Study On The Anticancer Active Components Of Scutellaria Baicalensis Based On Hollow Fiber Cell Capture And Liquid Phase Microextractio

Objective To establish a hollow fiber cell capture-HPLC(HFCC-HPLC)method for the screening and identification of the main anti-cancer active components in human hepatocellular carcinoma Hep G-2 cells by using hollow fibers as cell carriers;and to establish a hollow fiber liquid phase microextraction-HPLC(HFLPME-HPLC)method for the in vitro enrichment,concentration and extraction of the main active components of Vajra.Hollow fiber liquid phase microextraction-HPLC(HFLPME-HPLC)was established for the in vitro enrichment,concentration and extraction of the main active components of adamantine.Based on the optimized HFLPME-HPLC method,the pharmacokinetic characteristics of Vajra were studied in vivo in animals.Methods Hep G-2 cells were used as target cells,which were inoculated in the inner lumen of hollow fibers to construct the HFCC-HPLC method,and the morphological and activity changes of the cells before and after inoculation were observed.The screening experiments of anticancer active ingredients were conducted by immersing the hollow fibers inoculated with Hep G-2 cells into the extract of adamantine.The HFLPMEHPLC method was developed and optimized to enrich,extract and detect the content of the screened active ingredients in the extract of Vajra.The concentration of the active ingredients in the liver of KM mice was investigated by gavage of certain concentrations of adamantine extract,and the changes of the active ingredient concentration over time were studied by HFLPME-HPLC method with different time administration.Results In this paper,a hollow fiber cell capture-HPLC method HFCC-HPLC and a hollow fiber liquid phase microextraction-HPLC method HFLPME-HPLC were constructed:(1)the main anticancer active ingredients in the extract of Smilax china L.were screened as chlorogenic acid,lacoside,resveratrol,quercetin and kaempferol using the HFCC-HPLC method;(2)the active ingredients in the extract of Smilax china L.were screened as chlorogenic acid,lacoside,resveratrol,quercetin and kaempferol using the optimized HFLPME-HPLC method was used to determine the contents of the active ingredients in Vajra extract,which were 6.84 mg/g,19.33 mg/g,6.77 mg/g,12.42 mg/g and 0.54 mg/g,respectively;(3)the concentrations of the main active ingredients in the liver of KM mice after gavage of Vajra extract were determined using the HFLPMEHPLC method,and the pharmacokinetic parameters were calculated.The pharmacokinetic parameters were calculated as follows: the Cmax of chlorogenic acid,loxoside,resveratrol,quercetin and kaempferol were 0.927 μg/m L,0.789 μg/m L,0.285μg/m L,0.424 μg/m L and 0.817 μg/m L,respectively.t1/2 of the five compounds were in the range of 48-320 min,and the peak time Tmax were in the range of 30-180 min.Conclusion From the study,it can be concluded that HFCC-HPLC can be used as a convenient and reliable in vitro screening method for the anticancer active ingredients of traditional Chinese medicine,and the optimized HFLPME-HPLC can be used as a method with high enrichment index and high sensitivity for the detection of complex components of traditional Chinese medicine.Both methods can be used for the screening and pharmacokinetic study of the anticancer active ingredients of Smilax china L..The results of this study provide a reference for the refinement of the active substances in the quality standard of Smilax china L.herbal medicine,a basis for the elucidation of the antihepatocellular carcinogenic substance basis in Smilax china L.,and an idea for the expansion of the pharmacological activity study and clinical application of Smilax china L..