Determination of Cr2+ and Cd2+ in Centella asiatica using

Inductively Coupled Plasma Optical Emission

Spectrometry (ICP-OES)
Nurul A.Q Bahri1 , Nur H.D Ibrahim1 , Siti R.Z Iskandariah1, Nur A.M Azmi1

1Faculty of Applied Sciences, Universiti Teknologi MARA, Pahang Malaysia,

aqilah.sam95@gmail.com, nurhidayahaida94@gmail.com, razilahiskandar@gmail.com, amirah.azmi37@gmail.com.

Abstract Centella asiatica or commonly called as pegaga by the Malays is usually eaten as salad with the other main
dish. It is known as the great herbaceous plant that can act as rejuvenating agent and as a traditional medicine. However,
like the other plant, Centella asiatica may contain heavy metals from the soil because all plants absorb metal ions as its
food. On the other hand, Cadmium (Cd) and Chromium (Cr) ions are harmful to the human as it can affect our biological
system and disturbed the enzyme activity by cation exchange. Hence, ICP-OES was used to determine the Cd2+ and Cr2+
ions present in this particular herb plant as this spectroscopic technique is suitable for trace elements analysis. The samples
were dried in microwave for a day before undergoes acid digestion. HNO3 and H2O2 were used to digest the sample for a
complete decomposition of the matrix while avoiding contamination to the analyte. The acid digestions happen in
microwave again for next 20 minutes. Standard solutions for Cr were prepared from range 1 ppm, 2 ppm, 3 ppm, 4 ppm
and 5 ppm, while the standard solutions for Cd were from 0.2 ppm, 0.4 ppm, 0.6 ppm, 0.8 ppm and 1.0 ppm by using
standard addition method. Then, the sample was run by the ICP Model Perkin Elmer Optima 2100DV. The spiking
technique was used to determine unknown concentrations of metals ions in the sample. The result showed the 0.00 ppm for
Cd2+ and Cr2+ concentration in the sample. This proves that Cd2+ and Cr2+ not present in this wondrous herb plant and
safe to be ingested.

Keywords Centella asiatica, Cadmium, Chromium, ICP-OES, Spiking technique.

