Studies on the Optimal Process to Extract Flavonoids from Red

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Studies on the Optimal Process to Extract Flavonoids from Red

    Nature and Science, 3(2), 2005, Xu, et al, Optimal Process to Extract Flavonoids from Red-raspberry

    Studies on the Optimal Process to

    Extract Flavonoids from Red-raspberry Fruits

    Yaqin Xu, Rui Zhang, Hong Fu

    Science College, Northeast Agriculture University, Harbin, Heilongjiang 150030, China,

    Abstract: Effects of single factors such as temperature, time, material ratio and concentration of ethanol

    solution on the contents of flavonoids were investigated. On this basis, employing the orthogonal design,

    the optimum extraction conditions for flavonoids from raspberry fruits were determined by colorimetric

    estimation. Experiment results: the amount of flavonoids extract reached its maxima when extracted for 3

    h by using 95% ethanol solution at 80?C with the ratio of raw materials to ethanol solution of 1:10.

    [Nature and Science. 2005;3(2):43-46].

    Key words: red-raspberry; flavonoids; extract; single-factor experiments; orthogonal experiments

     3) orthogonal experiments after making analysis L9(3of variance and significance evaluation diagram of 1Introduction

    extracting process was: materials?extract ?filter

    Red-raspberry, which belongs to genus Rubus L. of ?enrichment ? extracted by using ethyl acetate?raw

    family to Rasaceae, is a kind of berry with tender and extract of flavonoids?confirm the volume by using juicy fruits. Fruits of red-raspberry are rich in vitamins 95% ethanol solution? measure. and minerals and especially abundant in caner- fighting,

    anti- aging substances such as SOD and gallogen 2.3Measuring of the contents of flavonoids (Häkkinen, 2000; Puupponen- Pimiä, 2005), therefore The contents of flavonoids was measured using famous as Fruit of Life in the US. Apart from those, colour comparing method of NaNO-Al(NO) and 233raspberry fruits contain significant flavonoids. It was calculated using the following formula (Xu, 2000): -reported that flavonoids could remove the O?in 2Relative contents of flavonoids (μg/g)=5YV/W human bodies, strengthen the natural disease-fighting Y= 75.944A- 0.3494 system, improve blood circulation and lower blood A: absorbance pressure (Fang, 1998; Liu, 2002; Wang, 1996). At W: the weight of raspberry fruits precisely measured:g present, studies on the extraction of flavonoids from V: the volume of extracting agent:mL. raspberry fruits have not been reported. In this study the Optimum conditions to extract Flavonoids from 3Results and Discussions Red-raspberry fruits were studied systematically in order to achieve scientific evidence for the processing 3.1Effects of single factor for flavonoids extraction and utilizing of raspberry fruits. 3.1.1The effect of the concentration of ethanol 2Materials and Methods solution on the content of flavonoids extracted

     The result of Figure 1 showed that the content of

    raw flavonoids extract increases with the concentration 2.1Materials

    of ethanol solution and the contents increased slowly The raspberry fruits were collected from the when the concent- ration was greater than 75%. Horticultural Center of Northeast Agricultural However, raw flavonoids extract had better solubility University and put into air-tight containers to store in a and the solvent was easily vaporized and reused in 95% refrigerator after instantly frozen and minced. ethanol solution, 95% ethanol solution was used.

     2.2Extracting process

    At first the main factors of the extraction 3.1.2The effect of temperature on the contents of

    temperature, the concentration of the extracting agent, flavonoids extracted extraction time and the materials ratio (the weight of Figure 2 was contents of raw flavonoids extracted fruits: volume of the extracting agent), which affect the at 50?C, 65?C, 80?C, 95?C under certain conditions. extraction of flavonoids were studied individually, and The contents of raw flavonoids extract tended to then the optimum extracting conditions of flavonoids increase gradually with the rise of temperature in range from raspberry fruits were determined by adopting of 50?C~80?C. It may be probable that the greater speed

    Nature and Science, 3(2), 2005, Xu, et al, Optimal Process to Extract Flavonoids from Red-raspberry

of the molecule movements in higher temperature so Therefore the optimum extraction time was 3 h.

    that flavonoids diffused more quickly from cell to

    extracting agent. But the flavonoids could be oxidized at 3.2Optimum conditions for flavonoid extraction temperature of surpassing 80? so that the contents of On the basis of the effects of single factor in above flavonoids extracted started to decrease gradually. 80?C chapters, adopting extracting agent of 95% ethanol was the optimum temperature. solution, the trials of tri- factor, tri-level Orthogonal

     design were conducted and the results were given in

    3.1.3The effects of the material ratio on the Table 1 and Table 2.

    contents of flavonoids extracted The results of orthogonal experiment were made an

    The result of Figure 3 showed that the contents of analysis of range, single-factor analysis of variance and flavonoids increased significantly with the rise of the the analysis of linear regression by SPSS software.

