Qualitative Control Technologies Of Lonicerae Japonicae Flos And Its Preparations

In recent years,prices of Lonicerae JaponicaeFlos stay high,as the medicinal sources specification in Chinese pharmacopoeia and increasing requirements for multiple industries,such as pharmaceuticals,health products,beverages,and tea industries.There have been some illegal phenomenons in production and sales of Lonicerae Japonicae Flos extractive and its preparations,mainly including mixed use of Lonicerae Japonicae Flos and Lonicerae Flos,illegal use of stems and leaves of Lonicera japonica Thunb.,and unauthorized change of extraction process to increase chlorogenic acid content.However,existing quality control technologies could not solve the above-mentioned problems,and are far from enough to meet the drugs supervision needs.In order to deal with the aforementioned issues,we have studied,established many technologies,and developed method groups of quality control technologies for Lonicerae Japonicae Flos,its extractive and preparations.The main research contents and results are as follows:1 The distinguishingtechnologiesof Lonicera Flosand its confusing varietiesLonicerajaponica Flos,and application of the technologies.1.1 Morphologicaland microscopic identification techniques of Lonicerae Japonicae Flos and its confusing varieties.Lonicerae Flosfrom different sources are main confusing varieties of Lonicerae Japonicae Flos.Lonicerae Japonicae Flos differs from Lonicerae Flos in some morphologicaland microscopic characteristics.Most of the existing literatures only describe the differencesin words,which is not intuitive.In this research,we take a lot of typical morphologicaland microscopic characteristic photographs,which showed the differences informatively and vividly,and provided more intuitivereferences for routine supervision and inspection.Furthermore,it is found that hair cases of corolla and calyxsurfaces and distribution of non-glandularhairs are important characteristics toidentify medicinal materials.This achievement would be included in the book of Practical Handbook of Traditional Chinese Medicinal Herbs Identificationcompiled by National Institutes for Food and Drug Control.1.2 Admixture of Lonicerae Flos in marketed Lonicerae Japonicae Flos,and related detection technologies.It is found thatLonicerae Flos have been added illegally in marketed Lonicerae Japonicae Flos for higher profits.Plant origin of Lonicerae Flos mixed in has been accurately identified as L.macranthoides Hand.-Mazz.,applying the molecular biology technique,combining with the morphologicaland microscopiccharacteristics,chemical constituent researches,and investigation of producing areas.The influence of processing methods on the appearance of the buds of L.macranthoides Hand.-Mazz.was explored in depth.To approach the appearance of Lonicerae Japonicae Flos,the mixed buds of L.macranthoides Hand.-Mazz.were slowly dried in mediumor normal temperature,which was proposed definitely for the first time.Considering the specificity,sensitivity,and cost of inspection methods,the chemical composition determination technology was adopted.An HPLC-ELSD method was finally established to inspect the admixture of Lonicerae Flos in marketed Lonicerae Japonicae Flos.In 41 batches of marketed Lonicerae Japonicae Flos,15 batches contained characteristic triterpenoid saponins of Lonicerae Flos,accounting for 36.6%of the total.In the problematic samples,weight percent of Lonicerae Flos ranges from 4%to 23%.Therefore,it is now a serious problem to add Lonicerae Flos in marketed Lonicerae Japonicae Flos.The plant origin,processing method,and detection technologies of the added Lonicerae Flos applied a strong technical guarantee and theoretical support.On the basis of the above researches,we completed the 2015 Edition of the Chinese Pharmacopoeia Specialized Research Subject on Multi-source Traditional Chinese Medicinal Herbs-Lonicerae Japonicae Flos and Lonicerae Flos from the Chinese Pharmacopoeia Commission,and the Shandong Province 2015 Drugs Planned Sampling Task of Lonicerae Japonicae Flos.1.3 Feeding with Lonicerae Flos to produce Lonicerae Japonicae Flos preparations,and related detection technologies.With characteristic triterpenoid saponins of Lonicerae Flos as the indicator components,HPLC-ELSD detection methods of Lonicerae Flos were established in six Lonicerae Japonicae Flos preparations and related extracts.In 121 batches of six Lonicerae Japonicae Flos preparations,77 batches contained characteristic triterpenoid saponins of Lonicerae Flos,accounting for 63.6%of the total;In 43 batches of Lonicerae Japonicae Flosextracts,13 batches contained characteristic triterpenoid saponins of Lonicerae Flos,accounting for 30.2%of the total.Thus,it is now a serious problem to produce extracts and preparations of Lonicerae Japonicae Flos feeding with Lonicerae Flos.Related enterprises have not strictly screened raw materials,or even deliberately feeded with Lonicerae Flos.Governments should intensify supervision and urge enterprises to strictly comply with the preparation prescriptions.The established inspection techniques of Lonicerae Flos in Yinhuang Tablets and Yinhuang Granules have been included in 2015 Edition of the Chinese Pharmacopoeia.The established inspection technique of Lonicerae Flos in Yinhuang Dropping Pills has been published in the website of the Chinese Pharmacopoeia Commission,and would be included in the supplement editionof the Chinese Pharmacopoeia.2 The distinguishing component of Lonicerae Japonicae FlosandLonicerae JaponicaeCaulis,and its application.Lonicerae JaponicaeCaulis is the stem of Lonicerae Flos,which