still high. This result suggests that not only the parent compound monoHER itself but also monoHER metabolites could be responsible for the observed cardioprotective effects in mice. Therefore, in the present study, we investigated the metabolism of monoHER in mice. Mice were administered 500 mg/kg monoHER intraperitoneally. At different time points after monoHER administration, bile was collected and analyzed for the presence of monoHER metabolites. The formed metabolites were identified by liquid chromatography-diode array detection-time of flight-mass spectrometry. Thirteen different metabolites were identified. The observed routes of monoHER metabolism are methylation, glucuronidation, oxidation of its hydroxyethyl group, GSH conjugation, and hydrolysis of its disaccharide. In line with other flavonoids, methylated monoHER and the monoHER glucosides are expected to have relatively high cellular uptake and low clearance from the body. Therefore, these metabolites might contribute to the observed protection of monoHER against doxorubicin-induced cardiotoxicity.
Subject(s)
Antioxidants/chemistry , Antioxidants/metabolism , Bile/metabolism , Cardiotonic Agents/metabolism , Hydroxyethylrutoside/analogs & derivatives , Animals , Antioxidants/analysis , Antioxidants/pharmacology , Cardiotonic Agents/chemistry , Cardiotonic Agents/pharmacology , Doxorubicin/toxicity , Glucuronides/chemistry , Glucuronides/metabolism , Glutathione/chemistry , Glutathione/metabolism , Hydroxyethylrutoside/chemistry , Hydroxyethylrutoside/metabolism , Hydroxyethylrutoside/pharmacology , Male , Methylation , Mice , Mice, Inbred BALB CABSTRACT
Comparative studies of enzymatic acylation of troxerutin by the alkaline protease from Bacillus subtilis under ultrasound and shaking were carried out in nonaqueous media. Using divinyl dicarboxylates (CH(2)CH-OOC-(CH(2))(n)-COO-CHCH(2), n=2, 3, 4, 7, 8, 11) featuring different chain length as acyl donors, troxerutin was regioselectively acylated at B ethoxyl group, whether under ultrasound or shaking. Ultrasonic treatment increased the reaction rate and led to high conversion. Several factors, such as pre-irradiation on the enzyme, the power and frequency of the ultrasound, operation manner, as well as the length of the acyl donors were investigated. Using enzyme pre-irradiated for 8 h, the conversion of troxerutin was increased to 87.3% compared with 56.3% obtained from the untreated enzyme. Experimental results also showed that continual ultrasound caused greater rate acceleration than interval ultrasound. Powers of 100, 150 and 200 W, frequencies of 40, 80 and 100 kHz all showed significant improvement on the transesterification, with the greatest effect observed at 150 W, 80 kHz. The acceleration effect increased as the chain length of the acyl donors decreased from C(13) to C(4).
Subject(s)
Bacterial Proteins/metabolism , Endopeptidases/metabolism , Esters/chemical synthesis , Hydroxyethylrutoside/analogs & derivatives , Ultrasonics , Bacillus subtilis/enzymology , Bacterial Proteins/chemistry , Endopeptidases/chemistry , Esters/chemistry , Hydroxyethylrutoside/chemistry , Solvents/chemistryABSTRACT
Antioxidants can scavenge highly reactive radicals. As a result the antioxidants are converted into oxidation products that might cause damage to vital cellular components. To prevent this damage, the human body possesses an intricate network of antioxidants that pass over the reactivity from one antioxidant to another in a controlled way. The aim of the present study was to investigate how the semi-synthetic flavonoid 7-mono-O-(ß-hydroxyethyl)-rutoside (monoHER), a potential protective agent against doxorubicin-induced cardiotoxicity, fits into this antioxidant network. This position was compared with that of the well-known flavonoid quercetin. The present study shows that the oxidation products of both monoHER and quercetin are reactive towards thiol groups of both GSH and proteins. However, in human blood plasma, oxidized quercetin easily reacts with protein thiols, whereas oxidized monoHER does not react with plasma protein thiols. Our results indicate that this can be explained by the presence of ascorbate in plasma; ascorbate is able to reduce oxidized monoHER to the parent compound monoHER before oxidized monoHER can react with thiols. This is a major difference with oxidized quercetin that preferentially reacts with thiols rather than ascorbate. The difference in selectivity between monoHER and quercetin originates from an intrinsic difference in the chemical nature of their oxidation products, which was corroborated by molecular quantum chemical calculations. These findings point towards an essential difference between structurally closely related flavonoids in their interplay with the endogenous antioxidant network. The advantage of monoHER is that it can safely channel the reactivity of radicals into the antioxidant network where the reactivity is completely neutralized.
