RESUMO
The inhalational administration of drugs is a practical and non-invasive approach with the potential to reduce side effects and with a quick onset of therapeutic activity. Perillyl alcohol (POH) is a monoterpene with antitumor activity that currently is undergoing clinical evaluation as an inhalational anticancer agent. A detection method was developed that will be applicable to pharmacokinetic studies of not only POH, but also its longer-lived main metabolite, perillic acid (PA), in lung tissue and plasma after inhalational delivery. The anticancer activity of POH was investigated in vitro with the use of various lung cancer cell lines. Toxicity was established by a standard MTT assay, and apoptosis markers were analyzed by Western blot. For the detection of POH and PA in lungs and plasma, albino Wistar rats were used that were exposed to POH inhalation. Tissues were subjected to chromatographic separation on an Agilent Zorbax Eclipse XDB C18 column, followed by detection of absorption in the ultraviolet (UV) range. In vitro, POH exerted cytotoxic activity against six different lung tumor cell lines, and apoptotic cell death was indicated by induction of active caspase 3 and cleavage of poly (ADP-ribose) polymerase 1 (PARP1). These results demonstrate that inhalational delivery of POH results in effective biodistribution and metabolism of POH in the systemic circulation. In addition, our study introduces a simple, rapid HPLC-UV method with high accuracy for simultaneous detection of POH and its metabolite PA in plasma, and for sensitive detection of PA in lung tissue, which should prove useful for applications in clinical studies.
Assuntos
Antineoplásicos/farmacocinética , Cicloexenos/metabolismo , Pulmão/metabolismo , Monoterpenos/metabolismo , Monoterpenos/farmacocinética , Administração por Inalação , Animais , Antineoplásicos/administração & dosagem , Antineoplásicos/sangue , Antineoplásicos/metabolismo , Linhagem Celular Tumoral , Cromatografia Líquida de Alta Pressão , Cicloexenos/sangue , Cicloexenos/farmacocinética , Monitoramento de Medicamentos , Humanos , Pulmão/efeitos dos fármacos , Neoplasias Pulmonares/tratamento farmacológico , Masculino , Monoterpenos/administração & dosagem , Monoterpenos/sangue , Ratos , Ratos Wistar , Distribuição TecidualRESUMO
Different types of hair were submitted to different milling procedures and their resulting powders were analyzed by scanning electron microscopy (SEM) and laser diffraction (LD). SEM results were qualitative whereas LD results were quantitative and accurately characterized the hair powders through their particle size distribution (PSD). Different types of hair were submitted to an optimized milling conditions and their PSD was quite similar. A good correlation was obtained between PSD results and ketamine concentration in a hair sample analyzed by LC-MS/MS. Hair samples were frozen in liquid nitrogen for 5min and pulverized at 25Hz for 10min, resulting in 61% of particles <104µm and 39% from 104 to 1000µm. Doing so, a 359% increment on ketamine concentration was obtained for an authentic sample extracted after pulverization comparing with the same sample cut in 1mm fragments. When milling time was extended to 25min, >90% of particles were <60µm and an additional increment of 52.4% in ketamine content was obtained. PSD is a key feature on analysis of pulverized hair as it can affect the method recovery and reproducibility. In addition, PSD is an important issue on sample retesting and quality control procedures.
Assuntos
Cabelo/química , Cabelo/ultraestrutura , Entorpecentes/análise , Tamanho da Partícula , Manejo de Espécimes , Cromatografia Líquida , Cocaína/análise , Humanos , Ketamina/análise , Lasers , Espectrometria de Massas , Microscopia Eletrônica de Varredura , Pós , Detecção do Abuso de Substâncias/métodosRESUMO
A preparative protein alkaline hydrolysis procedure, as part of a spectrophotometric collagen quantification method, is presented. The procedure is suitable for small amounts of fresh solid or liquid samples. Various aspects of the procedure, such as the NaOH concentration, time needed to hydrolyse different collagen contents, buffer strength of the reagent solution, pH control of the hydrolysate and spectrophotometric conditions, were evaluated. Compared to other procedures that use alkaline hydrolysis, the sensitivity of this procedure was increased by a factor of 5. Compared to the conventionally used Association of Official Analytical Chemists (AOAC) acid hydrolysis method, the reaction time was reduced from 16 h to 40 min and the amount of sample from 4 g to 3-20 mg, producing equivalent results when applied to porcine liver and sausage samples.
Assuntos
Colágeno/análise , Hidroxiprolina/análise , Produtos da Carne/análise , Animais , Bioensaio , Colorimetria/métodos , Jejum , Concentração de Íons de Hidrogênio , Hidrólise , Fígado/química , Sensibilidade e Especificidade , Hidróxido de Sódio , Soluções , Espectrofotometria/métodos , SuínosRESUMO
Information on natural concentrations or variability of secondary metabolites in marine organisms may be important both to ecological/evolutionary and applied approaches. A gas chromatographic procedure with an electron capture detector (GC-ECD) was developed to quantify the sesquiterpenoid elatol at the surface and within-thalli of 70 specimens of the red seaweed Laurencia obtusa. The concentration of elatol was highest within-thalli [9.89 mg g(-1) of L. obtusa, dry weight (d.w.)], compared to lower values found at the surface [0.006 mg g(-1) of L.obtusa (d.w.), or 0.5-10.0 ng cm(-2)]. This method provides a rapid and inexpensive quantification of small quantities of elatol, and probably may also be used to quantify other halogenated compounds usually found in red seaweeds.