| Objective To make a HPLC fingerprint analysis on 50% ethanol extract from Periploca forrestii Schltr of different growing areas and study their differences in quality; to make a HPLC fingerprint analysis on 95% ethanol extract, 50% ethanol extract, petroleum ether extract, chloroform extract, acetic ether extract, n-butyl alcohol extract and water extract of Periploca forrestii Schltr come from Xi Chang and determine the differences of compounds in these parts; to study the HPLC fingerprints of the cell sap samples of human lung cancer cell line A549 after being treated by 95% ethanol extract, 50% ethanol extract and chloroform extract of Periploca forrestii Schltr come from Xi Chang and compare the changes of anti-tumor components in different extracts of Periploca forrestii Schltr before and after treating tumor cells in hopes of discovering the anti-tumor compounds of Periploca forrestii Schltr and lay the foundations for studies on searching anti-tumor lead compounds, anti-tumor mechanism and drug absorption at cellular level.Method Adopted HPLC-UV method and Similarity Evaluation System for Chromatographic Fingerprint of TCM(Version 2004 A)to analyze the fingerprints of Periploca forrestii Schltr and various extracts from it, and analyzed the fingerprints of cell sap samples after being treated by various extracts from Periploca forrestii Schltr.Result The optimum preparation method of extracts from Periploca forrestii Schltr was obtained and it is: added 50% ethanol solution with the ratio of Periploca forrestii Schltr to ethanol solution is 1:30, ultrasonic extraction for one time at 50℃for 30min. Established determination method on HPLC fingerprint of Periploca forrestii Schltr, chromatographic method is: take Alltech Apollo C18(250 mm×4.6 mm, 5μm) as stationary phase and CH3OH-H2O as mobile phase; flow velocity: 0.5 ml/min; colum temperature:20℃; detective wavelength: 210 nm; optimum chromatography condition of gradient elution is: 0-5 min while methanol:water=7:93; 5-16 min while methanol:water=42:58; 16-29 min while methanol:water=82:18; 29-120 min while methanol:water=100:0. The analysis results by Similarity Evaluation System for Chromatographic Fingerprint of TCM(Version 2004 A) were obtained.The results of fingerprint analysis on 95% ethanol extract, 50% ethanol extract, petroleum ether extract, chloroform extract, acetic ether extract, n-butyl alcohol extract and water extract of Periploca forrestii Schltr indicated that the chromatographic peaks were quite different, every kind of extract has its characteristic absorption peak. The results of fingerprints of the cell sap samples of human lung cancer cell line A549 after being treated by 95% ethanol extract, 50% ethanol extract and chloroform extract from Periploca forrestii Schltr of Xi Chang indicated that the cell sap samples treated by the three extracts owned a common peak with the retention time around 13.7min, there was a same peak in the three extracts, but the peak area and ultraviolet absorption value peak in extracts from Periploca forrestii Schltr were larger and higher than those values in the cell sap samples. There was a chromatographic peak with the retention time of 7.67min existed in fingerprint of the cell sap samples, there was no same peak in fingerprints of other extracts. There were some chromatographic peaks in 95% ethanol extract, 50% ethanol extract and chloroform extract from Periploca forrestii Schltr not found in fingerprint of tumor cell sap samples.Conclusion Adopting HPLC-UV method to establish fingerprint of Periploca forrestii Schltr is simple, stable and with good reproducibility. The similarity results of Periploca forrestii Schltr from different growing areas indicated that quality of Periploca forrestii Schltr raised in Kun Ming, Chang Shou and Xi Chang region was relatively familiar, the Periploca forrestii Schltr raised in Ping Wu region was relatively different from Periploca forrestii Schltr raised in other regions, the quality is relevant to geographical position. The extracts by seven different solvents were quite different, and it indicated that each part owned their own compound peak group and their ingredients were distinguishable. There was a common peak with the retention time of 13.7min existed in 95% ethanol extract, 50% ethanol extract, chloroform extract and tumor cell sap treated by it from Periploca forrestii Schltr, it indicated these compounds were not metabolized in tumor cells. The common peaks with the retention time of 7.67 existed in cell sap samples were not found in extracts, it may be the metabolite of active ingredient in Periploca forrestii Schltr metabolized in tumor cells. The compounds, existed in extracts but not found in cell saps, may be metabolized by tumor cells, it indicated that those compounds in Periploca forrestii Schltr had relatively activity, however, further studies on whether all of these compounds own anti-tumor effects or not are still needed. Meanwhile, it is still the hot point that what kind of relationship exists between these compounds and compounds occurred in cell saps with the retention time of 7.67min. This experiment provided certain basis for studies on searching of anti-tumor compounds in Periploca forrestii Schltr, and absorption and metabolism at cellular level, and then made a foundation for the studies on absorption and metabolism within human body of anti-tumor compounds in Periploca forrestii Schltr.
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