Study On The Characteristic Quantitative Fingeiprint Of Ginkgo Biloba Leaf And Its Extract By Iiplc

Ginkgo biloba L,the only surviving species of Ginkgo biloba,is called living fossils by Scientists.Ginkgo biloba Extract refers to the enrichment of active ingredients containing 24flavonoids and 6%lactone from Ginkgo biloba,regulating blood lipid,scavenging free radicals,etc.A variety of ginkgo preparations and products containing Ginkgo biloba extract is full of market.In order to control the quality of Ginkgo biloba products,ginkgo flavone was usually used as the main evaluation index of Ginkgo biloba extract.Because of many kinds of flavonoids,their composition is not completely clear and it is difficult to obtain the reference substance,which makes it difficult to directly measure the content.Therefore,we usually adopt the method of acid hydrolysis to determine the content of flavonoids in standards and literature at home and abroad,quality control result is not accurate by indirectly determinate the quality of the Ginkgo biloba and its extract.This topic regarding the quality problem of the Ginkgo biloba extract production,Ginkgo biloba and its extract as the research object,a quality control method of Ginkgo biloba and its extract was established by multiple indexes,which is high content,high active flavonoids from Ginkgo biloba leaves and its extracts.The main research contents and results of this paper include the following parts:1.Ginkgo biloba and its extracts is analyzed by high performance liquid chromatography with diode array detector?HPLC-DAD?method,analysis by the Gemini C₁₈?4.6 x 250 mm,5microns?,temperature:35?,mobile phase:acetonitrile-0.1%formic acid aqueous solution,the flow rate of 0.8 ml/min,sample quantity 10 uL,detection wavelength of 360 nm.The high performance liquid chromatography?HPLC?of ginkgo biloba and its extracts was compared,the peak area is more than 4.0%of 12 common peak.The 12 compounds were identified as flavonoids by ultraviolet spectroscopic analysis,and they were formed the signature components of Ginkgo biloba.2.The Ginkgo biloba extract is separated and purificated by using SephadexLH-20 and the preparation high performance liquid chromatograph,and compounds separated was identificated by the physical and chemical properties,spectral information,and NMR spectrumanalysismethod,obtaining12flavonoids,quercetin3-O-?2'',6''-di-O-?-L-rhamnopyransosyl?-?-D-glueopyranoside,kaempferol3-O-?2'',6"-di-O-?-L-rhamnopyransoyl?-?-D-glueopyranoside,kaempferol 3-sophoroside,quercetin 3-O-?6"-O-?-L-rhamnopyransoyl?-?-D-glucopyranoside-7-O-?-D-glucopyranoside,laricitrin3-O-rutinoside,quercetin3-O-glucoside,quercetin3-O-?2''-O-?-D-glucopyranosyl-?-L-rhamnopyranoside?,kaempferol3-O-rutinoside,isorhamnetin-3-rutinoside,kaempferol 3-O-?-L-?2-O-?-D-glucopyranosyl?rhamnpyranoside,quercetin3-O-?-?6'''-p-coumaroyl-glucosyl-?-1,4-rhamnoside?,kaempferol3-O-?-?6'''-p-coumaroyI-glucosyl-?-l,2-rhamnoside?.3.Characteristic fingerprint is established by 12 flavonol glycosides of mixed standard,determining quercetin 3-O-glucoside as internal standard,establishing between the internal standard and other compounds of relative correction factor,calculating the content of other compounds,and the methodology validation.The linear relation?r2 is greater than or equal to0.9994?,the detection limit is 0.13-0.83,the quantitative limit is 0.43-2.76,and the RSD of relative correction factor is 0.22-1.34.Precision results show that the RSD of days is less than0.89%,the RSD of stability is 0.66%-1.92%,the RSD of repeatability is 0.67%-1.96%,RSD proved sample solution is good precision,repeatability and stability within 48 h.The results show that the method is simple,accurate and reliable,providing reference for the quality control of Ginkgo biloba and its extracts.4.The method of quality evaluation of Ginkgo biloba and its extract was applied,13different parts of Ginkgo biloba and the preparation of Ginkgo biloba extract was quantifed analysis by using external standard method and multi-components quantitive by one marker,respectively.Comparison of the different methods,difference between 0.02 to 0.8,the results showed that the method is accurate.Through the above research,the signature of the Ginkgo biloba flavonoids prototype compounds was mixed reference substance,establishing a kind of multiple indexes for quality control of Ginkgo biloba and its extract method.It can be more accurate to verify that may occur in the production of quality problem,making the method more simple,economic and accurate,and providing a scientific basis for the quality control of traditional Chinese medicine extract products.