Voltammetric Oxidation of Drugs of Abuse I. Morphine and Metabolites

A detailed study of the electrochemical oxidative behavior of morphine in aqueous solution is reported. Through the synthesis of several metabolites and derivatives, pseudomorphine, morphine N‐oxide, normorphine, dihydromorphine and 2‐(N,N‐dimethylaminomethyl)morphine, and their voltammetric study it was possible to identify the oxidation peaks for morphine. The anodic waves are related with the oxidation of phenolic and tertiary amine groups. It is also possible to verify that a poorly defined peak observable during morphine oxidation is not a consequence of further oxidation of pseudomorphine but due to formation of a dimer during phenolic group oxidation. The results obtained and especially those regarding the formation of a new polymer based on a C? O coupling could be useful for clarifying the discoloration phenomenon occurring during storage of morphine solutions as well as leading to a better understanding of its oxidative metabolic pathways.     

Species‐Specific Hydrolysis Kinetics of N‐Methylated Heroin Derivatives

The hydroxide‐catalyzed hydrolysis of 3,6‐diacetylmorphine (heroin) was shown to take place predominantly via its positively charged form. N‐Methylated quaternary derivatives of heroin bearing a permanent positive charge were synthesized, and thus, hydrolysis kinetics of these cationic species could be studied over a wide pH range. Specific rate equations were introduced to characterize either the simultaneous or the consecutive decompositions. The kinetic constants determined for the diester are distinctive for the site of hydrolysis. The rate of 6‐acetyl‐N‐methylmorphine was quantified in terms of microscopic kinetic constants of hydrolysis, in which the protonation state of the phenolic OH group had also been taken into account. The site‐specific data indicate that the 3‐acetoxy moiety is hydrolyzed 6 – 12 times faster than the 6‐acetoxy function. The latter, previously ignored minor pathway was shown to represent a non‐negligible 10% of the overall decomposition process. Protonation of the 3‐O⁻ site accelerates the rate of hydrolysis of the 6‐acetoxy moiety by a factor of 4, and replacement of the adjacent OH group by MeO or AcO substituents slows the rate of hydrolysis slightly, presumably due to the increased local hydrophobicity caused by the alkyl or acyl moiety.     

Voltammetric Oxidation of Drugs of Abuse II. Codeine and Metabolites

The oxidation of codeine on glassy carbon electrodes has been studied in detail using differential pulse voltammetry. The results obtained using a glassy carbon electrode clearly show a much more complex oxidation mechanism than that previously reported when platinum and gold electrodes were used. To clarify the codeine oxidative profile, several metabolites and analogues of this alkaloid, codeine N‐oxide, norcodeine, dihydrocodeine, acetylcodeine and 6‐chlorodesoxycodeine, were synthesized and studied. It was deduced that the anodic waves observed in codeine oxidation are related to the presence of methoxy, hydroxy and tertiary amine groups. Due to the similarity of potentials at which these oxidative processes take place, at some pHs an overlap of peaks occurs and only one anodic wave is observed.     

An investigation of the thermal and thermo‐oxidative degradation of polybenzoxazines with a reactive functional group

The thermal behavior of a series of polybenzoxazines based on 3‐aminophenylacetylene has been investigated. The effect of reactive amine on the thermal cleavage of the Mannich base is examined under both inert and oxidative environments. It has been shown that the thermal stability of polybenzoxazines is substantially improved by the reactive amine. Various biphenols are found to have insignificant effect on the thermal stability of this series of polybenzoxazines. These nitrogen containing phenolic resins are nonflammable polymers. © 1999 John Wiley & Sons, Inc. J Polym Sci B: Polym Phys 37: 647–659, 1999     

Group-selective antibodies based fluorescence immunoassay for monitoring opiate drugs

A novel carboxylic acid derivative of monoacetylmorphine (MAM-COOH) was synthesized and conjugated with bovine serum albumin (BSA) for generating polyclonal antibodies against the target molecule heroin and its major metabolites. The conjugate was characterized by fluorescence spectroscopy, polyacrylamide gel electrophoresis, and mass spectrometry to confirm the extent of haptenization of the carrier protein. A high titer (1:64,0000) of antibody was obtained by using the conjugate with an optimum protein/hapten molar ratio of 1:100. The generated antibody showed good binding affinity with heroin and its metabolites monoacetylmorphine (MAM) and morphine. The relative affinity constant (K (aff)) of the antibody was 3.1 x 10(7) l mol(-1), and the IC(50) values obtained for heroin, MAM, morphine, and codeine were 0.01, 0.013, 0.012, and 0.014 ng ml(-1), respectively. A fluorescence-based competitive inhibition immunoassay procedure was developed for the estimation of heroin and its major metabolites in standard and biofludic samples over a concentration range up to 0.01 ng ml(-1) with good signal reproducibility (p < 0.05). The method can be used as a convenient quantitative tool for the sensitive screening of major metabolites of heroin in biological samples.     