belongs to different parts of the same medicinal plant.There is a phenomenon of replacing Lonicerae Japonicae Flos with Lonicerae JaponicaeCaulis in extracts and preparations of Lonicerae Japonicae Flos.By comparing components of Lonicerae JaponicaeCaulis and Lonicerae Japonicae Flos,a characteristiccomponent of Lonicerae JaponicaeCaulis has been found,isolated,and identified as 5-O-[4’-O-(β-D-glucosylcaffeoyl)quinic acid.This compound was isolated from Lonicerae Japonicae Flos for the first time.,and this is the first report of complete NMRdata of 5-O-[4’-O-(β-D-glucosylcaffeoyl)quinic acid.According to the HPLCpeak area ratio of chlorogenic acid and 5-O-[4’-O-(β-D-glucosylcaffeoyl)quinic acid,a method to detect if Lonicerae Japonicae Flosextracts is produced with Lonicerae Japonicae Caulismaterials has been developed.In 48 batches of collected Lonicerae Japonicae Flos extracts,HPLCpeak area ratios of 19 batches,accounting for 39.6%of the total,were below the prescribed minimum value,which indicated the related extracts using Lonicerae JaponicaeCaulis as the raw material for production.Accordingly,it has been a serious problem to produce extracts of Lonicerae Japonicae Flos feeding with Lonicerae JaponicaeCaulis.Techniques forseparation,content determination,and application in adulteration identification of 5-O-[4’-O-(β-D-glucosylcaffeoyl)quinic acid have been authorized to obtain the invention patent(ZL2016103952621,ZL2016103950414,and ZL2016103947943)from State Intellectual Property Office,separately.3 Multi-components quality control technologies of Lonicerae Japonicae Flos preparations.Some enterprises have illegally changed the extraction process ofLonicerae Japonicae Flos to improve the chlorogenic acid content stipulated in the prepatation standard,so as to reduce the feeding amount of the raw material and save costs.However,types and proportion of caffeoyl quinic acids,main active components of Lonicerae Japonicae Flos extracts,are strikingly different with different extraction processes.Changes in the production processes are likely to affect curative effects of the related preparations.In our continuous research,the HPLC External Standard Method(ESM),Quantitative Analysis of Multi-components by Single-marker(QAMS),Specific Chromatogram,Reference Extracts techniques have been developed successively for qualitative and quantitative analysis of neochlorogenic acid,chlorogenicacid,cryptochlorogenic acid,3,4-O-dicaffeoylquinic acid,3,5-O-dicaffeoylquinic acid,and 4,5-O-dicaffeoylquinic acid in Lonicerae Japonicae Flos preparations and related extracts,which prevents enterprises from changeing the extraction process ofLonicerae Japonicae Flos to improve the chlorogenic acid content.Specific chromatogram techniques of Yinhuang Tablets,Yinhuang Granules,and related Lonicerae Japonicae Flos extracts have been included in 2015 Edition of the Chinese Pharmacopoeia.The specific chromatogram technique of Yinhuang Dropping Pills has been published in the website of the Chinese Pharmacopoeia Commission,and would be included in the supplement editionof the Chinese Pharmacopoeia.The composition and proportion of the prepared reference extract of Lonicerae Japonicae Flos are reasonable,the total content of the target components is over 70%,and the preparation method is simple and convenient.Thus,the extract has been authorized to obtain the invention patent(ZL2014100803166)from State Intellectual Property Office.The main innovations are as follows:1 The quality control system of Lonicerae Japonicae Flos extractive and its preparations was constructed,and the quality standards of Yinhuang series preparations and extracts were comprehensively upgraded,which were included in the Chinese Pharmacopoeia.The standards we made for Yinhuang series preparations have been applied to product quality control for more than 100 relevant pharmaceutical manufacturers in China.These standards have comprehensively improved the quality control level of Lonicerae Japonicae Flos and its preparations in an all-round way,and the illegal problems in LoniceraeJaponicae Flos industry have been effectively supervised and controlled,which provide strong technical supports for further enhancing the standards of Chinese medicinal materials,Chinese patent medicines,and drug safety supervision.25-O-[4’-O-,β-D-glucosylcaffeoyl)quinic acid was separated and determinated for the first time from the Lonicerae Japonicae Caulis.A method for the determination of the adulteration Lonicerae Japonicae Flos preparations and related extracts was established for the first time by the ratio of the above components to the peak of chlorogenic acid in the HPLC chromatogram.3 A Lonicera japonica Flosextract,in which the mass ratio ofneochlorogenic acid,chlorogenicacid,cryptochlorogenic acid,3,4-O-dicaffeoylquinic acid,3,5-O-dicaffeoylquinic acid,and 4,5-O-dicaffeoylquinic acidwas 0.2-0.4:1.0:0.3-0.5:0.2-0.4:0.1-0.3:0.2-0.4,and the total mass fraction of the above-mentioned compoents was more than 70%,were prepared for the first time.The controlled extract were applied to the identification of Loniceraejaponica Flosin Kanggan Granules.3 A method for the determination of six caffeoylquinic acids in Yinhuang series preparations by HPLC was established for the first time.The method was applied to the quality standard study of Yinhuang series preparations to achieve the simultaneous determination of multi-components and evaluate the whole quality4 The TLC and HPLC-ELSD methods of LoniceraeFlos in Qingre Jiedu Oral Liquid were established for the first time with characteristic triterpenoid saponins of Lonicerae Flos as the indicator components.