Pseudoephedrine is an over-the-counter drug commonly used as a decongestant, but also thought to have ergogenic effects. The World Anti-Doping Agency (WADA) has prohibited large doses (> 150 μg∙ml-1) of pseudoephedrine, while the National College Athletic Association (NCAA) does not include it on the banned substance list. The purpose of this study was to examine the effect of body weight dosing of pseudoephedrine on 800-m run times of NCAA female runners. Fifteen NCAA female track runners volunteered to participate in the randomized, double blind, crossover design. In trials that were a week apart, participants were given both 2.5 mg∙kg-1 pseudoephedrine and a placebo. Ninety minutes post-ingestion, participants completed an 800-m individual time trial on an indoor track. Finishing time was recorded with an automated video timing device. Heart rate and anxiety state scores were recorded immediately after each trial. Finally, a urine sample was taken from 5 participants about 2 hr post-ingestion. Placebo and pseudoephedrine running times were compared using a iv paired t test. Heart rate and anxiety state scores were also compared using a paired t test. Fourteen runners completed both trials and one was an outlier, giving thirteen participants used for statistical analysis. Despite being dosed (144 mg ± 17 mg) well above normal therapeutic levels, there was no significant difference (p = 0.92) in 800-m times between the placebo (2:39.4 ± 9.6) and pseudoephedrine (2:39.4 ± 9.6) trials, in post-exercise heart rate (p = 0.635, pseudoephedrine = 177.9 ± 14.5 beats∙min-1, placebo = 178.4 ± 18.5 beats∙min-1), or in anxiety state levels (p = 0.650, pseudoephedrine = 38.4 ± 11.6, placebo = 38.1 ± 8.8). A 2.5 mg∙kg-1 dose of pseudoephedrine had no effect on 800-m run times in NCAA female runners, and did not raise urine levels above 150 μg∙ml-1. This raises the question as to why pseudoephedrine is a specified prohibited substance by WADA. (49 pages)

Dose tolerance and pharmacokinetic studies of pseudoephedrine sustained action capsules were performed in thirty-three adult male subjects who received either 120 mg or 150 mg capsules every twelve hours for seven consecutive days in a double-blind parallel design study. Although only one subject in the 150 mg group was discontinued prematurely from this study, a large number of side effects typical of CNS stimulation were seen. A placebo effect might account for a portion of these complaints, however symtoms evaluated as being due to drug were significantly more severe and persistent in the 150 mg group. Pulse rates showed a persistent and significant increase while systolic and diastolic blood pressure fell from the baseline values in both groups. A pharmacokinetic analysis of the pseudoephedrine plasma concentration-time data provided estimates of half-life and the volume of distribution/availability ratio. The values obtained were in good agreement with values reported by others. Half-life was not influenced by urine pH probably as a result of the narrow range of urine pHs observed in the subjects. Calculations of relative bioavailability suggest that the 120 mg capsule formulation has a 30% greater bioavailability compared to the 150 mg capsule.

The production of methamphetamine in clandestine laboratories presents a particular hazard due to the environmental hazards it poses. In addition to the dangers associated with using caustic and reactive solvents, these clandestine laboratories also have to potential to cause a fire or explosion. This danger has caused some states to redefine arson to include fires caused by the illicit manufacture of drugs. Arson investigation can be challenging due to the destructive nature of the crime. Much of the evidence that existed prior the fire can be consumed and evidence that does survive can be difficult to identify in the rubble. Despite these difficulties, methods have been developed to determine the types of accelerants present in addition to identifying illicit substances such as methamphetamine and the precursor drug pseudoephedrine. This study was designed to determine if solid phase microextraction combined with gas chromatography/mass spectrometry could be used to analyze burned samples of wood to which pseudoephedrine had been applied. In addition, an experiment was designed to determine what concentration of pseudoephedrine must be present before a fire in a controlled laboratory setting, for a detectable amount to remain. Samples were created by adding pseudoephedrine hydrochloride, either in powder form or dissolved in methanol, to blocks of Douglas Fir and exposing the surface to a flame for two minutes. Additional samples were created by adding trace amounts, i.e. microliter quantities, of pseudoephedrine standard to blocks of wood before placing them in a fire for ten minutes. A thermal degradation product of pseudoephedrine was detected in samples containing more than 15 mg of the drug. To verify that the detected product was a result of thermal degradation, 10 mg of pseudoephedrine were heated at 200 °C for one hour. The product of the thermal degradation study and the product detected following two minutes of exposure to a flame had the same retention time and mass spectrum. Therefore, it was concluded that the detected thermal degradation product may be used to indicate the presence of pseudoephedrine in a fire.

