Modern Processing Research Of Bupleurum With Turtle Blood Pieces And Realgar Granules Pieces

Bupleurum with turtle blood pieces is traditional characteristics Chinese herbal pieces. It is a vital TCM drugs for relieving exterior syndrome and relieving shaoyang disorder. After being stir-frying with turtle blood,it is used for asthenia palpipation since QingDynasty. This chinese herbal pieces can also remission the inferiority of Bupleurum.However.modern studies about Bupleurum processed with turtle blood are very rare.Which also lack of Standardization research and quality standard.The mechanism of toxicity reducing and efficacy enhancing is still unknown.Realgar granules pieces were prepared by grinding into coarse powder particles.This processing method originated in the han dynasty, has longer history than elutriation pieces. However.modern studies about realgar granules pieces are very rare. Processing method and dosage is determine the quality of pieces and curative effect. Which also lack of Standardization research and quality standard.1. To optimize and establish a technology of processed Bupleurum with china turtle blood. With composite score of saikosaponin a.saikosaponin c,saikosaponin d.ethanol soluble extractives as index, orthogonal test was adopted to optimize the technology of processing by taking the amount of turtle blood,processing time, and processing temperature as factors. Contents of saikosaponin a, saikosaponin c, saikosaponin d were determined by HPLC, gradient elution with Mobile phase acetonitrile-water, detection wavelength was at 210nm. Temperature had significant impact on the processing technology. Optimal processing technology was:processing time of 10 min, amount of china turtle blood 0.05g·g-1 processing temperature at 150℃; under these conditions,contents of ethanol soluble extractives, saikosaponin a.saikosaponin c, saikosaponin d were 12.42%,0.8504%,0.2348, 0.8978%,respectively. This optimized processing technology is reasonable, reliable and suitable,which could guide produce of processed Bupleurum.2. In the aspect of chemical composition analysis:(1)To improve the standard of stir-frying Bupleurum with turtle blood by establishing a HPLC method for simultaneous quantitative of saikosaponin a, saikosaponin c, saikosaponin d.The optimized method is accurate and reproducible.and suitable for the determination of saikosaponin in stir-frying Bupleurum with turtle blood.(2) To analyze the chemical components of the volatile oil in raw Bupleurum and processed with turtle blood pieces by GC-MS. The volatile oil were obtained from PPFA by steam distillation. The amounts of the components from the volatile oil were determined by area normalization method. The separated components were identified by GC-MS.One hundred and fourteen components were separated and identified from the raw Bupleurum.One hundred and five components were separated and identified from the stir-frying Bupleurum with turtle blood pieces. Which included forty-seven common components. The research can be used to analyse the volatile oil.(3) To establish a method for simultaneous determination of 17 amino acids in in raw and processed Buplurum. The HPLC analysis was performed after derivatization by using phenyl isothiocyanate (PITC) as a derivative reagent. The sample was and analyzed on Prevail C18 column (4.6×250mm.5μm) with gradient elution of acetonitrile -water (4:1) and sodium acetate buffer solution (adjusted to pH 6.5)-acetonitrile(93:7). The detected wavelength was 254nm. Using this method,17 amino acids can be separated well in 32min.The concentration showing good linear relationship(r2=0.9917-1).The average recovery was between 93.36%-107.11%. The RSD was between 1.83%～3.91%. Compared with other pre column derivatization method for determination of the content of amino acid. this method is reliable with good accuracy and repeatability, which is useful for the determination of amino acids in raw and processed Buplurum.(4) To study the determination method of the polysaccharide components in raw Bupleurum and processed with turtle blood pieces by UV. The optimized method is accurate and reproducible. and suitable for the determination of polysaccharide components.3.In the aspect of pharmacological research for stir-frying Bupleurum with turtle blood pieces. we have done plenty of reserches as follow:(1) To clarify the mechanism of processing mechanism research for stir-frying Bupleurum with turtle blood pieces.A UPLC/Q-TOF-MS-based method was used to investigate the effect of raw Bupleurum and processed with turtle blood pieces on treatment of Dry yeast induced fever in rats.Research on the change of endogeous metabolites of plasma,to find the invasion pathogenesis in metabolic levels.Significant changes were found after the treatment of stir-frying Bupleurum with turtle blood pieces.Twelve biomarkers include lipids.fatty acid and amino acids were identified. This piece of work can promote clinical application and development of stir-frying Bupleurum with turtle blood pieces.