Investigation On Chemical Constituents And In Vitro Bioactivities Of Trapa Sp.pericarps

In this thesis,the ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-QTOF-MS)was used to study the chemical constituents of the ethyl acetate fraction of Trapa Sp.Pericarps.The UPLC fingerprints of 11 batches of Trapa Sp.Pericarps were established.In addition,the in vitro antiproliferative and antioxidant activities of the samples were also studied.Firstly,the extraction method and the UPLC analysis conditions of Trapa Sp.Pericarps were established and optimized.The extraction method was as follows:ultrasonic extraction,extracting solvent ethanol/water(V/V,60/40),extracting temperature 40 °C,extraction time 30 min.The crude extract was further extracted with water,petroleum ether and ethyl acetate twice respectively.The ethyl ester layer was obtained for further analysis.The UPLC chromatographic conditions were as follows: Waters Acquity UPLC HSS T3 column(2.1 mm × 100 mm,1.8 ?m);flow rate: 0.2 mL/min;injection volume: 1.0 ?L;column temperature: 30 °C;wavelength:280 nm.Secondly,the UPLC-QTOF-MS technology was used to identify chemical constituents in Trapa Sp.Pericarps extracts.A total of 39 compounds were detected,of which 35 compounds were identified.The compositions in this fraction were mainly tannins.A total of 13 kinds of gallotannins and 15 kinds of ellagitannins were identified,and the cleavage pattern of these two types of compounds were deduced.Thirdly,the UPLC fingerprints of 11 batches of the samples were established,and 30 common peaks were obtained.Through the similarity evaluation,principalcomponent analysis and hierarchical cluster analysis,a high similarity was shown among the 11 batches of samples,and all of these could be divided into two clusters.Three principal components including compound 10(tri-O-galloyl-D-glucose),20(tri-O-galloyl-HHDP-D-glucose)and 22(1,2,3,6-tetra-O-galloyl-?-D-glucose)were recognized as the highest contribution to the classification of the samples.In the antiproliferative activity of HepG2 cells in 11 batches of samples,it was found that the higher contents of gallotannins could contirbute to the lower antiproliferative activitiy while the higher contents of ellagitannins could contribute to the stronger antiproliferative activity with a dose range of 3.13-100.00 ?g/mL.It was speculated that the hexahydroxydiphenoyl(HHDP)played an important role in it.The antioxidant experiment to study the DPPH free radical scavenging ability showed that the stronger antioxidant capacity was combined with the higher chemical components of the samples.It was speculated that the number of hydroxyl groups in the chemical structures played a decisive role.This investigation could provide a further basis for the application and quality evaluation of Trapa Sp.Pericarps.