Pseudoephedrine is well established as a chiral auxiliary in the alkylation of amide enolates to form tertiary and quaternary carbon stereocenters. However, due to its facile transformation into the illegal narcotic methamphetamine, pseudoephedrine is either illegal or highly regulated in many countries, which limits its use in academic and industrial settings. To address this issue, pseudoephenamine has been developed as a replacement for pseudoephedrine in organic synthesis. This new auxiliary suffers no regulatory issues and exhibits several practical advantages over pseudoephedrine, including the high diastereoselectivities observed in alkylation reactions forming quaternary carbon stereocenters, the propensity for pseudoephenamine amides to be free-flowing crystalline solids, and the sharp, well-defined peaks that typically compose the 1H NMR spectra of these amides. / Chemistry and Chemical Biology

R-phenylacetylcarbinol (PAC) is used as a precursor for production of ephedrine and pseudoephedrine, which are anti-asthmatics and nasal decongestants. PAC is produced from benzaldehyde and pyruvate mediated by pyruvate decarboxylase (PDC). A strain of Rhizopus javanicus was evaluated for its production of PDC. The morphology of R. javanicus was influenced by the degree of aeration/agitation. A relatively high specific PDC activity (328 U decarboxylase g-1 mycelium) was achieved when aeration/agitation were reduced significantly in the latter stages of cultivation. The stability of partially purified PDC and crude extract from R. javanicus were evaluated by examining the enzyme deactivation kinetic in various conditions. R. javanicus PDC was less stable than Candida utilis PDC currently used in our group. A kinetic model for the deactivation of partially purified PDC extracted from C. utilis by benzaldehyde (0?00 mM) in 2.5 M MOPS buffer has been developed. An initial lag period prior to deactivation was found to occur, with first order dependencies of PDC deactivation on exposure time and on benzaldehyde concentration. A mathematical model for the enzymatic biotransformation of PAC and its associated by-products has been developed using a schematic method devised by King and Altman (1956) for deriving the rate equations. The rate equations for substrates, product and by-products have been derived from the patterns for yeast PDC and combined with a deactivation model for PDC from C. utilis. Initial rate and biotransformation studies were applied to refine and validate a mathematical model for PAC production. The rate of PAC formation was directly proportional to the enzyme activity level up to 5.0 U carboligase ml-1. Michaelis-Menten kinetics were determined for the effect of pyruvate concentration on the reaction rate. The effect of benzaldehyde on the rate of PAC production followed the sigmoidal shape of the Monod-Wyman-Changeux (MWC) model. The biotransformation model, which also included a term for PDC inactivation by benzaldehyde, was used to determine the overall rate constants for the formation of PAC, acetaldehyde and acetoin. Implementation of digital pH control for PAC production in a well-stirred organic-aqueous two-phase biotransformation system with 20 mM MOPS and 2.5 M dipropylene glycol (DPG) in aqueous phase resulted in similar level of PAC production [1.01 M (151 g l-1) in an organic phase and 115 mM (17.2 g l-1) in an aqueous phase after 47 h] to the system with a more expensive 2.5 M MOPS buffer.