(2)To clarify the mechanism of processing mechanism research for stir-frying Bupleurum with turtle blood pieces. The rats were caused fever by dry yeast. The cAMP of hypothalamus, AVP of ventral septal, IL-1β of serum were analyzed. Stir-frying Bupleurum with turtle blood pieces has good antipyretic effects.This piece of work can promote clinical application and development of stir-frying Bupleurum with turtle blood pieces.(3) To clarify the mechanism of processing mechanism research for stir-frying Bupleurum with turtle blood pieces. The liver injury in rats were caused by feeding carbon tetrachloride (CCl4) solution. The AS、ALT、LDH、TBA of serum were analyzed. Stir-frying Bupleurum with turtle blood pieces has good antipyretic effects.This piece of work can promote clinical application and development of stir-frying Bupleurum with turtle blood pieces.4.In the aspect of processing technology for raw realgar and granules piece. We have done plenty of reserches as follow:Carries on the change tradition to the realgar tablets ready for decoction to be used as medicine the form to make the realgar coarse particles; The formulation realgar coarse particles store the maintenance method and guarantee the nature time. This piece of work can promote clinical application and development of realgar granules pieces.Discovers in 10 different habitat realgars As (III) the content is different.Carries on the change tradition to the realgar tablets ready for decoction to be used as medicine the form to make the realgar coarse particles; The formulation realgar coarse particles store the maintenance method and guarantee the nature time.5. In the aspect of chemical constituents for raw realgar and granules pieces, we have done plenty of reserches as follow:To establish methods for determination of As2O3 and As2S2. The content of dissolve darsenic As(Ⅲ) in Realgar was tested by ultraviolet spectrophotometric method. By UV spectrophotometry in different areas of realgar content of arsenic trioxide. Methods:Using diethyl dithiocarbamate silver method(Ag-DDC method), At the 520nm UV-Vis spectrophotometry seven batches of realgar in the content of arsenic trioxide. the orthogonal test definite best craft reasonable, reliable, the duplication is good. The method simple, fast, accurate reliable, the duplication is good, around may concoct for the realgar raw material for medicine the quality control to provide the reference.The optimized method is accurate and reproducible, and suitable for the determination of effective composition and toxic component in realgar granules pieces.6.In the aspect of processing mechanism research Flies the realgar compared with the water and the coarse particles realgar different artillery product fills the stomach separately the acute toxicity response which appears to the medicine paired observation mouse.Methods:Makes separately water-treatment flies the realgar and the coarse particles realgar, according to the classics acute toxicity experimental technique, carries on the realgar different artillery product the acute toxicity experimental study, After paired observation for medicine, the km mouses acute toxicity response and the death situation, the empirical datum calculates LD50 with the Bills law. Results:Coarseparticles realgar component (LD50) the value is 10.072g.kg-l.d-1, Is equal in the clinical person amount used 1439.1 Time,95% limits is 8.1364-12.492 g.kg-l.d-1, Is equal in the clinical person amount used 1162.3-1784.6 time, the water flies the realgar component to give the dose and is unable for the medicine density limit to determine LD50, The MTD experimental result according to contains the crude drugs gauge to calculate is 24.3g.kg-l.d-1, Is equal in the clinical 65kg person each kilogram body weight everyday use quantity 3471.4 Time, Flies the realgar after to the water and the coarse particles realgar, discovered that the km mouse has the obvious acute toxicity performance, its body weight, the taking in food quantity, the behavior appear obviously exceptionally.