RESUMO
The present Special Issue is focused on developing and applying several sensors, biosensor devices, and actuators for the analysis of drugs, foods, and nutraceuticals. Some applications concern classical topics, such as clostridium determination in dairy products, flavouring material in foods like ethylvanillin, or the antioxidant properties of fruit juices, while other applications are more innovative, such as food safety analysis, artificial human senses (electronic nose, or tongue) development, or ethanol determination in pharmaceutical drugs, or forensic purposes using catalytic fuel cell; and lastly, new studies devoted to intelligent food packaging. Therefore, this Special Issue should interest both specialists in the sector and readers who are simply curious, or are simply interested in innovations in the field of food and drug analysis.
Assuntos
Técnicas Biossensoriais/métodos , Suplementos Nutricionais/análise , Drogas em Investigação/análise , Técnicas Biossensoriais/instrumentação , Nariz Eletrônico , Análise de Alimentos/métodos , Embalagem de Alimentos , Sistemas Automatizados de Assistência Junto ao LeitoRESUMO
PF-05231023, a long-acting fibroblast growth factor 21 (FGF21) analog, was generated by covalently conjugating two engineered [des-His1, Ala129Cys]FGF21 molecules to a nontargeting human IgG1 κ scaffold. The pharmacokinetics (PK) of PF-05231023 after i.v. and s.c. administration was evaluated in rats and monkeys using two enzyme-linked immunosorbent assays with high specificity for biologically relevant intact N termini (NT) and C termini (CT) of FGF21. Intact CT of FGF21 displayed approximately 5-fold faster systemic plasma clearance (CL), an approximately 2-fold lower steady-state volume of distribution, and at least 5-fold lower bioavailability compared with NT. In vitro serum stability studies in monkeys and humans suggested that the principal CL mechanism for PF-05231023 was degradation by serum proteases. Direct scaling of in vitro serum degradation rates for intact CT of FGF21 underestimated in vivo CL 5-fold, 1.4-fold, and 2-fold in rats, monkeys, and humans, respectively. The reduced steady-state volume of distribution and the bioavailability for intact CT relative to NT in rats and monkeys were compatible with proteolytic degradation occurring outside the plasma compartment via an unidentified mechanism. Human CL and PK profiles for intact NT and CT of FGF21 were well predicted using monkey single-species allometric and Dedrick scaling. Physiologically based pharmacokinetic models incorporating serum stability data and an extravascular extraction term based on differential bioavailability of intact NT and CT of FGF21 in monkeys improved accuracy of human PK predictions relative to Dedrick scaling. Mechanistic physiologically based pharmacokinetic models of this nature may be highly valuable for predicting human PK of fusion proteins, synthetically conjugated proteins, and other complex biologics.
Assuntos
Anticorpos Monoclonais Humanizados/farmacologia , Drogas em Investigação/farmacocinética , Fatores de Crescimento de Fibroblastos/química , Fatores de Crescimento de Fibroblastos/farmacologia , Hipoglicemiantes/farmacocinética , Hipolipemiantes/farmacocinética , Imunoglobulina G/química , Modelos Biológicos , Proteínas Recombinantes/farmacocinética , Substituição de Aminoácidos , Animais , Anticorpos Monoclonais Humanizados/administração & dosagem , Anticorpos Monoclonais Humanizados/sangue , Anticorpos Monoclonais Humanizados/química , Avaliação Pré-Clínica de Medicamentos , Drogas em Investigação/administração & dosagem , Drogas em Investigação/análise , Drogas em Investigação/química , Fatores de Crescimento de Fibroblastos/administração & dosagem , Fatores de Crescimento de Fibroblastos/sangue , Fatores de Crescimento de Fibroblastos/genética , Fatores de Crescimento de Fibroblastos/metabolismo , Meia-Vida , Humanos , Hipoglicemiantes/administração & dosagem , Hipoglicemiantes/sangue , Hipoglicemiantes/química , Hipolipemiantes/administração & dosagem , Hipolipemiantes/sangue , Hipolipemiantes/química , Imunoglobulina G/sangue , Imunoglobulina G/genética , Imunoglobulina G/metabolismo , Cadeias kappa de Imunoglobulina/sangue , Cadeias kappa de Imunoglobulina/química , Cadeias kappa de Imunoglobulina/genética , Cadeias kappa de Imunoglobulina/metabolismo , Injeções Intravenosas , Injeções Subcutâneas , Macaca fascicularis , Masculino , Taxa de Depuração Metabólica , Proteínas Mutantes/administração & dosagem , Proteínas Mutantes/sangue , Proteínas Mutantes/química , Proteínas Mutantes/farmacocinética , Fragmentos de Peptídeos/sangue , Proteólise , Ratos , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/sangue , Proteínas Recombinantes/químicaRESUMO
