# DETECTION OF HEAVY METALS IN HERBAL

PRODUCT BY USING ATOMIC ABSORPTION

SPECTROSCOPY
****HEMANT KULSHRESTHA,VINAY TYAGI,KIRTI
SHARMA,DEEKSHA SOLANKI.MONI SRIVASTAVA,
**RAJU V JOHN, ***SUSAN VERGHESE .P AND ***R.P
SINGH
****Head,Dept of chemistry,St.johns College ,Agra
***Asso.prof ,Dept of chemistry,St.johns College,Agra
**Reader,Dept of chemistry,StJohns College,Agra.
E mail-susan.jaison @yahoo.com

Abstract

The increasing prevalence of environmental pollution, especially soil

contamination with heavy metals has led to their uptake in the human food chains through
plant parts. Accumulation and magnification of heavy metals in human tissues through
consumption of herbal remedies can cause hazardous impacts on health. Thus this study
was aimed at determining the amount of heavy metals in selected herbal and plant leaves
samples. The ten heavy metals are Mn, Cr, Pb, Fe, Cd, Co, Cu, Zn, Ni and Hg in which the
chemical profiling of Cu, Ni, Pb was undertaken in samples of eight medicinal plants
(Turmeric, PudinHara, Aloe Vera, Black Pepper, CurryLeaves, Tomato, Carrot, Ginger).
The Atomic Absorption Spectrophotometer was employed for the estimation of heavy
metals Concentration levels (mg/l)of heavy metals was within permissible limits in the
samples.As a guideline, FAO/WHO defined the permissible limits (ppm) of the various
heavy metals in the consumed medicinal herbs for different countries as: chromium (2
ppm), manganese (44.6 to 339 ppm), iron (261 to 1239 ppm), cobalt (0.14 to 0.48 ppm),
nickel (1.63 ppm), copper (20 to 150 ppm), zinc (27.4 ppm), cadmium (0.3 ppm), mercury
(0.1 ppm) and lead (10 ppm).This study presents the status ofheavy metals and trace
elements in herbal plants and also provides a simple and convenient Atomic Absorption
Spectroscopic (AAS) method which can effectively be adopted at Industrial level for the
quality control and standardization of herbal cosmeticpreparations and other related
products.
INTRODUCTION
 Environmental pollution and exposure to toxic materials is a growing problem world-
wide. The unscientific use of hazardous metals in agriculture and industries and its dumping has
created a great risk for human, plant and animals. Traditional medicinal plants are the raw
material for many herbal formulations. The assimilation of heavy and toxic metals in plants is
evident. These metals have great tendency to accumulate in living organisms including
humanbeings. The presence of such metals above the permissible levels can cause various
consequences. Thus, both the deficiency and excess of essential micro minerals such as Fe,Zn,
Mn and Cu are harmful to the human health. Effects of toxic metals like Cd, Cr, Pb, Hg etc., on
human health andtheir interaction with essential trace elements many produce serious health
problems. The WHO recommends that medicinal plants which form the raw materials for
various medicines should be checked for the presence of different contaminants such as heavy
toxic metals.
Plants have been used in traditional medicine for several thousand years. The world
health organization (WHO) estimated about 75-80% of the world’s population use plant based
medicines. The traditional medicines, mostly plant drugs for their primary health care. The plant
has a reputation as an anti-aging agent in Ayurvedic medicine. Heavy metals, such as Cadmium,
Copper, Lead, Chromium and Mercury are important environmental pollutants.Heavy metals, in
general, are not biodegradable, have long biological half-lives and have the potential for
accumulationin different body organs leading to unwanted side effects.(1,6,7)
Accumulation of toxic industrial effluents in the soil is continuously increasing due to
fast urbanization and extensive pollution of the environment. Among these toxic substances,
presence of Heavy metals (HMs; atomic weights 63.5–200.6 g mol−1 and a specific gravity
greater than 5 g cm−3) , which are ubiquitous in nature, cause serious harmful effects on living
organisms. Some of the heavy metals are essential in very low concentrations for the survival of
all forms of life. Heavy metals such as iron, chromium, copper, zinc, cobalt, manganese and
nickel are essential metals since they play an important role in biological systems whereas
mercury, lead and cadmium are non- essential metals which can be toxic even in trace
amounts(2).
As a guideline, FAO/WHO defined the permissible limits (ppm) of the various heavy
metals in the consumed medicinal herbs for different countries as: chromium (2 ppm),
manganese (44.6 to 339 ppm), iron (261 to 1239 ppm), cobalt (0.14 to 0.48 ppm), nickel (1.63
ppm), copper (20 to 150 ppm), zinc (27.4 ppm), cadmium (0.3 ppm), mercury (0.1 ppm) and lead
(10 ppm).(3)
The present study is concerned with the assessment of the metal content of some
medicinally important herbsandplants. This study has been focused to estimate the
concentrations of some toxic or heavy metals ions in the medicinalplants.
MATERIALS AND METHOD
SAMPLING AREA:
The samplesof herbal plant were collected from different areas of Agra UP in India and
transferred in a sterile cloth bag and were transferred to the laboratory. Some fruit sample
were cut into smaller pieces and ground into fine particle by using Grinder at the
departmental research laboratory of St. John’s College, Agra.
INSTRUMENT:
 Flame Atomic Absorption Spectrometer Perkin Elmer AAnalyst 100 equipped with hollow
cathode lamp was used for determination trace metals and heavy metals. We used three
hollow cathode lamp of Cu, Ni (trace metal) and Pb (heavy metal) for the estimation of Cu,
Ni and Pb respectively. The flame used was Air/Acetylene and N2O/Acetylene. Nitrogen
was used as carrier gas.The hollow cathode lamps used were made by Perkin Elmer.