保护的几丁寡糖及结构类似物的合成：复杂氨基寡糖合成的通用方法

建立了逐步合成具有重要生物活性的2-脱氧-2-氨基葡萄糖寡糖链的通用方法。采用邻苯二甲酰基保护氨基、硫代苯基为还原末端的离去基团,以氨基葡萄糖为起始原料,几种保护的几丁寡糖及结构类似物被合成：3-O-乙酰基-4,6-O-亚苄基-2-脱氧-2-邻苯二甲酰亚氨基-b-D-吡喃葡萄糖-（1→4）-（3-O-乙酰基-6-O-苄基-2-脱氧-2-邻苯二甲酰亚氨基）-b-D-吡喃葡萄糖甲苷（4）、3-O-乙酰基-4,6-O-亚苄基-2-脱氧-2-邻苯二甲酰亚氨基-b-D-吡喃葡萄糖-（1→4）-（3-O-乙酰基-6-O-苄基-2-脱氧-2-邻苯二甲酰亚氨基-b-D-吡喃葡萄糖）-（1→4）-（3-O-乙酰基-6-O-苄基-2-脱氧-2-邻苯二甲酰亚氨基）-b-D-吡喃葡萄糖甲苷（6）、3-O-乙酰基-4,6-O-亚苄基-2-脱氧-2-邻苯二甲酰亚氨基-b-D-吡喃葡萄糖-（1→3）-（4,6-O-亚苄基-2-脱氧-2-邻苯二甲酰亚氨基）-b-D-吡喃葡萄糖甲苷（8）、3-O-乙酰基-4,6-O-亚苄基-2-脱氧-2-邻苯二甲酰亚氨基-b-D-吡喃葡萄糖-（1→3）-（4,6-O-亚苄基-2-脱氧-2-邻苯二甲酰亚氨基-b-D-吡喃葡萄糖）- （1→3）-（4,6-O-亚苄基-2-脱氧-2-邻苯二甲酰亚氨基）- b-D-吡喃葡萄糖甲苷（10）。所合成化合物通过核磁共振和质谱分析确证了其化学结构。     

A Convenient and Efficient Method for the Preparation of Unique Fluorophores of Lariat Naphtho‐Aza‐Crown Ethers

New naphtho‐aza‐crown ethers containing different phenolic side‐arms attached through the ortho‐position of the phenol have been prepared under solvent‐free conditions. The starting macrocyclic naphtho‐aza‐crown ether 2 was obtained by treatment of naphthalene dicarboxylic acid diester 1 with diethylenetriamine in EtOH at room temperature for two days without stirring in 77% yield (Scheme 1). Phenolic ligands ( 3 – 14 ) were synthesized by the Mannich reaction of the secondary macrocyclic amine 2 with the substituted phenols using nontoxic and inexpensive CaCl₂. This procedure was applied successfully for the synthesis of Mannich bases from simple secondary amines. The CaCl₂ powder can be reused up to three times after simple washing with dry acetone.     

A new kinetic spectrophotometric method for the determination of major metabolite of heroin in biological samples

This study describes a simple,rapid and selective catalytic kinetic spectrophotometric method for the determination of 6- monoacetylmorphine（6-MAM） as major metabolite of heroin in biological samples.The method is based upon the catalytic effect of 6-MAM on the oxidation of Janus Green by bromate in acid media.The reaction was followed spectrophotometrically by measuring the decrease in absorbance of Janus Green at 618 nm.The dependence of sensitivity on the reaction variables was studied.Under optimum conditions,two linear calibration curves over the range 0.1-1.0μg mL^-1 and 1.0-34.0μg mL^-1 of 6- MAM were obtained.The detection limit was 1.2×10²μg mL^-1 of 6-MAM.The relative standard deviations for six replicate determinations of 0.8 and 5.0μg mL^-1 of 6-MAM were 1.4 and 1.1%respectively.The effect of various species commonly associated with heroin in real samples was also investigated.The proposed method was successfully applied in human urine and serum samples with satisfactory results.     

