Correlation between the Skin Permeation Profile of the Synthetic Sesquiterpene Compounds, Beta-Caryophyllene and Caryophyllene Oxide, and the Antiedematogenic Activity by Topical Application of Nanoemulgels.

Sesquiterpene compounds are applied as permeation promoters in topical formulations. However, studies exploring their impact on nanostructured systems, changes in permeation profile, and consequently, its biological activity are restricted. This study aimed to investigate the correlation between the skin permeation of the major sesquiterpenes, beta-caryophyllene, and caryophyllene oxide from the oleoresin of Copaifera multijuga, after delivery into topical nanoemulgels, and the in vivo antiedematogenic activity. First, ten nanoemulgels were prepared and characterized, and their in vitro permeation profile and in vivo anti-inflammatory activity were evaluated. In equivalent concentrations, ß-caryophyllene permeation was greater from oleoresin nanoemulgels, resulting in greater in vivo antiedematogenic activity. However, an inverse relationship was observed for caryophyllene oxide, which showed its favored permeation and better in vivo anti-inflammatory effect carried as an isolated compound in the nanoemulgels. These results suggest that the presence of similar compounds may interfere with the permeation profile when comparing the profiles of the compounds alone or when presented in oleoresin. Furthermore, the correlation results between the permeation profile and in vivo antiedematogenic activity corroborate the establishment of beta-caryophyllene as an essential compound for this pharmacological activity of C. multijuga oleoresin.

Lisdexamfetamine and amphetamine pharmacokinetics in oral fluid, plasma, and urine after controlled oral administration of lisdexamfetamine.

Lisdexamfetamine (LDX) is a long-acting prodrug stimulant indicated for the treatment of attention-deficit/hyperactivity disorder (ADHD) and binge-eating disorder (BED) symptoms. In vivo hydrolysis of the LDX amide bond releases the therapeutically active d-amphetamine (d-AMPH). This study aims to describe the pharmacokinetics of LDX and its major metabolite d-AMPH in human oral fluid, urine and plasma after a single 70 mg oral dose of LDX dimesylate. Six volunteers participated in the study. Oral fluid and blood samples were collected for up to 72 h and urine for up to 120 h post-drug administration for the pharmacokinetic evaluation of intact LDX and d-AMPH. Samples were analyzed by LC-MS/MS. Regarding noncompartmental analysis, d-AMPH reached the maximum concentration at 3.8 and 4 h post-administration in plasma and oral fluid, respectively, with a mean peak concentration value almost six-fold higher in oral fluid. LDX reached maximum concentration at 1.2 and 1.8 h post-administration in plasma and oral fluid, respectively, with a mean peak concentration value almost three-fold higher in plasma. Intact LDX and d-AMPH were detected in the three matrices. The best fit of compartmental analysis was found in the one-compartment model for both analytes in plasma and oral fluid. There was a correlation between oral fluid and plasma d-AMPH concentrations and between parent to metabolite concentration ratios over time in plasma as well as in oral fluid.

Drug use and driving behaviors among drivers with and without alcohol-related infractions.

INTRODUCTION: Brazil is one of the countries with the highest rates of alcohol-related traffic infractions, but little is known about the profile of the drivers who commit them. Identifying the characteristics of impaired drivers is essential for planning preventive actions. OBJECTIVE: To compare drug use and driving behavior profiles of drivers with and without alcohol-related infractions. METHODS: 178 drivers stopped at routine roadblocks were assessed by traffic agents who conducted standard roadblock procedures (document verification; request of a breathalyzer test [BT]). Drug use and driving behavior data were collected through semi-structured interviews. Subjects were divided into three groups: drivers who refused the BT (RDs, n = 72), drivers who tested positive on the BT (PDs, n = 34), and drivers who had committed other infractions (ODs, n = 72). RESULTS: The proportion of alcohol use in the last year was higher among RDs (100%) than in the PD and OD groups (97.1% and 72.2% respectively, p < 0.001). Lifetime prevalence of cannabis and cocaine use for the overall sample was 44.3% and 18.2%, respectively. Fewer individuals in the OD group (31.5%) reported having been stopped at roadblocks in the previous year compared to the PDs (55.9%) and RDs (48.6%, p = 0.03). However, a higher proportion of RDs reported drunk driving in the same period (87.5%; PD 69.7%; OD 26.9%; p < 0.001). CONCLUSION: Essential differences among groups were observed. RDs had a higher proportion of alcohol use and drunk driving in the previous year; drivers who fit into this particular group may be unresponsive or less responsive to social deterrence and enforcement actions.

