RESUMEN
OBJECTIVE: To investigate the technological parameters and the process for separation and purification of pumiloside (PML) from Nauclea officinalis Pierrc ex Pitard by macroreticular resin. METHODS: PML was extracted by hot water, and the content of PML was determinated by HPLC method. The static capacity absorption, static elution ratios of five types of resin were studied respectively, and were compared to evaluate their absorption and desorption effect to PML. And then the absorption capacity, elution solution and elution volume of the AB-8 resin were researched to set up the optimum separation process for PML. Finally, PML was recrysallized from MeOH and identified by spectra analysis. RESULTS: The AB-8 resin had the best absorptive and separative properties to PML. The dynamic absorption ratio was 2.44 mg/g. The applicable process was as follows: the water extracted solution of Nauclea officinalis flow through the resin column repeatedly, after being eluted with 6BV of distilled water, the resin column was eluted with 4BV 30% ethanol, the 30% ethanol fraction was combined and the solvent was recovered in vacuum. The precipitation was filtered and recrysallized from MeOH to give pure PML. The yield of PML was 75.1%, and the product purity was up to 99.5%. CONCLUSION: AB-8 resin shows better comprehensive absorption property. It can be used for separation and purification of PML from Nauclea officinalis successfully.
Asunto(s)
Alcaloides/aislamiento & purificación , Camptotecina/análogos & derivados , Plantas Medicinales/química , Resinas Sintéticas/química , Rubiaceae/química , Tecnología Farmacéutica/métodos , Adsorción , Alcaloides/química , Camptotecina/química , Camptotecina/aislamiento & purificación , Cromatografía Líquida de Alta Presión , Medicamentos Herbarios Chinos/química , Medicamentos Herbarios Chinos/aislamiento & purificación , Etanol , Reproducibilidad de los Resultados , AguaRESUMEN
OBJECTIVE: To extract and identify semen-derived exosome using PEG6000. METHODS: Exosomes were extracted from semen specimens from 6 healthy volunteers with step-by-step centrifugations and ultracentrifugation prior to 8% PEG6000 enrichment. The extracted exosomes were characterized by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA) and Western blotting. RESULTS: The pellets obtained were round or elliptic membrane vesicles 30 to 150 nm in diameter with intact double membranes and contained low electron density material. The pellets expressed CD63, ALIX and TSG101 molecules but not calnexin that was expressed in sperm cells. CONCLUSION: The PEG6000-based method for extraction of exosomes from semen samples facilitates future studies of seminal exosomes.