    material ratio in a range of 1:5~1:10 and changed Table 2, Table 3 and Table 4 showed the results. unobviously when the ratio was greater than 1:10. This The results of all kinds of analysis showed the was because when the material ratio was between 1:5 degree of affecting the contents of flavonoid extract was and 1:10, flavonoids were extracted fully with the rise C>B>A. The material ratio had the greatest effect on the of volume the extracting agent so as to the contents of test results, time of extraction rank second and the flavonoids increased. Otherwise, when the material ratio temperature came last. When extraction temperature and ndreached a certain level, the extract has well soluted in extraction time were at the 2 level, the contents of the solution lead to the contents of extract become extract reached its maxima. When the material ratio was rdsteady and wouldnt increase significantly. The at the 3 level, the amount of extract reached its optimum material ratio was 1:10. maxima. However, to save the solvent, its not

     economical to adopt for the material ratio of 1:15 even

    it got the greatest contents of extract. Table 2 showed 3.1.4Effect of flavonoid extraction time ndthat the 2 level was suitable for all three factors The result of Figure 4 showed contents of stbecause of the greater contents of extract than the 1 flavonoids extracted for 3 h reached its maxima and

    level. then decreased with prolonging extraction time. This

    may increase amount of impurities with the rise of time.



    140 140

    110 110μg/gμg/g80 content of flavonol 80Content of flavonol50 5050658095110 35557595115 Temperature () Concentration of alcohol(%) Figure 2. Effect of different temperature Fig.1 Effect of different concentration of alcohol

    200200 170170 140140g/gg/g μμ110110 8080 Content of flavonolcontent of flavonol5050 12345510152025 Time (h)Material ratio(V:W)

    Fig.4 Effect of the different extraction time Fig.3 Effect of the material ratio

Nature and Science, 3(2), 2005, Xu, et al, Optimal Process to Extract Flavonoids from Red-raspberry

    Table 1. Factors and levels


    Level A. Temperature(?) B. Extraction time (min) C. Ratio of solid to liquid (W:V) 1 65 120 1 5 2 80 180 110 3 95 240 115

    Table 2. Experiment results and range analysis

    Item A B C Flavonoids (μg/g)

    1 1 1 1 80.2

    2 1 2 2 157.4

    3 1 3 3 164.2

    4 2 1 3 143.3

    5 2 2 1 107.2

    6 2 3 2 179.8

    7 3 1 2 142.2

    8 3 2 3 182.8

    9 3 3 1 100.0

    K 401.8 365.7 287.5 1

    K 430.3 447.4 479.3 2

    K 425.0 444.0 490.3 3

    k 133.9 121.9 95.8 1

    k 143.4 149.1 159.8 2

    k 141.7 148.0 163.4 3

    R 9.5 27.2 67.6

    Table 3. Single- factor analysis of variance

     Sum of Squares df Mean Square F P

    A 153.3 2 76.6 0.044 F2.2=99 0.01

    B 1423.0 2 711.5 0.467 F2.2=19 0.05

    C 8670.9 2 4335.4 13.788 F2.2=9 0.1

    Total 10247.2 6 F2.2=3 0.25

    Table 4. Analysis of variance in linear regression

    Model Sum of Squares df Mean Square F

    1 Regression 7965.613 3 2655.204 5.122

     Residual 2591.897 5 518.379

     Total 10557.510 8

    Nature and Science, 3(2), 2005, Xu, et al, Optimal Process to Extract Flavonoids from Red-raspberry


     References [1] Fang P. Biological Effects of Flavones. Journal of Dali medical college 1998;7(4):52~54. To sum up, the Optimum process to extract [2] Häkkinen SH, Kärenlampi SO, Heinonen IM, Törrönen AR. Flavonoids from raspberry fruits were obtained, namely Ellagic Acid Content in Berries: Influence of Domestic Extracted for 3 h by using 95% ethanol solution at 80?C Processing and Storage. European Food Research and Technology 2000;212(1):75~80. with the material ratio of 1:10 (W:V). [3] Liu M, Li X. Antioxidant and Antiprolife -rative Activities of Raspberries. J Agric Food Chem 2002;50:2926~2930. [4] Puupponen-Pimiä R, Nohynek L, Alakomi HL, Correspondence to: Oksman-Caldentey KM. Bioactive Berry Compounds-novel Yaqin Xu Tools against Human Pathogens. Applied Micro-biology and 59 Wood Street Biotechnology. 2005;67(1):8~18. [5] Xu Y, Sun Y, Fu H. Studies on the Optimal extraction Conditions Gongbin Road, Xifang District of Flavonoid in Leaves of Black Currant. Journal of Northeast Harbin, Heilongjiang 150030, China Agriculture University 2000;7(2):144~146. [6] Wang H, Cao G, Prior R. Total antioxidant capacity of fruits. J Telephone: 01186-451-5519-0732 Agric Food Chem 1996;44:701~705. Cellular phone: 01186-130-7453-5378


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