Study On The Chemical Constituents,Anti-flammatory Activity And Determination Of Total Flavonoids Of Nervilia FordⅡ(Hance) Schltr.

Objective:1. To study the chemical constituents of Nervilia fordii (Hance) Schlhtr.(QTK) and the anti-flammatory activity of the compounds which from this plant.2. To establish a method for determination of the content of Nervilia fordii (Hance) Schltr., and the quality of14batches of Nervilia fordii (Hance) Schltr. purchased from different markets are investigated. Methods:1. The cyclohexane extract and ethyl acetate extract of QTK. were isolated and purified by column chromatography.2. MTT assay is used for the determination of the toxicity of five compounds on the macrophage cell line RAW264.7, and the determination of RAW264.7cells on LPS-induced NO production.3. Rutin and AlCl3-HAc-NaAc (pH5.5) system are employed as reference substance and chromogenic reagent, respectively, the total flavonoids of Nervilia fordii (Hance) Schltr. are measured by UV spectrophotometry at the405nm. Results:The linear relationship of rutin was in the range of7.28-30.8mg·ml-1. The average recovery is100.73%(RSD=1.92%). The range of the amount of the total flavonoids of14batches of Nervilia fordii (Hance) Schltr. is1.25～3.12%. The average value of the total flavonoids of the three batches of Nervilia fordii (Hance) Schltr. from Guangdong province is2.69%, which is higher than that of the other samples. Conclusions:5compounds are isolated from Nervilia fordii (Hance) Schltr., the five compounds exhibited different LPS-induced production of NO inhibition, in which compound3showes strongest activity. This method is proved to be simple, quick, accurated, stable and reliable, and it could be used to determine the total flavonoids of Nervilia fordii (Hance) Schltr..