Water Testing and Subsequent Comparison of the methods Atomic Absorption/Atomic Emission Spectrophotometry Complimentary to Iron UV-visible Spectrophotometry

to Test for Calcium, Magnesium, Sodium, and Iron.

Jaden Smytherman IA-427 Summer 07-16-2011 Abstract Atomic absorption/atomic emission spectrophotometry (AA/AE) is complimentary to UVvisible spectrophotometry. The two methods were explored by performing elemental analysis of four water samples testing for calcium, magnesium, iron, and sodium concentrations. Atomic absorption showed trends in relative concentrations of calcium, magnesium, and sodium for all samples. The direct comparison of the two methods by testing for the presence of iron showed that UV-visible spectrophotometry was more precise by orders of magnitude compared with atomic absorption spectrophotometry for iron of the two instrumental methods. The water samples all test well within the limits of soft water, where the total sum of both calcium and magnesium concentrations together if greater than 1 mM is defined as hard water. Anything over 1 mM is considered hard water. The amount of iron in all four samples was tested and discovered that all of the local areas within and surrounding the Portland Oregon Metro Area have safe drinking water that has less than 0.3-ppm iron content. Comparison of the two methods showed that UV-visible spectrophotometry was the better method for iron quantitation only because of a poorly constructed iron standard calibration curve for atomic absorption. The atomic emission of sodium showed that elements that emit electromagnetic radiation when heated with a flame work best with atomic emission burner in the perpendicular orientation. The method of ion suppression was explored, but failed to live up to theory. Iron spectrophotometry showed that external calibration was the best calibration method for the UV-visible spectrophotometer. Comparison of the two methods showed few samples actually were measured at the limits of detection. However those samples at or slightly beyond the limit of detection were Kangen water sample or the Sandy well water, which was pre-filtered. The values at the limit of detection where taken to have at least something present, respectively. The two methods gave excellent results were calibration was performed properly, however the UV-visible spectrophotometry was preferred in the iron comparison. Introduction Four water samples from different sources were collected and prepared for analysis. The four sources are from Portland State University located in Portland Oregon, Gresham Oregon, Sandy Oregon (well water-pre-filtered), and Kangen from Gresham (processed). Water quality was determined by the relative concentrations of hard and toxic elements in the water samples. The two water-testing methods for quantitation were useful to give an

idea of which method was preferable. Atomic absorption/atomic emission spectrophotometry (AA/AE) was complimented by UV-visible spectrophotometry in testing the water samples for traces of minerals. The mineral concentrations analyzed were calcium, magnesium, sodium, and iron. These four minerals, depending on their relative concentrations, tell the health of the water samples. Softness or hardness of water was determined by whether or not the sum of the calcium and magnesium concentrations equaled one millimolar, or not for the water samples. The toxicity of the water samples was dependent upon whether or not the relative iron concentrations were equal to or greater than 0.3 ppm also. The method of atomic absorption spectrophotometry was based upon instrument sensitivity. Since transmittance was the parameter being measured the precision was thus based on the negative logarithmic ratio of transmittance of hollow-cathode lamp to the amount of radiation upon excitation. Absorbance equals this logarithmic ratio, respectively. Upon performing a run with the AA/AE the sample were aspirated into a flame ionization chamber with ignited acetylene and oxygen ionizing the sample into its constituent atoms. The hollow-cathode lamps specific for each element emitted electromagnetic-radiation at the same frequency as the sample atoms emitted, which in this case was the alkali earth metals and iron. Those augmented frequencies were detected by a monochromator and sent to the detector. The detector measured the amount of transmittance that was initially sent from the cathode light beam that was augmented by the atomic irradiance of the tested element ratioed with the irradiance of just the cathode itself. However, if no atoms are present to absorb heat they could not emit a frequency (electromagnetic radiation) and the hollow-cathode lamp frequency would be unaugmented resulting in no detection at all. This ratio, the negative logarithm of the ratio of the total irradiance to the hollow-cathode irradiance was quantitated. This logarithmic ratio called absorbance was used to produce calibration curves for atomic species with known concentrations by plotting the absorbance values versus the analytical concentrations. The unknown samples were compared with their respective plots and the relative atomic concentrations determined. This method did not require any external reagents to bring the analyte to a measured response, except when potassium chloride was added as an ion-suppressant for the perpendicular pathway of the sodium atomic emission spectrophotometry. Ion suppression created an atmosphere with excess electrons pushing sodium into a reduced neutral state with no charge, which was termed the atomic state. The flame excitation promoted the outer valence electrons to the 3p electron orbital so that the return to 3s electron orbital caused emission of electromagnetic radiation unique to each atoms quantum spectrum that the monochromator lamp detected. This gave the instrumental method for quantiation. The UV-visible spectrophotometer operated according to Beers law given below, A=bc 1 where, A is the absorbance of the species (-log10(I/I0)), is the molar absorptivity coefficient (1/M cm), b is the path length (1 cm), and c is the concentration (M). The block diagram of the UV-visible spectrophotometer consisted of an ultraviolet visible range light source that passed through a cuvette into the transducer that converts the light change into a current at the detector and that signal gets sent to the PC. The measured absorbance values

