This is a project report done by students of st. John's college agra under the supervision of Dr. Susan Verghese...

1. A PROJECT REPORT ON
DETERMINATION OF SOME HEAVY METAL LEVELS
IN SOFT DRINKS USING 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:
ANAITULLAH GANAIE
M Sc Final
Physical Chemistry
2013-14

2. CERTIFICATE
This is to certify that this project entitled “DETERMINATION OF
SOME HEAVY METAL LEVELS IN SOFT DRINKS USING 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
ANAITULLAH GANAIE 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.
ANAITULLAH GANAIE
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. INTRODUCTION
Soft drinks are the usual beverages used in every day life, most festivities and
celebrations in India. These celebrations include Marriages, Weddings, Naming
of babies and Funerals. Soft drinks, also called ready-to drink beverages are
sweetened water-based non-alcoholic beverages, mostly with balanced
acidity.The soft drinks are mostly carbonated usually prepared from a
concentrated syrup containing sugar, fruit juice or flavoring essence, citric acid
and preservative (sodium benzoate). Benzoic acid is commonly used as
preservative. 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 essential metal which perform important
biochemical functions and are 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, among others, while toxicity due to excessive
intake has been reported to cause liver cirrhosis, dermatitis and neurological
disorders. Lead and cadmium are two potentially harmful metals that have
aroused considerable concern. Impairment related to lead toxicity in humans
includes abnormal size and haemoglobin content of the erythrocytes,
hyperstimulation of erythropoiesis and inhibition of haeme synthesis.
Heavy metal contamination in foods and drinks has been an important topic.
Heavy metals contamination has become a matter of public health concern but
this has not received much research attention in India especially soft drinks
contamination through heavy metals. In the present study, levels of Cu, Cd, Cr,
Ni, Pb, and Ca of soft drinks bought from retail market in Agra, during
December 2013 was determined using Flame Atomic Absorption
Spectrophotometer (FAAS).

6. Review of Literature
Metals are present in foods (including drinks) 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 66 soft drink samples examined in Poland.
The research performed in England revealed that the heavy metal levels in the
non-alcoholic beverage 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 non-alcoholic beverage samples from
totally 100 samples (Maff 1998).
Ashraf et al. (2000) reported arsenic levels as 0.837 mg/l in 34 soft drinks in
Pakistan.
Maduabuchi et al. (2006) reported cadmium levels as 0.003–0.081 mg/l in
canned drinks and 0.006–0.071 mg/l in non-canned drinks. Also in this research,
the lead levels were 0.002–0.0073 mg/l in canned drinks and 0.092 mg/l in non-
canned drinks.
Onianwa et al. (2001) reported cadmium, copper, lead, and zinc levels in
carbonated soft drinks in Nigeria.

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

8. EXPERIMENTAL
MATERIALS AND METHODS
Sample Collection
Soft drink 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 eight (8) varieties 7up, appy fizz, coke, dew, fanta, limca, mazaa, and
thumpsup were collected. Sampling was done during four days in December
2013. The soft drink samples were then analyzed for Cd, Cr, Cu, Ni, Pb, and Ca.
Sample Preparation
Took 20 ml of soft drink in a 100 ml of volumetric flask, added 10 ml of HCl
and HNO3 then made upto the mark with distilled water, shaked well.
Sample 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
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.

9.  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.
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.

10. 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.
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.

11. 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.
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

12. 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.
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

13. 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 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.

14. Then feed sample solution to the atomizer to get the relative concentration. Wait
atleast for 30 s before taking the reading.

15. RESULTS AND DISCUSSION
The levels of chromium, copper, zinc, cadmium, and lead in soft drink samples
were determined. The respective values are shown in Table 1.
In the soft drink samples, maximum and minimum mean levels found were
0.0750 to 0.3389, -0.010 to 0.095, 0.046 to 0.381, -0.008 to 0.130, -0.073 to
0.088 and 28 to 50 for copper, nickel, lead, cadmium, chromium and calcium
respectively.
Our data revealed that the copper, nickel, lead, cadmium, chromium and calcium
levels found in all of the soft drink samples were within the RDI standard values.
Maduabuchi et al. (2006) reported cadmium levels as 0.003–0.081 mg/l in
bottled soft drinks. These cadmium contents are closer to those in our study.
Also in that research, the lead levels were 0.002–0.0073 mg/l in bottled drinks.
These lead levels were lower than those in our study.
The maximum concentration of lead detected in soft drink was 0.381 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 0.3389 mg/L for soft drink samples which is within the safe limit
set by WHO i.e., 3 mg/L. The copper level in soft drink samples does not pose a
threat to public health.
Cadmium DL is < 0.2, but it was found to be at the borderline.
Nickel DL is < 0.1 and it was found in accordance with DRI.
Chromium must be < 0.5 and it was found to be so.
Calcium was also under the DL.
Impacts of studied metals in biological system
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,
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,

16. 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.
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

17. 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).
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 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

18. 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.

19. TABLE-I
Metal concentration in mg/L studied in soft drinks during this project
Metals
Samples
Cu Ni Pb Cd Cr Ca
7 UP 0.1373 0.036 0.381 0.098 -0.026 50
Appy 0.0750 0.049 0.149 0.025 -0.073 40
Coca-cola 0.1200 -0.010 0.298 0.125 -0.026 47
Dew 0.2181 0.007 0.058 0.020 -0.011 39
Fanta 0.1595 0.013 0.307 0.102 0.054 32
Limca 0.1131 -0.002 0.279 0.130 0.062 49
Mazaa 0.3389 0.095 0.046 -0.008 0.088 28
Thumpsup 0.1311 0.019 0.142 0.025 -0.022 39

20. TABLE-II
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-III
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-IV
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-V
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-VI
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. CONCLUSIONS
Total number of 8 bottled soft drinks were collected from Agra and analysed.
The purpose of this study was to determine the levels of heavy metals in the
drinks commonly consumed in Agra and all over India. Quantitative
determination of heavy metals: Chromium, copper, cadmium, nickel and lead in
all samples was carried out by FAAS method. Our data revealed that copper,
zinc, cadmium, and lead mean levels found in all soft drinks, collected from
several regions in Agra India, were within the RDI values. However some metal
concentrations were at the borderline.
Facility modernization and quality manufacturing are required to prevent heavy
metal contamination in drinks and thus the possible health hazards to the
consumer. A long-term and/or excessive consumption of foods containing heavy
metals above the tolerance level has a hazardous impact on human health.
Because soft drinks are widely consumed, they contribute a large fraction to the
heavy metals intake and, therefore, strict control of these elements is advisable.
For this reason, the steps in all processes must be monitored for preventing the
contamination by heavy metals.

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