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Ginkgo biloba by High Performance Liquid Chromatography compound content of total flavonoids in Ginkgo biloba_478

By Lisa Hughes,2014-10-30 18:38
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Ginkgo biloba by High Performance Liquid Chromatography compound content of total flavonoids in Ginkgo biloba_478

    Ginkgo biloba by High Performance Liquid Chromatography compound content of total flavonoids in Ginkgo biloba

     Abstract Objective To establish a compound of Flavonoids in Ginkgo biloba Ginkgo biloba in the determination. Methods

    using high performance liquid chromatography, using Symmetry

     C18 column (250 mm × 4.6 mm, 5 μm), methanol 0.4%

    phosphoric acid (52:48) as mobile phase, flow rate of 1.0 ml * min -1, detection wavelength 360 nm, column temperature was 35 ?. Results The linear relationship was good, the average recovery was 98.35%, RSD of 1.17% (n = 5). Conclusion The method is simple, separation is good, accurate, specific and can be used as quality control of Ginkgo leaf compound.

     Key words compound Ginkgo biloba; flavonoids; quercetin;

    kaempferol; isorhamnetin; high-performance liquid

    chromatography

     Abstract: ObjectiveTo establish the determination method of flavonoids in compound Ginkgo biloba Leaves Tablets. MethodsThe analysis was performed on a Symmetry C18 column

    (4.6 mm × 250 mm, 5 μm) with methanol 0.4% phosphoric

    acid (52:48) as mobile phase at a flow rate of 1.0 ml * min-1,

    and at a column temperature of 35 ?. The detection wavelength

    was 360nm.ResultsThe linearity of this method was good.The average recovery of added samples was 98.35% with RSD 1.17%. (n = 5). ConclusionThe method is convient with a good separating degree for the determination of flavonoids and can be used to evaluate the quality of compound Ginkgo biloba Leaves Tablets.

     Key words: Compound Ginkgo biloba Leaves Tablets; Flavonoid; Quercetin; Kaempferol; Isorhamnetin; HPLC

     Ginkgo leaf is a compound of Ginkgo biloba extract and Guangxi Pueraria Mirifica extract, etc. made of compound preparation, with blood circulation Huayu, reduce blood fat,

    removing free radicals and anti-aging effects and the role of

    [1]. Ginkgo leaves contain quercetin, kaempferol, isorhamnetin and other flavonoid compounds, one of its main active ingredient. Ginkgo biloba leaves the determination of

    flavonoids in many ways, such as derivative spectrophotometry [2], near-infrared spectroscopy [3], spectrophotometry [4], polarography [5], fluorescence spectrometry [6] , coulometric titration [7], capillary electrophoresis [8] and high performance liquid chromatography [9]. And the ginkgo leaves and preparations of Pueraria both formulations Determination of Flavonoids in Ginkgo biloba has not been reported. In this paper, high-performance liquid chromatography in Ginkgo leaf compound quercetin, kaempferol and isorhamnetin content of

    compounds such as flavonoids, methods, fast and easy, separation result is good, a strong specificity. Ginkgo biloba can be used for quality control of compound.

     An instrument and reagent

     1.1 Instrument U.S. PerKinElmer Inc. Series 200 High

    Performance Liquid Chromatography (including the Series 200 UV detector, Series 200 binary pump, 600 Series interface and chromatography workstation).

     1.2 reagent quercetin, kaempferol and isorhamnetin reference substance (China Pharmaceutical and Biological Products Institution, batch, respectively 10081

    200406,110861 200405,110860 200407). Ginkgo leaf

    compound-based research group in Guangxi Science and

    Technology Agency Fund under the three developed new products.

    Chromatography of pure methanol, water, high pure water, phosphoric acid as the AR.

     2 Methods and Results

     2.1 Chromatographic conditions Symmetry C18 column

    (4.6 mm × 250 mm, 5 μm, the United States Waters

    Corporation), the mobile phase was methanol: 0.4% phosphoric acid solution (52:48), flow rate 1 ml * min-1, detection of

wavelength of 360 nm, column temperature 35 ?, injection

    volume 20 μl.

     2.2 Solution Preparation

     2.2.1 Preparation of reference substance solution

    precision that learn from phosphorus pentoxide drying overnight quercetin, kaempferol and isorhamnetin appropriate reference substance (15.6,17.0,10.0 mg) Purchase 100 ml Liang Ping, add methanol Micro-heat allows the dissolved and diluted to the scale, shake, spare.

     2.2.2 Preparation for the test product solution to take this product 10, except for coating, and research fine, fine, said appropriate amount (about containing Ginkgo flavonol glycosides 19.2 mg), cone-shaped bottle home with Cyprus,

    precision by adding methanol, 20 ml, Mesa, saying that given the weight, ultrasound treatment 30 min, let cool, and then said that given the weight by using methanol to complement the weight loss, shaking, filtration. Precise amount of filtrate continued to take 10 ml, set 100 ml conical flask, add

    methanol 10 ml, 25% hydrochloric acid 5 ml, shaken, heated in a water bath set back 30 min, rapidly cooled to room temperature, transferred to a 50 ml flask in methanol was diluted to scale, shake, with 0.45 μm microporous membrane

    filtration, the filtrate continued to take that too.

     2.2.3 Preparation of negative control solution, in addition to taking a prescription outside of Ginkgo biloba extract the remaining herbs and accessories, according to the

    provisions of Technology prepared into tablets, press materials for the test solution made from the negative control solution preparation method.

     2.3 The precise amount of the production of standard curve to take the above-mentioned reference substance solution

    1.0,2.0,3.0,4.0,5.0 ml, respectively, 10 ml Liangping in the home, using mobile phase was diluted to scale, shake, each sample 20 μl, according to the above color spectrum

    measurement conditions in order to reference substance concentration C (μg * ml-1) as the abscissa, the peak area A of the vertical axis, draw the standard curve as follows:

     Quercetin: A = 80 074C - 52 255, r = 0.999 4, linear

    range of 15.6 ~ 78.0 μg * ml-1