In this paper, an ex vivo model for the study of the transcorneal permeation of drugs, based on porcine tissues, was evaluated. The setup is characterized by ease of realization, absence of O2 and CO2 bubbling and low cost; additionally, the large availability of porcine tissue permits a high throughput. Histological images showed the comparability between porcine and human corneas and confirmed the effectiveness of the isolation procedure. A new de-epithelization procedure based on a thermal approach was also set up to simulate cornea permeability in pathological conditions. The procedure did not affect the integrity of the underlying layers and allowed the characterization of the barrier properties of epithelium and stroma. Six compounds with different physicochemical properties were tested: fluorescein, atenolol, propranolol, diclofenac, ganciclovir and lidocaine. The model highlighted the barrier function played by epithelium toward the diffusion of hydrophilic compounds and the permselectivity with regard to more lipophilic molecules. In particular, positively charged compounds showed a significantly higher transcorneal permeability than negatively charged compounds. The comparability of results with literature data supports the goodness and the robustness of the model, especially taking into account the behavior of fluorescein, which is generally considered a marker of tissue integrity.
Assuntos
Córnea/metabolismo , Avaliação Pré-Clínica de Medicamentos/métodos , Drogas em Investigação/metabolismo , Ensaios de Triagem em Larga Escala , Modelos Biológicos , Absorção Ocular , Matadouros , Administração Oftálmica , Animais , Fenômenos Químicos , Córnea/citologia , Substância Própria/citologia , Substância Própria/metabolismo , Drogas em Investigação/administração & dosagem , Drogas em Investigação/análise , Drogas em Investigação/química , Epitélio Corneano/fisiologia , Humanos , Interações Hidrofóbicas e Hidrofílicas , Técnicas In Vitro , Cinética , Permeabilidade , Especificidade da Espécie , Sus scrofaRESUMO
Sorafenib (Nexavar®) is currently the only FDA-approved small molecule targeted therapy for advanced hepatocellular carcinoma. The use of structural analogues and derivatives of sorafenib has enabled the elucidation of critical targets and mechanism(s) of cell death for human cancer lines. We previously performed a structure-activity relationship study on a series of sorafenib analogues designed to investigate the inhibition overlap between the major targets of sorafenib Raf-1 kinase and VEGFR-2, and an enzyme shown to be a potent off-target of sorafenib, soluble epoxide hydrolase. In the current work, we present the biological data on our lead sorafenib analogue, t-CUPM, demonstrating that this analogue retains cytotoxicity similar to sorafenib in various human cancer cell lines and strongly inhibits growth in the NCI-60 cell line panel. Co-treatment with the pan-caspase inhibitor, Z-VAD-FMK, failed to rescue the cell viability responses of both sorafenib and t-CUPM, and immunofluorescence microscopy shows similar mitochondrial depolarization and apoptosis-inducing factor release for both compounds. These data suggest that both compounds induce a similar mechanism of caspase-independent apoptosis in hepatoma cells. In addition, t-CUPM displays anti-proliferative effects comparable to sorafenib as seen by a halt in G0/G1 in cell cycle progression. The structural difference between sorafenib and t-CUPM significantly reduces inhibitory spectrum of kinases by this analogue, and pharmacokinetic characterization demonstrates a 20-fold better oral bioavailability of t-CUPM than sorafenib in mice. Thus, t-CUPM may have the potential to reduce the adverse events observed from the multikinase inhibitory properties and the large dosing regimens of sorafenib.
