The Studies On Quality Evaluation Method And Pharmacokinetics Of Hedera Helix

Hedera helix,also referred to as ivy,is a member of Araliaceae family.In addition to landscaping and greening,it is also an important herbal medicine.It can be used for the treatment of inflammatory diseases and asthma.Additionally,ivy leaves extracts also showed anti-elastase,anti-microbial,anti-spasmodic,and anti-fungal activities.This paper takes Hedera helix as the research object to study the inherent chemical constituents,the absorbed constituents of Hedera helix in rat plasma,the pharmacokinetics,tissue distribution,metabolization and excretion of three active saponins of Hedera helix in rats with quadrupole time-of-flight mass spectrometry(HPLC-Q-TOF-MS/MS)and ultra-high performance liquid chromatography coupled with tandem mass spectrometry(UPLC-MS/MS)techniques.A sensitive and reliable HPLC-Q-TOF-MS/MS method was established to separate and identify the inherent chemical constituents in Hedera helix extract.With the optimized conditions,a total of 30 compounds were identified or tentatively characterized by comparing the retention time and MS data with reference standards.Among them,22 compounds were identified as saponins,other compounds were identified as flavones,phenylpropanoids,and adenosine.Results indicated that saponins were major constituents in Hedera helix.A HPLC method was estabolished and validated for the fingerprint and quantitation of Hedera Helix.0.05%phosphoric acid solution and acetonitrile were as mobile phase in gradient mode.The detection wavelength was set at 210 nm.The similarity and clustering analysis results were calculated by traditional Chinese medicine fingerprint evaluation system and SPSS.21 common peaks in Hedera Helix was confirmed and the similarity was more than 0.85.The 8 contents of Hedera Helix in different batches ranged from 0.0100%?0.345%.A rapid,sensitive and high throughput UPLC-MS/MS method was established and validated to assay the concentration of hederacoside C,hederacoside D and ?-hederin in rat plasma,and the method was applied to the pharmacokinetic study of them in rats after oral administration of extract of saponins and mixture of saponins.Chromatographic separation was performed on a Thermo Hypersil GOLD C18column(2.1 mm×50 mm,1.9m)using a gradient mobile phase system of acetonitrile-water containing 0.1%formic acid.The intra-and inter-day precisions(as relative standard deviation)were less than 15%,and accuracy(as relative error)was between-9.04%?12.5%.The calibration curves were linear over a range of 1.05?1680 ng/mL,0.483?966 ng/mL and 1.08?1720 ng/mL for hederacoside C,hederacoside D and ?-hederin.Results showed that double peaks were evident on concentration-time profile for each of the three saponins.The difference in the pharmacokinetic characteristics of three saponins between a mixture of saponins and an extract of saponins from Hedera helix was found in rat,which would be beneficial for the preclinical research and clinical use of Hedera helixA rapid,sensitive and high throughput UPLC-MS/MS method was developed for quantifying three saponins in rat tissues.The tissue distribution of,hederacoside C,hederacoside D and ?-hederin in rats was investigated after oral admistration of 0.5g·kg-1 extract of saponins.The results showed that the saponins were fast and extensively distributed.The high levels of saponin in most tissues were observerd at 15 min after administration.The levels of saponins were almost elimination in 12 h except for intestine.The concentration of 3 saponins in intestine kidney and liver was higher than other tissues.The results indicated that liver and kidney were possiblely disposition organs,and the high level in lung may correspond to bronchial diseases.A HPLC-Q-TOF-MS/MS method was developed to investigate metabolic profiles of hederacoside C,hederacoside D and ?-hederin in rat blood,urine bile and feces after intragastric administration of the extract of saponins at a dose of 1.0 g·kg-1.Besides the parent drug,a total of 27 metabolites(24 phase I and 3 phase ? metabolites)were detected and tentatively identified.The phase I biotransformation pathways of saponins included deglycosylation,dehydrogenation and demethylation,and the phase ? metabolites of saponins were mainly glucuronides.A rapid,sensitive and high throughput UHPLC-MS/MS method was established to determine the concentration of hederacoside C,hederacoside D and ?-hederin in rat urine and feces.After intragastric administration of extract of saponins at a dose of 0.5g·kg-1 the cumulative amounts of hederasaponin C,hederacoside D and ?-hederin excreted amounts in rats urine(0?24 h)were 0.212%,0.167%and 0.164%of the dosage,respectively.The fecal extraction amounted to 0.562%,0.755%and 0.569%.The results showed low level of three saponins was excreted in urine and feces,which suggested three saponins mainly excreted in the form of metabolites and unchanged form were not major pathway.