Analysis Of Lignan Components From Valeriana Amurensis And Pharmacokinetics Study In Rats

Valeriana amurensis Smir.ex.Kom which plants belongs to the genus Valeriana L of Valerianaceae,in the use of traditional Chinese medicine which have the effect of tranquilizing,regulating qi,and analgesia,that often used with other herbs.In previous studies,we reported V.amurensis were possessed of the potential therapeutic effect on Alzheimer’s disease(AD)for the first time and we had also screened and determined the AD-effective fraction(50%EtOH fraction from AB-8 macroporous resin column of 95%EtOH extract,AD-EFV).and we found lignan element compositions with strong nerve cell protection function.To reveal the V.amurensis lignan of element compositions absorption and distribution characteristics of the reference data for drug quality control and clinical application,this research will for develop a determination and pharmacokinetics study of V.amurensis lignan of element compositions.The main research contents include:1.Set up a quantitative methods of lignans measuring researchTo research and establish the quantitative methods to determination of nine lignan components in V.amurensis with ultra high performance liquid chromatography(UPLC)for the extraction,used a hybrid triple quadrupole/LIT(linear ion trap)mass spectrometer for testing instrument.The UPLC conditions are as follows:the CSH Fluoro-Phenyl(150 mm x 2.1 mm,1.7 μm)chromatographic column,acetonitrile and water as mobile phase,gradient elution and 0.2 mL/min flow rate;Mass spectrometry conditions for:ESI source,negative ion scanning,multiple reaction monitoring(MRM).8,8’-hydroxypinoresinol(DH),prinsepiol-4-O-β-D-glucopyranoside(PG),(+)-medioresinol-4,4’-di-O-β-D-glucopyranoside(MDG),(+)-syringaresinol-4,4’-di-O-β-D-glucopyranoside(SG),(-)-massoniresino13a-O-β-D-glucopyranoside(MG),(+)-8-hydroxypinoresinol-4’-O-β-D-glucopyranoside(H4’G),(+)-8,8’-dihydroxypinoresinol-4,4’-di-O-β-D-glucopyranoside(DG),(+)-pinoresinol-4,4’-di-O-β-D-glucopyranoside(PDG),(+)-8-hydroxypinoresinol-4-O-β-D-gluc opyranoside(H4G),nine chemical compositions in the concentration range has a good linear relationship,precision of instrument(RSD)<4.91%,the repetition rate(RSD)is between 1.80%～2.94%,the recovery was 94%～104%.The method is fast,high efficiency,good linear,high sensitivity,detection limit and can meet the needs of the experiment.The developed method was used to determine 10 batches of different place of origin of V.amurensis grass lignan components content.2.Internal pharmacokinetic study of Valeriana amurensis in ratsTo establish an UPLC-MS/MS quantitative method for determination of the V.amurensis in plasma samples.The chromatographic column was CSH Fluoro-Phenyl(150 mm x 2.1 mm,1.7μm),the mobile phase was consist of acetonitrile-water contains 0.1%formic acid in gradient elution,the flow rate of 0.3 mL/min,10 min analysis time.(+)-8-hydroxypinoresinol-4’-O-β-D-glucopyranoside(HG),(+)-pinoresinol-4,4’-di-O-β-D-glucopyranoside(PDG),(-)-massoniresinol-3a-O-β-D-glucopyranoside(MG),prinsepiol-4-O-β-D-gluc opyranoside(PG),four compounds lignans analytes and 4-hydroxy benzoic acid ethyl ester(IS),Compound of HG,PDG,MG,PG and internal standard were monitored ions on 535.2/373.1,681.3/357.1,553.4/357.2,551.3/389.3,165.2/92.0 respectively.Methodology validation of four lignan compounds of standard curve of the linear range(ng/mL)were 0.39～154.0,0.62～244.7,0.50～134.5,0.34～198.7,the RSD in the intra day and inter day precision were less than 14.46%,the RSD in the accuracy between-5.87%～12.30,the analyte extraction recovery was more than 81.10%in the plasma of rats.The method is applied to the pharmacokinetics research of the rats after oral dosing V.amurensis extract.Drug HG,PG,PDG,MG of T1/2(h)between 2.13～3.00,Tmax(h)between 0.58～0.67,Cmax(ng/mL)were 27.33±5.32,33.54±6.18,9.85±1.88,24.95±7.05.3.Internal tissue distribution study of Valeriana amurensis in ratsTo establish UPLC-MS/MS quantitative methods for determination of V.amurensis in different tissue samples,chromatographic column was Waters ACQUITY UPLC HSS T3(100 mm × 2.1 mm,1.7μm),the mobile phase was consist of contains 0.1%formic acid in acetonitrile-water gradient elution,flow rate of 0.3 mL/min,10 min analysis time.Multiple reaction monitoring(MRM),negative ion mode,the source of electrospray ionization(ESI).The method was applied to the tissue distribution study of the rats after oral dosing V.amurensis extract.In 0.5,1,1gather the heart,liver,spleen,lung,kidney,and brain tissue samples,to.5,2,and 2.5 h after the treatment,detect the contents of drug HG,PG,PDG,MG concentration in each tissue organs.Results indicate that the four lignan element compositions in heart,liver,spleen,lung,kidney,and brain tissue were distributed,from the point of the whole distribution,the content was relatively high in the spleen tissue,followed by the liver and lung,drugs can through the blood brain barrier.The study of anti-AD in V.amurensis nine active ingredient in the establishment of a rapid,efficient multicomponent quantitative analysis and qualitative method,and used the method to determine lignan element compositions in 10 batch of V.amurensis medicinal materials,get the quality control method of medicinal materials.Established a rapid quantitative method of lignan element compositions in biological samples,and was applied this method to V.amurensis extracts in vivo pharmacokinetics research in rats,for drugs V.amurensis in anti-AD set of clinical applications to medicine,and drug using dosage and using method provide some reference data.