Drug use and driving behaviors among drivers with and without alcohol-related infractions

Abstract Introduction Brazil is one of the countries with the highest rates of alcohol-related traffic infractions, but little is known about the profile of the drivers who commit them. Identifying the characteristics of impaired drivers is essential for planning preventive actions. Objective To compare drug use and driving behavior profiles of drivers with and without alcohol-related infractions. Methods 178 drivers stopped at routine roadblocks were assessed by traffic agents who conducted standard roadblock procedures (document verification; request of a breathalyzer test [BT]). Drug use and driving behavior data were collected through semi-structured interviews. Subjects were divided into three groups: drivers who refused the BT (RDs, n = 72), drivers who tested positive on the BT (PDs, n = 34), and drivers who had committed other infractions (ODs, n = 72). Results The proportion of alcohol use in the last year was higher among RDs (100%) than in the PD and OD groups (97.1% and 72.2% respectively, p < 0.001). Lifetime prevalence of cannabis and cocaine use for the overall sample was 44.3% and 18.2%, respectively. Fewer individuals in the OD group (31.5%) reported having been stopped at roadblocks in the previous year compared to the PDs (55.9%) and RDs (48.6%, p = 0.03). However, a higher proportion of RDs reported drunk driving in the same period (87.5%; PD 69.7%; OD 26.9%; p < 0.001). Conclusion Essential differences among groups were observed. RDs had a higher proportion of alcohol use and drunk driving in the previous year; drivers who fit into this particular group may be unresponsive or less responsive to social deterrence and enforcement actions.

Comparison between counterfeit and authentic medicines: A novel approach using differential scanning calorimetry and hierarchical cluster analysis.

Erectile dysfunction medicines such as Cialis and Viagra are very popular worldwide and are between the most prevalent counterfeit medicines in Brazil. A range of analytical methods has been used to analyze Cialis and Viagra, such as ATR-FTIR, GCMS and UPLC-MS. Until now, there are no data available of DSC methods for analysis of counterfeit medicines of Cialis and Viagra. DSC is a thermal analysis that provides useful information of physico-chemical events, and however is almost not used for forensic purposes. In this study, thermal analysis of 25 counterfeit Viagra and Cialis seized by Brazilian Federal Police were performed by DSC and compared to their authentic medicines and analytical standards, along with chemometric tools. Authentic samples of Viagra displayed a similar thermal profile with the API, while Cialis were different with additional endothermic peaks, that could be related to excipients interference. Thermograms of Viagra counterfeit samples were similar to authentic samples, while Cialis showed an enlargement and displacement of endothermic peaks. Also, some Cialis counterfeit samples showed melting peaks attributed to sildenafil, the API of Viagra, instead tadalafil, confirming previous results obtained by UPLC-MS. Multivariate analysis with application of Hierarchical Cluster Analysis classified different groups of samples, including a cluster with counterfeit Cialis and Viagra, indicating the use of same API for both counterfeit medicines and possibly the same illicit production; and a cluster with authentic Viagra and counterfeit Cialis, confirming the addition of sildenafil instead tadalafil to Cialis counterfeit samples. Here for the first time we described the use of DSC for chemical profiling of Cialis and Viagra and showed that even when applied to a small group of samples, DSC along with chemometric tools can be considered as a good auxiliary method in forensic casework samples. DSC provided useful data to perform the identification of counterfeit and authentic medicines, with low cost and a simple method.

Optimization, validation and application of headspace solid-phase microextraction gas chromatography for the determination of 1-nitro-2-phenylethane and methyleugenol from Aniba canelilla (H.B.K.) Mez essential oil in skin permeation samples.

