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1.
J Pharm Pharmacol ; 53(6): 849-57, 2001 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-11428661

RESUMO

Alcoholic extracts of the roots and leaves of three Echinacea species (E. purpurea, E. angustifolia and E. pallida) were analysed for the presence of characteristic chemicals by HPLC directly coupled to ultraviolet absorbance and electrospray mass spectrometric detectors. The method permitted rapid characterization and tentative identification of a large number of caffeoyl conjugates and alkamides in all the samples investigated. The roots of the three species differed markedly in their contents of characteristic compounds. Cichoric acid and verbascoside predominated in extracts of E. purpurea root whereas cynarine and dodeca-2E,4E,8Z,10Z/E-tetraenoic acid isobutylamide were the major chemicals characteristic of E. angustifolia root extracts. Echinacoside and 6-O-caffeoylechinacoside predominated in extracts of E. pallida roots. Characteristic alkamides were also examined by electrospray tandem mass spectrometry (MS/MS) and these compounds provided characteristic fragmentation patterns. Extracts of the roots and leaves of all three species were found to have antioxidant properties in a free radical scavenging assay and in a lipid peroxidation assay.


Assuntos
Antioxidantes/farmacologia , Echinacea/química , Extratos Vegetais/farmacologia , Plantas Medicinais , Cromatografia Líquida de Alta Pressão , Sequestradores de Radicais Livres , Humanos , Peroxidação de Lipídeos , Espectrometria de Massas , Neuroblastoma/patologia , Extratos Vegetais/química , Folhas de Planta/química , Raízes de Plantas/química , Espectrofotometria Ultravioleta , Células Tumorais Cultivadas
2.
J Pharm Pharmacol ; 52(4): 451-9, 2000 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-10813558

RESUMO

The effects of Ginkgo biloba leaf extract on rat brain or livermonoamine oxidase (MAO)-A and -B activity, biogenic amine concentration in nervous tissue, N-methyl-D-aspartate (NMDA)- and N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4)-induced neurotoxicity and antioxidant activity was investigated to determine the effects of the extract on monoamine catabolism and neuroprotection. Ginkgo biloba leaf extract was shown to produce in-vitro inhibition of rat brain MAO-A and -B. The Ginkgo biloba extract was chromatographed on a reverse-phase HPLC system and two of the components isolated were shown to be MAO inhibitors (MAOIs). These MAOIs were identified by high-resolution mass spectrometry as kaempferol and isorhamnetin. Pure kaempferol and a number of related flavonoids were examined as MAOIs in-vitro. Kaempferol, apigenin and chrysin proved to be potent MAOIs, but produced more pronounced inhibition of MAO-A than MAO-B. IC50 (50% inhibition concentration) values for the ability of these three flavones to inhibit MAO-A were 7 x 10(-7), 1 x 10(-6) and 2 x 10(-6) M, respectively. Ginkgo biloba leaf extract and kaempferol were found to have no effect ex-vivo on rat or mouse brain MAO or on concentrations of dopamine, noradrenaline, 5-hydroxytryptamine and 5-hydroxyindoleacetic acid. Kaempferol was shown to protect against NMDA-induced neuronal toxicity in-vitro in rat cortical cultures, but did not prevent DSP-4-induced noradrenergic neurotoxicity in an in-vivo model. Both Ginkgo biloba extract and kaempferol were demonstrated to be antioxidants in a lipid-peroxidation assay. This data indicates that the MAO-inhibiting activity of Ginkgo biloba extract is primarily due to the presence of kaempferol. Ginkgo biloba extract has properties indicative of potential neuroprotective ability.


