A project report on fruit juices

1. A PROJECT REPORT ON
ANALYSIS OF SOME FRUIT JUICES FROM AGRA BY
FLAME ATOMIC ABSORPTION
SPECTROPHOTOMETER (FAAS)
SUBMITTED TO
DEPARTMENT OF CHEMISTRY, ST. JOHN’S COLLEGE, AGRA
FOR THE DEGREE OF MASTER OF SCIENCE (M Sc)
IN PHYSICAL CHEMISTRY (2013-2014)
UNDER THE SUPERVISION OF:
Dr. SUSAN VERGHESE .P
Associate Professor
Department of Chemistry
St. John’s College, Agra
SUBMITTED BY:
SHESHENDRA KUMAR
M Sc Final
Physical Chemistry
2013-14

2. CERTIFICATE
This is to certify that this project entitled “ANALYSIS OF SOME
FRUIT JUICES FROM AGRA BY FLAME ATOMIC ABSORPTION
SPECTROPHOTOMETER (FAAS)” submitted to St. John’s College,
Agra, for the fulfillment of the requirement for the Master degree
is a bona fide project work carried out by SHESHENDRA KUMAR
student of M Sc Final (PHYSICAL CHEMISTRY) under my
supervision and guidance during the session 2013-2014. The
assistance and help rendered during the course of investigation
and sources of literature have been acknowledged.
Dr. Susan Verghese .P
Associate Professor
Department of Chemistry
St. John’s College, Agra
(Supervisor)
Dr. Hemant Kulshreshtha
HEAD
Department of Chemistry
St. John’s College, Agra

3. ACKNOWLEDGEMENT
It is my proud privilege to express my profound sense of gratitude and sincere
indebtedness to honorable Dr Alexander Lal, Principal of St. John’s College,
Agra, for providing infrastructure for the completion of this project. I am
thankful to Dr Hemant Kulshreshtha, Head of the Chemistry Department; he
was always affectionate, pain taking and source of inspiration to me. I am
highly obliged to him for their guidance, constructive criticism and valuable
advice which they provided to me throughout the tenure of my project. The
project work could not have been possible without his worthy suggestions and
constant co-operation.
I am also thankful to my supervisor Dr Susan Verghese to guide me on the
various sides of this project and her help and guidance she provided to me for
the initiation of this project. My heart is filled with deep sense of thankfulness
and obeisance to my teachers Dr. R P Singh, Dr. H B Singh, Dr. P E Joseph, Dr.
Raju V John, Dr. Shalini Nelson, Dr. Mohd. Anis, Dr. Anita Anand, Dr. Padma
Hazra, and Dr. David Massey for their valuable suggestions and lively moral
boosting during the progress of this investigation.
I am also thankful to Ms. Nisha Siddhardhan (Instrumentation in-charge) for
their kind support during the project work. I also place my sincere thanks to
non-teaching staff for their support and co-operation.
I am highly grateful to my parents for their affectionate and moral support.
They have always been source of inspiration for me.
Above all, I thank The Almighty for giving me strength to complete this project.
Last but not the least I extend my sincere thanks to all those who have helped
me in one or the other way during my project work.
SHESHENDRA KUMAR
M Sc Final (Physical Chemistry)

4. ABBREVIATIONS
RDA = Recommended Dietary Allowance
AI = Adequate Intake
UL = Upper Limit
DDI = Daily Dietary Intake
DRI = Dietary Reference Intakes
MAL = Maximum Acceptable Limit
SAM = Standard Addition Method
AA = Atomic Absorption
FAAS = Flame Atomic Absorption Spectroscopy
HCL = Hollow Cathode Lamp
MIBK = Methyl isobutyl ketone
APDC = Ammonium pyrrolidine dithiocarbamate
ND = Non Detectable
PMT = Photomultiplier tubes
LPG = Liquefied petroleum gas
ppm = Parts per million
Cu = Copper
Cr = Chromium
Pb = Lead
Ni = Nickel
Na = Sodium
Fe = Iron
Ca = Calcium
Cd = Cadmium
UL = The maximum level of daily nutrient intake that is likely to pose no risk of
adverse effects. Unless otherwise specified, the UL represents total intake from
food, water, and supplements.
ND = Non detectable due to lack of data of adverse effects in this age group
and concern with regard to lack of ability to handle excess amounts. Source of
intake should be from food only to prevent high levels of intake.