New oxidative transformations of phenolic and indolic oxazolines: an avenue to useful azaspirocyclic building blocks

The oxidative cyclization of a phenolic amide to a spirolactam has long been regarded as an "impossible" reaction, because exposure of the substrates to a variety of oxidants results in formation of spirolactones with consequent loss of the amine segment. We recently communicated that this heretofore unknown transformation may be achieved by oxidation of oxazoline analogues of phenolic and indolic amides. Herein, we provide full details of our work.     

Online H/D exchange liquid chromatography as a support for the mass spectrometric identification of the oxidation products of melatonin

The hydrogen–deuterium exchange of protonated melatonin and its in vitro oxidation end‐products have been examined by liquid chromatography coupled with ion‐trap mass spectrometry. Specific H/D scrambling of protons in the C2 and C4 positions of the indole ring during gas‐phase fragmentation process was observed for both melatonin and its oxidation products. Collision‐induced dissociation spectra showed losses of variably deuterated NH₃, H₂O and CH₃CONH₂. In addition, a similar H/D scrambling behaviour was observed for the oxidation products, obtained from the opening of the indole ring by oxidative attack. Fragmentation pathways are proposed and H/D scrambling has been employed as a fingerprint, allowing identification of N¹‐acetyl‐5‐methoxykynurenin (AMK), N¹‐acetyl‐N²‐formyl‐5‐methoxykynurenin (AFMK), dehydro‐AFMK and hydroxymelatonin as the oxidation products of melatonin in vitro. Copyright © 2008 John Wiley & Sons, Ltd.     

Ultra‐high performance supercritical fluid chromatography of lignin‐derived phenols from alkaline cupric oxide oxidation

Traditional chromatographic methods for the analysis of lignin‐derived phenolic compounds in environmental samples are generally time consuming. In this work, an ultra‐high performance supercritical fluid chromatography method with a diode array detector for the analysis of major lignin‐derived phenolic compounds produced by alkaline cupric oxide oxidation was developed. In an analysis of a collection of 11 representative monomeric lignin phenolic compounds, all compounds were clearly separated within 6 min with excellent peak shapes, with a limit of detection of 0.5–2.5 μM, a limit of quantification of 2.5–5.0 μM, and a dynamic range of 5.0–2.0 mM (R² > 0.997). The new ultra‐high performance supercritical fluid chromatography method was also applied for the qualitative and quantitative analysis of lignin‐derived phenolic compounds obtained upon alkaline cupric oxide oxidation of a commercial humic acid. Ten out of the previous eleven model compounds could be quantified in the oxidized humic acid sample. The high separation power and short analysis time obtained demonstrate for the first time that supercritical fluid chromatography is a fast and reliable technique for the analysis of lignin‐derived phenols in complex environmental samples.     

Preparation of 5‐Aryl‐3‐Oxo‐δ‐Lactones by the Potassium Carbonate–Promoted Condensation of Aromatic Aldehydes and Ethyl Acetoacetate in Ethanol

5‐Aryl‐3‐oxo‐δ‐lactones (6‐aryl‐dihydro‐2H‐pyran‐2,4(3H)‐diones) were prepared by the potassium carbonate–promoted condensation of aromatic aldehydes and ethyl acetoacetate in absolute ethanol. Benzaldehyde and substituted benzaldehydes bearing an alkoxy group (2 or 3 position), a chlorine atom (2, 3, or 4 position), a nitro group (3 or 4 position), a cyano group (4 position), or an acetyl group (4 position) react in high yields under these conditions.     

Synthetic Study of Carthamin: Biomimetic Synthesis of Carthamin Model via Peroxidase‐Catalyzed Oxidative Decarboxylation of Its Precursor Model

A chiral carthamin model (3S,3′S)‐1‐[5‐acetyl‐2,6‐diketo‐3‐C‐β‐d ‐glucopyranosylcyclohex‐4‐enylidene]‐1′‐[5′‐acetyl‐3′‐C‐β‐d ‐glucopyranosyl‐2′,3′,4′‐trihydroxy‐6′‐oxocyclohexa‐1′,4′‐dienyl]methane, in which two cinnamoyl groups were replaced by an acetyl group, was synthesized by the dimerization of (S)‐2‐acetyl‐4‐C‐(per‐O‐acetyl‐β‐d ‐glucopyranosyl)cyclohexadienone with glyoxylic acid, followed by peroxidase‐catalyzed oxidative decarboxylation and de‐O‐acetylation, or de‐O‐acetylation and peroxidase‐catalyzed oxidative decarboxylation. The corresponding total yields were 12.5% or 17.1% from 3‐C‐(per‐O‐acetyl‐β‐d ‐glucopyranosyl)phloroacetophenone, and the reaction pathway was identical to the biosynthetic pathway.     