1. Introduction volunteers in sunny damp places like wetland edges,

Inductively coupled plasma optical emission
spectrometry (ICP-OES) is an analytical technique used for
the detection of trace metals. The samples are introduced
into the plasma in a process that desolvates, ionises, and
excites them. It is a type of emission spectroscopy that uses
the inductively coupled plasma to produce excited atoms
and ions that emit electromagnetic radiation at particular
wavelengths characteristic of an element. It is a flame
technique with a flame temperature in a range from 6 000 to
10 000K. The constituent elements within the sample can be
identified by their characteristic emission lines while the
concentration of the element can be quantified by the
intensity of the same lines.
Centella asiatica is the scientific name for
pegaga or commonly known as Asiatic pennywort or gotu
kola. It is a small creeping herb with shovel shaped leaves
emerging alternately in clusters at the stem nodes and it is
appeared as herbaceous native plant in Asia. The plants
roadside ditches, and soggy lawns and prefer sandy soils
enriched with organic matter. It can be grown around the
edges of a water feature, mixed species lawn or kept in a
well-irrigated corner of the herb garden.
Under the name "gotu kola", Centella asiatica is
referred as one of the great multi-purpose miracle herbs of
Oriental medicine. It has been in use for thousands of years
and also has been considered the most powerful
rejuvenating herbs and for promoting longevity as a
legendary healer, LiChing Yun who is said to have lived 256
years as a result of drinking gotu kola tea. In Indian
medicine, it is called "brahmi" meaning "greatest of the
great" and it is recommended for treatment of mental
disorders, immune system deficiencies, epilepsy, hair loss
and intestinal complaints.
This plant is well known for its benefit to the
human; henceforth it is ingested by people around the
world. In Malay cuisine the leaves of this plant are used for
 Determination of Cr2+ and Cd2+ in Centella asiatica using Inductively Coupled Plasma Optical Emission Spectrometry (ICP-
OES)
ulam, a type of Malay salad eaten with the main dish.
However, is this plant safe to eat and free of heavy metal?
In plants tissues, the present of Cadmium (Cd) in
low concentrations is most likely a normal constituent. The
concentration in the tissue is determined by the cadmium
concentration in the environment the inherent ability of the
plant species to absorb the ions. The cadmium content of
plant tissues tends to increase with increase
concentrations of soil cadmium above certain background
amounts. Cd gives numerous negative effects on the body
and can impact nearly all systems in the body including
cardiovascular, reproductive, the kidneys, eyes and even the
brain.
On the other hand, Chromium (Cr) is the seventh
most abundant metal in the earths crust and important
environmental contaminant released into the atmosphere
due to its huge industrial use. It is known to be a toxic metal
that can cause severe damage to plants and humans. High
chromium concentration can disturb the photosynthetic
process. Chromium can affect antioxidant metabolism in
plants. Antioxidant enzymes are found to be susceptibility
to chromium resulting in a decline in their catalytic
activities.
2. Materials and Method
2.1. Materials
The sample (plant) were obtained from the
laboratory in Universiti Teknologi MARA Pahang,
Malaysia. Deionized water was used to prepare all the
solutions. The apparatus used in this experiment were
250ml beaker, 100ml volumetric flask, pipette, filter paper,
filter funnel and microwave digestion.
The background correction methods were
performed using Inductively Couple Plasma Spectrometry
(ICP) Model Perkin Elmer Optima 2100DV, serial no.
080 N 80314401, location at 305 was used in the
experiments .
2.2. Sample Preparation
The plant sample was cut into small pieces and
50g of the sample was put in oven (110oC) to dry for 24
hours. 3 portions of the samples were weight ranging from
0.5g to 1.0g accurately and in microwave were place
digestion vessels separately. 7ml of concentrated HNO3 and
1ml of H2O2 was added into each vessel. Then, the samples
were digested in the microwave for 20 minutes. The vessels
then cooled down to room temperature before opening the
lid.
____________________________________________________________
*Corresponding author
aqilah.sam95@gmail.com (Nurul Aqilah Sham Shul Bahri)
Published Online
2.3. Preparation of Standard
1. Preparation of 10ppm of Standard Solution for ICP
5ml of 100ppm stock solution of Chromium/ cadmium was
pipette into 50ml volumetric flask was filled with distilled
water until the calibration mark. The volumetric flask
contained solutions were labeled as 10ppm Cr2+/ Cd2+
2.4. Chromium Series Standard
of 2. Preparation Series of Standard Solution for AAS
Standard series of Cadmium range from 1 ppm, 2 ppm, 3
ppm, 4 ppm and 5 ppm were made by using standard
addition method 1ml, 2ml, 3ml, 4ml and 5ml of 10 ppm
cadmium standard were pipetted and transfer into 50ml of
each volumetric flask. 10ml of water samples were added
into each volumetric flask contained standard series. All of
the volumetric flasks were then diluted with distilled water
until the calibration mark.
2.5. Cadmium Series Standard
Standard series of Cadmium range from 0.2 ppm, 0.4 ppm,
0.6 ppm, 0.8 ppm and 1.0 ppm were made by using
standard addition method 1ml, 2ml, 3ml, 4ml and 5ml of 10
ppm cadmium standard were pipetted and transfer into 50ml
of each volumetric flask. 10ml of water samples were added
into each volumetric flask contained standard series. All of
the volumetric flasks were then diluted with distilled water
until the calibration mark.
2.5. Inductively coupled plasma/optical emission
spectrometry (ICP-OES)
ICP-OES is a spectroscopic technique suitable for
trace elements analysis in several types of samples. The
technique is based on the unprompted emission of photons
from atoms and ions that have been excited in a
radiofrequency (RF) discharge. Samples are usually
introduced into the plasma in liquid form; thus, solid