5. Contents
INTRODUCTION
Review of Literature
EXPERIMENTAL
RESULTS AND DISCUSSION
CONCLUSIONS
REFERENCES

6. INTRODUCTION
Fruit juices are the usual beverages used in everyday life. Fruit juices that found
themselves in the retail markets are mostly derived from citrus fruits. After
expression in a reamer the juice is strained, flash pasteurized, filled into bottles
and sealed. Benzoic acid is commonly used as preservative. Many fertilizers are
used in fields, as a result of the soil, atmosphere, underground and surface
water pollution, our foods and beverages are contaminated with heavy metals.
Some essential metals are involved in numerous biochemical processes and
adequate intake of certain essential metals relates to the prevention of
deficiency diseases. Copper (Cu) is an essential metal which perform important
biochemical functions and is necessary for maintaining health throughout life.
Adult human body contains about 1.5-2.0 ppm of Cu which is essential as a
constituent of some metalloenzymes and is required in haemoglobin synthesis
and in the catalysis of metabolic oxidation. Symptoms of Cu deficiency in
humans include bone demineralization, depressed growth, depigmentation
and gastro-intestinal disturbances. Heavy metals contamination has become a
matter of public health concern but this has not received much research
attention in India especially fruit juice contamination through heavy metals. In
the present study, levels of Cr, Cu, Cd, Pb, Ni, Na and Ca of fruit juices bought
from retail market in Agra, in December 2013 were determined using Flame
Atomic Absorption Spectrophotometer (FAAS) and Flame photometer.

7. Review of Literature
Metals are present in fruit juices either naturally or as a result of human
activities such as agricultural practices, industrial emissions, car exhausts, or
contamination during manufacture. Food and beverage contamination may also
occur due to raw materials and water used.
In several countries, similar studies were previously reported concerning heavy
metals as is the case in the current study (Maff 1998; Onianwa et al. 1999;
Ashraf et al. 2000; Krejpcio et al. 2005; Maduabuchi et al. 2006).
Krejpcio et al. (2005) reported lead, cadmium, copper, and zinc levels as 0.020–
0.46 mg/l, 0.004–0.060 mg/l, 0.047–1.840 mg/l, and 0.063–3.39 mg/l,
respectively, in a total of 156 fruit juice samples examined in Poland.
Marshall Fiona, Ravi Agarwal, Dolf te Lintelo, D S Bhupal, Dr Rana P B Singh,
Neela Mukherjee, Chandra Sen, Dr Nigel Poole, Dr Madhoolika Agrawal, S D
singh, 2003. Heavy Metals Contamination of Vegetables in Delhi.
Lokeshwari, H, G.T. Chandrappa, 2006. Impact of Heavy Metal Contamination
of Bellandur Lake on Soil and Cultivated Vegetation; Current Science.
S. M. Dogheim; El M. M. Ashraf; S. A. G. Alla; M. A. Khorshid; and S. M.
Fahmy, 2004. Pesticides and heavy metals levels in Egyptian leafy vegetables
and some aromatic medicinal plants; Food Additives and Contaminants.
S. C. Barman, R. K. Sahu, S. K. Bhargava, C. Chaterjee, 2000. Distribution of
Heavy Metals in Wheat, Mustard, and Weed Grown in Field Irrigated with
Industrial Effluents;
The research performed in England revealed that the heavy metal levels in the
fruit juics samples were within the standard. In this study lead, arsenic, and
cadmium contents were determined as 0.02–0.05 mg/l, < 0.1 mg/l, and 0.0004–
0.001 mg/l, respectively, in fruit juice samples from totally 100 samples (Maff
1998).
Maduabuchi et al. (2006) reported cadmium levels as 0.003–0.081 mg/l in fresh
fruit juices.