Conselho Nacional de Desenvolvimento Científico e Tecnológico / Ebastine is a second generation antihistaminic clinically used for the treatment of allergic rhinitis and chronic urticaria. Pseudoephedrine hydrochloride is a direct- and indirectacting sympathomimetic, commonly combined with other drugs for their decongestant effect. In Brazil, this drugs association is available in capsules. In the literature, as well as in the official compendium, there are no methods for simultaneous analysis of ebastine and pseudoephedrine hydrochloride in pharmaceutical formulations. In the present work, liquid chromatography (HPLC) and derivative spectrophotometry (UVD) methods were developed and validated for quantification of this drugs association, in capsules. The HPLC analysis were performed on a C18 column, using a mobile phase composed of methanol: acetonitrile: ammonium acetate buffer pH 6.8 (85:5:15, v/v), run at a flow rate of 1,0 mL.min-1, with UV detection at 254 nm. In the UVD method, ebastine was quantified in the first derivative (dA/dλ), at 263.5 and 252.8 nm, respectively. All of them were validated in the following parameters: linearity, precision and accuracy. The specificity was evaluated in the HPLC assay by stress testing. The methods showed good linarity (r>0.99), precision (RSD<2%) and accuracy; the results, statistically compared, did not show significant difference (p>0,05). / A ebastina é um fármaco anti-histamínico de segunda geração utilizado clinicamente no tratamento da rinite alérgica e urticária crônica. O cloridrato de pseudoefedrina é um agente simpaticomimético de ação direta e indireta, muito utilizado em associação com outros fármacos pelo seu efeito descongestionante. No mercado brasileiro, encontra-se a associação desses dois fármacos disponível na forma de cápsulas. Na literatura, bem como em compêndios oficiais, não são descritos métodos para análise simultânea de ebastina e cloridrato de pseudoefedrina em formulações farmacêuticas. No presente trabalho, métodos por cromatografia líquida de alta eficiência (CLAE) e por espectrofotometria derivada (UVD) foram desenvolvidos e validados para quantificação simultânea desses fármacos, em cápsulas. As análises por CLAE foram realizadas em coluna C18, utilizando fase móvel composta de metanol: acetonitrila: tampão acetato de amônio pH 6,8 (80:5:15, v/v), eluída isocraticamente a 1,0 mL.min-1, com detecção UV em 254 nm. No método por UVD, a ebastina e o cloridrato de pseudoefedrina foram quantificados na primeira derivada (dA/dλ), em 263,5 e 252,8 nm, respectivamente. Ambos os métodos foram validados frente aos parâmetros de linearidade, precisão e exatidão. A especificidade do método por CLAE foi avaliada através do teste de estresse. Os métodos mostraram boa linearidade (r>0,99), precisão (DPR<2%) e exatidão. Os resultados obtidos por CLAE e UVD foram comparados estatisticamente e não apresentaram diferença significativa (p>0,05).

A successful supercritical fluid extraction method includes removal of the analyte from the matrix into the bulk fluid as well as trapping or concentration of the analyte prior to analysis. In the first phase of this research, the trapping capacities of three solid-phase traps (glass beads, 50/50 (w/w) glass beads/octadecylsilica), 50/50 (w/w) Porapak Q®/glass beads) were determined as a function of trap composition for a mixture of components varying in polarity and volatility. The Porapak Q®/glass beads mixture was found to be the most successful solidphase investigated exhibiting the highest trapping capacity. The use of the Porapak Q®/glass beads as a solid-phase trap was investigated in later extraction studies in this dissertation. The extraction of highly polar, multifunctional analytes may not be completely successful with modified carbon dioxide, therefore, a secondary modifier (i.e. additive) may be added directly to the extraction fluid in hopes of improving the recoveries. In the second phase of this research, the effect of secondary modifiers in the subcritical fluid extraction of lovastatin from in-house prepared tablet powder mixtures and MEVACOR® tablets was investigated. The effect of in-line methanol-modifier percentage, additive type (acidic, basic, neutral) to the in-line methanol, and additive concentration on the extraction efficiency were examined. The extraction recoveries of lovastatin from MEVACOR® tablets were shown to be highly dependent on methanol concentration and additive type. Isopropylamine was shown to be the most successful additive investigated. An optimized and reproducible extraction method was developed. The extraction of ionic compounds with carbon dioxide may be difficult due to the high polarity of the compounds. In the third phase of this research, the addition of ion-pairing additives to the matrix in hopes