Study On The Evaluation Of Quality Between Pericarpium Citri Reticulatae 'Chachi' And Other 5 Species Citri Reticulatae

Objective:Comparative study on traits and microscopic features,composition and corresponding percentage of essential oil,HPLC finger-print,the content of polysaccharide,hesperidin and other flavonoids between Pericarpium Citri Reticulatae 'chachi' and other 5 species Citri Reticulatae,build a complete and reliable quality control method,and then provide scientific basis on different levels and multiple angles for the Pericarpium Citri Reticulatae to develop quality standards.Method:1.Using traditional Chinese medicine identification method to study traits and microscopic features and find the difference between 6 species Citri Reticulatae.2.To get the essential oil by steam distillation and then find the difference of composition and corresponding percentage by using GC-MS.3.The chromatographic column was eluted with C18(5 ?m,4.6×250mm)and mobile phase was acetonitrile-0.1%formic acid gradient.The flow rate was 1.OmL/min,the column temperature was 30? and the detection wavelength was 283nm.Establish HPLC finger-print of Pericarpium Citri Reticulatae'chachi' and get the common pattern R,then compare the finger-print of Citri Reticulatae with R,know the corresponding similarity.Determine the content of Naringin,Hesperidin and Neohesperidin of 6 species Citri Reticulatae by HPLC.4.Single-factor experiment was first step for the extraction of Citri Reticulatae polysaccharide.On the basis of single-factor experiments,the relationship between main extraction conditions including extraction time,temperature and the ratio of water to raw material was modeled using a 3-factor,3-level Box-Behnken experimental design and studied the yield of polysaccharides.And then,the established model was analyzed by response surface methodology to obtain the optimum extraction conditions.Finally,verify the consistency between the actual experimental results and the theoretical mathematical model,and then extract polysaccharide of Citri Reticulatae on this condition.Result:1.Pericarpium Citri Reticulatae 'chachi' were different from other 5 species Citri Reticulatae on its odor in traits features and quantity and form of Hesperidin crystals in microscopic features.2.Experiment result shows that D-Limonene of the Pericarpium Citri Reticulatae 'chachi' essential oil were the lowest and ?-Terpinene were the highest between 6 species Citri Reticulatae.There were 9 and 5 species common ingredients between 3 species Pericarpium Citri Reticulatae and 6 species Citri Reticulatae respectively.And these essential oil contain unique chemical component:Pericarpium Citri Reticulatae 'chachi'contained 2-(methylamino)-benzoate,Pericarpium Citri Reticulatae'Unshiu' contained Cinnamic acid,Citrus reticulata 'Nian Ju 'contained Pseudo-limonene.3.Establish HPLC finger-print of Pericarpium Citri Reticulatae'chachi' and get the common pattern R,then compare the finger-print of Citri Reticulatae with R,know the corresponding similarity.The similarity of Pericarpium Citri Reticulatae 'chachi' were higher than 0.985,similarity of other species Citri Reticulatae were between 0.720?0.873,suggest that all of them were different from Pericarpium Citri Reticulatae'chachi'.The content of Hesperidin in Pericarpium Citri Reticulatae were higher then Citri Reticulatae,and there were no significant difference on the content of Naringin and Neohesperidin.4.The optimum extraction conditions were obtained as follows:temperature was 41?,extraction time was 32 min,and the ratio of water to raw material was 62:1(v:m,mL·g-1).Under these conditions,the observed values of maximal yield of polysaccharides was 15.54±0.26%(n=3).Actual value was consistent with the theoretical value.Conclusion:This study shows that there has a difference between 6 species Citri Reticulatae on the above aspects,which could be used to explane the geoherbalism of Pericarpium Citri Reticulatae 'chachi',which could be provided scientific basis on different levels and multiple angles for the Pericarpium Citri Reticulatae to develop quality standards.HPLC finger-print common pattern R could provide evidence for rapidly and accurately identifying Pericarpium Citri Reticulatae 'chachi'.The optimum extraction conditions for extracting polysaccharides that was obtained by using RSM were stable and reliable,which could be as reference for subsequent studies on extracting polysaccharides from other species Citri Reticulatae.