8. PERKIN ELMER AAnalyst 100 ATOMIC SPECTROPHOTOMETR
EXPERIMENTAL
SYSTRONICS 130 FLAME PHOTOMETER

9. EXPERIMENTAL
MATERIALS AND METHODS
Sample Collection
Fresh fruit juice samples were collected from Rajamandi, the main market of
Agra. Sampling was done at random from different retailers and vendors of this
market. A total of six (6) fruit varieties including oranges, pomegranates,
lemons, pineapple, apple, and mix juice were collected. Sampling was done for
a total of four days in December 2013. The fruit juice samples were then
analyzed for Cd, Cr, Cu, Ni, Pb, Na and Ca.
Sample Preparation
The collected fruit samples were thoroughly washed and rinsed with distilled
water. The samples were then sliced to small pieces and juice was prepared by
juicer.
Took 20 ml of juice in a 100 ml of volumetric flask, added 10 ml of HCl then
made upto the mark with distilled water. Shaked well, transferred to centrifuge
tube and filtered to remove solid particals.
Sample treatment and analysis
Apparatus
A Perkin-Elmer AAnalyst100 double beam atomic absorption
spectrophotometer (Perkin-Elmer
corp., CT) was used at a slit width of 0.7 nm, with hollow cathode lamps for
mineral measurements by FAAS. Samples were atomized for Cr, Cu, Cd, Ni, and
Pb. All analyses were performed in peak height mode to calculate absorbance
values.
SYSTRONICS Flame photometer 130 was used for the estimation of Ca and Na.
All solutions were prepared from analytical reagent grade reagents, for e.g.,
Commercially available 1,000 μg/mL Cu [prepared from Cu(NO3)2.3H2O in 0.5
M HNO3] were used. The water employed for preparing the standards for
calibration and dilutions was ultra pure water with a specific resistivity of 18
m_ cm-1 obtained by filtering double-distilled water immediately before use.
Calcium and sodium can be easily analysed by Flame Photometer. Standards
can be prepared as follows-
 Calcium – 1000 ppm

10. Dissolved 2.497 g CaCO3 in approx 300 ml glass distilled water and added 10 ml
conc. HCl diluted to 1 litre.
For calibration 20, 40, 60, 80 and 100 ppm solutions were prepared from the
stock solution.
 Sodium- 1000 ppm
Dissolved 2.5416 g NaCl in one litre of glass distilled water.
For calibration 20, 40, 60, 80 and 100 ppm solutions were prepared from the
stock solution.
Sample analysis of Heavy Metal content A Perkin Elmer Atomic Absorption
Spectrophotometer (AAS) model AAnalyst 100 with Air- C2H2 flame type of an
average fuel flow rate of between 0.8 to 4.0 L/min and the support gas flow
rate between 13.5 to 17.5 L/min was used.
INTRODUCTION/ BASIC PRINCIPLE
Spectroscopy is the measure and interpretation of electromagnetic radiation
absorbed, scattered or emitted by atoms, molecules or other chemical species.
When the electromagnetic radiation absorbed by atoms is studied, it is called atomic
absorption spectroscopy. This absorbance is associated with changes in the energy
state of the interacting chemical species since each species has characteristics
energy states. Atomic absorption spectroscopy (AAS) or atomic absorption (AA) or
atomic absorption spectrometry (AAS) uses the absorption of light to measure the
concentration of gas-phase atoms. Since samples are usually liquids or solids, the
analyte atoms or ions must be vaporized in a flame (such as air-acytelene flame) or
graphite furnace that contains the free atoms become a sample cell. The free atoms
absorb incident radiation focused on the from a source external to a flame and
reminder is transmitted to a detector where it is changed into an electrical signal and
displayed, usually after amplification, on a meter chart recorder or some other type
of read-out device.
The sample solution is introduced as an aerosol into the flame and atomized. A light
beam from the source lamp (hollow cathode lamp, HCL) composed of that element
(intense electromagnetic radiation with the wavelength exactly the same as that is
absorbed maximum by the atoms) is directed through the flame, into a
monochromator and onto a detector that measures the amount of the light
absorbed by the atomized element in the flame (Fig. 1). Because each metal has its
own characteristic absorption wavelength, the amount of energy at the
characteristics wavelength absorbed in the flame is proportional to the
concentration of the element in the sample over a limit concentration range.

