Determination of Cr2+ and Cd2+ in Centella asiatica usingInductively 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,
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[email protected]. 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 3 ppm.2.5. 2ml. 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. Cd gives numerous negative effects on the body Standard series of Cadmium range from 1 ppm. Specific wavelength of the photons can be used to 0. 4 ppm and 5 ppm were made by using standard and can impact nearly all systems in the body including addition method 1ml. Chromium can affect antioxidant metabolism in ppm cadmium standard were pipetted and transfer into 50ml plants. 2. The apparatus used in this experiment were radiofrequency (RF) discharge. Inductively coupled plasma/optical emission spectrometry (ICP-OES) 2. As the plasma 2. The cadmium content of 2. Materials ICP-OES is a spectroscopic technique suitable for The sample (plant) were obtained from the trace elements analysis in several types of samples. Materials and Method 2. 080 N 80314401. Samples are usually 250ml beaker. thus.0g accurately and in microwave were place identify the elements and the number of photons is directly digestion vessels separately.8 ppm and 1. Cadmium Series Standard due to its huge industrial use.2 ppm. the samples sample. selenium. 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. 4ml and 5ml of 10 process. 10ml of water samples were added into each volumetric flask contained standard series. filter paper. 2. The vessels in this technique such as nebulization.1. It is known to be a toxic metal that can cause severe damage to plants and humans. the kidneys. 2 ppm. Then. environmental contaminant released into the atmosphere 2. pipette. solid filter funnel and microwave digestion. Preparation of Standard However. Chromium Series Standard plant tissues tends to increase with increase of 2. and lid. High Standard series of Cadmium range from 0. 100ml volumetric flask. while gas The background correction methods were and liquid samples may be injected directly into the performed using Inductively Couple Plasma Spectrometry instrument. serial no. eyes and even the cadmium standard were pipetted and transfer into 50ml of each volumetric flask. Antioxidant enzymes are found to be susceptibility of each volumetric flask.4. 3 portions of the samples were weight ranging from photons. (ICP) Model Perkin Elmer – Optima 2100DV. Preparation of 10ppm of Standard Solution for ICP In plants tissues. sends into the centre of the plasma which maintains high atomization temperature of around 10. All of to chromium resulting in a decline in their catalytic the volumetric flasks were then diluted with distilled water activities. 3ml. A variety of sample introduction methods are used were digested in the microwave for 20 minutes. 4ml and 5ml of 10 ppm cardiovascular. Preparation Series of Standard Solution for AAS concentrations of soil cadmium above certain background amounts.4 ppm. 10ml of water samples were added brain. 3ml. reproductive. Chromium (Cr) is the seventh the volumetric flasks were then diluted with distilled water most abundant metal in the earth’s crust and important until the calibration mark.3. 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.sam95@gmail. technique is based on the unprompted emission of photons Malaysia. ____________________________________________________________ *Corresponding author aqilah. All of On the other hand. 0. 0.5g to 1.6 ppm. 7ml of concentrated HNO3 and proportional to the concentration of the element in the 1ml of H2O2 was added into each vessel. samples require acid digestion prior to injection. until the calibration mark. into each volumetric flask contained standard series. location at 305 was used in the The sample solution is converted to an aerosol then experiments .000 K. hydride generation then cooled down to room temperature before opening the (HG) for certain elements such as arsenic.com (Nurul Aqilah Sham Shul Bahri) Published Online . is this plant safe to eat and free of heavy metal? 1. Deionized water was used to prepare all the from atoms and ions that have been excited in a solutions. Determination of Cr2+ and Cd2+ in Centella asiatica using Inductively Coupled Plasma Optical Emission Spectrometry (ICP- OES) ‘ulam’. the present of Cadmium (Cd) in low concentrations is most likely a normal constituent. a type of Malay salad eaten with the main dish. introduced into the plasma in liquid form. 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.0 ppm were made by using chromium concentration can disturb the photosynthetic standard addition method 1ml.5. 2ml. 0. The laboratory in Universiti Teknologi MARA Pahang. among them Vitamin K. It also can help improve circulation and the rebuilding of Determination of the of Cr2+ and Cd2+ concentrations in the sample using spiking technique. Cadmium also even presence as non-essential element that possesses high toxicity and easily accumulate from the environment by organism. Contamination of soil and water by chromium (Cr) is of recent concern. The blank solutions was read first. This herb is and 1.com (Nurul Aqilah Sham Shul Bahri) Published Online .0ppm. It is also effective in reducing fevers. Toxicity of Cr to Table 1: result plants depends on its valence state. treating dysentery. fatigue various matrices. Besides that. we used pennywort as our and laser ablation. 2ppm. senility.00 ppm for cr2+ and cd2 Straits of Malacca the West region of Peninsular Malaysia. 3. Juice extracted that we got from the leaves is 2. it is also able to lower blood sugar The solutions of Cr2+ in different solvents were prepared. Next. the solutions of Cd2+ in different solvents were prepared by using 0. This wonder herb is used to instrumentation make possible the measurement of low treat venereal diseases. Cd. strokes and nervous system disorders.2ppm. Inductively coupled plasma/optical emission using this herb. 3ppm.4ppm. Determination of Cr2+ and Cd2+ in Centella asiatica using Inductively Coupled Plasma Optical Emission Spectrometry (ICP- OES) antimony. curve and determine the concentration of sample in the Vitamin B-Complex. It’s tarnish in air. The connective tissue. 