SAMPLE PREPARATION:
Preparation Standard solution
The stocks solution for Cu, Ni and Pb were made from a metal salt of Analar grade (purity
99.9%). The metal salt was dissolved in dil. HCl and diluted to the 1 litre mark using
distilled water. The required concentrations were 1000ppm, 100ppm, 10ppm, 1ppm, 3ppm,
5ppm prepared by serial dilution of stock solution.

PREPARATION HERBAL SAMPLE SOLUTION

(DIGESTION WITH Conc. HNO3):

Samples were digested by the wetdigestion method. 10 ml of nitric acid was added to 2 g
ofaccurately weighed dried sample in a 100 ml beaker and washeated on a hot plate at 95°C
for 15 min. The digest was cooledand 5 ml of concentrated nitric acid was added and
heated foradditional 30 min at 95°C. The last step was repeated and thesolution was
reduced to about 5 ml without boiling. The samplewas cooled and filtered through a
Whatman No. 42 filter paperand diluted in 100ml volumetric flask with deionized water.

RESULT AND DISCUSSION

Atmosphere and soil are continuously being polluted with chemicals and heavy metals due to
dynamic development of industries and motorization along with extensive use of pesticides
and fertilizers. The results analysis of the levels of heavy metal present in the selected herbs is
discussed in this section and the concentration of Cu, Ni, and Pb in the herbs is present in
(Table.1)

Copper: The highest concentration of Copper was found in Tomato (1.867

ppm), followed by Curry Leaves (1.725 ppm), Aloe Vera (1.537ppm), Carrot (1.004
ppm) and Neem (0.532 ppm) while Cu in the Turmeric, Pudin Hara, Black pepper
and Ginger samples is Not Detectable(ND). After comparison, metal limit in the
studied medicinal herbs with those proposed by FAO/WHO, it is found that all the
selected herb from AGRA region UP (west) have copper below the permissible limit
set by FAO/WHO in edible plants. However, for medicinal herbs the WHO limits not
yet been established for Copper. Although in medicinal plants, permissible limits for
Copper set by CHINA and SINGAPORE, were 20 ppm and 150 ppm respectively.
 The selected five herbs are found below the permissible limit given by China and
Singapore.

Nickel: Results obtained showed that maximum concentration of nickel was

found in Aloe Vera (96.00 ppm), Curry Leaves (20.05 ppm), Turmeric (4.024 ppm),
Pudin Hara (2.780 ppm), Neem (0.653 ppm), Carrot (1.004), Ginger (0.018 ppm)
while in Black pepper and Tomato sample Nickel is not detectable. In all nine
samples, there are only three samples Neem, Carrot and Ginger exist in permissible
limit for Nickel in Plant and medicinal herbs. According to WHO/FAO the
permissible limit is 1.63ppm, but here after seeing the result it is very surprise that
Aloe Vera have so high concentration of Nickel as well as Curry Leaves also. The
nickel toxicity in human is not a very common occurrence because its absorption by
the body is very low. The most common ailment arising from nickel is an allergic
dermatitis known as nickel itch, which usually occurs when skin is moist, further
more nickel has been identified as a suspected carcinogen and adversely affects lungs
and nasal cavities. Although nickel is required in minute quantity for body as it is
mostly present in the pancreas and hence plays an important role in the production of
insulin. Its deficiency results in the disorder of liver(4).