11. The atoms absorb ultraviolet or visible light and make transitions to higher electronic
energy levels. The analyte concentration is determined from the amount of
absorption. Applying the Beer-Lambert law directly in AAS is difficult due to the
variations in the atomization efficiency from the sample matrix, and non uniformity
of concentration and path length of analyte atoms (in graphite furnace AA).
Concentration measurements are usually determined from a working curve after
calibrating the instrument with standard of known solution.
ATOMIC TRANSITION THEORY
The probability that an atomic spectroscopic transition will occur is called the
transition probability or transition strength. This probability is determine the extent
to which an atom is absorb light at a resonance frequency, and the intensity of the
emission lines from an atomic excited state. The spectral width of a spectroscopic
transition depends on the widths of the initial and final states. The width of the
ground state is essentially a delta function and the width of an excited state depends
on its lifetime.
INSTRUMENTATION
Light source- The light source is usually a hollow cathode lamp of the element
that is being measured. Lasers are also used in research instruments. Since laser are
intense enough excite atoms to higher energy levels, they allow AA and atomic
fluorescence measurements in a single instrument. This disadvantage of these
narrow-band light sources is that only one element is measurable at a time.
Atomizer- AA spectroscopy requires that the analyte atoms be in the gas phase.
Ions or atoms in a sample must undergo desolvation and vaporization in a high
temperature source such as a flame or graphite furnace. Flame AA can only analyze
solutions, while graphite furnace AA can accept solutions, slurries or solid samples.
Flame AA uses a slot type burner to increase the path length, and therefore to
increase the total absorbance (see Beer-Lambert law).
Sample solutions are usually aspirated with the gas flow into a nebulizing/mixing
chamber to form small droplets before entering the flame.
The graphite furnace has several advantages over a flame. It is much more
efficient atomizer than a flame and it can directly accept very small absolute
quantities of sample. Samples are placed directly in the graphite furnace and the
furnace is electrically heated in several steps to dry the sample, ash organic matter,
and vaporize the analyte atoms.

12. Light separation and detection- AA spectrometers use monochromators and
detectors for UV and visible light. The main purpose of the monochromator is to
isolate the absorption line from background light due to interferences. Simple
dedicated AA instruments often replace the monochromator with a band pass
interference filter. Photomultiplier tubes (PMT) are the most common detectors for
AA spectroscopy.
AAS AT A GLANCE
Principle- It measures the decrease in light intensity from a source (HCL) when it
passes through a vapour layer of the atoms of an analyte element. The hollow
cathode lamp produces intense electromagnetic radiation with a wavelength, exactly
the same as that absorbed by the atoms, leading to high sensitivity.
Construction- It consists of a light source emitting the line spectrum of the element
(HCL), a device for the vaporizing the sample (usually a flame), a means of isolating
an absorption line (monochromator) and a photoelectric detector with its associated
electronic amplifying equipment.
Operating Procedure- HCL for the desired elements is installed in instrument and
wavelength dial is set according to the table and also slit width is set according to the
manual. Instrument is turned on for about 20 min to warm up. Air flow rate and
acetylene current are adjusted according to the manual. Standard solution is
aspirated to obtain maximum sensitivity for the element is adjusting nebulizer.
Absorbance of this standard is recorded. Subsequent determinations are made to
check the consistency of the instrument and finally the flame is extinguished by
turning off first acetylene flame and then air.
Lamps- Separate lamp (HCL) is used for each element since multi element hollow
cathode lamps generally provide lower sensitivity.
Vent- A vent is paced about 15-30 cm above the burner to remove the fumes and
vapours from the flame.
Determination of Heavy Metals-
Reagents-
1. Air- cleaned and dried through a filter air.
2. Acetylene- standard, commercial grade
3. Metal free water- all the reagents and dilutions were made in metal free water
4. Methyl isobutyl ketone (MIBK)- Reagent grade MIBK is purified by re-distillation
before use.

13. 5. Ammonium pyrrolidine dithiocarbamate (APDC) solution- 4 g APDC is dissolved in
100 ml water.
6. Conc. HNO3
7. Standard metal solutions: Five standard solutions of 0.01, 0.1, 1, 10 and 100 mg/L
concentrations of metals such as Cr, Mn, Fe, Ni, Cu, Zn, Cd and Pb for instrument
calibration and sorption study are prepared by diluting their stock solution of 1 g/l,
i.e., 1 ml = 1 mg metal.
Procedure-
a. Instrument operation- same as above. Solution is aspirated into flame after
adjusting the final burner position until flame is similar to that before aspiration of
solvent.
b. Standardization- five standard metal solutions in metal free water are selected for
the standardization of the instrument. Transfer standard metal solutions and blank
to a separatory funnel and added 1 ml APDC, 10 ml MIBK and was shaken vigorously.
Aqueous layer is drained off and organic extract was directly aspirated into the
flame.
c. Sample analysis- Atomizer (nebulizer) is rinsed by aspirating water saturated MIBK
and organic extracts obtained by above the method were directly aspirated into the
flame.
d. Calculation- concentration of each metal ion in milligrams per litre is recorded
directly from the instrumentation readout.