of forming an ion-pair complex of reduced analyte polarity was investigated. Therefore, a screening study consisting of a fractional-factorial design was performed in order to identify the factors which contribute most to the recovery of an anionic species, triphenylphosphinetrisulfonate (TPPTS), from a spiked-sand surface employing supercritical fluid extraction with carbon dioxide. The experimental parameters investigated were: type of ion-pairing additive (i.e. tetralkylammonium hydrogen sulfates) and its concentration, carbon dioxide density, extraction temperature, static extraction time, CO₂ mass used, liquid CO₂ flow rate, and the volume of methanol spiked into the matrix prior to extraction. Of the eight factors investigated, four factors were identified as significantly affecting the recovery of the anionic species. They were: 1) ion-pairing reagent added to the spiked sand surface and its concentration; 2) static extraction time; and 3) volume of methanol present in the extraction vessel. The experimental parameters and settings identified as influential by the statistical approach were later shown in concert to yield 100% recovery of TPPTS from the spiked-sand. In the fourth phase, the extraction of a cationic species, pseudoephedrine hydrochloride, from spiked-sand and Suphedrine tablets, with pure and methanol-modified CO₂ was examined. Once the extraction was shown to feasible, several strategies were compared in terms of their effectiveness in enhancing the analyte's extractability. The first strategy involved the addition of ion-pairing additives. Several sodium salts of alkylsulfonic acids varying in lipophilicity and concentration were investigated. The addition of 1-heptanesulfonic acid, sodium salt, in methanol, in a 5:1 mole ratio of reagent to analyte was shown to be the most useful in recovering the drug from the spiked-sand. The second strategy considered the influence of acids and bases and other modifier compositions such as a methanol/water mixture with or without 1-heptanesulfonic acid, sodium salt, on the pseudoephedrine recovery. The recoveries obtained from the drug spiked-sand were shown to comparable in the presence of a methanol/water solution, a tetrabutylammonium hydroxide in methanol solution, and a methanol solution with 1-heptanesulfonic acid, sodium salt. Next the extraction of pseudoephedrine hydrochloride from Suphedrine tablets was performed with pure and modified CO₂. Similar to the sand-spike studies, the effect of the addition of the ion-pairing reagent and other in-cell modifiers were examined. Once again, the recoveries obtained when the matrix was in the presence of a methanol/water mixture and a methanol solution containing 1-heptanesulfonic acid, sodium salt were similar. Finally, the identity of the extracted analyte was determined via IR analyses, and it was shown that pseudoephedrine hydrochloride was indeed extractable from the tablets with in-line modified CO₂ in the absence of any in-cell modifier. In the last phase of this research, a supercritical fluid chromatographic separation with evaporative light scattering detection was developed for the separation of five phospholipids varying in polarity and ionic characteristics. Several parameters were investigated and shown to be influential in the separation. They were: 1) stationary phase composition, 2) addition of an acidic additive and its concentration, 3) mobile phase ramp rate, and 4) column outlet pressure. / Ph. D.

Goals: Prepare a case study on the transition to the new OTC medicines groups with restrictions. To evaluate the positives and negatives, which brought the introduction of the group - with OTC restrictions. Pick up generally applicable principles and specify non-functional parts of the system. Method: For data processing method was applied theoretical research. Literature was searched in the database of the Czech National Library - Klementinum, WHO database, TRIBUNE and interfaces of News portals. The instrumental case study was processed for a detailed insight into the issues. The case study is also based on the analysis of the legal framework of the Czech Republic. The documents provided by SÚKL are used for financial analysis. As another method the stakeholder analysis was used - analysis of opinions of people involved in the drug policy. Results: The case study shows that switching medicines containing PSE to the limited group of OTC to prevent producers of drugs buing drugs in Czech pharmacies, but it did not reduce the amount of drugs produced in the country. Producers started to import precursors from abroad. The whole new system established in 2009 was correct in principle, but failed in practice. State Institute for Drug Control or other stakeholders in drug policy have not been able to resolve the ensuing complications.