Lead: Results obtained show that highest concentration of lead was found in
Aloe Vera (9.693 ppm) followed by Ginger (5.99 ppm) tomato (2.733 ppm) turmeric
(1.918 ppm), Curry leaves (1.780 ppm) and Neem (0.354 ppm) while inPudin Hara,
Black Pepper, Carrot Lead is not detectable. The WHO prescribed limit for
leadcontents in herbal medicine is 10 ppm while the dietary intake limit for lead is 3
mg/week. However, for medicinalherbs limit was 10 ppm set by China, Malaysia,
Thailand and WHO. Similarly plants with those proposed by WHO it was found that
all the selected six herbs have Lead below permissible limit set by China, Malaysia,
Thailand and FAO/ WHO. But here some interesting data for Aloe Vera because
their lead concentrations it just below the maximum point of permissible limit i.e.
9.693ppm and permissible limit is 10.00 ppm. Lead is not an essential element for
life and it is very toxic for the nervous system and for the kidneys.(5)

S.No. SAMPLE Cu Ni Pb pH CONDUCTIVITY

1 Turmeric ND 4.024 1.918 6.0 0.78

2 Neem 0.532 0.653 0.354

3 Pudinhara ND 2.780 ND

4 Aloe Vera 1.537 96.00 9.693 7.2 0.58

 5 Black pepper ND ND ND 6.7 0.78

6 curry leaves 1.725 20.05 1.780

7 Tomato 1.867 ND 2.733 4.4 0.88

8 Carrot 1.004 0.412 ND 5.3 1.13

9 Ginger ND 0.018 5.99 6.4 0.60

TABLE.1. Concentration (ppm) of Copper (Cu), nickel (Ni), lead (pb) in herbal samples
A total of three elements (Cu, Ni, and Pb) were determined in the samples of
medicinal plants using atomic absorption spectrophotometer (AAS). Graph.1, .2, .3, shows
the concentrations of various metals inthe analysed herbal plantsand tabular form of data of
metal concentration analysis is given in the (Table.1)

*ND= not detectable

 Graph.1 concentration of copper in samples
2 1.867
1.725
neem
1.8
1.537 aloe vera
1.6
kari leaves
concentration (in ppm)

1.4
tomato
1.2
1.004 carrot
1
0.8
0.6 0.532

0.4
0.2
0
Samples

Graph.2 concentration of nickel in samples

120

100 96
turmeric
concentration(in ppm)

80 neem
pudin hara
60
aloe vera
40 kari leaves

20.05 carrot
20
ginger
4.024 2.78
0.653 0.412 0.018
0
samples

Graph.3 concentration of lead in samples

12 turmeric
9.693
10 neem
concentration (in ppm)

8 aloe vera
5.99 kari leaves
6
tomato
4
2.733
1.918 1.78
ginger
2
0.354
0
samples
CONCLUSION
Medicinal plants are sources of a large number of active principles of herbal and modern
medicine. Indian people have a tremendous passion for medicinal plants and use them for a
wide range of health related applications from a common cold to cancer and treatment of
poisonous snake bites to a cure for genetic disorders like muscular dystrophy.The present
investigation clearly demonstrated the variation in heavy metal concentration.The
concentration (ppm) of heavy metals in plants extract was found to be as follows: Cu (0.532 to
1.867), Ni(0.018 to 96.00) and Pb(0.354 to 9.693). The maximum concentrations of copper
were found in tomato (1.867), Nickel in Aloe Vera (96) and lead in Aloe Vera (9.693).The
developed AAS technique is a precise, specific and accurate method for estimation of heavy
metal content in stem and leaf samples of these medicinal plants. From the comparison of the
results with the defined permissible concentration limits it was concluded that the levels of
heavy metals present in the Herbs fall in permissible range for consume medical herbs.