14. FLAME PHOTOMETER
Flame photometry is an atomic emission method for the routine detection of metal
salts, principally Na, K, Li, Ca and Ba. Quantitative analysis of these species is
performed by measuring the flame emission of solution containing the metal salts.
Solutions are aspirated into the flame. The hot flame evaporates the solvent,
atomizes the metal, and excites a valence electron to an upper state. Light is emitted
at characteristic wavelengths for each metal as the electron returns to the ground
state. Optical filters are used to select the emission wavelength monitored for the
analyte species. Comparison of emission intensities of unknown to either that of
standard solution, or to those of an internal standard, allows quantitative analysis of
the analyte metal in the sample solution.
Introduction- SYSTRONICS flame photometer 130 is an instrument with which it is
possible to estimate, with speed and accuracy, minute quantities of sodium (Na),
Potassium (K), Calcium (Ca) and Lithium (Li).
The principle of operation is simple. The fluid under analysis is sprayed as a fine
mist into a non-luminous (oxidizing or colorless) flame which becomes colored
according to the characteristic emission of the metal. A very narrow band of
wavelength corresponding to the element (Na: 589 nm, K: 768 nm, Ca: 622nm, Li:
671 nm) being analysed is selected by a light filter and allowed to fall on a photo-
detector whose output is measure of concentration of the element. The output of
photo-detector is connected to an electronic metering unit which provides digital
readouts. Before analyzing the unknown fluids, the system is standardized with
solutions of known concentrations of the element of interest.
The total system consists of two units-
1- Main unit,
2- Compressor unit. The main unit consists of an atomizer (for aspiration of solutions),
mixing chamber, burner, optical lens, light filters, photodetectors, control valves and
electronic circuit.
Compressed air (oil free) from the compressor unit is supplied to the atomizer. Due
to a draught of air at the tip of the atomizer, the sample solution is sucked in and
enters in the mixing chamber as a fine atomized jet. Liquefied petroleum gas (LPG)
or laboratory gas from a suitable source is also injected into mixing chamber at a
controlled rate. The mixture of gas and atomized sample is passed on to the burner
and is ignited. The emitted light from the flame is collected by a lens and is passed
through an appropriate filter (Selectable for different element). The filtered light is

15. then passed on to energize a sensitive photo-detector, the output of which is applied
to the electronic circuit for readout.
OPERATING PROCEDURE AND SAMPLE ESTIMATION
Once the burner is ignited and set, followed the steps described below-
Put on the mains supply to the unit. Digital display turned on.
Turned the SET F.S. COARSE and FINE controls in maximum clockwise position.
Select appropriate filter with the help of Filter Selector wheel (Na on the left side
and K on the right side).
Feed distilled water to the atomizer and wait atleast for 30 seconds.
Adjust the SET REF. COARSE and FINE controls for a zero readout as nothing
aspirated, for K only.
Aspirate 1 mEq/L of Na solution (or the standard 1.0 / 0.01 mEq/L of Na/K solution).
Wait atleast 30 s and then adjust the SET REF. COARSE and FINE controls for a
readout of 100 for, Na only.
Aspirate the standard mixed 1.7/0.85 mEq/L of Na/K solution and wait atleast for 30
s. Adjust SET F.S. control of the Na side for a readout of 170 and that of the K side for
a readout of 80. The unit stands calibrated.
For a recheck, aspirate the standard mixed solution of 1.0/0.01 mEq/L of Na/K. the
readout for Na and K should be close to 100 and 10 respectively.
Then feed sample solution to the atomizer to get the relative concentration. Wait
atleast for 30 s before taking the reading.

