ABSTRACT

Atomic Absorption Spectroscopy (AAS) instrument is used to measures the absorption of light at

specific wavelengths by elements in the Free State, such as calcium. The objective of this e periment is to detect calcium in the supplement tablets. Then, this e periment was conducted to compare the concentration of un!nown sample (commercial calcium supplement) with the amount of calcium that needed by human. "n!nown sample was prepared and the data obtained was used to construct calibration curve. The concentration values that were obtained from calibration curve are #$%.&#, ##'.'%, (().'%, $*.)& and #+.%, ppm which respect to the p- ', (, ,, %, and +. This showed that the larger the p- level, the less acidic a substance is. Since the relationship between the p- level and the concentration of the un!nown solution was determined, the effectiveness of the p- solubility and the digestion system in the human.

INTRODUCTION The e periment was conduct to determine the calcium content present in commercial tablets using Atomic Absorption Spectroscopy (AAS). .alcium is an essential mineral found in great abundance in the body and also essential for living organisms mostly in cell physiology. As a major material used in minerali/ation of bones and shells, calcium is the most abundant metal by mass in many animals and in humans. 0inety1 nine percent (##2) of all the calcium in the body is found in the bones and teeth. The remaining one percent ('2) is in the blood. .alcium plays important roles in nerve conduction, muscle contraction, and blood clotting. 3 tracellular calcium is also important for maintaining the potential difference across e citable cell membranes, as well as proper bone formation. 4f calcium levels in the blood drop below normal levels, calcium is transferred from the bones to the blood in order to maintain blood calcium levels. Therefore, it is important to consume sufficient calcium to maintain ade5uate blood and bone calcium levels. .alcium supplements are used to prevent and to treat calcium deficiencies. 6ost e perts recommend that supplements be ta!en with food and that no more than *)) mg should be ta!en at a time because the percent of calcium absorbed decreases as the amount of calcium in the supplement increases. 4t is recommended to increase doses throughout the day. 4n addition, they also recommended that daily calcium inta!e for adults7 ranges from '))) to '+)) mg and suggested to ta!e supplements with food to support in absorption. Atomic spectroscopy is the determination of elemental composition by its electromagnetic or mass spectrum. The study of the electromagnetic spectrum of elements is called 8ptical Atomic Spectroscopy. 3lectrons e ist in energy levels within an atom. These levels have well defined energies and electrons moving between them must absorb or emit energy e5ual to the difference between them. 4n optical spectroscopy, the energy absorbed to move an electron to a more energetic level and9or the energy emitted as the electron moves to a less energetic energy level is in the form of a photon. The wavelength of the emitted radiant energy is directly related to the electronic transition which has occurred. Since every element has a uni5ue electronic structure, the wavelength of light emitted is a uni5ue property of each individual element. As the orbital configuration of a large atom may be comple , there are many
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electronic transitions which can occur, each transition resulting in the emission of a characteristic wavelength of light as illustrated in Figure '.

Figure ': The transition resulting in the emission of a characteristic wavelength of light. The science of atomic spectroscopy has yielded three techni5ues for analytical use: '. (. ,. Atomic Absorption. Atomic 3mission. Atomic Fluorescence. The process of e citation and decay to the ground state is involved in all three fields of atomic spectroscopy. 3ither the energy absorbed in the e citation process, or the energy emitted in the decay process is measured and used for analytical purposes. Atomic Absorption Spectrophotometer (AAS) is designed to determine the amount (concentration) of an object element in a sample, utili/ing the phenomenon that the atoms in the ground state absorb the light of characteristic wavelength passing through an atomic vapor layer of the element. 4n addition, it also provides accurate 5uantitative analyses for metals in water, sediments, soils or roc!s. Atomic absorption units have four basic parts which are interchangeable lamps that emit light with element1specific wavelengths, a sample aspirator, a flame or furnace apparatus for volatili/ing the sample, and a photon detector. AAS has been used to study structure1function relationships in foods for both li5uid and solid to improve overall food 5uality, safety and sensory characteristics. Furthermore, it has also used to investigate fungal infections in plant materials li!e fruits and seeds or to study mobility of different chemical components in food materials. ;esides that, there are many applications of AAS in the industries such as in chemical, petrochemical and refineries industries,
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pharmaceutical industries, recycling and waste management, water analytics and water treatment and many more. 4n petrochemical and refineries industries, they apply AAS for determination of heavy metals, sulfur, nitrogen, and chlorine in oil, fuel, paraffin, fatty acids and polymers. 4n addition, they utili/e the AAS in identification of substance, 5uality control and sewages analysis. 4n pharmaceutical industries, AAS are use in screening of human body and for active ingredients. Additionally, they also apply for control of cleaning validation procedures and identification of substances.