ACKNOWLEDGEMENT
We are very grateful to Dr Hemant Kulshreshtha,Head , Department of chemistry St.
John’s College, Agra for the technical and material support and provision of laboratory space
for the extraction process, equipment to carry out project and for use of college Research
laboratory under supervision of Dr. Susan Verghese. My sincere gratitude goes to the St.
John’s College, Agra

REFERENCE
1. FAO/WHO., 1984. Contaminants. In Codex Alimentarius, vol. XVII, Edition 1. FAO/WHO,
Codex AlimentariusCommision,Rome
2. World Health Organization, 1998. Quality methods for medicinal plant materials. Published
by poisoning and also poses adverse effects on kidney, liver, WHO, Geneva.
3. WHO 2005. Quality Control Methods for Medicinal Plant Materials, Revised, Geneva.
4. Ernst E. Toxic heavy metals and undeclared drugs in Asianherbal medicines, Trends
Pharmacol. Sc. 23, 136-139 (2002)
5. WHO. Environmental Health Criteria: International Programme on Chemical Safety. World
Health Organization, Geneva. Pp 165 (1995)
6. Sharma D.K., Rai S., Arora S.S., Gupta P.M., Sharma R. and Chopra A.K. Study of the trace
elements in Aloe veraL. (Aloe barbandensisMiller) viz. Liliaceae and its biological and
environmental importance, J. Chem. Pharm Res., 3(3), 64 (2011)
7. Farr G., The hair tissue mineral analysis/why heavy metals are a hazard to your health,
Updated 31Dec., (2009).Available from:
(http://www.becomehealthynow.com/ebookprint.php?id=122)
8.
9.
 #PHYSICO-CHEMICAL EXAMINATION OF
SOIL IN AGRA REGION
*SUSAN VERGHESE.P, REKHA KUMARI,RITU RANA,SANDHYA YADAV,
NAMI JAIN,ARYAN VASHISHTH, SHILPA ANAND,SHALINI
NELSON**NELIA DAVID***& HEMANT KULSHRESHTHA****

****Head Dept of Chemistry,St. Johns College,Agra

***Asso.Prof.Dept of Geography, St. Johns College,Agra

*ASSO.PROF,DEPT OF CHEMISTRY, ST.JOHN’S COLLEGE, AGRA.

**READER DEPT. OF CHEMISTRY

E mail-susan.jaisan@yahoo.com

ABSTRACT
Soil is the most important parameter for the plants and human beings.
Soil provides a physic matrix, chemical environment and biological setting
for water in order to meet an ever increasing demand of production. The
soil is going to be polluted day by day. Soil contamination is caused by the
presence of xenobiotic chemicals or other alteration in the natural soil
environment. It is caused by industrial activity, agricultural activity,
chemicals or improper disposal of waste. The most common chemical
involved are petroleum, hydrocarbon,poly nuclear hydro carbons,
solvents, pesticides and heavy metals, Contamination is correlated with
the degree of industrialisation, and intensity of chemical usage. The
concern over soil contamination stems primarily from health risks from
direct contact with contaminated soil vapours from the contaminants and
from contaminated water supply within and underlying the soil. Heavy
metals are element that exhibit metallic properties such as ductility,
malleability, conductivity, cation stability, and ligand specificity. They are
characterized by relatively high density and high relative atomic weight
with an atomic number greater than 20. Some heavy metals such as Co,
Cu,Fe,Mn,Mo,Ni,V, and Zn are required in minute quantities by
organisms. However, excessive amounts of these elements can become
harmful to organisms. Other heavy metals such as Pb,Cd,Hg, and as(a
metalloid but generally referred to as a heavy metal) do not have a
beneficial effect on organisms and are thus regarded as the “main
threats” since they are very harmful to both plants and animals (Mellor
1999). Ten samples have been collected from different regions of Agra
during post winter season and have analysed for heavy metals apart from
basic parameters. Result indicates that in some region nickel and lead
exceeds the permissible limit.

Keywords: Atomic absorption spectrophotometer, soil, heavy metals,

farms and nurseries, physico- chemical properties.

INTRODUCTION
Soil is a multipart of physical and biological schemes, which give support
to the plants and supplies waterand essential nutrients to them. It is the
main pool of the nutrients essential for customary growth of the plants.
Soil is an essential constituent of the biosphere, but it must be
nurturedand cultivated to convey it to its highest potential (Olaniya
1998). The availability of minerals affects the specific enzymatic action
directly and eventually adversely affects plant growth. Due to plant
growth nutrient relationship and general importance the need of physico-
chemical parameter and heavy metals scrutiny of soil is essential with the
objective of using specific relationship as plant growth indicator.