16. RESULTS AND DISCUSSION
The concentrations of some heavy metals copper, chromium, cadmium, nickel,
lead, sodium and calcium in fruit juices are presented in Table V.
In the fruit juice samples, maximum and minimum mean levels found were
0.051 to 0.020, 4.811 to 0.280, 2.141 to 0.361, 0.267 to 0.073, 1.069 to 0.631,
44 to 18 and 62 to 39 for cadmium, copper, chromium, nickel, lead, sodium and
calcium respectively.
There is slight variation in the concentration of cadmium among all juice
samples.
Apple juice was found to be highest in copper concentration.
Our data revealed that the copper, nickel, lead, cadmium, chromium, sodium
and calcium levels found in all of the fruit juice samples were within the RDI
standard values.
The maximum concentration of lead detected in pomegranate juice was 1.069
mg/L respectively which is far above the safe limit of 0.01 mg/L recommended
by WHO;
Copper is an essential trace metal, the maximum concentration of copper
determined was 4.811 mg/L for in apple juice sample which is slightly crossing
the safe limit set by WHO i.e., 3 mg/L. The copper and lead level in juice
samples may pose a threat to public health of Agra.
Cadmium DL is < 0.2, was found to be extremely good in concentration.
Nickel DL is < 0.1, crossing the UL by WHO in pineapple juice and at the
borderline in others.
Chromium must be < 0.5, but it was crossing the UL in most of the samples
analysed.
Calcium was found to be under the DL proposed by WHO.
Impacts of studied metals in biological system
The effects and the functions of the metal determined during this project are given
below- According to DRI the DDI and UL of the same metals are listed in table 1-5.
Copper- copper is an essential constituent of many metallo-proteins and enzymes,
involved in electron transfer, oxygenation and oxidation processes. Hence,
deficiency of copper causes deactivation of these processes, leading to anaemia
(ceruloplasmin deficiency), and loss of hair pigment (Tyrosine deficiency).
Deficiency of Cu(II) containing enzyme, cytochrome C oxidase, causes reduced
arterial elasticity and stunted growth in adults and Meneke’s disease in children,

17. resulting in kinky hair, retarded growth, and respiratory problem, severely limiting
life span.
If synthesis of ceruloplasmin is hindered, the mechanism of the control of
copper level in the biological system is damaged. This leads to accumulation of
copper in liver, kidney and brain. Thus the central nervous system (CNS) is damaged,
leading to tremors, rigidity and abnormality of the brain. Accumulation of copper in
liver leads to Cirrhosis and ultimate death. This physical abnormality is called
Wilson’s disease.
External intake of small excess of copper causes gastro intestinal irritation and
vomiting. Serious toxic effect is observed, if more than one gram of copper is taken
at one time or there is continuous intake of 250 mg per day, for a period of time. The
toxic effect occurs because of strong affinity of Cu(II) for the –SH group of the
different enzyme proteins. The enzyme get deactivated, due to copper binding, and
thus specific biochemical activity are inhibited, leading to physical disorders.
Chromium- It is involved in the metabolism of glucose in the mammals. Cr (III) and
insulin both maintain the correct level of glucose in the blood.
Cadmium- Cadmium is an extremely toxic metal commonly found in industrial
workplaces. Environmental exposure to cadmium has been particularly problematic
in Japan where many people have consumed rice that was grown in cadmium
contaminated irrigation water. This phenomenon is known under the name itai-itai
disease.
Food is another source of cadmium. Plants may only contain small or moderate
amounts in non-industrial areas, but high levels may be found in the liver and
kidneys of adult animals.
Cigarettes are also a significant source of cadmium exposure. Although there is
generally less cadmium in tobacco than in food, the lungs absorb cadmium more
efficiently than the stomach.
Aside from tobacco smokers, people who live near hazardous waste sites or factories
that release cadmium into the air have the potential for exposure to cadmium in air.
However, numerous state and federal regulations in the United States control the
amount of cadmium that can be released to the air from waste sites and incinerators
so that properly regulated sites are not hazardous. The general population and
people living near hazardous waste sites may be exposed to cadmium in
contaminated food, dust, or water from unregulated releases or accidental releases.
Numerous regulations and use of pollution controls are enforced to prevent such
releases.

18. Some sources of phosphate in fertilizers contain cadmium in amounts of up to 100
mg/kg, which can lead to an increase in the concentration of cadmium in soil hence
in fruits.
Acute exposure to cadmium fumes may cause flu like symptoms including chills,
fever, and muscle ache sometimes referred to as "the cadmium blues." Symptoms
may resolve after a week if there is no respiratory damage. More severe exposures
can cause tracheo-bronchitis, pneumonitis, and pulmonary edema. Symptoms of
inflammation may start hours after the exposure and include cough, dryness and
irritation of the nose and throat, headache, dizziness, weakness, fever, chills, and
chest pain.
Inhaling cadmium-laden dust quickly leads to respiratory tract and kidney problems
which can be fatal (often from renal failure). Ingestion of any significant amount of
cadmium causes immediate poisoning and damage to the liver and the kidneys.
Compounds containing cadmium are also carcinogenic.
The bones become soft (osteomalacia), lose bone mineral density (osteoporosis) and
become weaker. This causes the pain in the joints and the back, and also increases
the risk of fractures. In extreme cases of cadmium poisoning, mere body weight
causes a fracture.
The kidneys lose their function to remove acids from the blood in proximal renal
tubular dysfunction. The kidney damage inflicted by cadmium poisoning is
irreversible. The proximal renal tubular dysfunction creates low phosphate levels in
the blood (hypophosphatemia), causing muscle weakness and sometimes coma. The
dysfunction also causes gout, a form of arthritis due to the accumulation of uric acid
crystals in the joints because of high acidity of the blood (hyperuricemia). Another
side effect is increased levels of chloride in the blood (hyperchloremia). The kidneys
can also shrink up to 30%. Cadmium exposure is also associated with the
development of kidney stones. Other patients lose their sense of smell (anosmia).
Sodium- People who regularly eat foods high in sodium risk having diseases such as
hypertension, Type II diabetes mellitus, respiratory complications, Dislipidemia,
Gallbladder disease, osteoarthritis and some cancers (endometrial, breast, colon).
Most of the daily sodium intake comes from salt.
The DRI Upper Limit (UL) for Sodium in adults is 2300 mg/day.
Calcium- The level of calcium in the body is usually controlled by vitamin D and
parathyroid hormones. But, if there is a metabolic imbalance of calcium regulation, it

