| "Xihuangcao" are common folk herbal medicine,and have been recorded by the"Chinese Pharmacopoeia" in 2010.It taste bitter,sweet,cold,liver,gallbladder,with heat clearing and dampness,jaundice,cooling blood and dissipating blood stasis.Ancient folk doctors put "xihuangcao" used as heat dampness.removing jaundice medicine.It can treatment acute jaundice hepatitis,acute cholecystitis?and are medicine for the prevention and treatment of common diseases of Iiver and gall.But its efficacy material basis is not clear,the lack of quality standards of reliability,has seriously hindered the further development and utilization of"xihuangcao" resource.Purpose:This artele started from lsodon lophanthotdey water exlract.The warer-solable compounds were lsolanted.The lsodon lophanthotdey watre-soluble flavonoids were enrichmented;HPLC fingerprint of water-soluble flavonoids were stablishmentedthe water-soluble components as index,an HPLC method for determination of different varieties source of "xihuangcao" were established.We still studied the dynamic changes of the water-soluble flavonoids of different harvest periods.The antitumor activity of water-soluble constituents were screened.For the stream of "xihuangcao"quality control processing,and provide a scientific basis for further development and ut(?)zation.Method:1.We separated the water extract of Isodon lophanthoides by using D101,polyamide,ODS,Scphadex LH-20 and preparative HPLC.The water-soluble components' structure were identificated by using modern spectroseopy like HPLC.UV.1H-nmr lechnology.13C-NMR and ESI-MS2.Enriching and purifying lsodon lophanthoides water-soluble flavonoids by asmg HPD-100 and polyamide.and to establish the HPLC fingerprint.3.Taking the isolated 8 water-soluble components as reference and adopting I-HPLC to establish the simultaneous determination of 8 water-soluble components of content determination method to evaluate the quality of the "xihuangcap" from different sourees.4.MTT method was used to measure the inhivitory components rate of water-soluble onHepG2 cell proliferation after administration of 24,48,72hours.We calculate the half inhibitory concentration IC50 with SPSS 20.0 software.Results:1.Structures of the isolated twelve water-soluble compounds are identified,namely:Vicexin ?(1),vicenin ?(2),isoschaftoside(3),schaftoside(4),vitexin(5),6,8-di-C-a-L-arabinosylapigenin(6),apigenin-7-O-glucuronide(7),apigenin 6-C-?-D-xylopyranosyl-8-C-a-L-arabinopyranoside(8),apigenin 6-C-?-L-arabinopyranoside-8-C-a-L-arabinopyranoside(9),caffeic acid(10),rosmarinic(11),rutin(12).2.The HPLC fingerprint of Isodon lophanthoides water-soluble total flavonoids were identified in 12 common peaks,and the water-soluble flavonoids common mode ware established.The fingerprints similarity of Lsodon lophanthoides water-soluble total flavonoid of 10 batch were in more than 0.90.3.we determinated 8 water-soluble components of "xihuangcao" from different varieties of source by HPLC.The linear ranges of caffeic acid,vicenin ?,vicenin?,isoschaftoside,schaftoside,vitexin,6,8-di-C-a-L-arabinosylapigenin and rutin were 0.09-2.28 ?g,0.13-3.11 ?g,0.06-1.38 ?g,0.18-4.54?g,0.08-1.97 ?g,0.07-1.71 ?g,0.06-1.49 ?g,0.05-1.31 ?g,respectively.The average recoveries(n=6)were 99.7%(RSD=0.86%),99.5%(RSD=1.8%),100.1%(RSD=1.8%),99.0%(RSD=1.7%),98.4%(RSD=2.0%),98.4%(RSD=1.2%),101.8%(RSD=1.2%),99.4%(RSD=1.4%),respectively.4.Screening results of antitumor activity on HepG2 cells showed that compound 1,6,11 and the water-soluble total flavonoids has certain inhibitory effect on the proliferation of HcpG2 cells.Conclusion:The water-extract of "Xihuangcao" contains a series of eases of flavonc C-glycosidcs that easily soluble in water,and some oxygen glycoside tlavonoids and phenolic compounds.In this paper,we research the water-soluble components of Isodon lophanthoides,and studied the antitumor activity of water-soluble components.The HPLC fingerprint of Isodon lophanthoides water-soluble flavonoids suggest that the basic situation of Isodon Iophanthordes water-soluble flavonoids.It is the firsr time to establishthe HPLC content determination method of 8water-soluble components of"xihuangcao".The method is simple,stable and reliable,and can be used for quality control of "xihuangcao" in different varieties of source. |
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