from the iron standards get plotted versus the analytical concentrations giving the iron standard calibration curve. All the water samples are then measured with the UV-visible instrument absorbance values and the iron standard calibration curve gives the concentrations. The chemical reaction that was required to bring the iron to a ferrous Fe 2+ reduced state from its ferric Fe 3+ oxidized state was the reaction of hydroxylamine with ferric iron. After that 1,10-phenanthroline indicator was used to complex with ferrous iron with 1,10-phenanthroline presented below. 1,10-phenanthroline

Fe2+ + 3 phen ----- Fe (phen)32+ Iron in the water standards and water samples was in an oxidized state of Fe3+. Thus, hydroxylamine was added to bring the ferric iron into the reduced state of Fe2+. The excess reducing agent drove the ferric iron into the Fe2+ state due to la Chateliers principle of chemical equilibrium. Also, it was necessary to remove the solutions from the open air by placing lids on all the standard stock solutions and samples to stop the ferrous iron from re-oxidizing. The ferrous iron was then complexed with 1,10-phenanthroline to produce iron complexed 1,10-phenanthroline that underwent color transitions when more of the 1,10-phenanthroline was complexed the ferrous iron. The maximal wavelength for 1,10-phenanthroline was at 504 nm and the resulting calibration and sample absorbencies were recorded at that maximal. The sodium acetate worked as the buffer mediating the pH at 3.5. This was because 1,10-phenanthroline cannot operate outside the pH range of 3-9, respectively. The best way of dealing with controlling the pH of 3.5 was to ensure that sodium acetate buffer was added in excess as was done. This set the control for the experiment and gave excellent results in the results and discussion section. Experimental Atomic Absorption/Atomic Emission Standard solutions of 0.02518 M calcium, 0.002506 M magnesium, 0.043635 M sodium, and 0.00051 M iron were prepared. Four water samples from different sources were collected and prepared for analysis. The four sources are from Portland State University located in Portland Oregon, Gresham Oregon, Sandy Oregon (well water-pre-filtered), and Kangen from Gresham (processed). To five dry beakers was added 250 ml of DI water then there was added 0.5 ml calcium stock solution to beaker two, 1 ml calcium stock solution to beaker three, 1.5 ml calcium stock solution to beaker four, and 2 ml of calcium stock

solution to beaker five. The concentrations of calcium in the standards for beakers two, three, four, and five were calculated and recorded. The atomic absorption spectrometer was setup and the absorbances of the standards were measured. The full set of standards and unknown samples were measured before switching to another element with the Shimadzu 6300 AAS AA/AE Spectrophotometer. Next any unknown samples with measured absorbance too large were diluted. 250 ml of DI water was added to five dry beakers. 250 ml of DI water was added to five dry beakers then 0.5 ml magnesium stock solution was added to beaker two, 1 ml magnesium stock solution was added to beaker three, 1.5 ml magnesium stock solution was added to beaker four, and 2 ml of magnesium stock solution was added to beaker five. The concentrations of magnesium were calculated in the standards for beakers two, three, four, and five and recorded. Next, 0 ml, 5 ml, 10 ml, 15 ml, 20 ml, and 25 ml standard iron solution was pipetted into a series of six 50 ml volumetric flasks then each was filled to a total volume of 50 ml. The absorbencies were measured by atomic absorption spectrophotometry. The concentrations of the iron standards were calculated and the absorbance values recorded. Next, atomic emission was setup on the instrument and the sodium standard solutions were prepared. 250 ml of DI was added to six 400 ml beakers and then was added 0.5 ml sodium stock solution to beaker two, 1 ml sodium stock solution to beaker three, 1.5 ml sodium stock solution to beaker four, and 2 ml of sodium stock solution to beaker five, and 2.5 ml of sodium stock solution to beaker six. The concentration of sodium was calculated in the sodium standards and the absorbance values were measured and recorded. The atomic emission was run in the parallel position with the standards as well as the samples and the absorbencies recorded. Next the atomic emission burner was turned to the perpendicular position and the standards were run as well as the samples and the absorbencies recorded. 35 ml of 1 M KCl was added to the new sodium standards that were prepared exactly as previously stated running the atomic emission in the perpendicular position and then running the samples and recording the absorbencies. Iron Spectrophotometry (UV-visible) 5.1 e-4 M of standard iron solution was prepared and 0 ml, 0.5ml, 1ml, 1.5ml, 2ml and 2.5 ml of the 5.1 e-4 M standard iron solution was pipetted into six 50 ml volumetric flasks. Next 1 ml of 0.29 M hydroxylamine hydrochloride was pipetted into each flask. Then 5 ml of 1.2 M sodium acetate was pipetted into each of the six flasks. Then 5 ml of 5e-3M 1,10phenanthroline was pipetted into each of the six flasks. Each flask was then filled to a total volume of 50 ml. The concentrations in each standard were calculated and recorded. The blank was measured first of all with the Biowave III WPA spectrophotometer. Next the standards and the samples were measured and their respective absorbencies were recorded. The standards were plotted on a calibration curve and the samples absorbencies determined and recorded. Results and Discussion Atomic Absorption Atomic absorption spectrophotometry was compared with Ultraviolet-Visible spectrophotometry to compare the limits of detection for the two quantitative methods, thus determining the better instrument for analysis of water samples. The results from the