O Brasil ocupa posição de destaque no consumo mundial de anfetaminas, contrariamente à tendência mundial de retração. Devido aos efeitos colaterais e ao alto potencial de abuso, a produção e comercialização de anfetaminas vêm sendo controladas no mundo inteiro. Com a restrição de uso, houve um retorno a procura pelos equivalentes naturais, especialmente as efedrinas presentes em diversas especialidades farmacêuticas, utilizadas no tratamento de doenças respiratórias. São componentes de vários compostos emagrecedores, suplementos alimentares e dietéticos utilizados para perda de peso e ganho de massa muscular. Face ao uso indiscriminado e a grande incidência de resultados falso-positivos nos testes de triagem para anfetaminas por imunoensaio enzimático homogêneo, fazem-se necessários testes confirmatórios. Neste sentido, este trabalho se propôs a desenvolver um método confirmatório simples e rápido para detecção, identificação e quantificação de efedrinas (efedrina/pseudoefedrina) em amostras de urina por por cromatografia a gás / espectrometria de massas-massas (CG/EM/EM), passível de ser adotado na rotina de laboratórios de análises toxicológicas. Devido à complexidade da matriz e as peculiaridades do analito, inicialmente procedeu-se o estudo do tratamento da amostra, considerando as etapas de derivatização, extração, pré-concentração e purificação, de modo a fornecer um extrato límpido, livre de impurezas, interferentes e com melhor sensibilidade, linearidade e seletividade analítica. Os métodos de extração usados foram extração líquido-líquido (ELL), extração em fase sólida (SPE) e microextração em fase sólida (SPME). Os resultados indicaram que o reagente de derivatização ciclohexanona foi o que apresentou melhor desempenho, menor custo e promoveu maior seletividade dos diasterômeros EF/PEF em colunas normais de CG. Sendo que o método mais apropriado para a detecção e identificação de efedrinas/anfetamina por CG/EM é a SPME levando em consideração características como simplicidade, rapidez, custo, recuperação e ausência de interferentes. Entretanto, considera-se valido o uso de SPE para a quantificação, devido à possibilidade de pré-concentração do analito. / Brazil is one of the biggest amphetamine consumers in the world, going against the worldwide retraction tendency. Due to serious adverse effects and high abuse potential, the production and commercialization of amphetamines has been controlled around the world. With the restriction of its use, there was a return in the search of natural equivalents, especially the ephedrines found in many medicines utilized in the treatment of respiratory diseases. Furthermore, they are components of dietary supplements used to lose weight and muscular mass gain. Because of the indiscriminate use and the high incidence of false-positive results in the amphetamines screening tests by enzyme immunoassay technique, it is necessary confirmatory tests. In this way, the aim of this work is to develop a confirmatory simple and quickly method for the detection and quantification of ephedrines (ephedrine and pseudoephedrine) gas chromatography / mass-mass spectrometry (GC/MS/MS), with possibility to be adopted in toxicological analyses laboratorial routine. Due to the complexity of the matrix and analyte peculiarities, initially proceeds the study of sample treatment, considering the derivatization, extraction, pre-concentration and purification steps, obtaining a limpidous extract, free of impurities, interferents and with better sensitivity, linearity and analytical selectivity. The extraction method used were liquid-liquid extraction (LLE), solid-phase extraction (SPE) and solid-phase microextraction (SPME). The results indicate that cyclohexanone was the derivatization agent with the best performance, lower price and good selectivity in diasteromers EF/PEF separation in normal GC columns. The most appropriate method for detection and identification of ephedrines/amphetamine by GC/MS is SPME, considering characteristics as simplicity, speed, cost, recovery and absence of interferents. However, the use of SPE must be considered to quantification, since it allowed analyte pre-concentration.