Assuntos
Antineoplásicos/farmacologia , Drogas em Investigação/farmacologia , Neoplasias/tratamento farmacológico , Niacinamida/análogos & derivados , Compostos de Fenilureia/farmacologia , Inibidores de Proteínas Quinases/farmacologia , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/antagonistas & inibidores , Quinases raf/antagonistas & inibidores , Administração Oral , Animais , Animais não Endogâmicos , Antineoplásicos/administração & dosagem , Antineoplásicos/sangue , Antineoplásicos/farmacocinética , Apoptose/efeitos dos fármacos , Disponibilidade Biológica , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Avaliação Pré-Clínica de Medicamentos , Drogas em Investigação/administração & dosagem , Drogas em Investigação/análise , Drogas em Investigação/farmacocinética , Meia-Vida , Humanos , Masculino , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Camundongos , Proteínas de Neoplasias/antagonistas & inibidores , Proteínas de Neoplasias/metabolismo , Neoplasias/sangue , Neoplasias/metabolismo , Neoplasias/patologia , Niacinamida/administração & dosagem , Niacinamida/sangue , Niacinamida/farmacocinética , Niacinamida/farmacologia , Compostos de Fenilureia/administração & dosagem , Compostos de Fenilureia/sangue , Compostos de Fenilureia/farmacocinética , Inibidores de Proteínas Quinases/administração & dosagem , Inibidores de Proteínas Quinases/sangue , Inibidores de Proteínas Quinases/farmacocinética , Fase de Repouso do Ciclo Celular/efeitos dos fármacos , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Quinases raf/metabolismoRESUMO
The prediction of brain extracellular fluid (ECF) concentrations in human is a potentially valuable asset during drug development as it can provide the pharmacokinetic input for pharmacokinetic-pharmacodynamic models. This study aimed to compare two translational modelling approaches that can be applied at the preclinical stage of development in order to simulate human brain ECF concentrations. A population-PBPK model of the central nervous system was developed based on brain microdialysis data, and the model parameters were translated to their corresponding human values to simulate ECF and brain tissue concentration profiles. In parallel, the PBPK modelling software Simcyp was used to simulate human brain tissue concentrations, via the bottom-up prediction of brain tissue distribution using two different sets of mechanistic tissue composition-based equations. The population-PBPK and bottom-up approaches gave similar predictions of total brain concentrations in both rat and human, while only the population-PBPK model was capable of accurately simulating the rat ECF concentrations. The choice of PBPK model must therefore depend on the purpose of the modelling exercise, the in vitro and in vivo data available and knowledge of the mechanisms governing the membrane permeability and distribution of the drug.
