Radiometric Study on Bioaccumulation of Gold by

Hesperidin Extracted from Orange peel

Abstract

With the advent of civilization and exponentially growing industries, earth is

being affected day to day. Mineral operating industries such as electronic device
manufacturing often contaminate our water by discharging chromium, copper, lead,
mercury. Many of these metals are highly toxic even at low concentrations and ultimately
taken by human via food chain, and some metals have high economic importance, like
gold. Thus the pre-concentration of useful metals from the polluted stream is urgent need.
This paper briefly summarizes the demand of present day science to find alternative
environmental friendly chemical methodologies even compromising with efficiency of
process using plant secondary metabolites which contains such as hesperidin as flavonoid

General terms:

Plant secondary metabolites: are organic compounds having origin from plants that are
not directly involved in the normal growth, development or reproduction of organism

Flavonides: the term flavonoid refers to class of plant secondary metabolites flavoniod
are most commonly known for their antioxidant activity flavonoid are also refered to as
bioflavonoid in the media

Preconcentration: Preconcentration falls under the general heading of sample

preparation techniques. Any analytical technique that requires a dissolved sample, and
there are many, often requires a lot of sample preparation.

Key words:
Hesperidin, pH dependent, bio-reagents

Introduction:
Now a day due to growing environmental awareness plant bio reagents are
getting renewed interest over the synthetic chemical reagents, as isolation of naturally
occurring plant bio molecules cause less environmental pollution than the synthesis of
chemical reagents. The use of spiked radio nucleoids to estimate the metal accumulation
by bio active plant materials is advantageous over the conventional methods because of
ease of measurements without the interference from other associated plant molecules and
lower detection limit. Studies have been made to examine the efficiency of various bio-
reagents as an alternative to conventional chemical reagents for preconcentration are
extraction of precious metals. The use of some biological reagents such as algae yeast etc
is efficient to pre concentrate different metal ions even in commercial scale. The present
paper involves the isolation of hesperidin which is bio flavoniod extracted from fruits of
orange peel. For ease of detection 198 Au (t1/2=2.696) has been spiked with macro
amount of gold. The studies revealed the efficiency of the hesperidin as a whole as an
extracting or pre-concentrating reagent for gold.
Hesperidin is a flavoniod which are polyphenols, occur as plant
secondary metabolites contain many hydroxyl groups and carbonyl groups capable of
forming complexes with metal ions suggest its potential use as adsorbents for removal of
metals including long lived radio active nuclide’s from natural waste water streams

Chemistry of and isotopic properties of gold:

Although gold is a noble metal, it forms many and diverse compounds.
Gold is attacked by and dissolves in alkaline solution of potassium or sodium cyanide,
and gold cyanide is used commercially in electro-plating of gold onto base metals. Gold
chloride solutions are used to make colloidal gold by reduction with citrate or ascorbate
ions. Gold chloride and gold oxide are used highly-valued cranberry or red-coloured
glass, which like colloidal gold sols contains evenly-sized spherical gold nanoparticles.
The oxidation state of gold in its compound ranges from – 1 to5+ but Au (1) and Au (III)
dominate. Gold (I), referred to as the aurous ion, is the most common oxidation state
with”soft” ligands such as thioethers, thiolates, and tertiary phosphates. Au (I)
compounds are typically linear. A good example is Au (CN) 2, which is the soluble form
of gold encountered in mining. Curiously, aurous complexes of water are rare. The binary
gold halides, such as AuCl, from zig zag polymeric chains, agin featuring linear
coordination at Au. Most drugs based on gold are Au (I) derivatives.
There is one stable isotope of gold, and 18 radioisotopes with 195Au
being the most stable with a half-life of 186 days. Gold has been proposed as a “salting”
material for nuclear weapons (cobalt is another, better-known salting material). A jacket
of natural gold, irradiated by the intense high-energy neutron flux from an expanding
thermonuclear weapon, would transmute into the radioactive isotope Au-198 with a half-
life of 2.697 days and produce approximately.411Mev gamma radiation, significantly
increasing the radioactivity of the weapons fallout for several days. Such a weapon is not
known to have ever been built, tested or used.

Methodologies for pre-concentration of elements:

The conventional principle metodologies applied in pre-concentration of elements are

a) Liquid - Liquid extraction
b) Ion Exchange chromatography

Liquid –Liquid Extraction:

Liquid –Liquid extraction is a technique in which solution is brought in to contact with a
second solvent, essentially imisible with the frst, in order to bring about a transfer of one
or more solutes in to second solvent. The separations that can be performed are simple,
clean, rapid and convenient. The technique is equally applicable to trace amounts of
materials.

Ion Exchange Process:

The term ion exchange is generally understood to mean the exchange of ions of like sign
between a solution and a highly

Bioflavonoid:
Flavonoids refer to a class of plant secondary metabolites. They are large family of
polyphenolic compounds synthesized by plants. Flavonoids share a common structure
consisting of 2 aromatic rings bound together by 3 carbon atoms that form an oxygenated
heterocycle, in which sixth carbon moiety is a benzene ring. Flavonoids are also
commonly reffered to as Bioflavonoid in the media the terms are equivalent and
interchangeable, for flavonoids are biological in origin. this group of polyphenolic
compounds is widely distributed through out the plant kingdom . To date about 3000
varieties of flavonoids are known.

Hesperidiin:
Hesperidin is a flavonone (flavonoid )(C28H34O15) found abundantly in citrus fruits .its
aglycone form is hesperetin . Hesperidin is believed to play a major role in the plant
defence . it acts as an antioxidant according to in vitro studies . In human nutrition it
contributes to the integrity of the vessel .