Functional group transformation from amines to aldehydes via IBX oxidation

IBX oxidation of secondary aryl amines has been applied towards their functional group transformation to aldehydes using a facile post-process with satisfactory yields. The oxidation of N-benzylmethylamine was used as a model substrate and suggested that the ratio of IBX oxidant to amine should be 2:1. Subsequently, several aryl amines were subjected to these standard conditions, which revealed that the oxidative activity depends on the electronic and steric structures of the substituent groups in the substrates. The oxidative selectivity to secondary amines was also discovered.     

Nucleophilic Addition of Thiaproline to Electrochemically Derived o‐Quinone,Application to the Sensitive Voltammetric Detection of Thiaproline

The mechanism of electrochemical behavior of catechol in the presence of thiaproline is investigated by cyclic voltammetry, controlled‐potential coulometry and spectrophotometric tracing of the reaction coordinate. The results indicate that thiaproline participate in with an ECEC mechanism in a nucleophilic (Michael) addition to o‐quinone. Effect of pH of buffer solution on reaction pathway is studied and showed that addition of thiaproline to the o‐quinone is performed only in solutions with pHs higher than 5. These results indicate that the addition of thiaproline is occurred first from amine functional group. In the second step, the addition of carboxylate group of thiaproline to C‐5 of catechol results the final product with a lactone ring in its structure. Observation of two isosbestic point in absorption spectrum during the progress of the electrolysis together with the FT‐IR results for final product can be presented as evidence for two step addition of thiaproline to catechol. Final product, due to the electron donor property of thiaproline, more easily oxidized respect to the former catechol and as a result, a new redox couple is obtained for this compound in lower potentials. The easier anodic oxidation of addition product (relative to catechol) caused to an increase in anodic current for catechol, which is proportional to the thiaproline concentration. The differential pulse voltammetry (DPV) is applied as a sensitive voltammetric method for the detection of thiaproline. A linear range from 5×10^?8 to 5×10^?6 M with a detection limit of 1×10^?8 M is resulted for thiaproline. With respect to the reversibility of the electrochemical reactions in the mechanism, and also more facile oxidation of the addition products, the square‐wave voltammetry is presented as a method with considerably more sensitivity for the detection of sub‐micromolar amounts of thiaproline. The advantageous properties of the voltammetric method for thiaproline detection lie in its excellent catalytic activity, sensitivity and simplicity.     

Preparative separation of the flavonoid fractions from Periploca forrestii Schltr. ethanol extracts using macroporous resin combined with HPLC analysis and evaluation of their biological activities

A preparative separation method using macroporous absorptive resin coupled with high‐performance liquid chromatography was developed for the separation of six fractions of the 80% ethanol extract of Periploca forrestii Schltr. The six ethanol fractions (5–95; A, B, C, D, E, and F) obtained were carefully analyzed to locate the corresponding peaks in the high‐performance liquid chromatography chromatogram of the total extract, which was established in a previous study. Furthermore, the biological activities, including antioxidant activities, acetyl cholinesterase inhibitory capacities, antihyaluronidase activities, and anti‐inflammatory effects, were evaluated in MH7A cells. The results demonstrated that fraction E could significantly prevent oxidation and inhibit hyaluronidase and acetyl cholinesterase. Finally, the main flavonoids in fractions A and E from P. forrestii Schltr. were purified, and the compounds were identified as chlorogenic acid, quercetin‐3‐O‐α‐L‐arabinopyranoside, and quercetin‐7‐O‐β‐D‐glucopyranoside. The chemical structures were confirmed by mass spectrometry and nuclear magnetic resonance spectroscopy. Furthermore, the inhibitory effects of these compounds against complete Freund's adjuvant‐induced secondary immune arthritis in rats were evaluated.     

