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Mothed of Analysis of Ginkgo Biloba Extract


Mothed of Analysis of Ginkgo Biloba Extract


Total flavonol glycosides, determine according to HPLC

Chromatography conditions and system suitability: using octadecylsilanebonded silica gel (C18)as filling agent; methanol : 0.4% phosphoric acidsolution ( 50:50) as mobile phase. Detection wavelength is 360nm and the numberof the theoretical plates of the column is not less than 2500.

Reference solution: accurately weigh reference quercetin, kaempferol andisorhamnetin previously dried overnight over phosphorous pentoxide, addmethanol to produce three solutions containing 0.03mg/ml. 0.03mg/ml,0.02mg/ml respectively as the reference solutions.

Test solution: accurately weigh 35mg of the test material, added the mixedsolution of methanol:25% hydrochloric acid (4:1), heated under reflux in waterbath for 2 hours, quickly cooling to room temperature and transfered to a 50mlvolumetric flask, diluted with methanol to the scale and shake well. Filterwith microporous membrane ( 0.5um), take the filtrate as testing solution

Procedure: accurately inject 10ul each of the above three referencesolutions and the test solution into the column separately, and determine thecontents of quercetin, kaempferol and isorhamnetin respectively. Calculated thecontent of total amount of flavonol glycosides using the following equation.

The content of total amount of flavonol glycolsides=( the content ofquercetin + the content of kaempferol + the content of isorhamnetin) X 2.51

Terpenoid lactones: carry out the test according to HPLC method ( appendix VID)

Chromatography conditions and system suitability: Using octadecylsilanebonded silica gel(C18)as filling agent and methanol:water(35:65) as mobile phase, detected by refrativeindex detector. The number of theoretic

al plate should not be less than 2500 calculated on the basis of terpenoiclactone peak and the separation degree between the peak of bilobalide and the Cpeak of ginkgolide should be greater than 1.5.

Reference solution: weigh accurately proper quantities of bilobalide,ginkgolide A, ginkgolide B and ginkgolide C, add methanol to produce per mlcontaining 2mg, 1mg, 1mg of mixed reference solution .

Test solution: weigh accurately about 150mg of the test materials, added10ml water and heated in water bath to dissolve, added 2 drops of 2%hydrochloric acid and extracted 4 times under shaking with ethyl acetate(15ml,10ml,10ml,10ml), combined the phase of ethyl acetate and washing with 20mlof 5% sodium acetate solution. Separately take sodium acetate solution and washagain with 10ml ethyl acetate. Combine ethyl acetate extract solution with washsolution and wash 2 times with water each 20ml. Separately take washed solutionand wash with 10ml ethyl acetate. Combine ethyl acetate solutions, recoverethyl acetate to dryness. Dissolve the residue with acetone and transfer to a5ml volumetric flask and add acetone to scale and shake well.

Procedure: Pipet accurately 10ul,15ul of reference solution and 10ul oftest solution respectively,

Inject into column and determine. Calculate the content of bilobalide,ginkgolideA, ginkgolide B and ginkgolide C respectively using the externalstandard two-point log equation method.

It contains terponid lactones not less than 6.0% in form of the sum ofbilobalide( C15H18O8), ginkgolide A( C20H24O9),ginkgolide B( C20H24O10), ginkgolide C( C20H24O11),calculated on the dried basis.

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