Comparative cytotoxicity study of nicotine and cotinine on MRC-5 cell line

Nicotine has several health hazards regarding carcinogenic potential. It also imparts increased risk for respiratory, cardiovascular, and gastrointestinal disorders. Several mechanisms have been proposed for the carcinogenic potential, including effects on cell proliferation, inducing oxidative stress, DNA mutation, or inhibition of apoptosis. The cotinine metabolite is generally thought to have effects similar to nicotine in some experimental systems. The purpose of this study was to assess the nicotine and cotinine cytotoxicity on MRC-5 lung fibroblasts. The pulmonary fibroblasts were treated with various concentrations of nicotine or cotinine (in the range 1 µM – 2 mM) for 24 or 48 h and analyzed for cell viability by MTT test. The results indicated that high nicotine concentrations (2 mM) induced marked cell death (about 50%) in MRC-5 cell line. Cotinine showed lower toxicity than nicotine on the MRC-5 cells. In contrast to nicotine treatment, cells treated with cotinine continued to proliferate after the 48h incubation period

Determination of Tramadol in human plasma by HPLC with fluorescence detection

Tramadol is a centrally acting analgesic, atypical opioid, and although it is generally considered as a medicinal drug with a low potential for dependence, there is growing evidence of tramadol abuse in some countries. The ultraviolet detection is not suitable for analysis of tramadol in plasma, due to the lack of sensitivity and selectivity. However, it was shown that tramadol has a weak fluorescence, and the latest techniques for determination of tramadol in plasma include liquid chromatographic methods with fluorescence detection (FL). The objective of the paper was to develop a HPLC-FL method applicable for quantification of tramadol in human plasma.The separation was achieved by reverse phase HPLC method, using as stationary phase C18 – Kromasil® column and a mobile phase consisted of acetonitrile:0.1% formic acid (20:80). The fluorescence detection has been applied with λex/em= 280/310 nm. A solid phase extraction procedure using C18 cartridge was carried out. The linearity of the method has been demonstrated in the range of both therapeutic and toxic plasma tramadol levels (concentrations of 0.100 – 1µg/mL). The selectivity, precision, and accuracy of the method have been demonstrated. The limit of detection (LOD= 0.010 µg/mL) and the limit of quantification (LOQ = 0.100 µg/mL) have been established.The proposed method can be used to assess tramadol levels in human plasma in pharmacokinetic studies, as well as in overdose cases. The utility of the method for the quantification of therapeutic levels of tramadol has been shown on the plasma samples from the patients with tramadol treatment as analgesic (doses ranging from 100 mg to 400 mg/day).The developed method is rapid, using simple experimental conditions and an accurate and short extraction procedure

Toxicological Analysis of Some Drugs of Abuse in Biological Samples

Consumption of drugs of abuse is a scourge of modern world. Abuse, drug addiction and their consequences are one of the major current problems of European society because of the significant repercussions in individual, family, social and economic level. In this context, toxicological analysis of the drugs of abuse in biological samples is a useful tool for: diagnosis of drug addiction, checking an auto-response, mandatory screening in some treatment programs, identification of a substance in the case of an overdose, determining compliance of the treatment. The present paper aims to address the needs of healthcare professionals involved in drugs addiction treatment through systematic presentation of information regarding their toxicological analysis. Basically, it is a tool that help you to select the suitable biological sample and the right collecting time, as well as the proper analysis technique, depending on the purpose of analysis, pharmacokinetic characteristics of the drugs of abuse, available equipment and staff expertise

Therapeutic Drug Monitoring and Methods of Quantitation for Carbamazepine

Carbamazepine is an early anticonvulsant still used today in the treatment of several forms of epilepsy. An active metabolite in the human body contributes to its pharmacological effect. Carbamazepine metabolism has high inter-individual variability, such that it is relatively difficult to establish a direct link between dose and concentration, or between concentration and pharmacological effect. Carbamazepine is thus a good candidate for therapeutic drug monitoring (TDM). Good UV specific absorbance and high plasmatic concentrations allow for the use of UV detection, which is often more accessible than other methods of detection. This paper presents several methods used for the detection of carbamazepine in plasma, methods that are capable of detecting drug and metabolites at adequate levels/ acceptance criteria. These methods have possible application not only in pharmacokinetic, bioequivalence, and permeability studies, but also in the therapeutic drug monitoring of carbamazepine

HPTLC assay of nicotine and cotinine in biological samples

This study presents the development of a simple high-performance thin layer chromatography (HPTLC) method for the determination of nicotine and its metabolite cotinine in human plasma and urine. The following mobile phases: methanol: ammonia (100:1.5, v:v), chloroform: acetone: ammonia (48.75: 48.75: 2.5, v:v:v), methanol: chloroform: ammonia (48.75: 48.75: 0.5, v:v:v) and glass plates precoated with silicagel 60 F254 (20x20) as a stationary phase were used. Densitometric scanning was performed at 263 nm. Two different extraction procedures have been applied: liquid-liquid extraction using dichloromethane at alkaline pH and solid-phase extraction using C18 cartridges. Preliminary tests in order to establish the system of solvents for development, as well as the range of linearity, were conducted. The best separation of nicotine and cotinine was obtained by using methanol: chloroform: ammonia (48.75: 48.75: 0.5, v:v:v) as the mobile phase. The liquid-liquid extraction technique led to better results than solid phase extraction. The regression curves were linear (with a corresponding correlation coefficient higher than 0.99) in the quantities range of 200 ng–1000 ng/spot for both nicotine and cotinine. The UV spectra confirm the identification of nicotine and cotinine both in the standards and in the extracts after liquid-liquid extraction. The proposed method can be applied for the simultaneous evaluation of nicotine and cotinine in biological samples at toxic/lethal levels. Thus, the method may be applicable in lethal nicotine intoxication cases in forensic toxicological analysis

