RESUMO
OBJECTIVE: To develop an approach to the determination of saponins in Radix Cynanchi Atrati, and to optimize the parameters for purified the preparation of total saponins by macroporous resin column chromatography. METHOD: Using cynanversicoside A as a reference, the determination of saponins was performed; according to the elution rate and the purity of the products, the preparation performance of total saponins by macroporous resin was investigated, and its parameters were optimized. RESULT: The saponins in Radix Cynanchi Atrati were successfully determined at 518 nm by vanillin-perchloric acid as spray reagent. The macroporous resin HP-20 showed static absorption ratio of 59. 3 mg x g(-1); the 70% ethanol extraction of Radix Cynanchi Atrati was eluted from column of macroporous resin HP-20 by water and 30% ethanol, and the saponins were concentrated in 90% ethanol solution. The content of saponin part eluted from HP-20 column was 77.62%. CONCLUSION: The proposed approach allows convenient and efficient preparation and purification of saponin in Radix Cynanchi Atrati.
Assuntos
Cynanchum/química , Resinas Vegetais/química , Saponinas/química , Saponinas/isolamento & purificação , Absorção , Benzaldeídos/química , Calibragem , Etanol/química , Percloratos/química , Porosidade , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
INTRODUCTION: Early diagnosis and treatment for thermal injury with septic complications continue to be a serious clinical problem. In this study, plasma biomarkers of rats in the burn and/or septic models were investigated with a metabolomic method. METHODS: Rat plasma samples were analyzed by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). Multivariate analysis, the principal components analysis (PCA), was used to validate metabolic changes. In addition, another multivariate method, the orthogonal partial least-squares analysis (OPLS), was used to profile potential biomarkers in models. RESULTS: Nine characteristic metabolites, including hypoxanthine, indoxyl sufate, glucuronic acid, gluconic acid, proline, uracil, nitrotyrosine, uric acid, and trihydroxy cholanoic acid were identified in models of thermal injury and/or sepsis. CONCLUSION: These biomarkers were mainly involved in oxidative stress and tissue damage, and might supply evidence for distinguishing burned septic patients from non-septic ones.
Assuntos
Queimaduras/sangue , Sepse/sangue , Aminoácidos/sangue , Animais , Biomarcadores/sangue , Ácidos Carboxílicos/sangue , Cromatografia Líquida de Alta Pressão/métodos , Modelos Animais de Doenças , Compostos Heterocíclicos/sangue , Masculino , Espectrometria de Massas , Metabolômica/métodos , Ratos , Ratos Sprague-Dawley , Sepse/diagnósticoRESUMO
Peripheral nerve injury, which gives rise to persistent chronic pain, has become an area of intense research activity, largely because it represents a disorder with a high unmet medical need. In this study, serum biomarkers of the spinal nerve ligation model were successfully investigated with the metabolomic method. The regulatory effect of gabapentin, a novel clinical antineuralgia drug, on biomarker levels in serum was also investigated. Rat serum extract samples were analyzed by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). A method of supervised multivariate analysis, the partial least squares-discrimination analysis (PLS-DA), was used to validate metabolic changes. In addition, another multivariate method, the orthogonal partial least-squares analysis (OPLS), was used to monitor the real biological variability and to detect potential biomarkers in the spinal nerve ligation model. The results demonstrated that the spinal nerve ligation model had several discriminating ions compared with the control model. Among the detectable metabolites, levels of norepinephrine were increased in the spinal nerve ligation model and were decreased to control levels by gabapentin.