Analysis Of The Major Activity Constituents In Easter Pear By High-Performance Liquid Chromatography

China is not only the cradle center of eastern pear but also the biggest county in pear's output of the world. During the lengthy process of growth and eating, some therapy function through biting and supping and many hygienically the ancients founded function. But the infrequent investigations and reports about the constituents with physiologic activity in pear fruit were founded by far. Our research team founded the upper level content of the constituents with physiologic activity for example arbutin and chlorogenic acid in eastern pear in initial stages. It was founded that Arbutin and chlorogenic acid should be the representative components with physiologic activity in eastern pear in fact by our test.Isocratic elution reversed-phase high-performance liquid chromatographic (HPLC) method and gradient elution reversed-phase high-performance liquid chromatographic (HPLC) method for the analysis of the main constituents with physiologic activity in pear fruit were developed with high-performance liquid chromatographic instrument which was made in china. Isocratic elution method make up of a reversed-phase column (4.6mmx200mm) filled with 5um HYPERSIL BDS as the stationary phase .The mobile phase was made of CH3OH-H2O (25: 75, v/v) dualistic system at pH3.20, which adjusted, by phosphoric acid. The flow rate of mobile phase was 0.6 ml / min. During 13 minutes the analysis was ended at 280nm wavelength with ultraviolet detector in room temperature. The linearity correlative coefficient are 0.99998 both arbutin and chlorogenic acid. The linear range is at 144-5000ng to arbutin and is at 200-5000ng to chlorogenic acid. The two components' lowest detected limit was 0.153 ng and 0.150 ng respectively. Gradient elution reversed-phase HPLC method using two pumps established to the main polyphenolic compounds for example arbutin, chlorogenic acid, catechin, epicatechin, caffeic acid, p-coumaric acid, quercetin, et, al in pear fruit. Gradient elution program (min/B%): 0/2, 3/2, 4/25, 11/50, 16/80, 26/80, 33/2, 45/2 .The mobile phase at pH3.20 adjusted by phosphoric acid and detected at 280nm wavelength. In this HPLC system, the main active components were separated within 30 minutes and chromatography column condition was renewably stabilized within 45 minutes and the next sample can be run. Dynamic changes in the main constituents with physiologic activity during the florescence and the fruits development stage of "Ya" Pear were monitored by the developed HPLC methods.The contents of the two constituents in different organs during the florescence areon upper level especially the content of arbutin .The contents of arbutin reached about 1.3% in floral bud and foliar bud while the contents of chlorogenic acid obtained about 3000u.g/g .The contents of the two polyphenolic compounds in fruit let stage during the process of fruit growing are the tiptop that reached 1.0% and 0.38% respectively . Then the content of the polyphenolic compounds decline swiftly and the content of the polyphenolic compounds reduce below 400ug/g in the maturation period of pear fruit. Twenty-two varieties include P. bretschnrideri Reh., P. pyrifolia Burm Nakai, P. ussuriensis Maxim., P. sinkiangensis Yu. of eastern pear system and five varieties of Pyrus communis L. was compared and analyzed. The content of arbutin in eastern pear fruit (the average content is 164ug/g of 17 varieties) were upper than the Pyrus communis L. (the average content is 83ug/g). Ya pear is one variety of P.bretschnrideri Reh. Which mostly planting variety in eastern pear system and its content of arbutin were uppest reaching 400(4ug/g. The content of chlorogenic acid was given prominence to Pyrus communis L. with 309(ug/g average and upper than the content of eastern pear (158ug/g). The content distribution of the components was prodigiously different in pear fruit. The tiptop content of arbutin was distributing in the fruit peel and obtained 3-5 times to fruit core and 10-40 times to fruit pulp. The content distribution of chlorogenic acid fruit core was upper than frui...