Assuntos
Encéfalo/metabolismo , Fármacos do Sistema Nervoso Central/farmacocinética , Drogas em Investigação/farmacocinética , Modelos Biológicos , Neurônios/metabolismo , Animais , Barreira Hematoencefálica/efeitos dos fármacos , Barreira Hematoencefálica/metabolismo , Encéfalo/efeitos dos fármacos , Permeabilidade da Membrana Celular/efeitos dos fármacos , Fármacos do Sistema Nervoso Central/administração & dosagem , Fármacos do Sistema Nervoso Central/análise , Fármacos do Sistema Nervoso Central/farmacologia , Ensaios Clínicos Fase I como Assunto , Simulação por Computador , Relação Dose-Resposta a Droga , Avaliação Pré-Clínica de Medicamentos/métodos , Drogas em Investigação/administração & dosagem , Drogas em Investigação/análise , Drogas em Investigação/farmacologia , Líquido Extracelular/química , Líquido Extracelular/efeitos dos fármacos , Líquido Extracelular/metabolismo , Humanos , Microdiálise , Neurônios/química , Neurônios/efeitos dos fármacos , Ratos , Software , Especificidade da Espécie , Distribuição Tecidual , Pesquisa Translacional Biomédica/métodosRESUMO
The identification of the no observed adverse effect level (NOAEL) is the key regulatory outcome of toxicity studies. With the introduction of "omics" technologies into toxicological research, the question arises as to how sensitive these technologies are relative to classical regulatory toxicity parameters. BASF SE and metanomics developed the in vivo metabolome database MetaMap®Tox containing metabolome data for more than 500 reference compounds. For several years metabolome analysis has been routinely performed in regulatory toxicity studies (REACH mandated testing or new compound development), mostly in the context of 28 day studies in rats (OECD 407 guideline). For those chemicals for which a toxicological NOAEL level was obtained at either high or mid-dose level, we evaluated the associated metabolome to investigate the sensitivity of metabolomics versus classical toxicology with respect to the NOAEL. For the definition of a metabolomics NOAEL the ECETOC criteria (ECETOC, 2007) were used. In this context we evaluated 104 cases. Comparable sensitivity was noted in 75% of the cases, increased sensitivity of metabolomics in 8%, and decreased sensitivity in 18% of the cases. In conclusion, these data suggest that metabolomics profiling has a similar sensitivity to the classical toxicological study (e.g. OECD 407) design.
Assuntos
Agroquímicos/toxicidade , Avaliação Pré-Clínica de Medicamentos , Drogas em Investigação/efeitos adversos , Metabolômica/métodos , Modelos Biológicos , Testes de Toxicidade , Agroquímicos/análise , Agroquímicos/farmacocinética , Animais , Bases de Dados de Compostos Químicos , Avaliação Pré-Clínica de Medicamentos/normas , Drogas em Investigação/análise , Drogas em Investigação/farmacocinética , Feminino , Alemanha , Guias como Assunto , Humanos , Legislação de Medicamentos , Masculino , Nível de Efeito Adverso não Observado , Medicamentos sem Prescrição/efeitos adversos , Medicamentos sem Prescrição/análise , Medicamentos sem Prescrição/farmacocinética , Medicamentos sob Prescrição/efeitos adversos , Medicamentos sob Prescrição/análise , Medicamentos sob Prescrição/farmacocinética , Ratos , Ratos Wistar , Reprodutibilidade dos Testes , Caracteres Sexuais , Testes de Toxicidade/normas , Toxicologia/legislação & jurisprudência , Toxicologia/métodosRESUMO
Accurate prediction of pharmacokinetics (PK) parameters in humans from animal data is difficult for various reasons, including species differences. However, chimeric mice with humanized liver (PXB mice; urokinase-type plasminogen activator/severe combined immunodeficiency mice repopulated with approximately 80% human hepatocytes) have been developed. The expression levels and metabolic activities of cytochrome P450 (P450) and non-P450 enzymes in the livers of PXB mice are similar to those in humans. In this study, we examined the predictability for human PK parameters from data obtained in PXB mice. Elimination of selected drugs involves multiple metabolic pathways mediated not only by P450 but also by non-P450 enzymes, such as UDP-glucuronosyltransferase, sulfotransferase, and aldehyde oxidase in liver. Direct comparison between in vitro intrinsic clearance (CL(int,in vitro)) in PXB mice hepatocytes and in vivo intrinsic clearance (CL(int,in vivo)) in humans, calculated based on a well stirred model, showed a moderate correlation (r² = 0.475, p = 0.009). However, when CL(int,in vivo) values in humans and PXB mice were compared similarly, there was a good correlation (r² = 0.754, p = 1.174 × 10â»4). Elimination half-life (t(1/2)) after intravenous administration also showed a good correlation (r² = 0.886, p = 1.506 × 10â»4) between humans and PXB mice. The rank order of CL and t(1/2) in human could be predicted at least, although it may not be possible to predict absolute values due to rather large prediction errors. Our results indicate that in vitro and in vivo experiments with PXB mice should be useful at least for semiquantitative prediction of the PK characteristics of candidate drugs in humans.