Aniba canelilla (H.B.K.) Mez is an aromatic plant from the Amazon region whose essential oil has 1-nitro-2-phenylethane (NP) and methyleugenol (ME) as major compounds. Despite of the scientifically proven antifungal and anti-inflammatory activities for these compounds, there is no report up to date about the topical permeation or quantification of NP and ME on skin samples. The aim of this study was the validation of an optimized bioanalytical method by solid-phase microextraction in headspace mode in gas chromatograph with flame ionization detector (HS-SPME-GC-FID) for the determination of NP and ME from the oil in different samples from permeation study, such as porcine ear skin (PES) layers (stratum corneum, epidermis and dermis) and receptor fluid (RF). For this propose polydimethylsiloxane fibers (100 µm) were used and HS-SPME extraction condition consisted of 53 °C, 21 min, and 5% w.v-1 NaCl addition. The wide range of the calibration curve (2.08-207.87 µg mL-1 for NP and 0.40-40.41 µg mL-1 for ME), the presence of matrix interferences and the intrinsic characteristics of HS-SPME required a data linearization using Log10. Thereby, data and the gained results presented homoscedasticity, normalization of residues and adequate linearity (r2 > 0.99) and accuracy for both compounds. In order to verify the applicability of the validated method, the HS-SPME-GC-FID procedure was performed to determine the amount of NP and ME permeated and retained in samples after Franz diffusion cell study from different dosages (20, 100 and 200 µL) of A. canelilla oil. Compounds permeation showed a progressive increase and penetration dependence based on the dosage applied. Furthermore, retention was in order receptor fluid >> dermis >> epidermis >> stratum corneum for both compounds, suggesting NP and ME could penetrate deep tissue, probably due to the partition coefficient, mass, size, and solubility of these compounds. In conclusion, the proposed method by HS-SPME-GC-FID to quantify 1-nitro-2-phenylethane and methyleugenol from Aniba canelilla essential oil was able to determine selectively, precisely and accurately these main compounds in skin permeation samples.

Anti-inflammatory Effect from a Hydrogel Containing Nanoemulsified Copaiba oil (Copaifera multijuga Hayne).

Copaiba oil is used as a popular medicine in the Amazonian forest region, especially due to its anti-inflammatory properties. In this paper, we describe the formulation of hydrogel containing copaiba oil nanoemulsions (with positive and negative charges), its skin permeation, and its anti-inflammatory activity in two in vivo models: mouse ear edema and rat paw edema. Three hydrogels were tested (Carbopol®, hydroxyethylcellulose and chitosan), but only Carbopol® and hydroxyethylcellulose hydrogels presented good stability and did not interfere with the nanoemulsions droplet size and polydispersity index. In skin permeation assay, both formulations, positively charged nanoemulsion (PCN) and negatively charged nanoemulsion (NCN), presented a high retention in epidermis (9.76 ± 2.65 µg/g and 7.91 ± 2.46 µg/cm2, respectively) followed by a smaller retention in the dermis (2.43 ± 0.91 and 1.95 ± 0.56 µg/cm2, respectively). They also presented permeation to the receptor fluid (0.67 ± 0.22 and 1.80 ± 0.85 µg/cm2, respectively). In addition, anti-inflammatory effect was observed to NCN and PCN with edema inhibitions of 69 and 67% in mouse ear edema and 32 and 72% in rat paw edema, respectively. Histological cuts showed the decrease of inflammatory factors, such as dermis and epidermis hyperplasia and inflammatory cells infiltration, confirming the anti-inflammatory effect from both copaiba oil nanoemulsions incorporated in hydrogel.

Determination of ß-caryophyllene skin permeation/retention from crude copaiba oil (Copaifera multijuga Hayne) and respective oil-based nanoemulsion using a novel HS-GC/MS method.

Copaiba oil is largely used in the Amazonian region for the treatment of inflammation, and recent studies demonstrated that one of the major components of the oil, ß-caryophyllene (CAR), is a potent anti-inflammatory. The nanoemulsification of this oleoresin, which has unctuous character, converts it in a more acceptable hydrophilic formulation and may improve CAR penetration through the skin due to the small droplet size and the high contact surface afforded by the nanoemulsions. This paper describes the validation of a novel, sensitive, practical and solvent free method that uses gas chromatography in headspace mode coupled with mass spectrometry to evaluate the skin permeation/retention of CAR from the crude copaiba oil and its nanoemulsion. Our results show that the bioanalytic method was fully validated, demonstrating linearity (r(2)>0.99), specificity (no peaks co-eluting with CAR retention time), precision (RSD<15%) and accuracy (recovery>90%) within the accepted parameters and that the copaiba oil nanoemulsion presented a better skin penetration compared to the crude oil, with CAR achieving the most profound layer of the skin, the dermis.