Assuntos
Flavonoides , Ginkgo biloba/química , Quempferóis , Inibidores da Monoaminoxidase/análise , Fármacos Neuroprotetores/farmacologia , Extratos Vegetais/farmacologia , Folhas de Planta/química , Plantas Medicinais , Quercetina/análogos & derivados , Ácido 3,4-Di-Hidroxifenilacético/metabolismo , Administração Oral , Animais , Encéfalo/efeitos dos fármacos , Encéfalo/enzimologia , Encéfalo/metabolismo , Células Cultivadas , Cromatografia Líquida de Alta Pressão , Dopamina/metabolismo , Relação Dose-Resposta a Droga , Ácido Hidroxi-Indolacético/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Fígado/efeitos dos fármacos , Fígado/enzimologia , Masculino , Camundongos , Monoaminoxidase/efeitos dos fármacos , Monoaminoxidase/metabolismo , Inibidores da Monoaminoxidase/farmacologia , N-Metilaspartato/farmacologia , Neurônios/citologia , Neurônios/efeitos dos fármacos , Norepinefrina/metabolismo , Extratos Vegetais/química , Quercetina/análise , Quercetina/farmacologia , Ratos , Ratos Sprague-Dawley , Serotonina/metabolismo , Células Tumorais Cultivadas/efeitos dos fármacos , Células Tumorais Cultivadas/metabolismo
3.
J Pharmacol Toxicol Methods ; 44(3): 461-5, 2000.
Artigo em Inglês | MEDLINE | ID: mdl-11395323

RESUMO

An electron-capture gas chromatographic procedure was developed for detection and quantification of p-hydroxymephenytoin (OHMEP), a metabolite of S-mephenytoin, in human liver microsomal preparations. OHMEP was derivatized with pentafluorobenzoyl chloride (PFBC) under basic aqueous conditions prior to analysis on a gas chromatograph equipped with a capillary column and an electron-capture detector. Dextrorophan was carried through the procedure as internal standard. The structure of the PFB derivative was confirmed using combined gas chromatography-mass spectrometry (GC-MS). The procedure is rapid and reproducible and produces a stable derivative that has excellent chromatographic properties. The limit of detection was less than 5 ng/ml, and the method was applied to extracts of human liver microsomes, which had been incubated with S-mephenytoin [a probe substrate for cytochrome P450 (CYP) 2C19].


Assuntos
Cromatografia Gasosa/métodos , Mefenitoína/análise , Anticonvulsivantes/metabolismo , Anticonvulsivantes/farmacologia , Dextrorfano/análise , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Mefenitoína/análogos & derivados , Mefenitoína/metabolismo , Mefenitoína/farmacologia , Microssomos Hepáticos/efeitos dos fármacos , Microssomos Hepáticos/metabolismo , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Fatores de Tempo
4.
J Chromatogr B Biomed Sci Appl ; 749(2): 275-9, 2000 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-11145064

RESUMO

A simple, rapid and sensitive procedure using gas chromatography with electron-capture detection to measure paroxetine levels in human plasma has been developed. The analyte was extracted from plasma with ethyl acetate after basification of the plasma and then derivatized with heptafluorobutyric anhydride before gas chromatographic separation. The calibration curves were linear, with typical r2 values >0.99. The assay was highly reproducible and gave peaks with excellent chromatographic properties.


Assuntos
Cromatografia Gasosa/métodos , Paroxetina/sangue , Inibidores Seletivos de Recaptação de Serotonina/sangue , Calibragem , Humanos , Controle de Qualidade
5.
Cell Mol Neurobiol ; 19(3): 325-54, 1999 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-10319191

RESUMO

1. The cytochrome P450 monooxygenases, CYP2D6, CYP2C19, and CYP2C9, display polymorphism. CYP2D6 and CYP2C19 have been studied extensively, and despite their low abundance in the liver, they catalyze the metabolism of many drugs. 2. CYP2D6 has numerous allelic variants, whereas CYP2C19 has only two. Most variants are translated into inactive, truncated protein or fail to express protein. 3. CYP2C9 is expressed as the wild-type enzyme and has two variants, in each of which one amino acid residue has been replaced. 4. The nucleotide base sequences of the cDNAs of the three polymorphic genes and their variants have been determined, and the proteins derived from these genes have been characterized. 5. An absence of CYP2D6 and/or CYP2C19 in an individual produces a poor metabolizer (PM) of drugs that are substrates of these enzymes. 6. When two drugs that are substrates for a polymorphic CYP enzyme are administered concomitantly, each will compete for that enzyme and competitively inhibit the metabolism of the other substrate. This can result in toxicity. 7. Patients can be readily phenotyped or genotyped to determine their CYP2D6 or CYP2C19 enzymatic status. Poor metabolizers (PMs), extensive metabolizers (EMs), and ultrarapid metabolizers (URMs) can be identified. 8. Numerous substrates and inhibitors of CYP2D6, CYP2C19, and CYP2C9 are identified. 9. An individual's diet and age can influence CYP enzyme activity. 10. CYP2D6 polymorphism has been associated with the risk of onset of various illnesses, including cancer, schizophrenia, Parkinson's disease, Alzheimer's disease, and epilepsy.


