Urinary Metabonomic Investigation into Acute Photo Damage of Rats Induced by Ultraviolet B Irradiation using Rapid Resolution Liquid Chromatography-Mass Spectrometry / 分析化学

ABSTRACT

A rapid resolution liquid chromatography quadruple time-of-flight mass spectrometric ( RRLC-Q-TOF/MS) method combined with multivariate statistical analysis was applied to investigate the changes of endogenous metabolites in murine urine before and after ultraviolet B ( UVB ) irradiation for the purpose of discussing the physiological mechanism of acute injury caused by UVB radiation. A narrow-band UVB ( NB-UVB) (TL-01, peak value 312nm) was used to establish the acute light damage model. The urine samples were centrifuged before four times dilution treatment, subsequently the diluted urine samples were separated on a Supelco Ascentis Express C18 column using water (0. 1% formic acid) and acetonitrile as mobile phase by gradient elution. The differences metabolites with major contribution for grouping were found out based on the metabolic profiling analysis of principal component analysis ( PCA) and cluster analysis ( CA) , which could illustrate their possible mechanism of actions by means of relevant pathways. A prediction model was built to investigate the forecasting ability of the acute photo damage induced by UVB irradiation through the partial least square discriminant analysis ( PLS-DA ) . The results of multivariate statistical analysis showed that the blank control group was separated from UVB model group quite well, 11 endogenous metabolites were identified as the potential biomarkers through comparison with the database, tandem mass spectrum data and standard substance, which indicated the UVB radiation may affect the sphingolipid metabolism, nucleic acid metabolism, linoleic acid metabolism and amino acid metabolism pathways. These different metabolites could be helpful for diagnosing the light damage induced by UVB radiation