atomic absorption showed that calcium, magnesium, iron and sodium where present in most samples except for the Sandy Oregon Well water, because this water sample had been filtered prior to being collecting at the tap. All the other samples showed measurable amounts of all elements just listed. The better method depends on how the calibrations turned out and evidence of any noticeable trends from both the atomic absorption and UVvisible spectrophotometric experiments. Thus, samples with the highest concentration of one element were highest in all other elements as well. This trend and good results determined the better method. The measurement of iron was the only measurement made with the UV-Visible spectrophotometer, and it was compared with the iron measurement made from the atomic absorption spectrophotometer allowing for direct comparison of the two methods, respectively. Below, the calibration of calcium and magnesium were plotted in figure 1. The calibration curve showed that the standards where properly prepared, because the R squared values were close to one. Sodium, calcium and magnesium showed the highest concentrations of all the elements tested, but iron tested the lowest concentration. The concentrations of the samples were determined within their respective calibration curves; however choosing the right curve for sodium was based on the best fit of the perpendicular curve without ion suppressant over the other curves. The three plots of the sodium data showed that the perpendicular method was the best method for quantitation for sodium.
Fig 1. Calcium and magnesium standard calibration curves.

Calcium and Magnesium Calibration curves Absorbance(Relativ e) 0.2 0.15 0.1 0.05 0 -5 -0.05 0 5 10 concentration (uM) 15 20 25

y = 0.0086x - 0.0018 R2 = 0.9994 Calcium Magnesium Linear (Calcium) Linear y = 0.0031x - 0.0147 (Magnesium) R2 = 0.9978

1.) Calcium and magnesium were made to have the same concentrations so as to coincide on the same graph for comparison. The absorption is far higher in calcium than magnesium possibly due to a weak calcium/magnesium lamp in the atomic absorption instrument. The two give close to the same absorbance, but magnesium is still lower in absorbing than calcium. The calcium-magnesium lamp is the same lamp and does not get changed when measuring calcium and then magnesium, or vice versa and the lamp emits at a lower energy wavelength for magnesium as was seen here, respectively. Data table 1.Calcium standard calibration curve error analysis for atomic absorption. Calcium slope intercept Parameter 0.008555 -0.00185 Std Dev 6.03E-05 0.000738 R^2 0.999355 0.001651STD Dev (y) 1.) This gives the error of the slope and intercept from the standard calibration curve, along with the R squared value that tells the precision of the standards preparation. Data table 2. Magnesium standard calibration curve error analysis for atomic absorption. Magnesium slope intercept Parameter 0.003051 -0.01467 Std Dev 4.49E-05 0.000615 R^2 0.997841 0.000869STD Dev (y)

1.) This gives the error of the slope and intercept from the standard calibration curve, along with the R squared value that tells the precision of the standards preparation.