O Brasil ocupa posição de destaque no consumo mundial de anfetaminas, contrariamente à tendência mundial de retração. Devido aos efeitos colaterais e ao alto potencial de abuso, a produção e comercialização de anfetaminas vêm sendo controladas no mundo inteiro. Com a restrição de uso, houve um retorno a procura pelos equivalentes naturais, especialmente as efedrinas presentes em diversas especialidades farmacêuticas, utilizadas no tratamento de doenças respiratórias. São componentes de vários compostos emagrecedores, suplementos alimentares e dietéticos utilizados para perda de peso e ganho de massa muscular. Face ao uso indiscriminado e a grande incidência de resultados falso-positivos nos testes de triagem para anfetaminas por imunoensaio enzimático homogêneo, fazem-se necessários testes confirmatórios. Neste sentido, este trabalho se propôs a desenvolver um método confirmatório simples e rápido para detecção, identificação e quantificação de efedrinas (efedrina/pseudoefedrina) em amostras de urina por por cromatografia a gás / espectrometria de massas-massas (CG/EM/EM), passível de ser adotado na rotina de laboratórios de análises toxicológicas. Devido à complexidade da matriz e as peculiaridades do analito, inicialmente procedeu-se o estudo do tratamento da amostra, considerando as etapas de derivatização, extração, pré-concentração e purificação, de modo a fornecer um extrato límpido, livre de impurezas, interferentes e com melhor sensibilidade, linearidade e seletividade analítica. Os métodos de extração usados foram extração líquido-líquido (ELL), extração em fase sólida (SPE) e microextração em fase sólida (SPME). Os resultados indicaram que o reagente de derivatização ciclohexanona foi o que apresentou melhor desempenho, menor custo e promoveu maior seletividade dos diasterômeros EF/PEF em colunas normais de CG. Sendo que o método mais apropriado para a detecção e identificação de efedrinas/anfetamina por CG/EM é a SPME levando em consideração características como simplicidade, rapidez, custo, recuperação e ausência de interferentes. Entretanto, considera-se valido o uso de SPE para a quantificação, devido à possibilidade de pré-concentração do analito. / Brazil is one of the biggest amphetamine consumers in the world, going against the worldwide retraction tendency. Due to serious adverse effects and high abuse potential, the production and commercialization of amphetamines has been controlled around the world. With the restriction of its use, there was a return in the search of natural equivalents, especially the ephedrines found in many medicines utilized in the treatment of respiratory diseases. Furthermore, they are components of dietary supplements used to lose weight and muscular mass gain. Because of the indiscriminate use and the high incidence of false-positive results in the amphetamines screening tests by enzyme immunoassay technique, it is necessary confirmatory tests. In this way, the aim of this work is to develop a confirmatory simple and quickly method for the detection and quantification of ephedrines (ephedrine and pseudoephedrine) gas chromatography / mass-mass spectrometry (GC/MS/MS), with possibility to be adopted in toxicological analyses laboratorial routine. Due to the complexity of the matrix and analyte peculiarities, initially proceeds the study of sample treatment, considering the derivatization, extraction, pre-concentration and purification steps, obtaining a limpidous extract, free of impurities, interferents and with better sensitivity, linearity and analytical selectivity. The extraction method used were liquid-liquid extraction (LLE), solid-phase extraction (SPE) and solid-phase microextraction (SPME). The results indicate that cyclohexanone was the derivatization agent with the best performance, lower price and good selectivity in diasteromers EF/PEF separation in normal GC columns. The most appropriate method for detection and identification of ephedrines/amphetamine by GC/MS is SPME, considering characteristics as simplicity, speed, cost, recovery and absence of interferents. However, the use of SPE must be considered to quantification, since it allowed analyte pre-concentration.