Extraction Method Optimization And Analyse Of HPLC Fingerprints Of Tripterygium Hypoglaucum (Levl.) Hutch (THH)

Objective :Tripterygium Hypoglaucum (Levl.) Hutch(THH)is a traditional Chinese medicine with anti-inflammatory, detoxification, and rheumatism effects. THH has been used clinically in rheumatoid arthritis, chronic nephritis and various kinds of skin diseases, and its pharmacognosy, chemical compositions, pharmacological effects and clinical applications have been studied by many researchers. THH's tablet preparation has been recorded in Chinese Pharmacopeia of 2005'edition. However, there is a great difference of active ingredient of THH with different locality of growth. In order to improve the quality control of THH, the extraction methods and content determination of Triptolide were studied in this paper. The HPLC fingerprint of THH from 11 different localities was analyzed besides determination the content of Triptolide.Methods: Extraction methods of THH were studied by methods of natural pharmacochemistry, such as lixiviation, abstraction and rotary evaporation,which then was optimized through the use of single factor experiments and orthogonal test. Content determination of Triptolide was performed by RP-HPLC (Reversed phase high-performance liquid chromatography). The fingerprint of THH from different area was determined by HPLC method.Results and Conclutions: (1) The optimal extraction parameters were as below: extraction solvent--ethyl acetate, 1:10 (g/ml) of solid to liquid ratio, and 5 hours reflux extraction. (2) There is a big difference of Triptolide in THH from 11 different area, which showed its necessary to study the quality of THH. (3) The fingerprint analysis by HPLC was follows: methanol-acetonitrile-water gradient elution; column: Agilent C18 column (5μm ,4.6mm×250mm); UV detection wavelength: 210nm; column temperature: 30℃; Injection volume: 20μl. (4)The fingerprints of THH form 11 area showed good similarity with big difference in content of ingredients. The similarity of THH form Dali Yangbi and Dali Binchuan were only 0.651 and 0.527. Our studies provided useful data for quality control of THH.