Research On Quality Control Standard Of Forsythia Suspensa

Forsythia,a traditional Chinese medicineis,is the dried fruit of the nymphal plant Forsythia suspensa(Thunb.)Vahl.Autumn fruit is harvested when it is green,removes impurities,steamed,dried,and is known as Green forsythia(GF).When the fruit is ripe,it is harvested,dried,and removed from impurities,and is known as Old forsythia(OF).The forsythoside A and Phillyrin are main active ingredients of Forsythia suspensa and forsythoside A is high in forsythia.In order to make the thin layer identification more accurate,forsythoside A was added as a standard control under the 2015 edition of the Chinese Pharmacopoeia for the thin layer identification.Due to the number of counterfeit and similar products of Forsythia,it is sometimes impossible to guarantee the authenticity of Forsythia by relying on thin layer identification.When using the fingerprint of traditional Chinese medicine,different origins,different collection times and different pharmaceutical companies will be under the same “fingerprint”.The control is difficult.So in order to better control the quality of Forsythia medicinal herbs,this paper identifies several specific chemical components of GF and OF by establishing the HPLC.In order to obtain the optimum method for content determination of Forsythia Fructus(FF),a variety methods for the sample preparation of FF were evaluated by the content determination methods of Chinese Pharmacopoeia.The method can achieve the best effect of simultaneously extracting forsythoside A and phillyrin.At the same time,it was found that the content of forsythoside A and phillyrin was significantly different in GF and OF.Therefore,it is recommended that the limit values of the two index components should be separately formulated.As a kind of commercial product,Forsythia has a great influence on the quality of Chinese patent medicine and clinical.In order to further control the quality of FF,this paper will combine the characteristic printing and content determination to study the classification standard of FF.That is divided into Green forsythia general goods(GFGG),Green forsythia selection goods(GFSG)And Old forsythia general goods(OFGG).The research content of this paper is as follows:1.I dentification of forsythia thin layerIn the 2015 edition of the Chinese Pharmacopoeia,the forsythia identification was added to increase forsythoside A as a standard control,and a series of different developingagents were screened.Finally,chloroform-methanol(8:2),ethyl acetate-formic acid-water(9:1.5:0.5)was used as the secondary development condition,and 10% sulfuric acid ethanol solution was used as the color developing agent for 3 batches of GF and 2 batches of OF.Simultaneous identification of forsythoside A and phillyrin.Complete the study of the specificity,reproducibility and durability of the method.2.Study on HPLC characteristic printing of Forsythia suspensaThe preparation methods of seven kinds of test samples were screened from the aspects of simple,rapid and separation of preparation methods,and 70% methanol ultrasonic for 30 min was selected as the preparation method of GF and OF.Due to the differences in the chemical composition of GF and OF,it was found that use same chromatographic conditions is difficult.Therefore,the gradient elution procedure of methanol-0.3% glacial acetic acid was used GF.The gradient elution procedure of acetonitrile-0.4% glacial acetic acid was used OF.According to the characteristic printing guiding principle,that is,the characteristic component specificity and the active ingredient,forsythiaside I,forsythoside A,(+)-rosin-β-D-glucopyranoside,rutin,phillyrin,forsythia sulphate as a characteristic peak of GF.RS-Forsythia suspensa methyl ether,rutin,forsythiaside A,phillyrin and forsythia sulphate are characteristic peaks of old warts.Through the HPLC characteristic printing of 17 batches of GFGG currency,15 batches of GFSG and 15 batches of OFGG,the relative retention time of each characteristic peak was less than 5%,which meets the requirements.There is no significant difference in chemical composition between GFGG and the GFSG,but there is a big difference in the peak area of each characteristic peak.There are significant differences in chemical composition and peak area between GF and OF.This study provides a good scientific basis for the Forsythia commodity specification grade standard.3.Determination of the content of active ingredients in Forsythia3.1 Improvement of preparation method of test articlesIn order to determine the content of forsythoside A and phillyrin in FF quickly,simply and efficiently,the method for preparing forsythoside A and phillyrin test solution separately from the pharmacopoeia is improved,That is,the forsythoside A and phillyrin were simultaneously prepared for the test solution,and the phillyrin test solution was notpretreated.By comparing the extraction methods of 8 kinds of samples including Pharmacopoeia,it is better to use 70% methanol for 30 min.Further research on whether the phillyrin extract was treated with a neutral alumina column.Comparing the results of GF and OF and the neutral alumina column treated by different manufacturers,the results showed that the area of phillyrin in GF and OF was small before and after neutral alumina treatment.The purity and resolution of the peaks treated without neutral alumina column reach the pharmacopoeia values.Thus an improved method of pharmaceutical preparation of the test is possible.3.2 Investigation of content determination methodIn the case of separate determination of the pharmacopoeia content,in order to carry out the rapid determination,various simple methods are used in this paper,and the two methods are used to study the content of GF and OF.Simultaneously we will determine the method of forsythoside A and phillyrin in GF by HPLC.Based on the investigation of different chromatographic conditions by Xu Jia et al.,the following methods were established: acetonitrile(A)-0.4% glacial acetic acid solution(B)gradient elution,0~33min,A-B(15:85)),33~43 min,A-B(15:85)→A-B(25:75),43~60 min,A-B(25:75);detection wavelengths are 277 and 330 nm.The method has high ultraviolet absorption intensity,good resolution and purity when measuring the content of forsythoside A and phillyrin in GFGG and GFSG.So the method is applied to the determination of GF.OF still uses the pharmacopoeia method for determination.Using the method and pharmacopoeia method of this paper,40 batches of GFGG,30 batches of GFSG and 30 batches of OFGG were determined.It was found that forsythoside A and phillyrin are different in GFGG,GFSG and OFGG.Especially the significant difference between GF and OF,and the content limit should be specified separately.The content of forsythoside A and phillyrin in GFGG not be less than 4.50% and 0.50%.The content of forsythoside A and phillyrin in GFSG not be less than 6.00% and 0.60%.The content of forsythoside A and phillyrin in GFGG not be less than 0.35% and 0.15%.This provides a scientific basis for the establishment of the Forsythia product specification grade standard.4.A preliminary study of harvesting period of the OFAt present,the content of forsythoside A in most OF is low,and the content ofphillyrin is not up to standard.The reason for the failure of the OF may be affected by the harvesting period.In this paper,according to the pharmacopoeia determination method,the content determination of 5 batches of OF shells in different harvesting periods was studied.The results showed that the content of two index components in OF was higher from mid-October to November 2017.As time went on,the chemical components of OF gradually decreased.We will initially determine that the harvest time of OF is mid-October to until November.