Assuntos
Sistema Enzimático do Citocromo P-450/genética , Sistema Enzimático do Citocromo P-450/metabolismo , Psicotrópicos/farmacocinética , Sequência de Aminoácidos , Dados de Sequência Molecular , Polimorfismo Genético , Psiquiatria
6.
J Pharmacol Toxicol Methods ; 41(4): 143-6, 1999 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-10691018

RESUMO

Dextromethorphan, a constituent of many over-the-counter cough syrups, is used as a probe drug for phenotyping subjects for their cytochrome P450 2D6 (CYP2D6) enzyme activity and for measuring CYP2D6 activity of preparations such as microsomes. In such studies, formation of the metabolite dextrorphan is used as indicator of the activity of this CYP enzyme. The present report describes an electron-capture gas chromatographic procedure developed for detection and quantification of dextrorphan in human liver microsomal preparations in vitro. After basification of the incubation mixture, dextrorphan was derivatized with pentafluorobenzoyl chloride under aqueous conditions prior to analysis on a gas chromatograph equipped with a capillary column, an electron capture detector, and a printer-integrator. Para-hydroxymephenytoin was carried through the procedure as internal standard. The procedure, which involves the derivatization of dextrorphan under aqueous conditions, is rapid and involves the use of the relatively economical procedure of electron-capture gas chromatography. The derivative is stable and possesses excellent chromatographic properties.


Assuntos
Benzoatos/química , Dextrorfano/análise , Microssomos Hepáticos/química , Cromatografia Gasosa , Estabilidade de Medicamentos , Humanos , Técnicas In Vitro , Mefenitoína/análogos & derivados , Mefenitoína/análise , Oxirredução , Solubilidade
7.
J Chromatogr B Biomed Sci Appl ; 698(1-2): 103-9, 1997 Sep 26.
Artigo em Inglês | MEDLINE | ID: mdl-9367198

RESUMO

An electron-capture gas chromatographic procedure was developed for the analysis of p-trifluoromethylphenol, an O-dealkylated metabolite of fluoxetine, in biological samples. A basic extraction of the biological sample was employed, followed by derivatization with pentafluorobenzenesulfonyl chloride. The internal standard, 2,4-dichlorophenol, was added to all samples used in the procedure to aid in quantitation. The practical limit of detection (signal-to-noise ratio>3) for p-trifluoromethylphenol was <5 ng/ml in human plasma samples, <10 ng/g of rat brain tissue, <25 ng/g of rat liver tissue and <25 ng/ml in human and rat urine samples. In the rat, the levels of free p-trifluoromethylphenol in the liver were 10-fold higher than those in the brain, and a substantial amount was excreted in the urine. Human urine samples contained levels of free p-trifluoromethylphenol approximately 30-fold higher than those found in human plasma samples. The procedure described is useful for the detection and quantitation of free p-trifluoromethylphenol in humans and rats treated with fluoxetine.


Assuntos
Cromatografia Gasosa/métodos , Fluoxetina/metabolismo , Fenóis/metabolismo , Inibidores Seletivos de Recaptação de Serotonina/metabolismo , Animais , Encéfalo/metabolismo , Humanos , Fígado/metabolismo , Masculino , Fenóis/sangue , Fenóis/urina , Ratos , Ratos Sprague-Dawley , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
8.
J Pharmacol Toxicol Methods ; 38(1): 27-32, 1997 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-9339413

RESUMO

A procedure for analysis of metoprolol-utilizing extraction followed by derivatization with pentafluoropropionic anhydride and analysis on a gas chromatograph equipped with a fused silica capillary column, an electron-capture detector and a printer/integrator is described. Propranolol was carried through the procedure as internal standard. The pentafluoropropionyl derivative of metoprolol yields a sharp peak on the gas chromatograph, and the structure of the derivative was confirmed using combined gas chromatography-mass spectrometry. The analytical method is linear, sensitive and reproducible and has been applied to analysis of metoprolol in brain and heart from rats treated with metoprolol intraperitoneally. Pretreatment of the rats with the antidepressant desipramine prior to metoprolol administration resulted in a marked increase in levels of metoprolol in both brain and heart, indicating a pharmacokinetic drug-drug interaction between desipramine and metoprolol.