Simultaneous analysis of amphetamine-type stimulants in plasma by solid-phase microextraction and gas chromatography-mass spectrometry.

Brazil is considered one of the countries with the highest number of amphetamine-type stimulant (ATS) users worldwide, mainly diethylpropion (DIE) and fenproporex (FEN). The use of ATS is mostly linked to diverted prescription stimulants and this misuse is widely associated with (ab)use by drivers. A validated method was developed for the simultaneous analysis of amphetamine (AMP), DIE and FEN in plasma samples employing direct immersion-solid-phase microextraction, and gas chromatographic/mass spectrometric analysis. Trichloroacetic acid 10% was used for plasma deproteinization. In situ derivatization with propylchloroformate was employed. The linear range of the method covered from 5.0 to 100 ng/mL. The detection limits were 1.0 (AMP), 1.5 (DIE) and 2.0 ng/mL (FEN). The accuracy assessment of the control samples was within 85.58-108.33% of the target plasma concentrations. Recoveries ranged from 46.35 to 84.46% and precision was <15% of the value of relative standard deviation. This method is appropriate for screening and confirmation in plasma forensic toxicology analyses of these basic drugs.

A comparison of alcohol and drug use by random motor vehicle drivers in Brazil and Norway.

BACKGROUND: A large proportion of road traffic crashes are related to driving under the influence (DUI) of alcohol or drugs. The aim of this study was to compare the use of alcohol, illegal drugs and psychoactive medicinal drugs among random drivers in Brazil and Norway, two countries with the same legal limit for drunk driving, but with marked differences in legislation history, enforcement and penalties for DUI, and to discuss any differences found. METHODS: Roadside surveys were conducted on Fridays and Saturdays between noon and midnight. Samples of oral fluid were collected for analysis of drugs, whereas alcohol was determined by breath testing or by analysis of oral fluid. RESULTS: High participation rates of 94-97% were obtained in both countries. The weighted prevalence of driving with alcohol concentrations in breath or oral fluid equivalent to blood alcohol concentrations (BAC) above 0.2g/L was 2.7% (95% CI 2.2-3.3) in Brazil and 0.2% (95% CI 0.0-0.5) in Norway. Stimulants (amphetamines or cocaine) were found in samples from 1.0% (95% CI 0.7-1.4) of drivers in Brazil and 0.3% (95% CI 0.1-0.7) in Norway. The prevalence of amphetamines was highest among Brazilian truck drivers (3.6%; 95% CI 2.0-6.4). Tetrahydrocannabinol was found in samples from 0.5% (95% CI 0.3-0.8) of drivers in Brazil and 1.0% (95% CI 0.6-1.5) in Norway, whereas benzodiazepines or zopiclone were found in 1.0% (95% CI 0.7-1.4) and 1.7% (95% CI 1.2-2.4) of the samples from Brazil and Norway, respectively. CONCLUSIONS: The difference in the prevalence of alcohol may be related to the fact that Norway has implemented steps to reduce drunk driving since 1936, whereas Brazil has attempted to do the same for only a few years. Differences for drugs may be related to different patterns in the use of stimulants, cannabis and medicines.

Drug use among drivers who drank on alcohol outlets from Porto Alegre, Brazil.

BACKGROUND: Driving under the influence of multiple substances is a public health concern, but there is little epidemiological data about their combined use and putative impact on driving in low and middle-income countries where traffic crashes have been clustering in recent years. The aim of this study is to estimate the prevalence of alcohol and drug use - as well as their associated factors - among drivers in the context of alcohol outlets (AOs). METHODS: A probability three-stage sample survey was conducted in Porto Alegre, Brazil. Individuals who were leaving AO were screened, with the selection of 683 drivers who met the inclusion criteria. Drivers answered a structured interview, were breathalyzed, and had their saliva collected for drug screening. Prevalences were assessed using domain estimation and logistic regression models assessed covariates associated with substance use. FINDINGS: Benzodiazepines 3.9% (SE 2.13) and cocaine 3.8% (SE 1.3) were the most frequently detected drugs in saliva. Among drivers who were going to drive, 11% had at least one drug identified by the saliva drug screening, 0.4% two, and 0.1% three drugs in addition to alcohol. In multivariable analyses, having a blood alcohol concentration (BAC)>0.06% was found to be associated with a 3.64 times (CI 95% 1.79-7.39) higher chance of drug detection, compared with interviewees with lower BACs. CONCLUSIONS: To drive under the influence of multiple substances is likely to be found in this setting, highlighting an association between harmful patterns of consume of alcohol and the misuse of other substances.

