Study On Purification Process,quality Control And Intestinal Absorption Of Tibetan Medicine Berberis Cortex Extracts

At present,the diabetic retinopathy?DR?is one of the most serious microvas cular complications of diabetes.It belongs to the disease category of Tibetan me dicine"Jing Ni Sa Ku disease".Tibetan Medicine Berberis Cortex?TMBC?has a very effective role for Tibetans in treating this diseases.TMBC is recorded in many Tibetan medicine classics such as The Four Medical Tantras and Lan Liu Li and so on,at the same time,this medicine is also included in the Ministerial Tibetan Medicine Standard in 1995 and Six provinces and districts Tibetan medi cine standard in 1979.Modern pharmacological studies have shown that the usi ng TMBC can significantly reduce blood glucose and expression leave of VEGF in DR model mice,and significantly improve damaged microvascular morpholog y of the retina too.Wu Qingxi and his team set the content of the total extract an d the total alkaloids as the target to investigate the optimization process of extract ion.Others study the optimal conditions of ethanol reflux extraction process of th e cortex of TMBC.This article will conduct a study of the quality standards and absorption performance of subsequent extract related study.This article includin g following contents:?1?Using acid dye colorimetry,Establish a method to determinating the tot al alkaloids in ethanol extracts of TMBC.?2?By using acid dye colorimetry and high performance liquid chromatograph y,the macroporous resin purification process for the alcohol extracts was selecte d.?3?Preparation TMBC'extracts for its study on quality standard and fingerpri nts.?4?Using everted gut sac rat model Carry out studies on the intestinal absorpt ion properties of TMBC.Results:?1?The final method of acid dye colorimetry like this:Accurately pipette an appropriate amount of samples,evaporate it,add it to a separating funnel cont aining acid dyes,shake well,and let stand for half an hour.Chloroform was a dded for four times,shaken and extraction.The chloroform layer was combine d,and was measured at 417 nm.Validated by methodology,this method is si mple,stable and feasible,and can be used as an accurate method for the deter mination of total alkaloids in the extraction of TMBC'alcohol.?2?TMBC'extraction purification process of macroporous reain:Using HP D100 macroporous reain,adjusting the sample solution concentration to 11 mg·mL^-1 and the loading liquid at 1 mL·min^-1,first eluting with 4BV water,and then using 9BV 30%ethanol at the same speed.Collecting the eluent,that is purified.The content of total alkaloids by using acid dye colorimetry greater than 50%,and the method is feasible.?3?Quality standard of TMBC'extraction:Purified material were prepared,concentrated and drying.Establish inspection methods and limits for the TLC identification,water content,ash content,heavy metals content,and use HPLC to establish a method for determining the content of the four main alkaloids.T he content of water,ash and heavy metals should be controlled within 8.3%,0.6%and 10ppm,respectively;the proposed extract magnoflorine,jatrorrhizine,p almatine and berberine content are varying from 20.0%³0.8%,1.5%².8%,0.71%¹.19%,14.1%²2.2%,respectively.And HPLC featureprint was established t o facilitate better control of the quality of the extraction product.?4?Intestinal absorption performance of TMBC'extraction:In everted gut s ac rat model,the absorption of berberine in different intestinal segments meets zero-order absorption.In the three bowel segments,the TMBC group had more absorption,and the absorption rate are increasing.Conclusion:A stable and feasible process for the purification of TMBC was es tablished.And establishing the quality standards and fingerprints of the extracti on.The results of intestinal absorption experiments showed that the extracts of TMBC has better absorption and increasing bioavailability of Berberine Hydroc hloride,to lay the foundation for the finding of drugs for the prevention and treat ment of diabetic retinopathy.