Agra is the most important tourism station so a great number of people

visit here but with the respect to the time the soil of Agra gets polluted
due to increments of the heavy metals. Sample of different region ofAgra
were studied during post winter season in chemistry laboratory of St.
John’s College, Agra.

Material and Methods

The soil samples were collected using a grab sample from sampling
station in Agra and transported to the laboratory. Sample preparation for
metal analysis involved procedure such as a drying sieving; crushing;
storage and samples were digested and prepared in the laboratory. Then
the samples were analysed on an ATOMIC ABSORBTION
SPECTROPHOTOMETER.

Result and Discussion

Environment pollution gives rise to direct and indirect effect of changes in
the energy pattern of physico-chemical composition of the environment
and abundance or diversity of organisms. There is considerably increase
in the level of heavy metal and other harmful organic compounds due to
the industrialization and agricultural activities, with increasing heavy
metal concentration genetic load on human population increased to its
maximum and its likely to create a distress in biological equilibrium.

The experimental data shown in table 1 that the lead (Pb) concentration
(mg/l) in soil ranges from 0.107 to 5.320mg/l similarly the Ni
concentrations (mg/l) in soil ranges from 0.893 to 3.216mg/l.These values
are lower than the permissible limits as shown in table 2. The distribution
pattern of nickel in agricultural soil understudy is clearly showing the
unsuitability of these soils for agricultural purpose since most of the
concentrations of the trace metals understudy were below the
permissible limit of trace metals in soil as recommended by various
agencies in the world the permissible limit. It has been realised that a
study of soil analysis is imperative in order to understand plant nutrition,
soil fertility as well as seasonal prototype of exchange between plant and
soil.
 TABLE-1

Concentration of heavymetal(ppm) in soil samples

Sample Location Ni Pb
1 BODLA 0.893 2.530
2 KHERIYA MOD 0.304 1.307
3 FATEHPUR -0.816 1.539
SIKARI
4 ST. JOHN’S 0.146 -0.810
5 MADHU NAGAR -0.540 0.107
6 TAJ GANJ 3.217 2.376
7 YAMUNA SOIL -0.955 1.628
8 PALIWAL PARK -5.590 2.077
9 DAYALBAGH -0.9023 5.0320

10 *TARGET VALUE 85 35
11 **INTERVENTION 550 210
VALUES OF SOIL
Where *target values are specified to indicate desirable maximum levels of
elements in unpolluted soils.
*
*
3.5 Concentration of heavymetal(ppm)
Ni in soil samples
I
3
n
t
e2.5
r
v2
e
n1.5 Ni

t
i 1
o
n
0.5

v
0
a 1 2 3 4
l
ue when remedial action is necessary.
 Concentration of heavymetal(ppm) in soil samples

Pb
6

3
Pb

0
1 2 3 4 5 6 7 8

CONCLUSION
The levels of these trace metal in soil were generally below the
permissible level of normal agriculture land particularly Pb and Ni. The
significant concentration of these metals in agricultural land is favourable
to the crops yield.

ACKNOWLEDGEMENT
Authors are grateful to school of Chemical Sciences Dept of chemistry, St
Johns College Agra for providing marvellous opportunity to carry out
project work and are highly thankful to Dr.HemantKulshreshtha (HOD of
Chemistry Dept), and Dr.SusanVerghese P. to give their valuable
instructions to carry out the whole work.

REFERANCES
1. Olaniya, M.S. et al. 1998. Heavy metal pollution of agricultural soil
and vegetation due to application of municipal soild waste –A case
study. I.J. enivromental health. 40(2): 160- 168.
2. Jackson, M.L. 1967. Soil chemical analysis. Prentice hall of indiapvt.
Ltd.., New Delhi .
3. Anderson, A. and Nilsson,K.O. 1973. Enrichment of trace element
from sewage sludge fertilizers in soils and plants. Ambio.I :176-179.
4. Sharma yogesh and totwat, K.L.2002. effect of municipal sewage
water on the soil properties and chemical composition of vegetables
grown on sandy soil of Bikaner (rajasthan). Proc. 11th national
symposium on environment. 320-323.
5. Mellor , A. and bevan, J.R. 1999. Lead in the soil and stream
sediments of an urban catchment in tyne side. U.K. water air soil
pollut. 112: 327-348.

10.
11.
12.
13.
14.
15.
16.
17.
18.
19.
20.
21.
22.
23.
24.
25.
26.
27.
28.
29.
30.
31.