Studies On Quality Control Of Ginkgo Dietary Supplement Tea

Ginko Biloba Extract(EGB),is the extract of ginkgo plants Ginkgo’s(Ginkgo biloba L.) dried leaves, fine yellow-brown powder, bitter and astringent. With blood-activiating and stasis-dissolving, relieve pain, meridians effect. For the treatment of obstruction of collaterals by blood stasis, asthma, high cholesterol and hyperlipidemia. Ginkgo dietary supplement tea is a kind of tea which with health care functions, its production process is : black tea used as the base tea,and the ginkgo biloba extract were added to it, not only keeping the tea original appearance and flavor, but also make red tea nutrition richer, more desirable. With improved memory, scavenging oxygen free radicals, improve the circulation of hearts and brains effects, because of less side effect, it can be long-term edibled. No reported for its quality control, so this study uses HPLC-DAD-IT/MSn and HPLC-DAD technology for quality control ginkgo dietary supplement tea study, research contents are as follows: 1. HPLC-DAD fingerprint establish ginkgo dietary supplement teaA fingerprint analysis method was developed for the quality control of ginkgo dietary supplement tea by high performance liquid chromatography coupled with DAD detector(HPLC-DAD). The samples were extracted by heating reflux extraction with water as solvent, and then reconstituted with methanol.HPLC-DAD separation was performed on a Agilent TC- C18(250 mm×4.6 mm, 5 μm)column by gradient elution with 0. 5% formic acid- acetonitrile as mobile phase, at the temperature of 30 ℃and a flow rate of 1.0 m L/min.320 nm wavelength were selected for the fingerprint analysis. A good separation of 28 common peaks was achieved in 126 min. Similarity evaluation was performed by a professional software named Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(Version 2004A),and the similarity among 11 batches of samples was no less than 0. 98, which confirmed that the quality of ginkgo dietary supplement tea was stable. The developed method was reliable, stable and efficient, and was suitable for the quality control of ginkgo dietary supplement tea. 2 HPLC-DAD-IT/MSn spectrum of the establishment of ginkgo dietary supplement tea qualitative componentTo analyze and identify the chemical constituents from the water extract of ginkgo dietary supplement tea by HPLC-DAD-IT/MSn. The analysis was performed on an Agilent TC- C18(250 mm×4.6 mm,5 μm). The mobile phase consisted of acetonitrile and 0.5% formic acid was used for gradient elution, the flow rate was 1.0 m L/min, the split ratio of the liquid chromatograph and mass spectrometer is 7:3 after column. Methods Ion trap of mass spectrometry was applied for the qualitative analysis under positive and negative ion modes and ESI ion source was used for mass spectra. The results indicated that 47 compounds from the water extract of ginkgo dietary supplement tea had been identified by direct comparison in both positive and negative ion mass data, and the data of the literature. The efficient separation ability of HPLC and high sensitive detection of MS are used in this study, which will provide the evidences for evaluating the quality of ginkgo dietary supplement tea, stabilizing the health effects, and explaining the mechanism. 3. The constituents absorbed into blood of ginkgo dietary supplement tea by HPLC-DAD-IT/MSnTo establish an high performance liquid chromatography coupled with tandem Ion trap of mass spectrometer detection( HPLC-DAD-IT/MSn) comprehensive quickly analysis method to clarify the the medicine serum after the rats were oraly administrated ginkgo dietary supplement tea. The effective constituents of Serum of rats administered ginkgo dietary supplement tea was anlysized by HPLC-DAD-IT/MSn compared with the blank serum, medicine in vitro and single ginkgo dietary supplement tea. The information on the total ion chromatogram, mass chromatogram and the mass spectrogram were synthetically analyzed to confirm the effective constituents absorbed into blood. 18 constituents of ginkgo dietary supplement tea were detected in the rats plasma post the intragastric administration of ginkgo dietary supplement tea, among which 5 were original consitunents came from ginkgo dietary supplement tea, and 13 might be metabolits of original consitutents. The findings abtained from the study can provide the useful information for the determination of health effects substances of the ginkgo dietary supplement tea. 4. Simultaneous determination of 16 flavonoids in the ginkgo dietary supplement tea by high performance liquid chromatography-tandem mass spectrometryA method for the determination of 16 functional components of ginkgo dietary supplement tea such as catechin, vitexin, Puerarin, isoflavoues Aglycone, silymarin, quercetin, luteolin, apigenin, naringenin, hesperitin dihydrochalcone, kaempferol, hesperitin, isorhamnetin, baicalein, nobiletin and tangeretin by high performance liquid chromatography-tandem mass spectrometry( HPLC-MS/MS) was proposed. The conditions of chromatography and mass spectrometry were optimized. The 16 flavonoids were separated on a C18 chromatographic column with acetonitrile and water( additional 0.1% formic acid) as mobile phases under gradient elution at a flow rate of 0.25 m L/min. The determination was conducted by tandem mass spectrometry in positive ESI mode under multiple reaction monitoring(MRM) mode. Good linearities for all the compounds, with correlation coefficients over 0.996, were acquired. The recoveries were in the range of 70.9% to 100.0( n=6), while the relative standard deviations( RSDs) were less than 10%. The results showed that nine flavonoids, which were kaempferol, quercetin, hesperitin, vitexin, luteolin, catechin, apigenin, naringenin and isorhamnetin, are higher in content among the 16 flavonoids, and they constituted up 99.6% of the total flavonoids. The contents of these nine flavonoids can be considered as quality control index of the Ginkgo dietary supplement tea. The method proved to be rapid, selective, sensitive and stable, and it can be applied to the quality administration of the Ginkgo dietary supplement tea.