Ginkgo biloba polysaccharide extraction, purification and the determination of
【Abstract】 Objective To study the extraction and
purification of polysaccharides of Ginkgo biloba and determination. Methods of extraction of different frequency,
solid-liquid ratio and extraction time of experiment, and the study of purified polysaccharide to establish conditions for various factors, using phenol - sulfuric acid method
polysaccharide content determined. The results of various factors, conditions of extraction temperature 80 ?,
extraction time 3 h, solid-liquid ratio 1:30, extracted two
times, purified with activated carbon. Ginkgo biloba may result in Polysaccharide yield was 12.2%, purified polysaccharides yield of 9.8%. Determination of polysaccharide
extracted crude polysaccharide content of 35.7%, purified polysaccharide content of 43.3%, polysaccharide determination of the relative standard deviation less than 3.0%. Conclusion This method is a good purification effect measured with good
reproducibility obtained in the polysaccharide content of Ginkgo biloba 4.3%.
【Key Words】 Ginkgo biloba polysaccharide determination of extraction and purification of polysaccharides
Abstract: ObjectiveThe extraction and purification methods for polysaccharide in Gingko leaves were studied and the content of polysaccharide in it was determined.MethodsThe
condition of each factor for extraction and pruification from the leaves of Gingko was established through the experiments of extraction times, different ratios of material to extraction solvent and extraction time.The method of phenol-
sulfuric acid was applied to determination of the content of the polysaccharide.ResultsThe conditions of the factors were
extraction temperature 80 ?, extraction time 3h, material to
extraction solvent 1:30, extraction 2 times and activated carbon for purification.Raw polysaccharide in gingko leaves
was 12.2%, the polysaccharide after prurified was 9.8%, the content of polysaccharide in the raw polysaccharide was 35.67%, and in the pure one was 43.32%, ConclusionThe purification method is efficient and stable. The content of
polysaccharide in gingko leaves is 4.3%.
Key words: Ginkgo leaves; Polysaccharide; Extraction; Purification; Determination of polysaccharide
Polysaccharide also known as polysaccharides
(polysaccharide), the four major life activities constitute
one of the basic material, not only the body primarily for energetic materials, but also has a wide range of biological functions in life activities involved in the cells activities. Currently known active polysaccharides have anti-tumor, anti-
virus, enhance immunity, anti-aging, blood fat, lowering blood
sugar, detoxification, anti-radiation effects [1 ~ 5], its
role increasingly attention.
Ginkgo (Ginkgo) is the period of survival in glacial relict plant on the planet, known as a "living fossil," said.
China is the birthplace of Ginkgo biloba and its resources accounted for about 70% of the world total [6,7]. At present, the Ginkgo biloba extract from the medicinal pure gradually applied to the food and beverage, health care products and
other industries, including Ginkgo biloba extract hair care, hair growth, skin care cosmetics such as the development has made some progress. Ginkgo leaves contain an important role in physiological and pharmacological function of the active ingredient, but on the polysaccharides in Ginkgo biloba is currently very little coverage because people are concentrating their efforts are concentrated in the leaves of the Ginkgo Flavonoids research on [7 ~ 12], the neglect of polysaccharides. The experiments of extracting polysaccharide
from Ginkgo biloba in the influencing factors, purification carried out a series of studies to determine the conditions for extraction and purification. At the same time on the extraction and purification of the polysaccharide content were
measured to determine the polysaccharide content in the leaves of the Ginkgo biloba. This is a comprehensive development and utilization of Ginkgo biloba leaves of great significance.
1 Materials and methods
1.1 Instrument and reagents constant temperature water
bath, centrifuge, constant temperature drying oven, vacuum drying, UV - visible spectrophotometer.
95% ethanol, anhydrous ethanol, chloroform were of analytical grade; glucose as biochemical reagents.
1.2 Sample source and pretreatment of the samples of Ginkgo biloba, in 2005 11 taken from the Dalian
Nationalities University campus. Will be mined by the Ginkgo biloba washed with tap water, then washing with distilled water, splashing water, after dry, with 70 ? constant
temperature drying oven.
1.3 extraction and purification methods of Ginkgo biloba polysaccharide extraction, purification steps are as follows:
a) Ginkgo biloba treatment: before the experiment with the 70 ? constant temperature drying oven to bake another 1
of Ginkgo biloba, and then use high-speed grinder grinding,
over 80 mesh sieve, placed in brown bottle to save.
b) Extraction temperature: according to a certain solid-
liquid ratio by adding water, extracted in constant temperature water bath temperature.
c) centrifugation: centrifuge extracted later to 5 000 r / min centrifugation 20 min, collect supernatant, residue re-
extracted one time with the former, merging two supernatant.
d) purification: the supernatant obtained after
centrifugation, purified with activated carbon.
e) Concentration: vacuum drying of the purified liquid for centrifuge enrichment.
f) alcohol precipitation: Add 5 times the volume of 95% ethanol, placed in the refrigerator put it aside for 24 h.
g) centrifugation: The alcohol precipitation of a white floc sedimentation centrifuge polysaccharide.
h) washing: With the polysaccharide with anhydrous ethanol and chloroform followed by washing and then placed a small beaker, evaporated in the water bath after the pot placed in 80 ? oven drying.
2 Results and Conclusion
2.1 Experimental Factors Affecting Polysaccharide extraction
2.1.1 solid-liquid ratio on the yield of crude
polysaccharide obtained five copies of 5 g of Ginkgo biloba
samples were placed in 250 ml flask with plug in, according to solid-liquid ratio 1:10,1:20,1:30,1 : 40,1:50 were added 50,100,150,200,250 ml of distilled water, pH 7, at 80 ? water
bath extraction 3 h, then centrifuged, concentrated with
ethanol precipitation, in the refrigerator put it aside for 24 h, re-centrifugation, crude polysaccharide yield shown in Figure 1.
Can be seen from Figure 1, solid-liquid ratio the bigger
the better, but to carry out two times extraction, therefore,
solid-liquid ratio not too big. As can be seen from Figure 1, solid-liquid ratio of 1:30 after the crude polysaccharide yield increases slowly. Solid-liquid ratio of 1:30, the crude
polysaccharide yield was 10.4%, solid-liquid ratio of 1:40,
when crude polysaccharide yield was 10.5%, only an increase of 0.1%; solid-liquid ratio of 1:45, the coarse polysaccharide yield was 10.7% over the solid-liquid ratio of 1:40 increased
by 0.2% over the solid-liquid ratio of 1:30 to increase 0.3%. The solid-liquid ratio of 1:20, the crude polysaccharide yield was 9.4%, solid-liquid ratio of 1:30 compared to 1:20 and the yield of crude polysaccharide