0. recorded in the computer. Cr (VI) is highly toxic and mobile whereas Cr(III) is less toxic. followed by 1 levels. minor burns. as well as electro thermal vaporization (ETV) Based on the result. the result ICP-OES has been used for the determination of showed that there is no presence of cr2+ and cd2+. relate ailments including skin ulcer also can used pennywort.1. and As in marine fish and shellfish from the result (sample 4) showed the 0. The plotting calibration curve then was also able to greatly benefit the central nervous system. calcium. chromium is considered a serious environmental pollutant. the objective from this experiment is to familiarize with spiking technique in unknown concentration determination.2 Determination Chromium Series Standard Chromium also has a bad effect in plant.sam95@gmail. The Hg. of Pennywort contains many of minerals and ppm. There spectrometry (ICP-OES) are many advantages of pennywort. Those suffering from leg cramps and phlebitis can benefit from 3. 3. Then. skin impurities. making it effective in the treatment of ADD. Pb. they are considered superior and stress. asthma. assisting in the prevention of premature plant sample were labelled and keep plant in lab 306 for ageing. Fresh leaves have been known to be used in tools in elemental analysis compared to other methods of element detection. plant sample. It is sample. level of concentrations with good accuracy and precision in hepatitis. 0. stomach disorders and bronchitis. Therefore. zinc and manganese.8ppm also able to help protect the body from toxins. sodium. 4ppm and 5ppm to obtain calibration vitamins. Due to its wide industrial use. People who suffer from eczema and other skin ICP determination. The modern spectroscopic analytical methods and concentration in the sample. skin ulcers. After we run the sample using ICP. It also highly toxic element and has been describe as one of the most dangerous trace element in food and environment of human. Since plants lack a ____________________________________________________________ *Corresponding author aqilah. high blood pressure. If pennywort contain higher of cadmium it is dangerous because cadmium is a very permeable metal.6. varicose veins.6ppm. magnesium. Results and Discussions epilepsy. 0. Determination of Cr2+ and Cd2+ in Plant Tissue highly beneficial in promoting the purification of the blood. contaminated soil. stems Mohd Izuan Effendi H. the spiking solution is usually 50 -100 times higher than the 5) Brown RJ. Öktem AB. 2017 from http://umpir. Heavy metals concentration in staff and the lecturer Madam Khadijah Khalid of UiTM four commercially valuable marine edible fish species Jengka Branch. 19(1):135- relations and mineral nutrition.pdf accumulate or to stabilize Cr compounds for bioremediation 3) Beaty RD. The spike should not increase the http://floridata. (1972). cadmium and chromium elements in our optical-emission-spectroscopy-icp-oes sample which is pennywort plant (Centella asiatica) are not 8) Nurul Izyatulikma Binti Yusoff (2012. Pahang. pp 8-12 3. 1993. Azrina A. Analytical techniques for trace concentration found in the unspiked sample. PerkinElmer Shelton CT. Veerasingam S. Suresh G. Malaysia for their assistance in this from Parangipettai Coast.ump. Determination of Cr2+ and Cd2+ in Centella asiatica using Inductively Coupled Plasma Optical Emission Spectrometry (ICP- OES) specific transport system for Cr.edu. The concentration of the analyte in 2010. significantly change the sample volume. Czech Journal of Food Sciences standard that is chosen. Metabolic alterations by Cr 2) Applications Tip of the Week Matrix Spiking – Why exposure have also been described in plants either by a Spike and How to Do It. 2017 direct effect on enzymes or other metabolites or by its from https://static. 1(1):10-14. The authors wish to extend their gratitude to the Venkatachalapathy R. Conclusions Retrieved from https://www. Marichamy G. (2000).fishersci. 24(3): 266-274. Trends Analyt Chem concentration chosen to ensure that the spike does not 2005. Int Food Res J 2012. T. 7) Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES Analysis).com/cmsassets/downloads/ ability to generate reactive oxygen species which may cause segment/Scientific/pdf/WaterAnalysis/Log112tipMatrix oxidative stress. Retrieved Jun 1. it must be a/769 taken into account. Cr also causes deleterious effects on plant selected marine fish and shellfish along the Straits of physiological processes such as photosynthesis.pdf 9) Raja P. arsenic. Retrieved Jun 1. C. Concepts Instrumentation and of Cr contamination has gained interest in recent years.sam95@gmail. Determination of some heavy metal levels in soft drinks from Turkey In preparing a matrix spike.lucideon. 28(3): 213-216.com (Nurul Aqilah Sham Shul Bahri) Published Online . Centella asiatica. South East Coast of India.). Er B. ____________________________________________________________ *Corresponding author aqilah.com/testing- characterization/techniques/inductively-coupled-plasma- In our experiment. germination process as well as in the growth of roots. L. This element analysis: an overview. This simplifies the 6) Duever. a spiking solution is a using ICP-OES method. As a conclusion. Toxic effects of Cr on REFERENCES: plant growth and development include alterations in the 1) Alina M.d. (n. J Anim Vet Adv 2009. USA.3 Concentration Of The Analyte 4) Bıngöl M.com/Plants/Apiaceae/Centella%20asiatic sample volume by more than 5%. it is taken up by carriers of essential ions such as sulfate or iron. January). Int experiment. Retrieved from dilution of the matrix. In Floridata calculations and avoids introducing unknown effects from Plant Encyclopedia. Mohd Zakiuddin S. Otherwise. 4. Heavy and leaves. 10) Shacklette. Muhammad Rizal R. Milton MJ. 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