| Turkish galls(TG),with inducing astringency,astringency,eliminating dampness,hemostasis and anti-inflammation effects,is the major material of "Wei" ethnic medicine preparation "kui-jie-an".The preparation is widely used for the treatment of ulcerative colitis disease.It is called for an efficient and reliable method for the quality control of TG because of its increasing clinical application and qualityproblems.However,there was no any suitable and selective method reported for the quality control of TG,so far,which limits the further research and utilization of TG.ObjectiveTo make a series of studies about qualitative analysis,metabolite analysis and pharmacokinetics study of gallic acid,methyl gallate and 1,3,6-tri-O-galloyl-?-D-glucose of TG for providing scientific basis for the further research and utilization of TG.Methods1.Qualitative analysisThe rapidly profiling,identifying and determining of chemical constituents in TG was achieved by using a high performance liquid chromatography-electrospray ionization-ion trap-time-of-flight-tandem mass spectrometry(HPLC-ESI-IT-TOF-MS/MS),for the qualitity control of this herb.2.Metabolite analysisThe rapidly profiling,identifying and determining of metabolites in TG was achieved by using a high performance liquid chromatography-electrospray ionization-ion trap-time-of-flight-tandem mass spectrometry(HPLC-ESI-IT-TOF-MS/MS).3.Pharmacokinetics(PK)study of gallic acid,methyl gallate and 1,3,6-tri-O-galloyl-?-D-glucoseIn this study,a novel LC-MS/MS method was developed and validated for the simultaneous quantifcation of gallic acid,methyl gallate and 1,3,6-tri-O-galloyl-?-D-glucose in rat plasma,which are the major bioactive compounds of TG,MS was operated in negative mode.And the method was applied in a PK study of gallic acid,methyl gallate and 1,3,6-tri-O-galloyl-?-D-glucose after a single oral administration of TG extract to rats.Results1.Qualitative analysisA total of 21 compounds including 4 phenolic acid-types?15 Turkish gallotannin-types and other types were identified or tentatively characterized by HPLC-ESI-IT-TOF-MS/MS based on standard components comparison,UV absorption curve and MS spectra data in negative ion mode,13 of which were firstly reported from TG.2.Metabolite analysisA total of 21 metabolites were identified by HPLC-ESI-IT-TOF-MS/MS based on UV absorption curve and MS spectra data in negative ion mode,13 metabolites of which were firstly reported from TG.3.Pharmacokinetics(PK)study of gallic acid,methyl gallate and 1,3,6-tri-O-galloyl-?-D-glucoseThis method was linear between 0.1 and 50 ?g/ml for gallic acid,0.04 and 100?g/ml for methyl gallate and 0.05 and 5 ?g/ml for 1,3,6-tri-O-galloyl-?-D-glucose.The MS/MS ion transitions monitored were m/z 169.0?125.0,m/z 183.0?124.0,m/z 635.1?169.1 and m/z 179.2?135.0 for gallic acid,methyl gallate,1,3,6-tri-O-galloyl-?-D-glucose and caffeic acid(IS),respectively.Linearity,accuracy,precision and extraction recovery of three analytes were all satisfactory.The PK study indicated that the PK characteristics of gallic acid,methyl gallate,and 1,3,6-tri-O-galloyl-?-D-glucose have been proposed to be similar to each other and they absorbed quickly and eliminated very slowly after administration.ConclusionIn this study,the newly developed HPLC-ESI-IT-TOF-MS/MS method was effective and comprehensive and could act as an useful tool for qualitative assessment and quality control of both of TG and its related medicinal products in terms of selectivity,detection levels and structural information;the PK study of gallic acid,methyl gallate,and 1,3,6-tri-O-galloyl-?-D-glucose has filled the blank of PK study of TG and it will be helpful to better understand the pharmacological and clinical effects of TG. |
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