Atomic absorption with iron showed problems in the beginning with improperly prepared iron standards that made the iron calibrations questionable when measuring concentrations at the limit of detection. All the samples showed concentrations at the lower end of the calibration. The reliability of the iron calibration curve for atomic absorption in figure two was questionable due to the R squared value being fairly less than one. The iron calibration curve showed that there was a 0.0027 standard deviation in absorbance of the yintercept. That correlates to a 21 uM concentration. When the iron values were analyzed in data table four, the iron concentrations were high and low for replicate measurements. That was due either to improper flame positioning of the atomic absorption instrument, or improper standard preparation. When the negative of the standard deviation was taken the concentration became zero for each sample and when the positive deviation was taken the concentration became extremely high. That means the data was highly unreliable even with such a high R squared value because the concentrations were on the low end of the calibration curve as well as the limit of detection for the atomic absorption instrument.
Fig 2. Standard iron calibration curve for atomic absorption.

Fe Calibration Curve 0.15 Absorbance 0.1 0.05 0 -0.05 0 50 100 150 200 250 300

y = 0.0004x - 0.0057 R2 = 0.9766 Series1 Series2 Linear (Series1)

Fe concentration (uM)
1.) Calibration curve was constructed to determine the amount of iron in the water samples, however the limit of detection for the atomic absorption instrument made concentration determination difficult. The deviation from linearity caused a standard deviation in the slope of 1.73e-5 and 0.002689 for the intercept. Data table 3. Standard iron calibration curve error analysis for atomic absorption. Iron slope intercept Parameter 0.000447 -0.00566 Std Dev 1.73E-05 0.002689 R^2 0.976617 0.006358STD Dev (y) 1.) This gives the error of the slope and intercept from the standard calibration curve, along with the R squared value that tells the precision of the standards preparation. Data table 4. Iron, calcium, and magnesium concentrations in their respective water samples. 1.) Iron concentration (uM) Std Dev in samples (uM) Kangen water 18.25 2.136000936 Kangen water 16.5 Kangen water 20.75 PSU Tap Water 18 1.127312438 PSU Tap Water 15.75 PSU Tap Water 17 Gresham Tap water 25.25 5.507570547 Gresham Tap water 20.25 Gresham Tap water 14.25 Sandy Well water 15 3.905124838 Sandy Well water 22

Sandy Well water 2.) Calcium Kangen water Kangen water Kangen water PSU Tap Water PSU Tap Water PSU Tap Water Gresham Tap Water Gresham Tap Water Gresham Tap Water Sandy Well water Sandy Well water Sandy Well water concentration (uM)

21.5 Std Dev in samples (uM) 38 0.577350269 37 38 18 1 19 20 16 0.577350269 16 15 5 1.154700538 5 3

3.) Magnesium concentration (uM) Std Dev in samples (uM) Kangen Water 53.2257 0.290250013 Kangen Water 53.5161 Kangen Water 52.9356 PSU Tap Water 30.5162 0.331001289 PSU Tap Water 30.7096 PSU Tap Water 31.1612 Gresham Tap Water 24.3226 0.985471995 Gresham Tap Water 25.613 Gresham Tap Water 26.258 Sandy Well Water 2.32258 0.409733939 Sandy Well Water 3.03226 Sandy Well Water 3.03226 1.) Iron showed concentrations that were equal in all samples with the atomic absorption instrumental. This was due to large error in the iron standard preparation that resulted in an R squared value of 0.97 that was not the 0.99 value in the calcium and magnesium standard calibrations. The iron concentrations show that the highest deviations occur for itself in comparison to the other elements, calcium and magnesium. 2.) Calcium showed the highest concentrations in the kangen water and the lowest concentrations in the filter Sandy well water. The standard deviations are not nearly as high due to R squared value of 0.99 for calcium. 3.) Magnesium showed the highest concentrations in the kangen water and the again the lowest concentrations in Sandy well water. The standard deviations are not as high due to the R squared value of 0.99 for magnesium.

The measured values for the atomic absorption concentrations of calcium, and magnesium in the samples showed a trend of Kangen, Portland State University (PSU), Gresham, and then Sandy well water in that order from highest relative concentrations to lowest relative concentrations. The iron standard calibration was poorly conducted where at the initial region of the iron calibration curve the concentrations are limited by the standard error in the curve itself. The limit of detection was reached despite the error analysis, since the calibration curve showed significant error, thus the concentrations were still unknown. Water hardness was a measure of the concentrations of both calcium and magnesium in the water samples where the sum of these two minerals, if equal to or greater than one millimolar within a given water sample, the water sample was considered hard. [2] Anything below the value of 1 mM was considered soft water. The calculated hardness of the water samples was tabulated in data table five below.
Data table 5. The measured water hardness of the water samples. 1.) [Ca]+[Mg]>1mM then have hard water Hardness or Softness of the water samples Kangen PSU Gresham concentration (mM) Hardness 0.091soft water 0.05soft water 0.041soft water