Profiling counterfeit Cialis, Viagra and analogs by UPLC-MS.

In this work, the chemical profile of 43 commercial samples of tablets for male erectile dysfunction (Viagra, Cialis, Lazar, Libiden, Maxfil, Plenovit, Potent 75, Rigix, Vimax, Pramil 75 and Pramil) and 65 counterfeit samples (Viagra and Cialis) were obtained from UPLC-MS data. Methanol extracts of crushed tablets were investigated by ultra performance liquid chromatography (UPLC) with diode array detection (DAD) coupled with eletrospray ionization in the positive ion mode (ESI(+)) quadrupole time-of-flight (Q-Tof) mass spectrometry (MS). A validated method was employed for the simultaneous determination of sildenafil citrate (SLD) and tadalafil (TAD). The ultra-chromatograms obtained with method provide high resolution of MS, and are a quick (less to 1.5 min) and reliable tool in the distinction between authentic and counterfeit tablets. It was observed in most cases the presence of other active pharmaceutical ingredients (APIs) than specified on the package (TAD and SLD). Additionally, high concentrations of TAD and SLD were detected in counterfeit samples when compare with observed values for a typical commercial product. Chemometric methods were employed and the samples were grouped in five groups as function of API content.

Trends in counterfeits amphetamine-type stimulants after its prohibition in Brazil.

Brazil is one of the world's highest users of anorectic drugs, mainly diethylpropione, fenproporex and sibutramine. The present work focuses on physical and chemical characteristics of 17 counterfeited capsules containing amphetamine-type stimulants (ATS) from three seizures conducted by Brazilian Federal Police. The physical profile was useful in indicating forgery, bring complementary information, but the use of this data singly was not sufficient to distinguish between authentic and counterfeited medicines. The chemical analysis revealed that the seizures capsules labeled as Desobesi-M (fenproporex 25mg), actually contained the active pharmaceutical ingrediente (API) sibutramine. The amount of this API ranged from 1/3 to 2 times the amount of drug found in commercial product, may reach twice the recommended daily dose. Multivariate analysis with application of principal component analysis on data from spectroscopy attenuated total reflectance Fourier transform infrared classified the samples according to their similarities, indicating that two seizures had common origin. This study represents the first step in the elucidation of falsification of ATS in Brazil. Considering the forensic intelligence these information are valuable in order to develop and establish a database that enables correlate samples from different locations and/or suppliers and to map the profile and trends of trafficking.

Citotoxic activity evaluation of essential oils and nanoemulsions of Drimys angustifolia and D. brasiliensis on human glioblastoma (U-138 MG) and human bladder carcinoma (T24) cell lines in vitro

The species Drimys angustifolia Miers and D. brasiliensis Miers, commonly known as "casca-de-anta", have in their leaves essential oils that can confer cytotoxic effects. In this study, we evaluated the citotoxic effects of the volatile oils from these two species. We also proposed a nanoemulsion formulation for each of the species and assessed the in vitro cytotoxicity on U-138 MG (human glioblastoma) and T24 (human bladder carcinoma) cell lines. The plant chemical composition was evaluated by gas chromatography coupled to mass spectrometer. Furthermore, the nanoemulsions were prepared and characterized. Our results showed that; bicyclogermacrene (19.6%) and cyclocolorenone (18.2%) were the most abundant for the D angustifolia oil and D brasiliensis oil, respectively. Both nanoemulsions, D angustifolia and D brasiliensis appeared macroscopically homogeneous and opalescent bluish liquids, with nanometric mean diameters of 168 nm for D brasiliensis and 181 nm for D angustifolia. The polydispersity indices were below 0.10, with an acid pH of 4.7-6.3, and negative zeta potentials about -34 mV. The results of transmission electron microscopy showed that droplets are present in the nanometer range. Only the D brasiliensis oil was efficient in reducing the cell viability of both U-138 MG (42.5%±7.0 and 67.8%±7.8) and T24 (33.2%±2.8, 60.3%±1.6 and 80.5%±8.8) cell lines, as assessed by MTT assay. Noteworthy, similar results were obtained with cell counting. Finally, D brasiliensis oil incubation caused an increase of annexin-V and propidium iodite population, according to evaluation by cytometry analysis, what is characteristic of late apoptosis. The results presented herein lead us to consider the potential therapeutic effects of the essential oils and nanoformulations as novel strategies to inhibit tumor growth.