Development of a Carbon Paste Electrode Modified with Reduced Graphene Oxide and an Imidazole Derivative for Simultaneous Determination of Biological Species of N‐acetyl‐L‐cysteine,Uric Acid and Dopamine

In this work, the modified carbon paste electrode (CPE) with an imidazole derivative 2‐(2,3 dihydroxy phenyl) 4‐methyl benzimidazole (DHPMB) and reduced graphene oxide (RGO) was used as an electrochemical sensor for electrocatalytic oxidation of N‐acetyl‐L‐cysteine (NAC). The electrocatalytic oxidation of N‐acetyl‐L‐cysteine on the modified electrode surface was then investigated, indicating a reduction in oxidative over voltage and an intensive increase in the current of analyte. The scan rate potential, the percentages of DHPMB and RGO, and the pH solution were optimized. Under the optimum conditions, some parameters such as the electron transfer coefficient (α) between electrode and modifier, and the electron transfer rate constant) k_s) in a 0.1 M phosphate buffer solution (pH=7.0) were obtained by cyclic voltammetry method. The diffusion coefficient of species (D) 3.96×10^?5 cm² s^?1 was calculated by chronoamperometeric technique and the Tafel plot was used to calculate α (0.46) for N‐ acetyl‐L‐cysteine. Also, by using differential pulse voltammetric (DPV) technique, two linear dynamic ranges of 2–18 µM and 18–1000 µM with the detection limit of 61.0 nM for N‐acetyl‐L‐cysteine (NAC) were achieved. In the co‐existence system of N‐acetyl‐L‐cysteine (NAC), uric acid (UA) and dopamine (DA), the linear response ranges for NAC, UA, and DA are 6.0–400.0 µM, 5.0–50.0 µM and 2.0–20.0 µM, respectively and the detection limits based on (C=3s_b/m) are 0.067 µM, 0.246 µM and 0.136 µM, respectively. The obtained results indicated that DHPMB/RGO/CPE is applicable to separate NAC, uric acid (UA) and dopamine (DA) oxidative peaks, simultaneously. For analytic performance, the mentioned modified electrode was used for determination of NAC in the drug samples with acceptable results, and the simultaneous determination of NAC, UA and DA oxidative peaks was investigated in the serum solutions, too.     

Matrix‐assisted laser desorption/ionization time‐of‐flight mass spectroscopic analysis of telechelic polythiourethanes obtained by the cationic ring‐opening polymerization of six‐membered cyclic thiourethane

A matrix‐assisted laser desorption/ionization time‐of‐flight (MALDI‐TOF) mass spectroscopy analysis of polythiourethanes obtained by the cationic ring‐opening polymerization of a six‐membered cyclic thiourethane [3‐benzyltetrahydro‐1,3‐oxazine‐2‐thione (BTOT)] is described. A MALDI‐TOF mass spectrum of a polymer obtained by the polymerization of BTOT with boron trifluoride etherate (BF₃OEt₂) as the initiator in nitrobenzene at 50 °C for 24 h followed by an end‐capping reaction with diethyldithiocarbamic acid diethylammonium salt showed a series of well‐resolved signals that were assignable to polythiourethanes possessing an amino group at the initiating end and a diethyldithiocarbamate group at the terminating end. In comparison with the MALDI‐TOF mass spectra of polymers obtained by polymerization with trifluoromethanesulfonic acid or methyl trifluoromethanesulfonate, the plausible initiating species in the polymerization with BF₃OEt₂ was estimated to be a proton, which successively eliminated carbonyl sulfide to produce a secondary amine group at the initiating end. The secondary amine group in the obtained telechelic polymer was converted to a tertiary amine group by a reaction with benzyl bromide in the presence of triethylamine, and this was confirmed by MALDI‐TOF mass spectroscopy. Furthermore, a telechelic polymer with a pyrrole end group was successfully synthesized by the end‐capping reaction of the growing species in the polymerization of BTOT with sodium 1‐pyrrolecarbodithioate. © 2006 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 44: 4281–4289, 2006     

Inhibited oxidation of a polyethylene melt

The oxidation of linear polyethylene inhibited by an effective antioxidant of the phenolic type, viz. 2,2′-methylene-bis (4-methyl-6-tert. butylphenol), has been studied over the temperature range 170–210. There is a critical concentration of antioxidant above which the rate of its consumption is directly proportional to its concentration. Deviations from this dependence are due to chain initiation by direct oxidation of the polymer and to reactions involving the products derived from the antioxidant: these products can also retard the oxidation. Similar results have been obtained using the amine type antioxidant, N-phenyl-N′-cyclohexyl-p-phenylenediamine. Some features of the mechanism of polyolefin oxidation are discussed.