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

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

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)

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.

Contents INTRODUCTION Review of Literature

EXPERIMENTAL RESULTS AND DISCUSSION CONCLUSIONS

REFERENCES

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.

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.

PERKIN ELMER AAnalyst 100 ATOMIC SPECTROPHOTOMETR

EXPERIMENTAL

SYSTRONICS 130 FLAME PHOTOMETER

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

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.

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.

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.

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.

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

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.

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,

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.

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

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.

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

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

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

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

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

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

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.

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