Assuntos
Sistema Enzimático do Citocromo P-450/metabolismo , Avaliação Pré-Clínica de Medicamentos/métodos , Drogas em Investigação/metabolismo , Drogas em Investigação/farmacocinética , Fígado/metabolismo , Animais , Células Cultivadas , Pré-Escolar , Quimera , Sistema Enzimático do Citocromo P-450/genética , Drogas em Investigação/análise , Feminino , Meia-Vida , Hepatócitos/citologia , Hepatócitos/enzimologia , Hepatócitos/metabolismo , Hepatócitos/transplante , Humanos , Separação Imunomagnética , Fígado/citologia , Fígado/enzimologia , Masculino , Taxa de Depuração Metabólica , Camundongos , Camundongos SCID , Proteínas Recombinantes/metabolismo , Especificidade da Espécie , Ativador de Plasminogênio Tipo Uroquinase/genéticaRESUMO
Reliably predicting pharmacokinetic behavior in humans from preclinical data is an important aspect of drug development. The most widely used technique in this regard is allometric scaling. In this review, various approaches developed for predicting pharmacokinetic parameters in humans using interspecies scaling are introduced and discussed. Methods to predict plasma concentration-time profiles in humans after intravenous and oral administration are also reviewed. The reliable prediction of human pharmacokinetics with regard to investigational drugs is aimed, ultimately, at selecting the first in-human dose with which to begin clinical studies. Approaches for the selection of the first in-human dose are also reviewed. Although there have been many trials to compare and optimize interspecies scaling methods, no firm conclusions have been reached. Because interspecies scaling methods are still highly empirical, further effort is needed to improve the reliability of predicting human pharmacokinetics by interspecies scaling.
Assuntos
Drogas em Investigação/farmacocinética , Modelos Biológicos , Animais , Tamanho Corporal , Ensaios Clínicos como Assunto/métodos , Avaliação Pré-Clínica de Medicamentos/métodos , Indústria Farmacêutica/métodos , Drogas em Investigação/análise , Humanos , Especificidade da EspécieRESUMO
Innovative attempts have been made to overcome nonproductivity and high expenditure in the clinical stages of new drug development. Microdosing studies using subpharmacological doses provide early insight into the body's disposition toward candidate compounds, and are innovative exploratory trials that can promote productivity in drug development. Highly sensitive analytical technology is crucial in microdosing studies that employ qualitative and quantitative assays of target materials in humans. Accelerator mass spectrometry (AMS) has facilitated the adoption of a human microdosing study in the early phase of clinical drug development. Results derived from AMS microdosing studies using labeled compounds can provide various types of information for candidate selection, including pharmacokinetic characteristics and metabolic profiles of candidate compounds. The applicability of microdosing studies is currently expanding into absolute bioavailability and mass balance studies. Although it remains uncertain whether microdosing adequately predicts the pharmacokinetics of therapeutic doses, further development of microdosing studies using AMS may benefit the field of new drug development and could pose a new challenge to researchers. The use of advanced technology in candidate selection will contribute to improved productivity and competitiveness in pharmaceutical research and development. The introduction of microdosing studies using AMS in Korea will present a newly applicable method for innovative clinical trials and contribute to development potential in global competition.