OBJECTIVE '. To determine the commercial calcium supplement by Atomic Absorption

Spectrophotometry. (. To compare the concentration of un!nown sample (commercial calcium supplement) with the amount of calcium that needed by human.

THEORY Atomic absorption units have four basic parts which are interchangeable lamps that emit light with element1specific wavelengths, a sample aspirator, a flame or furnace apparatus for volatili/ing the sample, and a photon detector. The techni5ue atomic absorption spectroscopy typically ma!es use of a flame to atomi/e the sample, but other atomi/ers such as a graphite furnace are also used. Three steps are involved in turning a li5uid sample into an atomic gas: '. <esolvation = the li5uid solvent is evaporated, and the dry sample remains. (. >aporisation = the solid sample vaporises to a gas. ,. Atomi/ation = the compounds ma!ing up the sample are bro!en into free atoms. 4f light of just the right wavelength impinges on a free, ground state atom, the atom may absorb the light as it enters an e cited state in a process !nown as atomic absorption. This process is illustrated in the Figure (. Atomic absorption ta!es place when une cited atoms absorb energy and become e cited atoms. 3lectrons can e ist in one of two states: '. ?round State: 4n this state, the electron contains the least energy possible, orbiting as close as it can to the nucleus. (. 3 cited State: 4n this state, the electron contains more energy than in its ground state, orbiting further from the nucleus of the atom. Absorption therefore is carried out by une cited atoms, whereas emission arises from e cited atoms.

Figure (: The atomic absorption process

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Atomic absorption measures the amount of light at the resonant wavelength which is absorbed as it passes through a cloud of atoms. As the number of atoms in the light path increases, the amount of light absorbed increases in a predictable way. ;y measuring the amount of light absorbed, a 5uantitative determination of the amount of analyst element present can be made. The use of special light sources and careful selection of wavelength allow the specific 5uantitative determination of individual elements in the presence of others. The atom obscure re5uired for atomic absorption measurements is produced by supplying enough thermal energy to the sample to dissociate the chemical compounds into free atoms. Aspirating a solution of the sample into a flame aligned in the light beam serves this purpose. "nder the proper flame conditions, most of the atoms will remain in the ground state form and are capable of absorbing light at the analytical wavelength from a source lamp. The ease and speed at which precise and accurate determinations can be made with this techni5ue have made atomic absorption one of the most popular methods for the determination of metals. The use of light absorption measurements to determine concentrations of atoms is called atomic absorption spectroscopy (AAS). Although this phenomenon has been studied since early in the nineteenth century, it was about '#*) when the techni5ue became part of the analytical repertoire, in large part as a conse5uence of wor! by @alsh in Australia. ;ecause of the simplicity of the electronic structure of an atom (compared with a molecule), and the absence of vibration and rotational transitions, atomic spectra are simpler than molecular spectra. 4nstead of the band spectra characteristic of molecular absorption in the "> and visible regions, atoms e hibit line spectraA that is, the absorption pea!s are so narrow that they are essentially lines. Some subtle effects are responsible for producing lines of finite width, but they are e tremely narrow nevertheless, with line widths of ).)'1).'A being typical. As a conse5uence, AAS is capable of high specificity, since the narrow lines minimi/e the overlapping absorption so common in molecular band spectra. As in any form of absorption spectroscopy, the light source must generate radiation that includes the fre5uency that is absorbed by the sample. 4n AAS, e treme specificity is achieved by using as the light source a lamp whose emitting element is identical with the element to be determined. The most common AAS flame gas mi ture is air1 acetylene, which generates a flame temperature of (,))B.. Instrumentation of Atomic absorption spectroscop
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Figure ,: Schematic of an atomic1absorption e periment The e5uipment used in AAS is basically the same as for other spectroscopic absorption methods. 4t consists of: Ra!iation source The light source is usually a hollow1cathode lamp of the element that is being measured. Casers are also used in research instruments. Since lasers are intense enough to e cite atoms to higher energy levels, they allow atomic absorption and atomic fluorescence measurements in a single instrument. The disadvantage of these narrow1band light sources is that only one element is measurable at a time. "onoc#romator The function of the monochromator is to select radiation of the correct wavelength and eliminate other radiation from the light path. For many elements such as sodium, potassium, and copper the spectrum is simple and only low resolution monochromator is re5uired. -owever, for certain other elements, particularly the transition elements, high resolution is necessary to prevent unabsorbable emission lines. Detectors Dhotomultipliers are used e clusively on commercial e5uipment. 4n practice, it is the function of the detector to measure the intensity of radiation before and after absorption by the sample. From this one can calculate how much radiation has been absorbed from the intense beam.
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Optica$ s$it s stem Two slits are included in the optical system, an entrance slit and an e it slit. The entrance

