Components Analysis Of Y-Oryzanol Tablets And Improve The Quality Standards

Objective:To establish the HPLC method for content determination of y-Oryzanol tablets,while by using liquid chromatography mass spectrometry to analyze the structure of the components of it,and by using the fingerprint similarity evaluation system to analyze the similarity among these tablets.To establish the ICP-MS method for examining the content of trace elements which may exist in y-Oryzanol.Methods:1.Components' structure analyzing:using the LC-Q-TOF with a C18 column(5?m,4.6x250mm),while the mobile phase was composed of acetonitrile and methanol(containing 5mmol/L ammonium formate)(50:50),with a flow rate at 1.0ml/min,column temperature at 30?.Using electrospray ionization(ESI)ion source,and collecting data in the negative ion mode.2.Content determination of HPLC method:using C18 column(5?m,4.6x250mm),with the detection wavelength at 326nm.The mobile phase was composed of acetonitrile,methanol,methylene chloride and glacial acetic acid(50:50:0.3:0.3),and the flow rate was 1.0ml/min,with column temperature at 30?.3.Similarity analyzing:using the fingerprint similarity evaluation system of 2009 to analyze the similarity of y-Oryzanol tablets according to the components and relative contents.4.Contents of trace elements determination of ICP-MS method:using ICP-MS to examine the contents of those trace elements,including Cr,Mn,Ni,Cu,As,Cd,Hg and Pb,with Sc,Ge,In and Bi as internal standards.Results:1.Molecular weight of the corresponding compounds was obtained by MS1,cleavage fragments information of the corresponding compounds was obtained by MS2.Uniting correlation literature to speculate fragmentation pathway of each component,and preliminary identified structures of 8 components of y-Oryzanol tablets.2.Established the method for content determination:y-Oryzanol was in a good linearity within the range of 20.10-100.50?g/ml(r=1).The average recovery was 101.2%with RSD at 0.4%accordingly.3.Results of similarity:domestic products showed a high similarity of the composition,however,with a quite difference of relative contents.Besides,they also showed a great difference of the chromatographic peak in characteristics to those product by Otsuka Pharmaceutical Co.,Ltd.of Japan.4.Established the method for trace elements contents determination:each element in all was in a good linearity(r>0.999),with the recovery from 89.0%to 117.0%(RSD<3.0%accordingly).The detection limit was 0.001-0.009mg/kg.Conclusions:This experiment preliminary identified structures of 8 components in y-Oryzanol tablets,established and verified the HPLC method for determination of y-Oryzanol tablets,developed and verified the inductively coupled plasma mass spectrometry method for the determination of trace elements content in oryzanol;analysed the similarity of the chemical composition and relative content of domestic oryzanol tablets;provided scientific theory basis for improving the quality of y-Oryzanol tablets standard.