Research On The Quality Of Arctiin And The Evaluation Of ARC-SMEDDS In Vitro And In Vivo

Arctiin (ARC) is an active ingredient extracted from the arctium which isa traditional Chinese medicine,and it belongs to the lignans.The modernpharmacology shows that it has many functions to our body,such as anti-inflammatory,lowering blood sugar, anti-viral,anti-tumor and so on.ARC haslittle solubility in water,so that it always accompanied with low oralabsorption and low bioavailability,those are also limiting it’s clinicalapplication.Self-microemulsifying drug delivery system (SMEDDS) has greatadvantages in improving the solubility, bioavailability and reducedgastrointestinal side effects,therefore,being prepared as ARC-SMEDDS canincrease it’s solubility, improve the solubility and bioavailability.This research have used the HPLC method to determine the content andrelated substances of ARC raw material medicine,prepared the SMEDDS ofARC, using the HPLC method to determine the content of ARC-SMEDDS,and evaluated it’s performance in vitro and in vivo.Specific research contentmainly includes the following three parts:1. Determination of the content and related substances of ARC raw materialmedicine 1.1Established an HPLC method for determination the content of ARCraw material medicine.0.152~1.220mg/mL concentration range had goodlinearity,the regression equation was A=5199.3452C-4.0592(r=0.9999);recovery rate was98%~102%range;both the inter-day and intra-dayprecision were less than1.0%.Idicating that the method had highprecision, good accuracy, and suitable for the determination of ARC rawmaterial medicine.Content of three batches of samples were above91.5%.1.2Established an HPLC method for determination the relatedsubstances of ARC raw material medicine.Developed the factors and forceddegradation experiment on related substances.The main peak and impuritypeak were separated,the degree of separation was more than1.5;Both theinter-day and intra-day precision were less than1.0%;the detection limitationwas1.04ng.Idicating that the method had good specificity,high sensitivity,andsuitable for the determination of related substances of ARC raw materialmedicine.The sum of peak area from each impurity was less than2%controlsolution’s peak area.1.3Setting a quality standard for ARC raw material medicine.The qualitystandard was that the content of ARC raw material medicine should more than90%,and the sum of peak area from each impurity should less than2%controlsolution’s peak area,if there were impurities in the chromatogram of samples.2. Preparing for ARC-SMEDDS and study on in vitro evaluation2.1The ARC-SMEDDS was prepared successfully by prescription screening and optimized experiment.The compositions of ARC-SMEDDSwere selected by solubility assay and pseudo-ternary phase diagramsanalysis,the formulation was further optimized by central composite designcombined with response surface methodology.The optimum formulation ofARC-SMEDDS was composed of19%ODO,54%RH40,27%Transcutol-P.The content of ARC-SMEDDS was40mg/g,the average particle size was26.65nm.2.2Developed an HPLC method for determination the content ofARC-SMEDDS.2~200μg/mL concentration range had good linearity,theregression equation was A=11.4221C+24.8058(r=0.9996);recovery rate was98%~102%range;The inter-day precision was less than2.0%and theintra-day precision was less than5.0%.Idicating that the method has highprecision, good accuracy, and suitable for the determination of ARC-SMEDDS. The content of ARC-SMEDDS was40.09mg/g.2.3The release of ARC-SMEDDS was measured by using the dialysisbag.The result showed that different kinds of release medium and releasemethods had no significant effect on it’s release,and it’s release was fasterthan the raw material medicine in vitro.3. Study on in vivo evaluation of ARC-SMEDDSTo study the absorption of ARC-SMEDDS in rat intestine by developinga in situ recirculation model.Using the HPLC method for determination ofARC and ARG from intestinal fluid.Both ARC content determination method in2~200μg/mL concentration range and ARG content determination methodin1~100μg/mL concentration range had good linearity;recovery rate were95%~105%range.At the concentration range from10to50μg/mL,the drugabsorption percent was less than25%.The ARC-SMEDDS has someabsorption in rat intestine but started to hydrolysis ARG after30minutes.