19. gets deposited in the tissues, leading to their calciferation. Formation of stones
cataract are due to calcium salt deposition.
Nickel- it is an essential trace element for several hydrogenases and ureases
enzymes. Its deficiency in food slows down the functioning of the liver in chicks.
It is highly toxic to plants and moderately toxic to mammals. It is carcinogenic if
present in higher concentrations in biological systems.
It causes skin and respiratory disorders. It can produce bronchial cancer. It
deactivates cytochrome C oxidase and also the enzymes, assisting dehydrogenation
process, and thus inhibits biochemical processes.
Lead- It has no known biological function. It is highly toxic to plants and is a
cumulative poison for mammals. It inhibits the synthesis of hemoglobin in mammals
and is highly toxic for central nervous system. Lead tertraethyl used in gasoline as an
antiknock and lead pigments are serious health hazard.
Lead gets deposited in the softer tissues. From there, the reversibly fixed lead passes
to the blood stream. Like transition metals, lead has strong affinity for the –SH group
of the enzymes and hence it gets bound to the enzymes strongly and deactivates
them. In the blood stream, lead is known to inhibit the activity of several enzymes,
involved in the synthesis of heme.
Excess lead lowers the formation of delta amino levulinic acid, its conversion to
porpho-bilinogen and also the conversion of protoporphyniogen to protoporphyrin
IX. Thus the biosynthesis of heme is inhibited, leading to anemia.
Lead also affects the biosynthesis of bones, because, divalent lead replaces calcium
in bone. Deposition of lead in brain results in its reduced activity, leading to
depression, nervousness and lack of concentration. Excess lead leads to damage of
kidney, liver and intestinal track, with consequent loss of appetite, muscle and joint
pain, weakness and tremors. Excess lead also causes dental carries and
abnormalities in female reproductive system.

20. TABLE-I
Nutrient Life Stage
Group
RDA/AI
(μg/d)
UL
(μg/d)
Copper
Males
14-18 y
19-50 y
Females
14-18 y
19-50 y
Pregnancy
19-30 y
31-50 y
Lactation
19-30 y
31-50 y
890
900
890
900
1000
1000
1300
1300
8,000
10,000
8,000
10,000
10,000
10,000
10,000
10,000

21. TABLE-II
Nutrient Life Stage
Group
RDA/AI
(μg/d)
UL
(μg/d)
Chromium
Males
14-18 y
19-50 y
Females
14-18 y
19-50 y
Pregnancy
19-30 y
31-50 y
Lactation
19-30 y
31-50 y
35
35
24
25
30
30
45
45
ND
ND
ND
ND
ND
ND
ND
ND

23. TABLE-III
Nutrient Life Stage
Group
RDA/AI
(mg/d)
UL
(mg/d)
Calcium
Males
14-18 y
19-50 y
Females
14-18 y
19-50 y
Pregnancy
19-30 y
31-50 y
Lactation
19-30 y
31-50 y
1,300
1,000
1,300
1,000
1,000
1,000
1,000
1,000
2,500
2,500
2,500
2,500
2,500
2,500
2,500
2,500

24. TABLE-IV
Nutrient Life Stage
Group
RDA/AI
(mg/d)
UL
(mg/d)
Nickel
Males
14-18 y
19-50 y
Females
14-18 y
19-50 y
Pregnancy
19-30 y
31-50 y
Lactation
19-30 y
31-50 y
ND
ND
ND
ND
ND
ND
ND
ND
1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0