slit serves to obtain a narrow, parallel beam of light from the source. The e it slit is used to select radiation of the correct wavelength after it emerges from the monochromator. Atomi%er The function of the atomi/er is to convert the combined atoms of the li5uid sample into free atoms. The most common atomi/er is the flame. 4n practice the li5uid sample is introduced into the flame in the form of a droplet. The droplets evaporate and leave a solid residue. The residue, which contains the same atoms, is decomposed by the flame and free atoms are liberated. These free atoms absorb the radiation which is measured in this procedure. Ana$ tica$ &ariab$es The 5uantitative application of atomic absorption spectroscopy is based on two relationships. First, the degree of absorption is a function of how many free atoms are present in the light path. Second, the number of free atoms is a function of the concentration of the metal on the original sample. The process of atomi/ation depends on several variables. This are1 a) ;urner design b) Sample flow rate c) Type of flame d) Solvent e) @avelength choice

'ROCEDURES A stoc! solution with +)ppm concentration is prepared using '))mC volumetric flas!. Then a series of standard solution of (+, '(.+ and *.(+ppm is to be prepared using the stoc! solution )f +)ppm. The solutions then are to be used in calibration of the spectrometer. Five different sample of p- is to be ta!en to be measured using Atomic Adsorption Spectrometer (AAS). -aving the value of the standard solution, then the calibrating curve is plotted to determine the concentration of the sample.

RESU(TS AND CA(CU(ATION

i) Standard Solutions Sample 0ame ' ( , % + Sample .oncentration (ppm) ).)) *.(+ '(.+ (+.) +) Adsorption ).))), ).)(#* ).)%'( ).)+*# ).)#,%

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ii)

Sample Solutions Sample 0ame p- ' p- ( p- , p- % p- + Sample .oncentration (ppm) #$%.&# ##'.'% (().*% $*.)& #+.%, Adsorption '.%))% '.%)#, ).,,)* ).'%(( ).'++,