Counterfeit Cialis and Viagra fingerprinting by ATR-FTIR spectroscopy with chemometry: can the same pharmaceutical powder mixture be used to falsify two medicines?

This paper proposes a direct and efficient method to discriminate between counterfeit and authentic Cialis and Viagra samples by combining attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy with multivariate techniques. The chemical profile of 53 commercial samples (Viagra(®), Cialis(®)) and 104 counterfeit samples (Viagra and Cialis) from distinct seizures were obtained from ATR-FTIR data derived from 10mg of crushed core tablets. Principal component analysis (PCA) technique was employed to classify samples based on the fingerprint region mid-infrared spectra (1800-525 cm(-1)) using OMNIC v.7.2 software; PCA enabled categorizing samples in groups with different chemical profiles, successfully distinguishing between authentic and counterfeits samples in forensic routine. The existence of active pharmaceutical ingredients (API) and technological adjuvant others than specified on the medicine package were also detected in counterfeit samples. In addition, we applied the similarity match (SM) method to demonstrate that a mixture of pharmaceutical powders deriving from a common origin may have been used to manufacture both counterfeit Cialis and Viagra tablets from distinct seizures.

Amphetamine-type medicines: a review of pharmacokinetics, pharmacodynamics, and toxicological aspects.

Amphetamine-like drugs are sympathomimetic agents with marked central and peripheral stimulant properties. Despite the street illegal drugs such as amphetamine and ecstasy, some amphetamine-like compounds are also legally marketed under medical prescription in the treatment of attention deficit-hyperactivity disorder (methylphenidate) and obesity/overweight (fenproporex and diethylpropione). However, similar with what happens with their illicit analogues, therapeutic amphetamine-like drugs also share important toxicological risks. Although methylphenidate is considered the first choice in the treatment of attention deficit-hyperactivity disorder, its high popularity among teenagers and children is raising concern in the medical community. Regarding weight-loss purposes, the use of amphetamine-like compounds are very controversial, though. Thus, the present review will address pharmacokinetic, pharmacodynamic, and toxicological aspects of amphetamine-like compounds used with therapeutic aims.

Isoflurane-loaded nanoemulsion prepared by high-pressure homogenization: investigation of stability and dose reduction in general anesthesia.

Isoflurane is a halogenated ether which is used for general anesthesia. To stabilize a new formulation in order to evaluate the potential to reduce the dose required for general anesthesia, an isoflurane-loaded nanoemulsion was proposed. A high-pressure homogenization technique was used to develop drug-loaded nanoemulsions which presented droplet size of 150 +/- 0.78 nm with a narrow size distribution and low polydispersity index (0.08 +/- 0.01). The zeta potential was -18 +/- 2.4 mV and pH was 6.03 +/- 0.04. Rheological analysis showed Newtonian behavior for the formulations, whose physical stability was confirmed by multiple light scattering. It was verified that isoflurane volatilization did not occur in these formulations. The preclinical evaluation, carried out via the end-tidal isoflurane concentration, showed that the dose required for anesthetic maintenance significantly decreased when the nanostructured formulation was administered compared to inhaled isoflurane. There was no significant difference (p < 0.05) between experimental groups (inhaled isoflurane and intravenous isoflurane-loaded nanoemulsion) in terms of the cardiac rate, oxygen hemoglobin saturation, and arterial blood pressure, as well as the biomarkers of renal, hepatic and skeletal muscle system functionalities. Slight tachypnea, edema, and erythema were observed after isoflurane-loaded or unloaded-nanoemulsion. The stability and significant dose reduction observed for drug-loaded nanoemulsion render this formulation a promising option for intravenous delivery of isoflurane.

