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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 roadside ditches, and soggy lawns and prefer sandy soils
spectrometry (ICP-OES) is an analytical technique used for enriched with organic matter. It can be grown around the
the detection of trace metals. The samples are introduced edges of a water feature, mixed species lawn or kept in a
into the plasma in a process that desolvates, ionises, and well-irrigated corner of the herb garden.
excites them. It is a type of emission spectroscopy that uses Under the name "gotu kola", Centella asiatica is
the inductively coupled plasma to produce excited atoms referred as one of the great multi-purpose miracle herbs of
and ions that emit electromagnetic radiation at particular Oriental medicine. It has been in use for thousands of years
wavelengths characteristic of an element. It is a flame and also has been considered the most powerful
technique with a flame temperature in a range from 6 000 to rejuvenating herbs and for promoting longevity as a
10 000K. The constituent elements within the sample can be legendary healer, LiChing Yun who is said to have lived 256
identified by their characteristic emission lines while the years as a result of drinking gotu kola tea. In Indian
concentration of the element can be quantified by the medicine, it is called "brahmi" meaning "greatest of the
intensity of the same lines. great" and it is recommended for treatment of mental
Centella asiatica is the scientific name for disorders, immune system deficiencies, epilepsy, hair loss
pegaga or commonly known as Asiatic pennywort or gotu and intestinal complaints.
kola. It is a small creeping herb with shovel shaped leaves This plant is well known for its benefit to the
emerging alternately in clusters at the stem nodes and it is human; henceforth it is ingested by people around the
appeared as herbaceous native plant in Asia. The plants 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-
ulam, a type of Malay salad eaten with the main dish.
2.3. Preparation of Standard
However, is this plant safe to eat and free of heavy metal?
1. Preparation of 10ppm of Standard Solution for ICP
In plants tissues, the present of Cadmium (Cd) in
low concentrations is most likely a normal constituent. The 5ml of 100ppm stock solution of Chromium/ cadmium was
pipette into 50ml volumetric flask was filled with distilled
concentration in the tissue is determined by the cadmium water until the calibration mark. The volumetric flask
concentration in the environment the inherent ability of the contained solutions were labeled as 10ppm Cr2+/ Cd2+
plant species to absorb the ions. The cadmium content of
2.4. Chromium Series Standard
plant tissues tends to increase with increase of 2. Preparation Series of Standard Solution for AAS
concentrations of soil cadmium above certain background
amounts. Cd gives numerous negative effects on the body Standard series of Cadmium range from 1 ppm, 2 ppm, 3
ppm, 4 ppm and 5 ppm were made by using standard
and can impact nearly all systems in the body including addition method 1ml, 2ml, 3ml, 4ml and 5ml of 10 ppm
cardiovascular, reproductive, the kidneys, eyes and even the 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
On the other hand, Chromium (Cr) is the seventh the volumetric flasks were then diluted with distilled water
most abundant metal in the earths crust and important until the calibration mark.
environmental contaminant released into the atmosphere
2.5. Cadmium Series Standard
due to its huge industrial use. It is known to be a toxic metal
that can cause severe damage to plants and humans. High 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
chromium concentration can disturb the photosynthetic standard addition method 1ml, 2ml, 3ml, 4ml and 5ml of 10
process. Chromium can affect antioxidant metabolism in ppm cadmium standard were pipetted and transfer into 50ml
plants. Antioxidant enzymes are found to be susceptibility of each volumetric flask. 10ml of water samples were added
into each volumetric flask contained standard series. All of
to chromium resulting in a decline in their catalytic the volumetric flasks were then diluted with distilled water
activities. until the calibration mark.

