The Quality Of Radix Paeoniae Alba Formula Particles And In Vitro Anti HepG2 Cell Proliferation Activity

Objective: Radix paeoniae alba formula particles is the root of herbaceous peony yinpian, concentration, spray after boiled, add right amount particles made of dextrin, as a new form of slices instead of traditional Chinese medicine yinpian for clinical prescription. This paper aims to the quality of radix paeoniae alba formula particles and initially the antitumor activity in vitro, the further development and clinical application of radix paeoniae alba formula particles to provide the reference.Methods:1 The quality of researchIn 3 batch of pilot products as the research object, from the aspects of thin layer identification and content determination, determine the quality of radix paeoniae alba formula particles.1.1 Thin layer identification methodBy thin layer chromatography, respectively for the preparation of test sample solution method, developing solvent system, examining method etc were studied.1.2 Content determination methodEstablish a HPLC method for determining the content of paeoniflorin in radix paeoniae alba formula particles, a study on methodology and, ultimately for radix paeoniae alba formula particles content determination method and limit, as the important basis to evaluate the quality of the radix paeoniae alba formula particles.2 HepG2 cells in vitro proliferation activity of the preliminary studyIn vitro human hepatocellular carcinoma HepG2 cells, radix paeoniae alba formula particles determined by MTT method to observe the different concentration(1.6, 0.8, 0.4, 0.2, 0.1 g/L) influence on HepG2 cell proliferation,calculate the inhibition rate(inhibition rate % = 1- OD/OD controls).To observe the effects of Baishao Peifang granules on the apoptosis of HepG2 cells by Hoechst33342 staining.Results:1 The quality of the results of the study1.1 Qualitative identification resultsSet up the thin layer identification method of radix paeoniae alba formula particles.1.2 High performance liquid chromatography(HPLC) method for determining the content of paeoniflorinUsing high performance liquid chromatography(HPLC) method to establish the determination method of the content of paeoniflorin in radix paeoniae alba formula particles. Methodology test showed that paeoniflorin sample quantity within the scope of 0.0596 to 0.5960 mu g, had good linear relationship, the regression equation is: Y = 1.3612 + 1.9410 x 104 x 106 x, r= 0.9998; Que paeoniflorin average sample recovery rate was 98.30%(n = 6).The RSD was 1.59%, less than 2.0%, show that the method is accurate and reliable, good repeatability, which can realize to control the content of paeoniflorin in radix paeoniae alba formula particles.2 The influence of radix paeoniae alba formula granules on HepG2 cell proliferation0.2 ~ 1.6 g/L radix paeoniae alba formula granule can obviously inhibit HepG2 cell proliferation(P<0.01), the inhibition rate was concentration dependent, 0.01 mg/L, radix paeoniae alba formula particles incubation cells reached the maximum inhibition rate, about 80.5%.Fluorescence staining showed that the nucleus was dense and dense.And is accompanied by nuclear fragmentation,cell apoptosis and other typical apoptosis changes.Conclusion: This study to establish a TLC identification method for radix paeoniae alba formula granules paeoniflorin and the content of paeoniflorin in HPLC method; And found the root of herbaceous peony formula granule can obviously inhibit human hepatocellular carcinoma HepG2 cells proliferation,induce apoptosis, significant antitumor activity, related research results for the quality control of radix paeoniae alba formula particles and provides the experimental basis for clinical application.