samples require acid digestion prior to injection, while gas
and liquid samples may be injected directly into the
instrument.
The sample solution is converted to an aerosol then
sends into the centre of the plasma which maintains high
atomization temperature of around 10,000 K. As the plasma
free atoms in the gaseous state are generated and adequate
energy is often available to convert the atoms to ions then
promote the ions to excited states. The ionic excited state
species may then return to the ground state via emission of
photons. Specific wavelength of the photons can be used to
identify the elements and the number of photons is directly
proportional to the concentration of the element in the
sample. A variety of sample introduction methods are used
in this technique such as nebulization, hydride generation
(HG) for certain elements such as arsenic, selenium, and
 Determination of Cr2+ and Cd2+ in Centella asiatica using Inductively Coupled Plasma Optical Emission Spectrometry (ICP-
OES)
antimony, as well as electro thermal vaporization (ETV)
and laser ablation.
ICP-OES has been used for the determination of
Hg, Cd, Pb, and As in marine fish and shellfish from the
Straits of Malacca the West region of Peninsular Malaysia.
The modern spectroscopic analytical methods and
instrumentation make possible the measurement of low
level of concentrations with good accuracy and precision in
various matrices. Therefore, they are considered superior
tools in elemental analysis compared to other methods of
element detection.
2.6. Determination of Cr2+ and Cd2+ in Plant Tissue
Determination of the of Cr2+ and Cd2+
concentrations in the sample using spiking technique. The
plant sample were labelled and keep plant in lab 306 for
ICP determination.
The solutions of Cr2+ in different solvents were
prepared. The blank solutions was read first, followed by 1
ppm, 2ppm, 3ppm, 4ppm and 5ppm to obtain calibration
curve and determine the concentration of sample in the
sample. Next, the solutions of Cd2+ in different solvents
were prepared by using 0.2ppm, 0.4ppm, 0.6ppm, 0.8ppm
and 1.0ppm. The plotting calibration curve then was
recorded in the computer.
3. Results and Discussions
3.1. Inductively coupled plasma/optical emission
spectrometry (ICP-OES)
Table 1: result
____________________________________________________________
*Corresponding author
aqilah.sam95@gmail.com (Nurul Aqilah Sham Shul Bahri)
Published Online
Based on the result, we used pennywort as our
plant sample. After we run the sample using ICP, the result
showed that there is no presence of cr2+ and cd2+. The
result (sample 4) showed the 0.00 ppm for cr2+ and cd2
concentration in the sample. This wonder herb is used to
treat venereal diseases, skin impurities, high blood pressure,
hepatitis, varicose veins, skin ulcers, minor burns, fatigue
and stress. Fresh leaves have been known to be used in
treating dysentery, asthma, stomach disorders and
bronchitis. Juice extracted that we got from the leaves is
highly beneficial in promoting the purification of the blood.
It also can help improve circulation and the rebuilding of
connective tissue, assisting in the prevention of premature
ageing. People who suffer from eczema and other skin
relate ailments including skin ulcer also can used
pennywort. Besides that, it is also able to lower blood sugar
levels. of Pennywort contains many of minerals and
vitamins, among them Vitamin K, calcium, sodium,
Vitamin B-Complex, magnesium, zinc and manganese. It is
also able to help protect the body from toxins. This herb is
also able to greatly benefit the central nervous system,
making it effective in the treatment of ADD, senility,
epilepsy, strokes and nervous system disorders. Those
suffering from leg cramps and phlebitis can benefit from
using this herb. It is also effective in reducing fevers. There
are many advantages of pennywort. If pennywort contain
higher of cadmium it is dangerous because cadmium is a
very permeable metal. Its tarnish in air. It also highly toxic
element and has been describe as one of the most dangerous
trace element in food and environment of human. Cadmium
also even presence as non-essential element that possesses
high toxicity and easily accumulate from the environment
by organism. Then, the objective from this experiment is to
familiarize with spiking technique in unknown
concentration determination.
3.2 Determination Chromium Series Standard
Chromium also has a bad effect in plant. Due to its
wide industrial use, chromium is considered a serious
environmental pollutant. Contamination of soil and water
by chromium (Cr) is of recent concern. Toxicity of Cr to
plants depends on its valence state. Cr (VI) is highly toxic
and mobile whereas Cr(III) is less toxic. Since plants lack a
 Determination of Cr2+ and Cd2+ in Centella asiatica using Inductively Coupled Plasma Optical Emission Spectrometry (ICP-
OES)
specific transport system for Cr, it is taken up by carriers of
essential ions such as sulfate or iron. Toxic effects of Cr on
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3.3 Concentration Of The Analyte
4)
In preparing a matrix spike, a spiking solution is a
standard that is chosen. The concentration of the analyte in
the spiking solution is usually 50 -100 times higher than the
5)
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7)
4. Conclusions
In our experiment, cadmium and chromium elements in our
sample which is pennywort plant (Centella asiatica) are not
8)
identified. As a conclusion, pennywort is free from heavy
metal elements.
ACKNOWLEDGEMENTS
9)
The authors wish to extend their gratitude to the
staff and the lecturer Madam Khadijah Khalid of UiTM
Jengka Branch, Pahang, Malaysia for their assistance in this
experiment.
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*Corresponding author
aqilah.sam95@gmail.com (Nurul Aqilah Sham Shul Bahri)
Published Online
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____________________________________________________________
*Corresponding author
aqilah.sam95@gmail.com (Nurul Aqilah Sham Shul Bahri)
Published Online