The Establishmentof Folium Syringae Extract HPLC Fingerprint And Determination Of Protocatechuic Acid Component

Folium Syringae, having a broad spectrum of antibacterial and antiviral effect, was dry leaves of Syringa oblata Lindl, Syringa vulgaris L, Syringa diatata Nakai being Oleacea Syringa, which was proved as the most development potential of Chinese medicine through its plant resource, chemical constituents, pharmacological action and clinical application research report. Active ingredient being contained by traditional Chinese medicine was the material basis of traditional Chinese medicine efficacy and any a kind of active ingredients could not reflect the overall efficacy of clinical medicine of traditional Chinese medicine. TCM fingerprint was a multi-index quality control model, which not only comprehensive reflected the amount and speicies of traditional Chinese medicine(TCM) but also effectively reflected the Chinese native medicine ingredient integrity and comprehensive effect, that to better evaluate the quality of traditional Chinese medicine and its preparations. We could not effectively control quality and ensure the bioequivalence only to analyze a single component index because of the differences between the species and content of Folium Syringae active ingredients by the source, origin, ecological environment, collecting time and so on factors of medicinal materials.Folium Syringae was the research object of this experiment, with C18column being solid phase extraction column for pretreatment. HPLC-diode array detector(DAD) method was established to detect protocatechuic acid which analyzed the Folium Syringae effective substances to guarantee the feasibility, accuracy and precision of experimental method. The results are as follows:HLB column was determined as SPE column through the selection of SPE column by comparing C18column and HLB adsorption recovery with90.48%of HLB and88.79%of C18. Pure methanol was determined as elution solvent by comparing recovery of eluting target samples of different elution solvents like methanol, acetone, ethyl acetate elution. Elution curve was established to determine the target substance elution volume of5ml which optimized the sample purification process to select an ideal pretreatment method. The experimental results show that the method has the advantage of fast extraction and good purification effect, easy operation, short analysis time, test number, high accuracy.Protocatechuic acid in Folium Syringae extract detection technology was researched and developed, with HPLC reversed phase chromatographic technology being main means of analysis by comparing column column efficiency of XDB-C18, ASB-C18, ODS C18, mobile phase selection, acidity. Folium Syringae extract SPE-HPLC fingerprint was established by determining to use ODS-C18(250*4.6mm,5microns) column, diode array detector, mobile phase of methanol-20mmol/L potassium dihydrogen phosphate(20:80, v:v), acidity of pH=2, detection wavelength of260nm, sample quantity of20μL, flow rate of1.0mL/min, column temperature of30℃. This method of precision, stability, repeatability was good through RSD of Folium Syringae extract precision, repeatability, stability being less than3%. Protocatechuic acid was determined as better-stability characteristic peak, which was evaluated adopting national pharmacopoeia committee issued the " Traditional Chinese medicine(TCM) chromatographic fingerprint similarity evaluation system "(2004a) with the similarity results of10batches of sample being above0.9, which conformed to the rules. The quantitative fingerprint characteristic was treated with cluster analysis and principal component analysis with the help of SPSS20.0software, which proved to be consistent with the similarity evaluation. We could effectively evaluate the Folium Syringae extract fingerprint.HPLC analysis method established was evaluated through the linearity, precision, accuracy, and recovery with standard curve of y=75.757x-9.3676, R2=0.9997and protocatechuic acid had good linear relationship in2.58～83.32μg/mh with precision RSD less than1%that instrument precision was good. The average recovery of each component was96.79%, which indicated good accuracy. Experiments showed that the method was simple, stable and good reproducibility. Content determination of samples collected from Qiqihaer, Zibo, Weifang, Daqing, Beijing, Changchun, Shenyang, HuLin, Jixi, Harbin was in29.67～78.87μg/g with Harbin and Jixi sample content higher than other provinces.