Subject(s)
Antioxidants/metabolism , Flavonoids/metabolism , Hydroxyethylrutoside/analogs & derivatives , Quercetin/metabolism , Antioxidants/chemistry , Antioxidants/pharmacology , Ascorbic Acid/chemistry , Ascorbic Acid/metabolism , Ascorbic Acid/pharmacology , Flavonoids/chemistry , Flavonoids/pharmacology , Glutathione/metabolism , Humans , Hydrogen Peroxide/chemistry , Hydrogen Peroxide/metabolism , Hydrogen Peroxide/pharmacology , Hydroxyethylrutoside/chemistry , Hydroxyethylrutoside/metabolism , Hydroxyethylrutoside/pharmacology , Models, Chemical , Molecular Structure , Oxidants/chemistry , Oxidants/metabolism , Oxidation-Reduction/drug effects , Quercetin/chemistry , Quercetin/pharmacologyABSTRACT
Flavonoids protect against oxidative stress by scavenging free radicals. During this protection flavonoids are oxidized. The oxidized flavonoids formed are often reactive. Consequently, protection by flavonoids can result in the formation of toxic products. In this study the oxidation of 7-mono-O-(beta-hydroxyethyl)rutoside (monoHER), which is a constituent of the registered drug Venoruton, was studied in the absence and presence of glutathione (GSH). MonoHER was oxidized by horseradish peroxidase/H(2)O(2). Spectrophotometric and HPLC analysis showed that in the presence of GSH, a monoHER-GSH conjugate was formed, which was identified as 2'-glutathionyl monohydroxyethylrutoside by mass spectrometric analysis and (1)H NMR. Preferential formation of this glutathione adduct in the B ring at C2' was confirmed by molecular quantum chemical calculations. This conjugate was also detected in the bile fluid of a healthy volunteer after iv administration of monoHER, demonstrating its formation in vivo. These results indicate that in the process of offering protection against free radicals, monoHER is converted into an oxidation product that is reactive toward thiols. The formation of this thiol-reactive oxidation product is potentially harmful. Thus, the supposed beneficial effect of monoHER as an antioxidant may be accompanied by the formation of products with an electrophilic, toxic potential.
Subject(s)
Glutathione/chemistry , Hydroxyethylrutoside/analogs & derivatives , Adult , Bile/chemistry , Body Fluids/chemistry , Computer Simulation , Horseradish Peroxidase/metabolism , Humans , Hydrogen Peroxide/chemistry , Hydroxyethylrutoside/administration & dosage , Hydroxyethylrutoside/analysis , Hydroxyethylrutoside/chemistry , Infusions, Intravenous , Magnetic Resonance Spectroscopy , Male , Mass Spectrometry , Models, Chemical , Oxidation-Reduction , Quantum Theory , Reference Values , Solid Phase Extraction , Spectrophotometry, Ultraviolet , Time FactorsABSTRACT
The voltammetric responses of troxerutin were investigated at polyvinylpyrrolidone (cross-linked) (PVP) modified carbon paste electrode (CPE) in 0.1 mol/L KCl by several electrochemical techniques. A well-defined oxidation peak was observed at about 0.97 V. Compared with poor responses of troxerutin at bare electrode that at this modified electrode has been greatly improved. It is PVP that enhances the adsorption of troxerutin to electrode surface based on their hydrophobic property. Under some optimized experimental conditions, a simple and sensitive electroanalytical method was developed for the quantitative analysis of troxerutin. A very low detection limit of 5.0 x 10(-9)mol/L was obtained for 5 min accumulation at open circuit (S/N=3). This proposed method was successfully applied to the detection of troxerutin in pharmaceutical dosage forms and satisfying results had been obtained.
Subject(s)
Carbon/chemistry , Hydroxyethylrutoside/analogs & derivatives , Povidone/chemistry , Adsorption , Calibration , Dosage Forms , Electrochemistry , Electrodes , Electrolytes , Hydroxyethylrutoside/analysis , Hydroxyethylrutoside/chemistry , Molecular Structure , Oxidation-Reduction , Potassium Chloride/chemistry , Sensitivity and SpecificityABSTRACT
The interaction of rutin and venoruton (troxerutin), with alpha-, beta- and gamma-cyclodextrin (CD), hydroxypropyl-beta-cyclodextrin (HP-beta-CD) and methyl-beta-cyclodextrin (M-beta-CD) was investigated by reversed-phase thin layer chromatography on polyamide plates. A mobile phase consisted of NH(4)OH; NH(4)Cl buffer solution containing various CD concentrations (pH = 9.7, 20 degrees C) was used as mobile phase. The equilibrium constants (K(f)) and the retention factor (R(f)) were determined and used to study the inclusion process. The in fluence of CDs on the solubility of rutin and venoruton was characterized by R(M) values and the increasing hydrophilicity of drugs. The results show that the inclusion capacity of cyclodextrins follows the order HP-beta-CD > M-beta-CD > beta-CD > gamma-CD, and rutin is more easily included by the studied cyclodextrins than venoruton. In addition, the thermodynamic parameters (Delta H, Delta S) for the formation of complexes were obtained from the van't Hoff equation, displaying the enthalpy-entropy compensation effect.