Comparative cytotoxicity study of nicotine and cotinine on MRC-5 cell line

Nicotine has several health hazards regarding carcinogenic potential. It also imparts increased risk for respiratory, cardiovascular, and gastrointestinal disorders. Several mechanisms have been proposed for the carcinogenic potential, including effects on cell proliferation, inducing oxidative stress, DNA mutation, or inhibition of apoptosis. The cotinine metabolite is generally thought to have effects similar to nicotine in some experimental systems. The purpose of this study was to assess the nicotine and cotinine cytotoxicity on MRC-5 lung fibroblasts. The pulmonary fibroblasts were treated with various concentrations of nicotine or cotinine (in the range 1 µM – 2 mM) for 24 or 48 h and analyzed for cell viability by MTT test. The results indicated that high nicotine concentrations (2 mM) induced marked cell death (about 50%) in MRC-5 cell line. Cotinine showed lower toxicity than nicotine on the MRC-5 cells. In contrast to nicotine treatment, cells treated with cotinine continued to proliferate after the 48h incubation period

Integrating Ecosystem Services in Historically Polluted Areas: Bioremediation Techniques for Soils Contaminated by Heavy Metals

Bioremediation of soils contaminated by heavy metals is based on the use of specially selected plants able to reduce the hazards of toxic metals. Depending on the mode of action on the heavy metals existing in the soil and the place where the action takes place, the following mechanisms for soil phytoremediation are distinguished: phytostabilization, phytoextraction, phytoimobilization, rhizofiltration, or evapotranspiration. These mechanisms are complex and include the plant ability to reduce the mobility and bioavailability of heavy metals and other pollutants, to extract large amounts of heavy metals from the soil or to evaporate water together with various pollutants already reached in the rhizosphere. Decontamination of polluted soils by using bioaccumulative plants is proposed as an environmental-friendly alternative to the traditional physicochemical methods, being a sustainable method with a great potential in the terms of environmental protection and cost management

Methadone Treatment for Heroin Dependence

In substitution therapy for treatment of heroin addiction, methadone is the synthetic opioid agonist of first choice. Methadone doses vary depending on addict profile established by repeated evaluation. It studied a group of 82 patients both male and female, aged between 19 and 47 years, residing in Bucharest, with diagnosis of heroin addiction. They were voluntarily submitted in the methadone substitution treatment at a specialized treatment center for addiction in Bucharest. The study group was characterized in detail, taking into account demographic, comorbid and addiction characteristics, heroin use history, treatment history, and clinical and paraclinical evaluation. The outcomes resulting from the study design on 82 heroin addict patients enrolled into a methadone maintenance program highlighted: lowering of the onset age of heroin use, HVC infection comorbidity, and the extension of the treatment period due to the relapses. The results obtained by clinical, laboratory, and psychological complex evaluations in a correlative approach is essential both in initiating methadone treatment and monitoring the detox period but also in the supervision of methadone maintenance treatment

HPTLC assay of nicotine and cotinine in biological samples

This study presents the development of a simple high-performance thin layer chromatography (HPTLC) method for the determination of nicotine and its metabolite cotinine in human plasma and urine. The following mobile phases: methanol: ammonia (100:1.5, v:v), chloroform: acetone: ammonia (48.75: 48.75: 2.5, v:v:v), methanol: chloroform: ammonia (48.75: 48.75: 0.5, v:v:v) and glass plates precoated with silicagel 60 F254 (20x20) as a stationary phase were used. Densitometric scanning was performed at 263 nm. Two different extraction procedures have been applied: liquid-liquid extraction using dichloromethane at alkaline pH and solid-phase extraction using C18 cartridges. Preliminary tests in order to establish the system of solvents for development, as well as the range of linearity, were conducted. The best separation of nicotine and cotinine was obtained by using methanol: chloroform: ammonia (48.75: 48.75: 0.5, v:v:v) as the mobile phase. The liquid-liquid extraction technique led to better results than solid phase extraction. The regression curves were linear (with a corresponding correlation coefficient higher than 0.99) in the quantities range of 200 ng–1000 ng/spot for both nicotine and cotinine. The UV spectra confirm the identification of nicotine and cotinine both in the standards and in the extracts after liquid-liquid extraction. The proposed method can be applied for the simultaneous evaluation of nicotine and cotinine in biological samples at toxic/lethal levels. Thus, the method may be applicable in lethal nicotine intoxication cases in forensic toxicological analysis

Psychological and psychiatric characterization of various groups of drugs users

Aim. We aimed to assess the differences among various groups of drugs users, especially in the psychiatric and psychological domains. Materials and Methods. A retrospective study was carried out in collaboration with C.E.T.T.T `St. Stelian` Institute from Bucharest. There were analyzed the medical records of 604 hospitalized patients with heroin or polydrug addiction. Results. Significant differences in diagnosis at submission among groups were outlined (personality and behavior disorders, p-value = .04298, psychotic disorders, p-value = .004274, schizophrenia, p-value = .000141) as well as significant differences among psychiatric parameters: perception (legal highs, opiates), attention (cannabis), consciousness (legal highs), thinking (legal highs), and, instinctive life (legal highs). Conclusions. Personality and behavioral disorders have been particularly linked to opiate use, the psychotic disorder was related to cannabis and legal highs intake, while schizophrenia was related to legal highs intake