Assuntos
Química Encefálica , Cromatografia Gasosa/métodos , Metoprolol/análise , Miocárdio/química , Animais , Desipramina/administração & dosagem , Cromatografia Gasosa-Espectrometria de Massas , Masculino , Metoprolol/química , Ratos , Ratos Sprague-Dawley
9.
Clin Rheumatol ; 16(3): 291-5, 1997 May.
Artigo em Inglês | MEDLINE | ID: mdl-9184268

RESUMO

OBJECTIVE: To assess the frequency of adverse drug reaction in patients with fibromyalgia in relation to medications prescribed for this condition. To evaluate the potential role of the P450IID6 phenotype in the pathogenesis of these adverse drug reactions. METHODS: Thirty-five patients with fibromyalgia were assessed using a structured questionnaire with demographic and clinical data and perceived adverse drug reactions. A sample of 60 patients with rheumatoid arthritis and 62 patients with localized back pain served as controls. The P450IID6 phenotype was determined for each of the fibromyalgia patients. RESULTS: Overall, 141 patients had used NSAID and 79 (56%) of them reported adverse effects. Antidepressant drugs were used by 68 patients and 35 (51%) patients had adverse effects. Muscle relaxant drugs were used by 48 patients and 15 (31%) of them reported side effects. Analgesics were used by 122 patients and 22 (18%) had experienced adverse effects. Statistical differences in the frequency of adverse effects were found with antidepressant drugs in the fibromyalgia group, compared with rheumatoid arthritis (p=0.01) and back pain (p=0.02). Four of the 35 patients (11.4%) had a metabolic ratio (M.R.) greater than 0.30 (log M.R.= -0.52) indicative of the poor metabolizers (PM) phenotype. M.R. varied from 0.005 (log M.R. = -2.30) to 4.99 (log M.R. = 0.70). CONCLUSIONS: The problem of adverse drug reactions in fibromyalgia patients does not appear to correlate with the PM phenotype of the P450IID6 oxidative enzyme. It also is unlikely that altered xenobiotic detoxification attributable to this PM phenotype would have a significant role in the development of fibromyalgia.


Assuntos
Analgésicos/efeitos adversos , Anti-Inflamatórios não Esteroides/efeitos adversos , Antidepressivos/efeitos adversos , Citocromo P-450 CYP2D6/genética , Debrisoquina/metabolismo , Fibromialgia/tratamento farmacológico , Polimorfismo Genético , Adulto , Analgésicos/uso terapêutico , Análise de Variância , Anti-Inflamatórios não Esteroides/uso terapêutico , Antidepressivos/uso terapêutico , Antirreumáticos/efeitos adversos , Antirreumáticos/uso terapêutico , Artrite Reumatoide/tratamento farmacológico , Artrite Reumatoide/enzimologia , Dor nas Costas/tratamento farmacológico , Dor nas Costas/enzimologia , Canadá , Citocromo P-450 CYP2D6/metabolismo , Debrisoquina/análise , Feminino , Fibromialgia/enzimologia , Fibromialgia/genética , Humanos , Pessoa de Meia-Idade , Fenótipo , Prevalência
11.
J Chromatogr B Biomed Appl ; 682(2): 283-8, 1996 Jul 12.
Artigo em Inglês | MEDLINE | ID: mdl-8844421

RESUMO

An electron-capture gas chromatographic procedure was developed for the analysis of 4-(4-chlorophenyl)-4-hydroxypiperidine (CPHP), a metabolite of haloperidol. The assay involved basic extraction of this metabolite from the biological samples, followed by back-extraction with HCl. After basification of the acid phase, extractive derivatization with pentafluorobenzoyl chloride in toluene was conducted. The pentafluorobenzoyl derivative was quantified on a gas chromatograph equipped with a fused-silica capillary column, an electron-capture detector and a printer-integrator. N-(3-Trifluoromethylphenyl)piperazine was carried through the procedure as an internal standard and calibration curves were determined for each assay run. The procedure was demonstrated to be linear and reproducible and was utilized to detect and quantify CPHP in urine, plasma, brain and liver samples from rats treated with haloperidol. The structure of the derivatized metabolite was confirmed by gas chromatography-mass spectrometry.