Toxicological effects of a mixture used in weight loss products: p-synephrine associated with ephedrine, salicin, and caffeine.

p-Synephrine is an adrenergic amine found in Citrus aurantium L. fruits and has been used for weight loss in dietary supplements. There are commercial products containing this substance associated to caffeine, salicin, and ephedrine. The aim of this study was to evaluate the acute toxicity of this mixture in mice of both sexes. The significative results observed after acute oral administration to male and female mice of 300, 350, and 400 mg/kg total of p-synephrine, ephedrine, salicin, plus caffeine in a 10:4:6:80 w/w ratio included a reduction in locomotor activity and ptosis in all treated groups for both sexes. Seizures were also observed in male (400 mg/kg) and female groups (350 and 400 mg/kg). Gasping and tearing were observed in males. Salivation (400 mg/kg), agitation (350 and 400 mg/kg), and piloerection (all treated groups) were significantly observed only in females. Deaths occurred in males at 350 and 400 mg/kg treated groups and the necropsy showed cardiopulmonary hemorrhage. A reduction in locomotor activity was confirmed through the spontaneous locomotor activity test, in which the number of crossings considerably decreased (P < .01) in all treated groups. The rotarod test showed a decrease in motor coordination at 400 mg/kg. Body temperature decreased significantly (P < .01) in all treated groups compared to controls. The results suggested clear signs of toxicity of p-synephrine, ephedrine, salicin, and caffeine association; this toxicity augments the attentiveness on commercial products containing this mixture, given the expressive number of adverse events related to its utilization.

Optimization of headspace solid-phase microextraction for analysis of ß-caryophyllene in a nanoemulsion dosage form prepared with copaiba (Copaifera multijuga Hayne) oil.

Recent studies have shown the anti-inflammatory activity of Copaiba oils may be addressed to the high content of ß-caryophyllene, the most common sesquiterpene detected, especially in the Copaifera multijuga Hayne species. In the present study, nanoemulsions were proposed as a delivery system for copaiba oil in view to treat locally inflamed skin. This article describes the optimization and validation of a stability-indicating SPME-GC method, for ß-caryophyllene analysis in the nanoemulsions produced by high pressure homogenization. SPME methods are performed with PDMS (polydimethylsiloxane) fiber (100 µm). Three SPME parameters were evaluated by a three-level-three-factor Box-Behnken factorial design as potentially affecting the technique efficiency. According to the results obtained, the best conditions to extract ß-caryophyllene were: (i) sampling temperature of 45°C, (ii) sampling time of 20 min and (iii) no NaCl addition. Results coming from the forced degradation tests showed a reduction of ß-caryophyllene peak area when both caryophyllene methanolic solution and nanoemulsions were exposed to acid hydrolysis, UV-A irradiation, oxidative (H(2)O(2)) and thermolitic (60°C) conditions. Such reduction occurred in lower extent in the nanoemulsions, suggesting a protective effect of the formulation to ß-caryophyllene content. Since no degradation products were detected in the same retention time of ß-caryophyllene, the specificity of the method was demonstrated. The method was linear in the range of 0.14-0.68 µg mL(-1) of ß-caryophyllene (r(2)>0.999), and was also validated for precision (R.S.D.≤5.0%), accuracy (97.85-101.87%) and robustness. Finally, the method was applied to quantification of ß-caryophyllene content in the developed formulations.

Which amphetamine-type stimulants can be detected by oral fluid immunoassays?

INTRODUCTION: The use of oral fluid for monitoring drug consumption on roads has many advantages over conventional biological fluids; therefore, several immunoassays have been developed for this purpose. In this work, the ability of 3 commercial immunoassays to detect amphetamine-type stimulants (ATSs) in oral fluid was assessed. In addition, it was reviewed the main controlled ATSs available worldwide, as well as the oral fluid immunological screening tests that have been used for identifying ATSs in drivers. MATERIALS AND METHODS: The analytical specificity of amphetamine direct enzyme-linked immunosorbent assay (ELISA), methamphetamine direct ELISA (Immunalysis Corporation), and Oral-View saliva multidrug of abuse test (Alfa Scientific Designs) was evaluated using ATS-spiked oral fluid. Legislation and published articles that report the use of immunological screening tests to detect ATS consumption in conductors were reviewed, including the kit's technical information, project reports, police and drug databases. RESULTS AND DISCUSSION: Even at high concentrations, the tested assays were not able to detect methylphenidate, fenproporex, or diethylpropion, controlled ATSs legally marketed in many countries. CONCLUSIONS: This evidences the need to develop new kits that enable one to control the misuse of prescription ATSs on roads through oral fluid immunoassays.