25. TABLE-IV
LEAD
For Whom Amount Known To
Cause Health Problems
(μg/d)
FDA’s Recommended Safe
Daily Diet Lead Intakes
(μg/d)
For children under age 6 60 6
For children 7 and up 150 15
For Adults 750 75

26. TABLE- V
Metal Concentration in mg/L Studied in Fruit Juices during This Project
Metals
Samples
Cadmium Copper Chromium Nickel Lead Sodium Calcium
Pomegranate 0.031 3.624 1.739 0.073 1.069 21 39
Apple 0.051 4.811 2.141 0.116 0.631 44 46
Lemon 0.040 1.062 1.187 ND ND 18 62
Pineapple 0.020 0.580 0.539 0.267 0.640 25 46
Orange 0.038 1.860 0.361 0.109 0.890 22 48
Mix juice 0.044 0.280 1.196 ND ND 26 55

27. CONCLUSIONS
Juice prepared from the fruits purchased from the retail market in Agra posed a
health risk based on the concentration of trace metals analysed in the present
work. Copper posed the greatest risk as its level far exceeded WHO safe limit.
Lead, Nickel and chromium levels were out of the limit set by WHO and
therefore may pose threat to public health of Agra. There were variations in the
level of trace metals analysed. Cadmium, sodium, and calcium level was under
the limit.
Fruit juices (from Rajamandi, Agra), which are supposed to be healthy and safe,
an important part of our daily diet were not found to be crossing the limit
concentration of heavy metals.
A long-term and/or excessive consumption of foods containing heavy metals
above the tolerance level has a hazardous impact on human health. Fruit juices
are widely consumed and supposed to be pure and a healthy diet. For this
reason, the vegetation of fruits must be in the less polluted area, for preventing
the contamination by heavy metals. The water must be free from heavy metals
and natural fertilizers must be used.

28. REFERENCES
1. AOAC. Official methods of analysis of AOAC International, 18th Edition.
AOACInternational, Gaithersburg, Maryland, USA; 2005.
2. Cabrera C, Lorenzo ML, Lopaz MC. Lead and Cadmium contamination in dairy
product and its repercussion on total dietary intake. Journal of Agricultural and
Food Chemistry. 1995;43:1605-1609.
3. http://creativecommons.org/licenses/by/3.0
4. http://www.sciencedomain.org
5. Eyong EU, Eteng MU, Eyong UO. Comparative analysis of some brand of
juices consumed in southern Nigeria. Nig. J. Biochem. Mol. Biol. 2010;25:36-39.
6. FAO/UN. Requirements of Vitamin A, iron, folate, and Vatamin B12. Report
of a Joint WHO/FAO Expert consultation. Rome, Food and Agriculture
Organization of the United Nations. FAO Food and Nutrition series. 1998;23.
7. Krejpocio Z, Sionkowskis, Bartela J. Safety of fresh fruits and juices available
on the Polish market as determined by heavy metal residues. Polish Journal of
Environmental studies. 2005;14:877-881.
8. Kirk, Ronald S. Pearson’s composition and analysis of foods, 9th Edition.
Longman Singapore; 1991.
9. Ma J, Betts NM. Zinc and Copper intakes and their major food sources for
older adults in the 1994-96 continuing survey of food intakes by individuals
(CSF- II). Journal of Nutrition. 2000;130:2838-2843.
10. Senesse P, Meance S, Cottet V, Faivre J, Boutron-Ruault MC. High dietary
iron and copper and risk of colorectal cancer: a case-control study in Burgundy,
France. Natural Cancer. 2004;49:66-71.
11. Silvestre MD, Lagarda MJ, Farre R, Martineze-Costa C, Brines J. Copper,
Iron, and Zinc determination in human milk using FAAS with microwave
digestion. Food Chemistry. 2000;68:95-99.
12. Venugopal B, Luckey T. Toxicity of non-radioactive heavy metals and their
salts. In Heavy Metal Toxicity, Safety and Harmology, Ed. F. Coulston. Academic
Press, George Thieme, Stuttgart, New York; 1975.
13. WHO. World Health Organization Guidelines for drinking water quality
recommendations. WHO, Geneva. 1985;1:130.
14. Bhattacharya P. K. a book of bioinorganic chemistry.
15. www.nap.edu
16. www.wikipedia.com