Studies On Determination Of The Content And The Characteristic Chromatogram Of Ephedra Formula Granules By HPLC

Ephedra is the dried herbaceous stems from Ephedra sinica Stapf,Ephedra intermediate Schrenk et C.A.Mey.or Ephedra equisetina Bge.,of the Ephedraceae.The Ephedra Formula Granules are made from Ephedra Pieces conforming to Pharmacopoeia regulation and manufactured by modern technology of extract,concentration,spray drying and granulation.The advantages of Ephedra Formula Granules are no need of decoction,of small volume,and easy for carrying,transportation and storage.In order to control the quality of Ephedra Formulation Granules and to ensure the safety and effectiveness of clinical medication,in this thesis,the content of Ephedrine and Pseudoephedrine in Ephedra Formula Granules was determined,and the HPLC characteristic chromatogram of Ephedra Formula Granules were also studied.Part 1 Determination of Ephedrine and Pseudoephedrine in Ephedra Formula GranulesObjective: To establish a method for determination of Ephedrine and Pseudoephedrine in Ephedra Formula Granules by HPLC.Methods: The analysis was carried out by a Wondasil C18 column(4.6×250mm,5μm)with acetonitrile and 0.1% phosphoric acid(5:95)as the mobile phase.The detection wavelength was set at 210 nm.The flow rate was set at 1.0ml/min.The column temperature was kept at 30℃.The sample volume was 10μl.Results: The linear equation: Ephedrine Y=26.6967X-0.0631;Pseudoephedrine Y=29.2241X+0.0593.The linear range: Ephedrine 0.0826μg ~ 0.8256μg(r=0.9999);Pseudoephedrine 0.0404μg ~ 0.4044μg(r=0.9998).The average recovery: Ephedrine 99.52%(RSD=1.22%,n=9);Pseudoephedrine 98.66%(RSD=1.71%,n=9).Conclusions: The method is stable,reproducible and simple.Hence it is an ideal method for the determination of Ephedrine and Pseudoephedrine in Ephedra Formula Granules.Part 2 Study on the Characteristic Chromatogram of Ephedra Formula Granules by HPLCObjective: To establish the HPLC characteristic chromatogram of Ephedra Formula Granules.Methods: The analysis was carried out by a Wondasil C18 column(4.6×250mm,5μm)with acetonitrile and 0.1% phosphoric acid as the mobile phase in a gradient elution mode.The detection wavelength was set at 210 nm.The flow rate was set at 1.0ml/min.The column temperature was kept at 30℃.The sample volume was 10μl.The data collection time was 60 min.Results: The HPLC characteristic chromatogram of the Ephedra Formula Granules were obtained.The chromatograms of the 10 batches of the samples were basically the same.All the chromatograms possessed 15 same chrmatographic peaks in common.The components of Ephedrine and Pseudoephedrine were identified in the 15 peaks.The similarity of HPLC characteristic chromatograms of Ephedra Formula Granules was calculated by using the similarity evaluation system of traditional Chinese medicine chromatographic fingerprints(2012 version).The similarity of the 10 batches of samples was more than 0.98.Conclusions:The method has good stability and reproducibility.The HPLC characteristic chromatograms can provide references for the identification and quality evaluation of Ephedra Formula Granules.