Hesperidin

Experimental:

Chemical reagents:
Methanol, petroleum ether, ethanol, acetic acid

Isolation of hesperidin from orange peel:

Hesperidin was first isolated by leberton in 1828 from albedo (the spongy inner
portion of peel) of orangaes of the family Hesperides and was given the name hesperidin.
Hesperidin may be extracted from orange peel by the procedure below. It is important to
have good quantity of albedo and not just the outer orange colored part of the peel.
Sun dried peel 200g is powdered in a disintegrator. Powdered dry orange peel (50 mg)
and petroleum ether (250ml) are placed in a round bottomed flask for one hour . While
hot; the mixture is filtered through a Buchner funnel. The powder is allowed to dry at
room temperature. The dry powder is returned to flask and 20ml of methanol is added.
The contents are heated under reflux for 3 hours and then filtere hot and washed with hot
methanol. The filterate is concentrated under reducing pressure, leaving a syrupy residue.

Re-crystallization of hesperidin:
The crude hesperidin is added to dimmethylfarmamide (7ml per g of syrup) . the mixture
is stirred vigorously at room temperature for 15 min , and then filterate to remove any
insoluble material , in a fume hood , the filterate is added drop wise with stiring to a
boiling solution of water (20ml per gold hesperidin) and acetic acid ( 0.5 ml per g of
hesperdin) . Allow the mixture to cool. Where possible, longer term cooling of the
mixture in the refrigerator may produce higher yields of product. the precipitated
hesperidin is collected by suction filtration and washed with a little cold water .

Flow chart for hesperidin isolation:

Analysis of isolated product:

Colour: Yellowish white

Nature: Amorphous
Solubility: product found insoluble in water, ether and was soluble in ethanol, boiling
acetic acid and methanol
Melting point: 242 degree Centigrade - 24 degree centigrade (acceptable value from
literature data)

UV Spectroscopy:
The obtained white powder is dissolved in ethanol and UV spetrum of the sample was
taken against pure ethanol baseline.
Spectrum was found to contain 2 peaks, one at 285nm and other at around 219nm is
agreeable with literature data of hesperidin, and another peak at 219nm may be attributed
to aromatic rings and conjugated dienes present in the flavonoid structure.

Observed data Literature data

Melting 243 245
point
UV 219 nm 285 nm
Absorption
Peak

Accumulation of Au ( 198) tracer by range peel and hesperidin: Variation f HCL

concentration:

0.1g of powdered orange peel was taken in to six sets of experiments.

Different concentrations of 4 ml HCL (10 M to 1M) was added separately to 0.1ml Au
(198) tracer (0.2 ml BRIT Solution + 1ml TDW) was added to each set and was shaken or
10 min. Centrifuge and allow to settle. The supernatant was removed and assayed for Au
(198) radionuclide by means of gamma-spectroscopy an was compared to the initial Au
(198) counts. Similarly, gold accumulation by hesperidin was measured at different HCL
concentrations.

Desorption of Au (198) from hesperidin:

Three chemicals were used to study the desorption properties of Au (198)
radionuclide from hesperidin. After desorption the supernatant liquid was removed and
the hesperidin sample is washed with deionized water till pH7. For all desorption
experiments, Au (198) bound hesperidin was suspended in different chemicals for 10 min
was shaken at room temperature. Then the hesperidin samples were subjected to gamma
spectroscopy. Percentage desorption was calculated as the percentage release of Au (198)
nuclides initially bound to the hesperidin. Chemicals used for desorption studies were 0,1
HCL .0.1 M NaNO2 , 0.1M Thiourea in pH 1 .
Accumulation of 198Au:
The accumulation of 198Au tracer by orange peel at different
concentrations of HCl has been presented in fig. it is evident from the figure that gold
accumulation occurs at 10-1M concentration. It has been found that the accumulation if
198Au tracer by hesperidin derived from orange peel is also pH dependent and maximum
accumulation of gold is observed at 10-2 HCl concentration. Therefore, it may be
concluded that accumulation of 198Au by orange peel is perhaps mainly due to
hesperidin which may form complex with 198Au

Fig: Bioaccumulation obtained with orange peel

Fig: Bioaccumulation obtained with orange peel

Fig: Bioaccumulation obtained with hesperidin

100
Desorption of 198Au:
Desorption of 198Au from hesperidin was done with reagents such as dilute

80
mineral acid, complexing agent or reducing agent, was presented in fig. nearly
rption, %

94%desorption of 198Au from hesperidin was found when 0.1M NaNo2 solution was
used as desorbing agent.

60
100
Conclusion:
The orange peel as well as alkaloid derived from it may be used as an
efficient bioreagent for extraction of gold. The potential of natural resource to
preconcentrate precious metal will help to exclude the use of hazardous chemicals in
similar processes and is a step forward to green chemistry. The developed method may
also find useful application for recovery of gold from the waste effluent of electroplating
and jewelery farm.

References:

1. D.NAYAK, M.NAG, S.LASKAR, S.LAHARI, j.radional. chem.., 268(2006) 337

2. D.NAYAK,S.LAHIRI,j.RAdional.Nuel.Chem.,267(2006)
3. D.NAYAK, K. M. HAZRA, S. LAKSHAR, S.LAHIRI, 6th intern.Conf. Nucl.
Radiochem. Symp.
4. B.CHOUDHRY, B.P.DAS, Curr. Sci., 52(1983)1130.

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