Determination Of The Content Of The Active Ingredient In Traditional Chinese Medicine By HPLC

This thesis is composed by two parts of the review and research reports. Reviewof part mainly introduces the development of Chinese medicine、the characteristics ofhigh performance liquid chromatography、 application of HPLC to content oftraditional Chinese medicine and the Index components selection of determination ofthe content of Chinese herbal medicines. Research reports of part establishes thedetermination of the content of the active ingredient in fuyankang granules、xiaoyanlidan granules、yanbianning granules、piyanxiao granules、fufangshenghuagranules by HPLC.Research reports composed of five parts as follows.1. Determination of Seven Effective Constituents in Fuyankang Granules by HPLCTo establish a method for the simultaneous determination of Protocatechuicacid、 Protocatethualdehyde and andrographolide, dehydroandrographolide,cryptotanshinone, tanshinone Ⅰand tanshinone ⅡA in Fuyankang granules byHPLC. Agilent ZORBAX SB-C₁₈column（4.6mm×250mm，5μm）was used. Themobile phase were methanol^-1%acetic acid（25:75） and water（A）-methanol（B）,gradient elution（0¹5min,58%B;15³5min,80%B;35⁴0min,58%B）；The flow rate were1.0mL min^-1and1.2mL min^-1；the detection wavelengthwere280nm and254nm；the column temperature was28℃.The linear ranges ofProtocatechuic acid、 Protocatethualdehyde、 andrographolide、dehydroandrographolide,、cryptotanshinone、 tanshinone Ⅰand tanshinone ⅡAwere1.408⁷04、1.096⁵48、1.0976⁷84，0.6888⁴92，1.0136⁷20.4，0.5824⁴16，0.5152³68μg·mL^-1,respectively. The average recoveries （n=5） of Protocatechuicacid、 Protocatethualdehyde、 andrographolide, dehydroandrographolide,cryptotanshinone, tanshinone Ⅰand tanshinone ⅡA were102.3%、102.1%、 100.2%、99.97%、98.93%、100.83%、99.38%，RSD were0.86%、0.70%、1.34%、2.57%、1.53%、2.42%、2.64%.The method seems simple,accurate,it can be used asquality control in Fanyankang granules.2. Determination of Five Effective Constituents in Xiaoyanlidan Granules by HPLC.To set up a determination method of content of five active ingredient inXiaoyanlidan granules. The HPLC method was established for Xiaoyanlidan granules.The analysis was performed on an Agilent ZORBAX SB-C₁₈（4.6mm×250mm，5μm） column. Polydatin: The mobile phase was composed of acetonitrile and water（25:75）with at a flow rate of1.0mL·min-1. theUV detection wavelength was306nm.The linear ranges of polydatin was0.24⁸4.8μg·mL-1（r=0.9999）, respectively.Resveratrol：The mobile phase was composed of acetonitrile and4%glacial aceticacid（27:73）with at a flow rate of1.2mL·min-1. theUV detection wavelength was308nm. The linear ranges of resveratrol was2.44²44μg·mL-1（r=0.9998）,respectively.Paeoniflorin：The mobile phase was composed of methanol and2%phosphate（40:60）with at a flow rate of1.0mL·min-1. theUV detection wavelength was230nm. Thelinear ranges of paeoniflorin was1.008¹68μg·mL-1（r=0.9998）, respectively.Rhein and Emodin: The mobile phase was composed of methanol and2%glacialacetic acid （80:20）with at a flow rate of1.0mL·min-1. theUV detection wavelengthwas254nm. The linear ranges of rhein and emodin were0.054¹80and0.0612²04μg·mL-1（r=0.9999）, respectively. This method is simple accurate and reproduceable which can be used for the quality control of Xiaoyanlidan granules.3. Determination of Catalpol,Cinnamicacid and Chlorogenic acid in YanbianningGranules by HPLCTo establish a method for determination of catalpol,cinnamicacid andchlorogenic acid in Yanbianning granules. Catalpol,cinnamicacid and chlorogenicacid were determined by HPLC on C₁₈column at212nm,278nm and327nm,withwater-methanol-acetonitrile（95:4:1）,methanol^-1%ice acetic（50:50） and methanol-3%ice acetic（15:85） as mobile phase.The flow rate were1.0mL·min-1,1.2mL·min^-1and1.0mL·min-1.Under each condition,the calibration curves of catalpol ,cinnamic acid and chlorogenic acid were linear in the ranges of3.36¹68、0.156¹04、0.3544¹77.2μg·mL^-1, respectively.And the each average recoveries ofthe method were102.09%for catalpol （RSD1.09%,n=5）,102.1%for cinnamicacid（RSD0.99%,n=5）,100.7%for chlorogenic acid（RSD1.14%,n=5）. The method israpid and reliable for qulity control of yanbianning granules4. Determination of Paeonoland and Baicalin in Piyanxiao Granules by HPLCTo set up a determination method of content of paeonoland and baicalin inPiyanxiao granules. Paeonoland and baicalin were determined by HPLC on C₁₈column at274nm and278nm.with methanol-water（67:33） and methanol-0.2%phosphate（55:45） as mobile phase.The flow rate were1.0mL·min-1and1.2mL·min-1. The linear ranges of paeonoland and baicalin were0.136²72、1.428⁴76μg·mL^-1.The each average recoveries of the methods were100.22%for paeonoland（RSD1.39%）,100.52%for baicalin（RSD1.33%）.The method is simple,rapid and is agood method for paeonoland and baicalin.5.Determination of Naringin and Hesperidin in Fufangshenghua Granule by HPLCTo establish the method for determination of naringin and hesperidin inFufangshenghua granule. The column was Agilent ZORBAX SB—C₁₈（4.6mm×250mm,5μm）,The flow rate was1.0mL·min-1.The detection wavelength was277nm. The content of naringin ranged from1.64to116.9μg·mL^-1and that of hesperidinfrom1.06to42.4μg·mL^-1.The method is simple, reproducible, more accurate results.It can be used for determine quality control of Fufangshenghua granule.