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2.2 %mg(w/w)
97.8 %mg(w/w)
Comparison of Different Methods and Storage Conditions for Anthocyanin Extraction from Black Goji Berry
(Lycium ruthenicum)
Supamas Kruawan1, Vichulada Janchom2 and Apinun Kanpiengjai1*
1Division of Biochemistry and Biochemical Technology, Department of Chemistry, Faculty of Science,
Chiang Mai University, Chiang Mai 50200, Thailand. 2Detox (Thailand) Company Limited
[1] Dhar, P. Tayade, A., Ballabh B., Chaurasia, O.P., Bhatt, R.P., and Srivastava, R.B. (2011). Lycium ruthenicum Murray: A less-explored but high-value medicinal plant from Trans-Himalayan cold deserts of Ladakh, India. Plant
Archieves. 11(2): 583-586.
[2] Martin J., Navas M.J., Jimenez-Moreno A.M., and Asuero A.G. Anthocyanin pigments: Importance, sample preparation and extraction. In: Soto-Hernandez M. (ed). Phenolic compounds-Natural sources, importance and
applications. IntechOpen; 2017: p.117-152.
[3] Islam T., Yu X., Badwal T. S., and Xu B. (2017). Comparative studies on phenolic profiles, antioxidant capacities and carotenoid contents of red goji berry (Lycium barbarum) and black goji berry (Lycium ruthenicum). Chemistry
Central Journal, 11(1): 59.
[4] Kupina S., Fields C., Roman, M. C., and Brunelle, S. L. (2019). Determination of Total Phenolic Content Using the Folin-C Assay: Single-Laboratory Validation, First Action 2017.13. Journal of AOAC International, 102(1), 320-321.
[5] Lee J., Durst R., and Wrolstad R. (2005). AOAC official method 2005.02: total monomeric anthocyanin pigment content of fruit juices, beverages, natural colorants, and wines by the pH differential method.
Official methods of analysis of AOAC International, 37, 37.
Acknowledgements➢ Advisor : Dr. Apinun Kanpiengjai
➢ Ms. Vichulada Janchom
➢ Division of Biochemistry and Biochemical Technology, Department of Chemistry,
Faculty of Science, Chiang Mai University
➢ Detox (Thailand ) company Limited.
ABSTRACT
Black goji berry is one of the most popular berries that is widely used in traditional Chinese medicine. It contains a water-soluble pigment, anthocyanin, as the major bioactive
compounds, thus a natural color enhancer and health benefits. The objectives of this study were to determine different methods for extraction of anthocyanin from black goji berry and
evaluate storage stability of anthocyanin. A total of three different extraction solvents including water, 60% ethanol and the mixture of ethanol-acetic acid-water in the ratio of 50:2:48
(v/v/v) were applied to the black goji berry and the extraction was performed at 30ºC and 50ºC. The extracted anthocyanin content was determined by the spectrophotometric pH
differential method. The results revealed that monomeric anthocyanin content of 180.74±20.9 mg/100 g black goji berry was obtained when the mixture of ethanol-acetic acid-water
was applied with the extraction temperature at 50ºC. This amount was of higher than those extracted by other conditions. Moreover, storage of the anthocyanin extracted by this
condition at -20ºC for 24 h resulted in 85% residual anthocyanin content. With regard to the extraction by 60% ethanol at 50ºC, the anthocyanin content of 151.88±12.5 mg/100 g
black goji was obtained, and this amount was retained with 100% residual anthocyanin content after the storage at both 4 and -20ºC for 24 h. Therefore, this extraction method and
storage condition is considered an appropriate protocol for production of anthocyanin from black goji berry.
Lycium ruthenicum
Lycium ruthenicum or black goji
berry is an important medicinal plant. It is
also known as black wolfberry that is native
of the Himalayan Mountains in Central Asia
where it is revered for its health benefits.
Black Goji fruits are a rich source of
carotenoids , anthocyanins , vitamins A, C,
B1 and B2 ; trace mineral like iron, zinc,
selenium and alkaloid betaine that helps to
produce choline [1].
Prevention of diabetes
Prevention of cardiovascular
and cerebrovascular diseases
Lowering blood pressure
Anti-inflammatory, anti-bacterial
and antioxidant activities.
Reduction of cancer cell
proliferation [3]
Anthocyanin
Fig.1 Structure of the major
anthocyanins-3-O-glucoside
❖ Anthocyanins are the largest
group of phenolic pigments and
the most important group of water-
soluble pigments in goji berry [2].
❖ Among the anthocyanin pigments,
cyanidin-3-glucoside is the major
anthocyanin found in most of the
plants.
References
Goji berry sample
3 g of dry ground goji berry
Extracted twice with 30 mL of different
extraction solvents
Placed in 30 ºC
water bath for 24 h
7,500 rpm for 5 min.
Collected supernatant
- Total phenolic content (TPC) [4]
- Monomeric anthocyanin content [5]
Ground with pestle and mortar
Storage at 4˚C in
dark condition
Storage at -20˚C in
dark condition
Extraction with
60%ethanol
Extraction with ethanol/
acetic acid / water
(50:2:48)
Extraction
with water
1 2 3
Solvent
The mixture of ethanol-acetic acid-water (50:2:48 v/v/v) in combination with
extraction temperature of 50ºC was the best anthocyanin extraction condition.
The best conditions for storage of anthocyanin was in 60% ethanol and
stored at 4 and -20ºC for 24 h.
CONCLUSIONS
Fig. 2 Anthocyanin content of black goji berries after extraction at 30ºC
and 50ºC with water, 60%ethanol and ethanol-acetic acid-water
Fig. 3 Effect of temperature on stability of
anthocyanins extracted from various organic
solvents at 30ºC
EXTRACTION
Fig. 4 Effect of temperature on stability of
anthocyanins extracted from various organic
solvents at 50ºC
Anthocyanin
Other phenoliccompounds
STORAGE CONDITIONS
Anthocyanin content
in Black Goji berry
Preparation
Placed in 50 ºC
water bath for 24 h
% Residual anthocyanin
Fig. 5 Anthocyanin obtained after
extracted the black goji berry by
ethanol-acetic acid-water (50:2:48) at 50ºC
0
50
100
150
200
30 ºC 50 ºC
Anth
ocyanin
conte
nt(
%m
g)
Temperature (ºC)
Water 60%Ethanol Ethanol : Acetic acid : Water
0
40
80
120
160
4 ºC - 20 ºC
%R
esid
ual A
nth
ocyanin
Temperature (ºC)
Water 60%Ethanol Ethanol : Acetic acid : Water
0
40
80
120
160
4 ºC - 20 ºC
%R
esid
ual A
nth
ocyanin
Temperature (ºC)
Water 60%Ethanol Ethanol : Acetic acid : Water
INTRODUCTION RESULTS
METHODOLOGY
Spectrophotometric pH differential method