Anthocyanin Extraction and Purification

This assignment focuses on anthocyanins, natural pigments found in plants with antioxidant properties. It examines different methods for extracting and purifying anthocyanins, including aqueous two-phase extraction. The document references various studies highlighting the importance of anthocyanins in human health, their role as food colorants, and the challenges associated with their purification.

INTRODUCTION
Under environmental stress human bodies produce reactive oxygen
species (ROS), it is produced because our bodies do not have the ability to
detoxify the excess free radicles produced by ROS [1]. The excess free radicals
damage membranes, proteins, nucleic acids and lipids causing cancer,
Alzheimer’s and other diseases [2]. Antioxidants could neutralize the free
radicals produced by ROS but they have to be supplied through our diet [2]. Most
of the antioxidant rich food items could reduce diseases as per reports and
herbal medicines which have almost negligible side effects are used extensively
for supplying antioxidants [3]. Anthocyanins are part of flavonoid group and it
possesses antioxidant capabilities because it scavenges free radical. It helps in
fighting DNA cleavage, estrogen activity, lipid peroxidation and so on [4].
It is also used as anti-carcinogenic medicine but the exact behavior of
anthocyanins as carcinogenic medicine is still unclear [5]. Sixteen anthocyanins
have been tested against lung, skin and colon cancer [5]. Dacryodes rostrata is
mainly found in Sarawak, Malaysia and is indigenous to this region [6]. It is rich
in natural antioxidants, protein, oil, and minerals [6]. Aqueous two phase
extraction (ATPE) is an effective method for extraction and purification of a
variety of colorants like anthocyanins and it is widely used for recovering
biological products due to its short processing time, low cost, high capacity and
ease of use [7]. Therefore, in this study ATPE is used for the extraction and
purification of Anthocyanins from Dacryodes Rostrata Peel.
LITERATURE REVIEW
Cancer prevention agents are atoms that impedes oxidation that produces
free radicals harming cells in the human body. Manufactured cancer prevention
agents like butylated hydroxytoluene (BHT) cause damage to the liver and
cancer-causing driving individuals to investigate more on normal cell
reinforcements from plants [3]. At present, the utilization of the therapeutic plant
is quickly developing the world over because of its low symptoms,
notwithstanding that restorative plants demonstrate an equivalent measure of
advantages as that of engineered cancer prevention agent [4].
A few distinctive phenolic classes have been accounted for in the natural
products in high sums [6]. Among these mixes, anthocyanins are one of the
fundamental polyphenols exhibit, particularly in the peels of the natural products

[7]. Anthocyanins are the biggest and most essential gathering of water-solvent
and vacuolar shades in nature [8]. They are glycosylated polyhydroxy and
polymethoxy subsidiaries of 2-phenylbenzopyrylium cation i.e., anthocyanidin
gathering [9]. Anthocyanins are of unique intrigue on the grounds that their cell
reinforcement action and their potential use in the sustenance business as
normal colorants and saving operators [10].
Extraction of polyphenols from the product of the jambul is typically led
utilizing techniques that were adjusted from various specimen networks [8]. Be
that as it may, the specimen grid can assume a noteworthy part in extraction
energy and impact extraction productivity since associations amongst
polyphenols and proteins are known to exist [11]. In this manner, it is important
to assess the extraction of polyphenols for each specimen sort and, if
conceivable, utilizing distinctive procedures and techniques. All in all, ordinary
extraction procedures, for example, blending/shaking or Soxhlet extraction have
long extraction time and utilize high measure of dissolvable [8]. The long
extraction times are somewhat because of the utilization of non-particular
strategies utilizing extraction methods that outcome in low extraction efficiencies
and can prompt the debasement of some polyphenols [9].
Box-Behnken response surface experimental design (BBD) was utilized to
contemplate and improve the procedure factors, for example, extraction
temperature, time and solid– fluid proportion on the extraction yield of
anthocyanin and shading from jamun organic product mash and was watched
that, the extraction conditions effectsly affect the extraction yield of colors and
shading [8]. ANOVA demonstrated that the impacts of factors like solid– fluid
proportion, extraction time and Extraction temperature were critical and
quadratic models were created for foreseeing the reactions. Ideal arrangement
of the autonomous factors was acquired by derringer's coveted capacity
philosophy to remove the most extreme levels of shade and shading [8]. The
ideal conditions were observed to be: extraction temperature of 44 ˚C, extraction
time of 93 min and solid– fluid proportion of 1:15 g/ml separately [8].
ATPE can possibly accomplish the coveted refinement and grouping of the
item in a solitary advance. In the division of the colors, polyethylene glycol
(PEG)/(NH4)2SO4 framework has been utilized to isolate betalains from rough
beetroot extricate [9]. Notwithstanding, polymers are exceptionally costly and
difficult to recuperate. As of late, a novel fluid twophase framework (ATPS) made

out of short chain liquor and salt has been utilized to separate common mixes
because of its minimal effort and simple recuperation of dissolvable by
dissipation. For instance, ethanol/(NH4)2SO4 framework has been utilized to
isolate piceid, resveratrol and emodin from Polygonum cuspidatum [10], and n-
propanol/phosphate framework has been utilized for the refinement of salvianolic
B from rough concentrate of Salvia miltiorrhiza by counter-current
chromatography (CCC) [11]. Energized by the past outcomes, ATPE is believed to
be reasonable for the extraction of common colors.
When partition behavior and stability of anthocyanins were studied in
ATPE using ethanol and other phase forming salts, it was observed that the
stability of anthocyanins were affected by the pH of the system [7]. When the pH
of the solution was above 8 the pyran ring structure of anthocyanin opens up [7].
The effects of ATPE strategies on recovery and partition coefficients of
anthocyanins were studied so as to optimize ATPE [7]. Other parameters like
yield of anthocyanins and extraction time were also studied to optimize ATPE [7].
Thus it was observed that ATPE with hydrophilic solvent and an inorganic salt like
ammonium sulfate or ethanol was suitable for extraction of anthocyanins [7].
The study thus focusses on the purification and extraction of anthocyanins from
Dacryodes Rostrata Peel using ATPE.
REFERENCES
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