2. Materials and Method 2.5. Inductively coupled plasma/optical emission

spectrometry (ICP-OES)
2.1. Materials
ICP-OES is a spectroscopic technique suitable for
The sample (plant) were obtained from the
trace elements analysis in several types of samples. The
laboratory in Universiti Teknologi MARA Pahang, technique is based on the unprompted emission of photons
Malaysia. Deionized water was used to prepare all the from atoms and ions that have been excited in a
solutions. The apparatus used in this experiment were radiofrequency (RF) discharge. Samples are usually
250ml beaker, 100ml volumetric flask, pipette, filter paper, introduced into the plasma in liquid form; thus, solid
filter funnel and microwave digestion. samples require acid digestion prior to injection, while gas
The background correction methods were and liquid samples may be injected directly into the
performed using Inductively Couple Plasma Spectrometry instrument.
(ICP) Model Perkin Elmer Optima 2100DV, serial no.
080 N 80314401, location at 305 was used in the The sample solution is converted to an aerosol then
experiments . sends into the centre of the plasma which maintains high
atomization temperature of around 10,000 K. As the plasma
2.2. Sample Preparation free atoms in the gaseous state are generated and adequate
energy is often available to convert the atoms to ions then
The plant sample was cut into small pieces and promote the ions to excited states. The ionic excited state
50g of the sample was put in oven (110oC) to dry for 24 species may then return to the ground state via emission of
hours. 3 portions of the samples were weight ranging from photons. Specific wavelength of the photons can be used to
0.5g to 1.0g accurately and in microwave were place identify the elements and the number of photons is directly
digestion vessels separately. 7ml of concentrated HNO3 and proportional to the concentration of the element in the
1ml of H2O2 was added into each vessel. Then, the samples sample. A variety of sample introduction methods are used
were digested in the microwave for 20 minutes. The vessels in this technique such as nebulization, hydride generation
then cooled down to room temperature before opening the (HG) for certain elements such as arsenic, selenium, and
*Corresponding author
aqilah.sam95@gmail.com (Nurul Aqilah Sham Shul Bahri)
Published Online
Determination of Cr2+ and Cd2+ in Centella asiatica using Inductively Coupled Plasma Optical Emission Spectrometry (ICP-
antimony, as well as electro thermal vaporization (ETV) Based on the result, we used pennywort as our
and laser ablation.
plant sample. After we run the sample using ICP, the result
ICP-OES has been used for the determination of showed that there is no presence of cr2+ and cd2+. The
Hg, Cd, Pb, and As in marine fish and shellfish from the result (sample 4) showed the 0.00 ppm for cr2+ and cd2
Straits of Malacca the West region of Peninsular Malaysia.
The modern spectroscopic analytical methods and concentration in the sample. This wonder herb is used to
instrumentation make possible the measurement of low treat venereal diseases, skin impurities, high blood pressure,
level of concentrations with good accuracy and precision in hepatitis, varicose veins, skin ulcers, minor burns, fatigue
various matrices. Therefore, they are considered superior
and stress. Fresh leaves have been known to be used in
tools in elemental analysis compared to other methods of
element detection. treating dysentery, asthma, stomach disorders and
bronchitis. Juice extracted that we got from the leaves is
2.6. Determination of Cr2+ and Cd2+ in Plant Tissue highly beneficial in promoting the purification of the blood.
It also can help improve circulation and the rebuilding of
Determination of the of Cr2+ and Cd2+
concentrations in the sample using spiking technique. The connective tissue, assisting in the prevention of premature
plant sample were labelled and keep plant in lab 306 for ageing. People who suffer from eczema and other skin
ICP determination. relate ailments including skin ulcer also can used
pennywort. Besides that, it is also able to lower blood sugar
The solutions of Cr2+ in different solvents were
prepared. The blank solutions was read first, followed by 1 levels. of Pennywort contains many of minerals and
ppm, 2ppm, 3ppm, 4ppm and 5ppm to obtain calibration vitamins, among them Vitamin K, calcium, sodium,
curve and determine the concentration of sample in the
Vitamin B-Complex, magnesium, zinc and manganese. It is
sample. Next, the solutions of Cd2+ in different solvents
were prepared by using 0.2ppm, 0.4ppm, 0.6ppm, 0.8ppm also able to help protect the body from toxins. This herb is
and 1.0ppm. The plotting calibration curve then was also able to greatly benefit the central nervous system,
recorded in the computer. making it effective in the treatment of ADD, senility,
3. Results and Discussions epilepsy, strokes and nervous system disorders. Those
suffering from leg cramps and phlebitis can benefit from
3.1. Inductively coupled plasma/optical emission using this herb. It is also effective in reducing fevers. There
spectrometry (ICP-OES) 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
Table 1: result
plants depends on its valence state. Cr (VI) is highly toxic
and mobile whereas Cr(III) is less toxic. Since plants lack a
*Corresponding author
aqilah.sam95@gmail.com (Nurul Aqilah Sham Shul Bahri)
Published Online
Determination of Cr2+ and Cd2+ in Centella asiatica using Inductively Coupled Plasma Optical Emission Spectrometry (ICP-
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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relations and mineral nutrition. Metabolic alterations by Cr 2) Applications Tip of the Week Matrix Spiking Why
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3.3 Concentration Of The Analyte
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the spiking solution is usually 50 -100 times higher than the
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concentration chosen to ensure that the spike does not
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significantly change the sample volume. This simplifies the
6) Duever, L. C. (2000). Centella asiatica. In Floridata
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taken into account.
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4. Conclusions Retrieved from https://www.lucideon.com/testing-
In our experiment, cadmium and chromium elements in our
sample which is pennywort plant (Centella asiatica) are not
8) Nurul Izyatulikma Binti Yusoff (2012, January). A pre
identified. As a conclusion, pennywort is free from heavy
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staff and the lecturer Madam Khadijah Khalid of UiTM four commercially valuable marine edible fish species

Jengka Branch, Pahang, Malaysia for their assistance in this from Parangipettai Coast, South East Coast of India. Int

experiment. J Anim Vet Adv 2009; 1(1):10-14.

10) Shacklette, H. T. (1972). Cadmium in Plants. United
States Government Printing Office, Washington.

*Corresponding author
aqilah.sam95@gmail.com (Nurul Aqilah Sham Shul Bahri)
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Determination of Cr2+ and Cd2+ in Centella asiatica using Inductively Coupled Plasma Optical Emission Spectrometry (ICP-
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*Corresponding author
aqilah.sam95@gmail.com (Nurul Aqilah Sham Shul Bahri)
Published Online