The Study Of The Quality Evaluation Of Pericarpium Citri Reticulatae And Pericarpium Citri Reticulatae Viride From The Same Plant Source

Pericarpium Citri Reticulatae (PCR) and Pericarpium Citri Reticulatae Viride (PCRV), harvested at different growth stage, are from the Citrus reticulata Blanco and its cultivars of fruit. For varied harvesting period and processing methods, PCRV were divided into two commodities specifications:Ilex M acropoda and Four petal ilex; For being used as different medicine, PCR is divided into two commodities specifications:Chenpi and GuangChenpi. GuangChenpi, which comes from the dry tangerine peel of varied C. chachiensis Hort, is taken as the authentic medicine of Pericarpium Citri Reticulatae. As a result from the same plant source, PCR and PCRV have some similarities in the terms of ingredients, efficacy and so on. However, due to the different harvest period, the accumulated changes of secondary metabolites make them different. For example, they are both featured as "regulate qi medicine", but for the clinical application, PCR has a better effect on regulating qi and strengthening spleen whereas PCRV is strong in soothing liver and breaking qi. (2010 edition) takes hesperidin as quality control index for PCR and PCRV without reflecting their effect similarities and characteristics in the quality evaluation system.This study takes PCR and PCRV as the research object, on the basis of traditional Chinese medicine theory, and around this core objective:to clarify their related scientific content of "variety - quality - efficiency". The main contents will include the following:quantitative determination for index and representative components of varied medicines from different harvest times and different processing methods and different using; overall study of chemical profiles charicteristcs; combined with pharmacodynamic experiments, study on building the quality evaluation system of PCR and PCRV from the same plant source.Research and experimental results are as follows:1. Comparative study on chemical components of PCR and PCRV1.1 Flavonoids componentsBy the HPLC technology, this study takes the lead in building the method for determining simultaneously the contents of hesperidin, naringin, narirutin, naringenin, hesperetin, nobiletin and tangereti.The results show that the Chenpi and Four petal ilex which both belong to Citrus peel medicine contain hesperidin, narirutin, nobiletin and tangereti., and all contents of which in Four petal ilex is higher than those in Chenpi (p<0.01). This result shows the law:content of flavonoids decreases as the fruit maturity increases. Ilex M acropoda, of which the landing young fruit can be used for medicine, contains not only the four kinds of flavonoids metioned above but also naringin, trace naringenin and hesperetin.Guang Chenpi's contents of hesperidin si lower than that of Chenpi (p<0.01) whereas other contents of Flavonoids are nearly equal to those of Chenpi (p> 0.05). Clustering results shows that taking the hesperidin, rutin naringin, nobiletin, and tangeretin as indicators has a better effect on comprehensively reflecting the the characteristics of medicine than taking only total flavonoids or hesperidin as index.1.2 AlkaloidsPCR and PCRV contain alkaloids ingredients represented by synephrine. Synephrine content of PCRV is higher than that in PCR (p<0.01), which shows that the content decreases with the increase of fruit maturity levels; Synephrine content of GuangChenPi is significantly lower than that in Chenpi (p<0.01). Some scholars have pointed drug safety issues for herbs with synephrine.It still need further study on how to ensure safe and effective clinical treatment and wether we should set up rules for synephrine content limitation in the quality evaluation system of such kind medicine.1.3 Volatile oilThis study has extracted volatile oil of PCR and PCRV by the method of volatile oil determination metioned in the Appendix XD of(2010 edition), and analized the volatile oil components using the GC-MS method. The results show that volatile oil components of PCR and PCRV are limonene and linalool, but becase of the different origin, different using, different harvest times and different processing methods and the instability of volatile oil, varied medicines are quite different in the composition and content of volatile oil.1.4 HPLC fingerprint By the HPLC technology, this study takes the lead in comparing the fingerprint of methanol extraction from PCR and PCRV.Acoording to the visual analysis of chromatograms, Chenpi and Four petal ilex have less differences, there is no specific or indicative peak, the similarity with the standard fingerprint is more than 0.95; Ilex M acropoda is quite different from fruit peel medicine in the number of peaks and peak area, and the similarity with standard fingerprint in the range of 0.8 to 0.9; the similarity of the fingerprint of GuangChenpi and Chenpi with the standard fingerprint is more than 0.97. Cluster analysis showes that Chenpi and Four petal ilex from the same species can be categored into a class; Ilex M acropoda harvested in different stages and processed in different methods can be takens as a class; GuangChenpi and Four petal ilex both from the C. chachiensis Hort can be taken as a calss.2. Comparison Study about the effect of Pericarpium Citri Reticulatae and Pericarpium Citri Reticulatae Viride on the rat's intestinal propulsion, gastric emptying and rabbit's intestinal muscle in vitroPCR and PCRV are commonly used as medicine of regulating qi, mainly used for qi stagnation and qi inversion which manifest as abdominal distention and pain, nausea and vomiting, constipation or diarrhea, etc., such sysdroms are all related to gastrointestinal movement. This study has compared the effect of PCR and PCRV on the role of rat's intestinal propulsion and gastric emptying. The results show that under the high doses, they are both equally functional in normal rat's gastric emptying, but under the low doses, they do not promote the trend of gastric emptying of normal rats; at different doses, they both can promote the normal rat's intestinal propulsion, and at the same dosage, the effect of PCR on the role of promoting the normal rat's intestinal propulsion is stronger than that of PCVR.To further explore the different parts of two herbs on the role of rerugating qi, this study takes the rabbit's intestinal muscle in vitro as objective comparing the functional differences in terms of water extract, volatile oil and flavonoids of PCR and PCRV. The results show that different parts all take effect on inhibiting the rabbit's intestinal muscle in vitro, which conforms to the fact that PCR is used as clinical medicine for relieving the spasm pain resulting from stomach qi stagnation; water extract and flavonoids of PCR have a stronger effect on inhibiting the rabbit's intestinal muscle in vitro whereas volatile oil is weaker than that of PCRV. Suggested from the differences of the varied components'inhibition functions, we can infer that the functional differences of PCR and PCRV are the result of combined effects of multi-component rather than the result of a single component.3. On building the quality evaluation system of Pericarpium Citri Reticulatae and Pericarpium Citri Reticulatae Viride.from the same plant sourceThis is the first time to carry out the comparative study on PCR and PCRV from the same plant source by the starting point of comprehensive and systematic differences in chemical composition. The results suggest, the quality assessment system construction should focus on the the flavonoids content differences of PCR and PCRV. Taking hesperidin, rutin naringin, nobiletin, tangeretin as the evaluation indicators leads to better effect of comprehensively reflecting the characteristics and exclusive of medicine; it still need further study on how to ensure safe and effective clinical treatment and wether we should set up rules for synephrine content limitation in the quality evaluation system of such kind medicine. Becase of the havest environment and its instability, volatile oil is not recommended into the quality evaluation system. Meanwhile, in the construction of quality assessment system, it is suggested that Ilex M acropoda and Four petal ilex, Chenpi and Guangchenpi should be separately listed.