DETERMINATION of ELLAGIC ACID and RESVERATROL
in BLUEBERRY SAMPLES GROWN in TURKEY
Süleyman Seyhan a, Güler Yalçın b, Serap Ayaz Seyhan
b
a. Marmara University Institute of Health Sciences
02-05 November 2013
Antalya TURKEY
b. Marmara University Faculty of Pharmacy Department of Analytical Chemistry, Haydarpasa – Istanbul / Turkey
[email protected]
INTRODUCTION
Epidemiological studies suggest that consumption of fruit and vegetables contributes to a reduced risk
of certain types of human cancer and cardiovascular diseases. Berries are popular because of their good
taste and well-known nutritional value.
Vaccinium berries such as blueberries (Vaccinium corymbosum L.)
(Fig. 1) contain high amounts of sugars and acids as well as
phenolic compounds which display potential health-promoting
effects (1).
Blueberries are grown in large scale in Black Sea Region of Turkey
(Fig. 2) and they are currently being promoted as a rich source of
antioxidants.
Figure 2. Blueberry gardens
Ellagic acid (Fig. 3) and Resveratrol (Fig. 4) have been detected in some blueberry cultivars. Interest in
Ellagic acid and Resveratrol has been increased during the past decade due to their possible
antimutagenic, anticarcinogenic, and antioxidative effects (1, 2, 3).
DAD1, 17.443 (23.8 mAU,Apx) of SS000077.D
DAD1, 32.943 (137 mAU, - ) of SS000077.D
mAU
In this study, a chromatographic method was developed for Ellagic Acid
and Resveratrol by high performance liquid choromatography and diode
array detector.
mAU
20
100
80
60
40
15
10
5
METHOD
20
0
0
300
400
500
600
200
nm
Figure 3. The spectrum of ellagic acid
600
nm
32.796
33.517
31.525
30.508
0
-5
An ASE-100 extractor was used for the extraction of samples. The
extraction cell was filled with 2 g lyophilized samples mixed with
diatomaceous earth.
The extraction was performed employing methanol %100 as the solvent,
in three cycles, at 60 0C.
They were evaporated to dryness. The dry residue was dissolved in
methanol–water (1:1, v/ v).
Chromatographic conditions;
Mobile phase: 5 mM Potassium dihydrogen phosphate / ACN.
Gradient elution was used.
Column : Luna ODS-2 RP- C18 (5μ m, 4.6 × 250 mm i.d.),
Detector: DAD Detector, Wavelenght for Ellagic Acid λ : 260 nm, for
Resveratrol λ : 310 nm (Band width: 4 nm),
Flow Rate : 1 mL/min., injection volume: 20µL, pressure: 162 bar.
28.175
26.779
27.390
27.894
28.498
28.844
29.551
25.864
10
19.338
3.588
4.067
2.613
2.821
20
18.169
15
17.221
30
15.356
20
13.797
40
12.725
25
10.690
17.442
30
9.192
50
6.849
7.294
mAU
0
500
DAD1 A, Sig=260,4 Ref=off (SS\SS000031.D)
mAU
35
5
400
Figure 4. The spectrum of resveratrol
DAD1 A, Sig=260,4 Ref=off (SS\SS000077.D)
10
300
2.078
2.218
2.405
2.620
3.047 2.812
3.483
3.690
4.135 3.875
4.368
4.622
5.040
5.307
5.615
200
Figure 1. Blueberry (Vaccinium corymbosum L. ) plant,
flower, underripe and ripe fruit (4)
-10
30
35
min
Figure 5. The chromatogram of ellagic acid, DAD:Figure 6. The chromatogram of blueberry sample
260 nm
DAD: 260 nm.
RESULT AND DISCUSSION
DAD1 B, Sig=313,4 Ref=off (SS\SS000030.D)
mAU
35
mAU
35
30
30
25
25
20
20
15
10
5
Ellagic Acid and Resveratrol in four different blueberry varieties, grown in
the Black Sea Region of Turkey (Bluecrop, Brigitta, Darrow and Bluejay) were
analyzed for the first time in one run (Fig. 6, Fig. 8). We achieved good
resolution for the Ellagic acid and Resveratrol in the chromatogram.
The newly developed HPLC method in this study can assist for the
determination of the phenolic acids in berries. Besides, the findings will be
useful for cancer research on blueberries.
ACKNOWLEDGEMENTS
Blueberry samples were provided by NUHOĞLU VAKFI.
The study was supported by M. Ü. Bilimsel Araştırma Projeleri Birimi as SAG-C-DRP270109-0002 numbered project.
DAD1 B, Sig=313,4 Ref=off (SS\SS000031.D)
15
10
0
5
0
-5
-5
-10
-10
0
32.798
25
31.521
20
26.776
27.386
27.894
28.176
28.845
15
25.877
10
18.166
5
19.337
0
15.344
min
10.255
10.689
25
7.303
20
6.777
15
3.690
3.843
4.369
4.621
5.053
5.614
10
2.262
2.417
2.626
2.803
5
32.635
0
2.667
2.802
2.970
3.591
4.048
-10
5
10
15
20
25
30
35
min
0
5
10
15
20
25
30
35
min
Figure 7. The chromatogram of resveratrol, DAD: Figure 8. The chromatogram of blueberry sample
313 nm
DAD: 313 nm.
REFERENCES
1. Milivojevic J., Maksimovic V., Ercişli S., (2012). A Comparison of major taste and health related compounds of
Vaccinium berries. Turk J. Biol 36, 738-745.
2.Wang C.Y., Chen C. T., (2008). Fruit Quality, Antioxidant Capacity and Flavonoid Content of Organically and
Conventionally Grown Blueberries. J. Agric. Food Chem. 56, 5788-5794.
3. Sellaphan S., Akoh C.C., Krewer G., (2002). Phenolic Compounds and Antioxidant Capacity of Georgia-Grown
Blueberries and Blackberries. J. Agric. Food Chem 50, 2432-2438.
4. Çelik H. (2006). Karadeniz Bölgesindeki Asitli Topraklar için Mükemmel Bir Meyve, Likapa (Yaban Mersini). Ziraat
Odası Yayın Organı-Çiftçi Dünyası, 2(2).
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