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DISCUSSION From the data obtained, the concentrations of calcium in the un!nown solution were calculated via calibration curve. The calibration curve plays an important role as the reference to determine the un!nown solutions. The calibration curve was measured using the standard solution which had been prepared. The data was then accumulated in present of graph in term of the absorbance versus the concentration. From the graph obtained, the e5uation ofA y E ).))'% F ).)'(& is used to determine the concentration of the un!nown solutions. The un!nown solution of p- ' to p- + is to be determined. From the graph plotted, the correlation coefficient value is ).##$. For the second graph the value of the concentration is calculated using the first e5uation obtain from the calibration graphA y E ).))'% F ).)'(&. The concentration for p- ' was #$%.&#ppm with an adsorption of '.%))%. Thus the second p- of p- ( was accumulated to have a concentration of ##'.'%ppm. 4t adsorption value for p- ( was '.%)#,. The concentration of p- , was (().*%ppm. The adsorption for the p- , was ).,,)*. The concentration of p- % was $*.)&ppm. The adsorption for the p- % was ).'%((. The concentration of p- + was $*.)&ppm. The adsorption for the p- % was ).'%((ppm. The adsorption for the p- + was ).'++,. <ue to the mista!e in calculating the mass need to be measured to prepared the stoc! solution and the series of standard solution, the calibrating is giving a very slightly value for the edge. The edge with , decimal point ().))') giving the concentration to be highly concentrated. Although the value was in range ).'+ to '.%, the highest concentration was turn to almost ')))ppm. This is the error related to the second graph measurement for AAS. 3ven the calibration is slightly wrong, the pattern of the concentration for the second graph is still can be manually calculated as the value of the adsorption doesn7t correspondent to the calibration value. As shown on the second graph, the highest concentration stated was in p- (. This indicated that the more acidic the solution, the higher concentration of the calcium. This pattern related that the acidic solution have the more calcium to dissolved. There are relationship between the p- and the concentration of the calcium hydro ide. The same scenario happens in the human stomach. The digestion process been divided into some part. The acidic part was when the gastric acid is release from cells lining the stomach, which are coupled to systems to increase acid production when needed. ?astric acid is a digestive fluid, formed in
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the stomach. 4t has a p- of ' to ( and is composed of hydrochloric acid (-.l) (around ).+2, or +))) parts per million), and large 5uantities of potassium chloride (G.l) and sodium chloride (0a.l). The acid plays a !ey role in digestion of proteins, by activating digestive en/ymes, and ma!ing ingested proteins unravel so that digestive en/ymes can brea! down the long chains of amino acids. The acidic p- gain from the gastric acid have the possibility to dissolve the calcium and other nutritious that the body needed. The same ways goes for the e periment where the calcium hydro ide is strongly reacted with the lower p-. This indicated that at the lower p-, the calcium hydro ide can dissolved very well comparing to the higher value of p- or the basic solution.

CONC(USION 4n the nutshell, the analytical techni5ue which Atomic Adsorption Spectroscopy (AAS) was used was to determine the concentration of the calcium inside the un!nown solution. The relation of the p- and the concentration was then investigated and to be determine to which p- has the highest concentration. The relation between the concentration and the p- can be determining from the calibration curve. As from the calibration curve, the e5uation of y E ).))'% F ).)'(& was obtained. Thus, from the calculation of the concentration of the calcium content was related to each of the p- investigated. . The concentration values which correspond to p- ', (, ,, %, + and * are #$%.&#, ##'.'%, (().*%, $*.)&, and #+.%,ppm respectively. This had proven the e istence of the relationship between the p- and the concentration of the calcium. . 6a imum amount of calcium that recommended is *))mg9C per day. 4n our stomach, gastric juice appears at p- ' to p- , in order to digest our food. The concentration of un!nown sample from p- ' to p- , is suitable for human body. The total concentration from p- ' to p- , can be supported by stomach. ;esides, this showed the concentration for p- in the range of ' to , is ade5uate by blood and bone calcium levels.

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RECO""ENDATION '. 6a!e sure the sample preparation which is the dilution is conducted by a proper procedure. (. "se digital pipette to ensure the e periment easy to run and more accurate. ,. 4t is better to have the e5uipment chec!ed and maintenance to prevent any error. %. "se the e act cathode lamp to determine the correct composition. +. Hinse and dried the apparatus to get a better calibration curve.

RE)ERENCES http:99dietary1supplements.info.nih.gov9factsheets9calcium.asp http:99www.calciuminfo.com9about9foodsources.asp S!oog, <. A., -oller, F. I., J 0ieman, T. A. ('##$). Principles of Instrumental Analysis. ;roo!s .ole. S!oog, <. A., @est, <. 6., -oller, F. I., J .rouch, S. H. (())%). Fundamentals of Analytical Chemistry. ;roo!s .ole.

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