Assuntos
Drogas em Investigação/administração & dosagem , Drogas em Investigação/análise , Microquímica , Animais , Disponibilidade Biológica , Biotransformação , Ensaios Clínicos como Assunto/métodos , Aprovação de Drogas , Avaliação Pré-Clínica de Medicamentos/métodos , Drogas em Investigação/farmacocinética , Guias como Assunto , Humanos , Espectrometria de Massas , Microquímica/instrumentação , Aceleradores de Partículas , República da Coreia , Distribuição TecidualRESUMO
The option, for practical and ethical reasons, to replace animal plasma with human plasma for calibration standards was successfully applied to 73 analytical methods developed in our laboratory during the last years. The animals used for obtaining blank plasma could then be reduced with a number corresponding to about 25% of mice or 5% of rats in ordinary one-month toxicology studies. This is of important public concern and also in accordance with the 3R-strategy. The methods were successfully validated for determination of drug concentrations in plasma from rat, dog, mouse, rabbit and cynomolgus monkey. Reproducibility of study samples from dosed animals was established, showing a mean accuracy of 100.8% with a CV of 7.2% (n=1339). The purpose of this paper is to present a scientific basis for the alternative approach to adopt human plasma matrix for calibration standards, which will reduce animal use, without compromising the quality of appropriately validated assays. Additional advantages are cheaper and simplified plasma maintenance and the possibility to validate methods for several species in the same analytical batch.
Assuntos
Alternativas aos Testes com Animais , Avaliação Pré-Clínica de Medicamentos/métodos , Drogas em Investigação/análise , Preparações Farmacêuticas/sangue , Animais , Calibragem , Cromatografia Líquida de Alta Pressão , Avaliação Pré-Clínica de Medicamentos/economia , Estabilidade de Medicamentos , Humanos , Reprodutibilidade dos Testes , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas em Tandem , Testes de Toxicidade/economia , Testes de Toxicidade/métodosRESUMO
During a submission procedure, the validity of a few dietary toxicity studies was questioned because low levels of the drug were detected among control toxicokinetic samples. Although several lines of reasoning suggested that these findings arose from ex vivo contamination, the Regulatory Authority stated that it was not possible to establish a no-effect-level in any of the studies and so the submission was withdrawn. In response, Novartis conducted a thorough review and modification of the procedures involved in the collection and analysis of toxicokinetic samples to minimize such contamination in future studies. Ongoing monitoring of contamination in toxicology studies has subsequently revealed that although it was not possible to completely eliminate the problem, the new procedures together with an increasing awareness of the issue have considerably reduced the incidence of contamination. The process of contamination and its control was also modeled in a feeding study in mice. This provided good evidence that the detection of drug in control samples in the previous studies originated from external sources and not from in vivo exposure.
Assuntos
Drogas em Investigação/toxicidade , Testes de Toxicidade/métodos , Animais , Contaminação de Medicamentos/prevenção & controle , Avaliação Pré-Clínica de Medicamentos/métodos , Avaliação Pré-Clínica de Medicamentos/normas , Indústria Farmacêutica/normas , Drogas em Investigação/análise , Drogas em Investigação/farmacocinética , Contaminação de Equipamentos/prevenção & controle , Fidelidade a Diretrizes/normas , Guias como Assunto/normas , Cabelo/metabolismo , Cooperação Internacional , Mucosa Intestinal/metabolismo , Jejuno/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos ICR , Controle de Qualidade , Reprodutibilidade dos Testes , Distribuição Tecidual , Testes de Toxicidade/normasRESUMO
An approach is described with turbulent flow on-line extraction liquid chromatography/electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) for GLP quantitative bioanalysis of a drug candidate. Two systems were built in-house with standard laboratory parts and equipments. One system consisted of one gradient HPLC pump, one isocratic pump, one ten-port valve, two turbulent flow columns, one analytical column, one autosampler and one mass spectrometer. Using this system, an injection-to-injection cycle time of 0.8 min was achieved. By adding an additional valve, another analytical column and an isocratic pump, the injection-to-injection cycle time decreased to 0.4 min. Validation results from the two systems showed that precision and accuracy were acceptable for GLP quantitative analyses. The system was utilized to support sample bioanalysis of a drug candidate in a first-time in-human clinical trial.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Técnicas de Laboratório Clínico , Avaliação Pré-Clínica de Medicamentos/métodos , Drogas em Investigação/análise , Espectrometria de Massas por Ionização por Electrospray/métodos , Animais , Técnicas de Laboratório Clínico/normas , Ensaios Clínicos como Assunto/métodos , Avaliação Pré-Clínica de Medicamentos/normas , Humanos , Reprodutibilidade dos Testes , Testes de ToxicidadeRESUMO