Assuntos
Cromatografia Gasosa/métodos , Haloperidol/metabolismo , Piperidinas/análise , Animais , Masculino , Piperidinas/sangue , Piperidinas/urina , Ratos , Ratos Sprague-Dawley , Reprodutibilidade dos Testes
12.
J Pharmacol Toxicol Methods ; 34(3): 143-7, 1995 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-8573764

RESUMO

The combination of fluoxetine (FLU) and desipramine (DMI) has been reported to be useful for the treatment of depression, and these drugs are also known to undergo a metabolic drug-drug interaction because of their effects on cytochrome P-450 2D6. A procedure that separates these two drugs and norfluoxetine (NFLU), the N-demethylated metabolite of FLU, and that allows simultaneous quantification of their levels would be of value and has been developed in our laboratories. The procedure involves an initial extraction into ethyl acetate after basification of the homogenate. The organic phase is retained and taken to dryness; the residue is reconstituted in water and acetylated with acetic anhydride under slightly basic conditions. Ethyl acetate is then used to extract the acetylated compounds from the aqueous medium. The organic layer is taken to dryness and the residue reconstituted in toluene. An aliquot of the solution in toluene is injected directly into a gas chromatograph equipped with a nitrogen-phosphorus detector, a fused silica capillary column, and an integrator/printer. Maprotiline is added to the initial homogenate and carried through the procedure as the internal standard. The assay is rapid and sensitive and has been applied successfully to liver and brain tissue taken from rats treated with FLU, DMI, or the combination.


Assuntos
Antidepressivos/isolamento & purificação , Química Encefálica , Desipramina/isolamento & purificação , Fluoxetina/análogos & derivados , Fluoxetina/isolamento & purificação , Fígado/química , Animais , Antidepressivos/administração & dosagem , Antidepressivos/metabolismo , Cromatografia Gasosa , Desipramina/administração & dosagem , Desipramina/metabolismo , Interações Medicamentosas , Fluoxetina/administração & dosagem , Fluoxetina/química , Fluoxetina/metabolismo , Masculino , Espectrometria de Massas , Nitrogênio/química , Fósforo/química , Ratos , Ratos Sprague-Dawley , Padrões de Referência , Distribuição Tecidual
14.
J Pharmacol Toxicol Methods ; 31(3): 141-8, 1994 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-8068975

RESUMO

Acetylation with acetic anhydride, under both aqueous and anhydrous conditions, has been utilized to derivatize various biogenic amines and psychotropic drugs for subsequent analysis by gas chromatography (GC) or gas chromatography-mass spectrometry (GC-MS). Under basic aqueous conditions, acetic anhydride derivatizes phenols and amines but not alcohols; under anhydrous conditions, all three functions are acetylated. Primary amines, once derivatized with acetic anhydride, can be further derivatized with other reagents; these diderivatives have proven useful for subsequent analysis by GC or GC-MS. Examples of applications of derivatization with acetic anhydride to analysis of biogenic amines, antidepressants, antipsychotics, and some of their metabolites are presented.


Assuntos
Anidridos Acéticos , Aminas Biogênicas/análise , Líquidos Corporais/química , Psicotrópicos/análise , Acetilação , Animais , Cromatografia Gasosa , Humanos , Espectrometria de Massas , Psicotrópicos/metabolismo
15.
Biochem Pharmacol ; 44(9): 1894-7, 1992 Nov 03.
Artigo em Inglês | MEDLINE | ID: mdl-1449543

RESUMO

A novel assay procedure has been developed that allows for the separation and quantification of the enantiomers of the monoamine oxidase inhibitor tranylcypromine (TCP) in brain and liver of rats. The analytical method involves extraction of the drug from rat tissue with an organic solvent. TCP is then derivatized with S-(-)-N-(trifluoroacetyl)-prolyl chloride to allow gas chromatographic analysis of the resulting diastereoisomers. Conditions for analysis by a gas chromatograph equipped with a nitrogen-phosphorus detector and a capillary column are described. The method has been applied to the separation and quantification of the enantiomers of TCP in samples of brain and liver of rats that had been injected with this drug alone and after pretreatment with iprindole, a drug known to block aromatic ring hydroxylation.


