Rapid Identification And Integrated Pharmacokinetic Study Of Panax Notoginseng Saponins From Compound Xueshuantong Capsules

AIM:Fufang Xueshuantong capsule(FXT)is a kind of pure traditional Chinese medicine which is jointly developed by the Department of Ophthalmology center of Zhongshan and Guangdong Zhongsheng Pharmaceutical Co.,Ltd FXT has been regarded as a preferred Chinese herbal formula for the treatment of Diabetic retinopathy(DR)and Diabetic nephropathy(DN).The basic research of FXT is relatively weak.In this subject,a novel and efficient strategy was developed and used for the screening and identification of notoginsenosides in vivo and in vitro using high performance liquid chromatography combined with LTQ-Orbitrap mass spectrometry(HPLC-LTQ-Orbitrap).It was to study the metabolic and tissue distribution of FXT in Beagle dogs.And,a novel integrated pharmacokinetic was used to study the FXT.This study was to provide a foundation for the material basis and clinical application of the FXT.METHODS:According to the structural characteristics of notoginsenosides,the data mining techniques,the mass defect filtering(MDF)and the characteristics ion filter(CIF),were adopted for the rapid screening of notoginsenosides from complicated peaks,with the high resolution mass spectrometry.The samples,which were plasma at different time points,urine and feces within 12h,eye and kidney at 2h,were collected after beagle dogs were given the FXT for 3 days by oral administration.Then,the notoginsenosides in the samples were identified by the above identification strategy.A simple,the triple quadrupole mass spectrometer method was developed for the simultaneous determination of five notoginsenosides(notoginsenoside Rl,ginsenoside Rgl,ginsenoside Re,ginsenosideRbl,ginsenoside Rd of FXT)in beagle dog plasma.After oral administrated with different does(30 grain/10kg,15 grain/10kg,8 grain/10kg),the beagle dog plasma was collect in different times.The pharmacokinetic parameters of five notoginsenosides were calculated through the DAS software.An approach of self-defined weighting coefficient has been created to the holistic pharmacokinetic profiles of protopanaxatriol(PPT)and protopanaxadiol(PPD)in FXT.A novel approach of self-defined weighting coefficient based on the area under the curve from zero to infinity(AUC0-?)has been created to obtain the holistic pharmacokinetic profiles of FXT.The classified and integrated synthetic concentrations were obtained,and then the pharmacokinetic parameters of PPT and PPD were calculated from non-compartmental model analysis.RESULTS:A rapid screening strategy based on HPLC-LTQ-Orbitrap combined with multiple data mining techniques(MDF and CIF)has been established for the identification of notoginsenosides.By using this strategy,46 notoginsenosides were totally screened and identified in vivo,and 18,48,15 notoginsenosides were respectively screened and identified in plasma,feces,urine,and 13,18 notoginsenosides were respectively screened and identified in eye and kidney after oral administration of FXT in beagle dogs.The pharmacokinetic characteristics of PPT and PPD were significant differente.The Tmax,t1/2,MRT0-t,MRT0-?,CLz/F,AUC0-?,Cmax of PPT(notoginsenoside R1,ginsenoside Rg1,ginsenosideRe)were 1-2.5h,3.5-12h,4-5h,6-16h,1.5-10 L/h/kg,200-2800ng/mL*h,30-400 ng/mL respectively.The Tmax,t1/2,MRT0-t,MRT0-?,CLz/F,AUC0-?,Cmax of PPD(ginsenosideRbl,ginsenoside Rd)were 5-12h,40-65h,65-90h,0.05-1L/h/kg,2000-400000ng/mL*h,30-4500 ng/mL repectively.The pharmacokinetic characteristics of PPT and PPD were significantly different.The PPT had faster absorbtion,longer retention time,and faster elimination than PPD.Based on the self-defined AUC weighting coefficients integration approach,the Tmax,t1/2,MRT0-?,CLz/F,AUC0-?,Cmax of the holistic pharmacokinetic profiles of PPT were 1-1.5h,3-11h,6-14h,5-8 L/h/kg,600-2000 ng/mL*h,80-320 ng/mL respectively.The Tmax,t1/2,MRT0-?,CLz/F,AUC0-?,Cmax of the holistic pharmacokinetic profiles of PPD were 5-8h,50-60h,73-90h,73-90h,0.05-0.15/h/kg,39000-400000 ng/mL*h,520-4100 ng/mL,respectively.The classified and integrated pharmacokinetic was taken into account the pharmacokinetic of each monomer notoginsenoside.CONCLUTION:This subject laid the foundation for the material basis research of FXT and provided a new strategy for rapid identification of notoginsenosides.The notoginsenosides metabolism of FXT in beagle dogs has been explored.The notoginsenosides in plasma and urine were less than in feces.It showed that the bioutilization of the notoginsenosides was low,and most of the non-absorbed blood was excreted directly from the body.The notoginsenosides in plasma were similar in urine.The main glycine replaces the saponins,which have a small number of glycosides,which are three to four saponins.It may be that the samll notoginsenosides were easy to penetrate the membrane into the blood The distribution of FXT in beagle 's eye and kidney was explored The notoginsenosides in beagle 's eye were less than in kidney due to blood-eye barrier.The main glycine replaces the saponins,which are one to three saponins in eye and three to five saponins in kidney.This subject laid the foundation for the effective ingredients for the treatment of DR and DN.A classified and integrated pharmacokinetic approach to describing the holistic pharmacokinetic properties has been successfully developed using FXT as a model traditional Chinese compound recipe.