Study On The Quality Evaluation Methods Of Sophora Flavescentis Radix And Eriobotryae Folium

Traditional Chinese Medicine(TCM)is an important part of the Chinese medicine system.Its quality has always been one of the important foundations for Chinese medicine development.,and affects the safety and effectiveness of the clinical medicine.Since the implementation of the TCM modernization development strategy,a series of modern analytical techniques such as spectroscopy and chromatography,has improved the TCM quality control evaluation system greatly with indicator components as the core.But this still cannot meet the ever-increasing quality control requirements.This research is financially supported by the National Key Research and Development Program "Research on Modernization of Traditional Chinese Medicine"(2018YFC1707900),subproject No.3 "Key technologies for qualitative identification based on overall quality control and formulation of international quality standards".After collecting the medicinal materials of Sophora flavescens Radix(SFR)and Eriobotryae Folium(EF)from different origins,the quality control methods for thin layer chromatography,HPLC fingerprint and HPLC content determination of SFR and EF were established,thus to provide a technical support for improving the quality evaluation standards of SFR and EF.1.Study on the quality evaluation methods of SFRSFR is the dry root of the leguminous plant Sophora flavescens Ait,which was firstly recorded in Shennong Materia Medicina Classic.It has a long history with functions of clearing away heat and dampness,killing insects and diuretics.(1)The TLC of SFR was established after investigating different developing system,different kinds of thin layer plate,different developing height and other factors.The thin plate is Merck GF254;Extraction method:To 2 g of SFR powder,add 2 mL concentrated ammonia and 10 mL trichloromethane,ultrasound for 30 minutes and filtered.Evaporate the filtrate to dryness,dissolve the residue in 2 mL of methanol as the test solution.Use a mixture of trichloromethane,methanol and concentrated ammonia(5:6:0.1)as the mobile phase.Spray with potassium iodobismuthate successively.Three orange spots in the chromatogram obtained with the test solution correspond in position and colour to the spots obtained with the reference solution.(2)For the HPLC fingerprint,the extraction conditions of SFR were determined after investigation of different extraction methods,and the chromatographic conditions for fingerprint were developed as:using an Agilent Poroshell HPH-C18(3 mmx 100 mm,2.7 μm)column with acetonitrile-0.05 mol/L potassium dihydrogen phosphate(2 mL/L triethylamine)as mobile phase and gradient elution.The detection wavelength was set at 210 nm and the flow rate was 0.5 mL/min.A total of 14 common peaks were detected and 4 of them were identified by comparison with references.The similarity of common peaks in 40 batches of SFR ranged from 0.890 to 0.996.(3)For the content control of SFR,different preparation methods and chromatographic conditions were investigated,and the method was decided as:using Phenomenex Luna-C18 column(4.6 mm × 250 mm,5 μm),and acetonitrile-[0.01 mol/L ammonium acetate solution(concentrated ammonia test solution adjusted to pH 8.1)](3:2)as mobile phase A,0.01 mol/L ammonium acetate solution(concentrated ammonia test solution adjusted to pH 8.1)as mobile phase B,with gradient elution.The contents of matrine and oxymatrine were 0.94%-3.36%in 40 batches of SFR,indicating that the quality was different among different producing areas.2.Study on the quality evaluation methods of EFEF is the dried leaf of Eriobotrya japonica(Thunb.)Lindl,with effects of clearing the lung,relieving cough,relieving nausea and relieving nausea.In this study,a total of 24 batches of EF were collected from Zhejiang,Fujian,Jiangsu,Guangxi,Guangdong and other places.Through sorting and analyzing the chemical constituents,pharmacological effects and quality markers of EF,this research provides a reference for the establishment of relevant quality standards.(1)Thin layer chromatography(TLC)of EF was established after investigating different developing systems,different brands of thin layer plates and different developing heights.Merck GF254 plate is selected.The extraction method is:ultrasound with 20 times methanol for 30 minutes;Use the mixture of cyclohexane-ethyl acetate-glacial acetic acid(8:4:0.1)as the mobile phase,After developing and removal of the plate,dry in air,spray with a 10%solution of sulfuric acid in ethanol,and heat at 105℃ to the spots clear.The spots in the chcromatogram obtained with the test solution correspond in position and colour to the spots in the chromatograms obtained with the reference drug solution and reference solution,Moreover,it can be used to distinguish EF with those confused products such as photinia serrulata,dillenia turbinata and Magnolia lotosa readily。(2)For the fingerprint,the extraction conditions of EF were determined after investigating of different extraction methods,and the chromatographic conditions for fingerprint were developed as:using an Agilent ZORBAX SB-C18(4.6 mm × 250 mm,5 μm)with gradient elution using methanol-0.1%formic acid as mobile phase and ELSD detector at the flow rate of 0.8 mL/min.Six common peaks were detected and 4 of them were identified by comparison with references.The similarity of common peaks in 24 batches of EF was 0.818-1.000.(3)For the content control of EF,different chromatographic conditions were investigated and the method was decided as:using Agilent ZORBAX SB-C18(4.6 mm × 250 mm,5 μm)chromatographic column with methanol-0.5%ammonium acetate as the mobile phase for isocratic elution,and ELSD detector was selected.The contents of 24 batches of EF was determined,and the total content of ursolic acid and corosolic acid was 0.61%～1.03%.