Assuntos
Tranilcipromina/isolamento & purificação , Animais , Encéfalo/metabolismo , Química Encefálica , Cromatografia Gasosa/métodos , Cromatografia Gasosa-Espectrometria de Massas , Iprindol/farmacologia , Fígado/química , Fígado/embriologia , Masculino , Ratos , Ratos Sprague-Dawley , Reprodutibilidade dos Testes , Estereoisomerismo , Fatores de Tempo , Tranilcipromina/metabolismo
16.
J Chromatogr ; 579(1): 99-106, 1992 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-1447355

RESUMO

An electron-capture gas chromatographic procedure was developed for the simultaneous analysis of the enantiomers of fluoxetine and norfluoxetine. The assay involves basic extraction of these enantiomers from the biological samples, followed by their conversion to diastereoisomers using the chiral derivatizing reagent (S)-(-)-N-trifluoroacetylprolyl chloride. The method was utilized to detect and measure the quantity of these enantiomers in plasma and urine of patients and in liver and brain tissue of rats treated with (R,S)-fluoxetine.


Assuntos
Química Encefálica , Cromatografia Gasosa/métodos , Fluoxetina/análogos & derivados , Fluoxetina/análise , Fígado/química , Animais , Feminino , Fluoxetina/sangue , Fluoxetina/urina , Humanos , Masculino , Ratos , Ratos Sprague-Dawley , Estereoisomerismo
17.
J Chromatogr ; 573(2): 313-7, 1992 Jan 17.
Artigo em Inglês | MEDLINE | ID: mdl-1601966

RESUMO

An electron-capture gas chromatographic procedure for the simultaneous determination of amphetamine and N-methylamphetamine in biological samples is described. The method employs extraction with the ion-pairing reagent bis(2-ethylhexyl)phosphoric acid, and back-extraction with 0.5 M hydrochloric acid. The hydrochloric acid phase is basified, and the amphetamines and the internal standard benzylamine are derivatized with pentafluorobenzenesulfonyl chloride prior to analysis on a gas chromatograph equipped with a capillary column. Levels of amphetamine and N-methylamphetamine have been determined in the urine and liver of rats treated chronically with (-)-deprenyl.


Assuntos
Anfetamina/metabolismo , Fígado/química , Metanfetamina/metabolismo , Anfetamina/urina , Animais , Cromatografia Gasosa , Elétrons , Cromatografia Gasosa-Espectrometria de Massas , Masculino , Metanfetamina/urina , Ratos , Ratos Endogâmicos , Selegilina/administração & dosagem
18.
Res Commun Chem Pathol Pharmacol ; 70(1): 115-24, 1990 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-2263759

RESUMO

A rapid, sensitive procedure has been developed for the assay of gamma-aminobutyric acid (GABA) in brain tissue. The method involves sequential reaction with isobutyl chloroformate and pentafluorophenol under aqueous conditions and subsequent separation and quantitation of the derivative on a gas chromatograph equipped with a capillary column and an electron-capture detector. The method produces a derivative with good stability and provides for simultaneous analysis of GABA and five other aliphaticamino acids in very small volumes of supernatant from brain homogenates.


Assuntos
Química Encefálica , Cromatografia Gasosa/métodos , Espectrometria de Massas/métodos , Ácido gama-Aminobutírico/análise , Aminoácidos/química , Animais , Masculino , Ratos , Ratos Endogâmicos
19.
J Neurosci Methods ; 32(2): 105-9, 1990 May.
Artigo em Inglês | MEDLINE | ID: mdl-2366572

RESUMO

A rapid procedure has been developed for the analysis of phenylalanine in brain tissue. Perchloric acid extracts of brain tissue were made basic, and benzoyl chloride was added to derivatize the amine function. The aqueous layer was retained and made slightly acidic. To derivatize the carboxylic acid group, a solution of pentafluorobenzyl alcohol was added in the presence of the coupling agent dicyclohexylcarbodiimide and chloroform. After shaking for 15 min, the organic phase was retained and taken to dryness. The residue was taken up in toluene, washed, and an aliquot used for analysis on a gas chromatograph equipped with an automatic injector, a capillary column and an electron-capture detector. The procedure has been utilized for analysis of phenylalanine in brains of rats treated with vehicle or phenylalanine.


Assuntos
Química Encefálica , Encéfalo/metabolismo , Cromatografia Gasosa/métodos , Fenilalanina/análise , Animais , Masculino , Ratos , Ratos Endogâmicos
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