A generic high-performance liquid chromatography (HPLC) system interfaced with an atmospheric pressure photoionization (APPI) source and a tandem mass spectrometer was developed for the quantitative determination of small molecules in plasma in support of exploratory in vivo pharmacokinetics. This report summarizes the effects of variations in reversed-phase mode HPLC conditions such as mobile-phase flow rate, solvent composition, organic modifier content, and nebulizer temperature on the photoionization efficiency of both clozapine and lonafarnib. The matrix ionization suppression effect on this method was investigated using the postcolumn infusion technique. The procedure was used to quantitate plasma levels following oral administration of 42 drug discovery compounds to rats. The pharmacokinetic results of 42 drug discovery compounds in rats evaluated by both APPI and atmospheric pressure chemical ionization interfaces were found to be well correlated.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Clozapina/sangue , Drogas em Investigação/análise , Espectrometria de Massas/métodos , Piperidinas/sangue , Piridinas/sangue , Administração Oral , Animais , Pressão Atmosférica , Clozapina/química , Avaliação Pré-Clínica de Medicamentos/métodos , Drogas em Investigação/farmacocinética , Fotoquímica , Piperidinas/química , Piridinas/química , RatosRESUMO
A method using zirconia-based column high-performance liquid chromatography (HPLC) interfaced with an atmospheric pressure photoionization (APPI) source and a tandem mass spectrometer (MS/MS) was developed for the quantitative determination of new chemical entities in rat plasma in support of pharmacokinetics studies. The ionization suppression resulting from endogenous components of the biological matrices on the quantitative zirconia-based column HPLC/APPI-MS/MS method was investigated using the post-column infusion technique. The analytical results for 'rapid rat pharmacokinetics' for 12 drug discovery compounds, obtained by both silica-based phase (S-phase) and zirconia-based phase (Z-phase) chromatographic separation, are in good agreement in terms of accuracy. The application of a Z-phase column for high-temperature fast HPLC/MS/MS methods was explored to reduce the analysis time from 3 min to 30 s for column temperatures of 25-110 degrees C, respectively. The chromatographic retention times and peak responses of all analytes were found to be reproducible under high-temperature conditions following 100 continuous injections, with %CV less than 0.4 and 5, respectively.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Avaliação Pré-Clínica de Medicamentos , Drogas em Investigação/análise , Espectrometria de Massas/métodos , Zircônio/química , Animais , Pressão Atmosférica , Avaliação Pré-Clínica de Medicamentos/métodos , Drogas em Investigação/farmacocinética , Fotoquímica , RatosRESUMO
A new parallel liquid chromatography/tandem mass spectrometry (LC/MS/MS) system has been developed, in which the mass detector was shared between two staggered parallel chromatographic runs. Since the chromatography for biofluids assay generally requires good analyte retention and thus tends to leave large blank chromatographic windows, this parallel system allowed the efficient use of the mass detector during these blank windows, resulting in significantly improved sample throughput. Also, in order to remove the bottleneck in sample extraction for this parallel separation system, a high-flow extraction device was used to perform on-line extraction. This allowed for the direct injection of biofluids onto the system. The performance and capability of this system was evaluated in tests that contained a single analyte (oxazepam) and multiple analytes (12-in-1). The results indicated that the data generated from this system were comparable to those obtained on a conventional single-column system. An application of the system for high-throughput pharmacokinetic screening of drug candidates was also demonstrated.