Sandy

0soft water

2.) If concentraion greater than 0.3mg/L(ppm), toxic. Iron Toxicity concentration (mg/L) Toxicity Kangen 1Toxic PSU 0.95Toxic Gresham 1.12Toxic Sandy 1.01Toxic 1.) The sum of the determined concentrations of the calcium and magnesium from atomic absorption spectroscopy defines whether or not water is hard or soft dependent upon if the sum of the calcium and magnesium concentrations is higher than one millimolar. This experiment showed that all water samples fall below the definition of hardness and are thus considered soft water. This is because Kangen had a concentration of 0.091mM compared to if it may have been found to be 1mM and the same for the rest of the water samples. 2.) Iron toxicity is defined by an iron concentration greater than 0.3 mg/L in a given sample of water. Atomic absorption spectroscopy showed that the relative concentrations of iron in all samples is extremely toxic. The values are listed above. However, consideration of the calibration curve thus far renders this iron data useless, respectively.

The water hardness results reflected a trend in all samples. Kangen water had the highest value but was still soft water as were all the other samples. This was determined from the calcium and magnesium concentrations from data table four above. The iron concentrations determined from iron atomic absorption spectrophotometry located table four showed that all the samples were toxic, or over the 0.3-ppm regulation limit. [2] However, if that were true the water would not be used. The propagation of error or uncertainty within the iron atomic absorption measurements deserved a full treatment with a least squares analysis with production of confidence intervals. The values are listed below in data table six for each water sample.
Data table 6. Fe Atomic Absorption 50% and 90% confidence intervals. Gresham 50% level 17.739+ 0.765*11.45 Gresham 95% level 17.739+2.306*11.45 Kangen Kangen Sandy Sandy 50% level 95% level 50% level 95% level 16.47+ 0.765*11.47 16.47+ 2.306*11.47 17.37+ 0.765*11.45 17.37+ 2.306*11.45 26.5,9 44.1427,-8.67 25.3,7.7 42.92,-10 26.13,8.61 43.77,-9.034

PSU 50% level 15.05+ 0.765*11.486 23.84,6.26 PSU 95% level 15.05+ 2.306*11.486 41.54,-11.44 1.) These intervals were calculated by taking the measured mean value of y for the replicates measurements made for each water sample and plugged into a excel spreadsheet that produced the derived X and the Sx (standard deviation in X). The values of the confidence intervals were taken from a table in reference [2] that can be found in the references. The least squares spread sheet is found in the appendix under data table 6.

The confidence intervals were calculated by using a least squares analysis with associated student t values to compute the confidence intervals. [1] The method produced reasonable values that reflect those seen in figure two, iron atomic absorption calibration curve that showed the intervals and the confidence in trusting those intervals. The Gresham water sample produced a value of ~17.7 uM of Fe with a 50% confidence interval ranging from 26 uM to 5.9 uM. That means the concentration varies quite significantly and at the limit of detection for the atomic absorption instrument the true concentration of iron was anybodies guess. This was seen for the Kangen, Sandy, and PSU water samples as well. Another method of quantitation was in order. Comparing the atomic absorption spectrophotometry and UV-visible spectrophotometry quantitation methods for determining iron concentrations showed that the UV-visible method was superior. The UV-vis spectrophotometer showed reasonable values that were within the allowed limits for iron concentrations in all the water samples. The highest

concentration of iron determined by spectrophotometry was for the Gresham water sample. It had a concentration of ~0.2 ppm and the regulated limit is 0.3 ppm. In terms of trends of the healthy desirable calcium, magnesium, and sodium levels in our water iron does not follow that same trend, where Kangen was first, PSU second, Gresham third, and then Sandy. The trend here for iron contamination was Gresham had the highest relative concentration, PSU, Sandy, and the lowest was the Kangen water.
Data table 7. Comparison of the Atomic Absorption spectroscopy to UV-visible spectrophotometry. If concentraion greater than 0.3mg/L, toxic. Fe Spectrophotometry Kangen Water PSU Tap Water Gresham Tap Water Sandy Well Water Ave concentration (uM) Concentration (mg/L) -0.005225 2.9275 3.4985 0.037733 toxicity -0.000292safe 0.163486safe 0.195374safe 0.002107safe

Atomic Aborption Fe Ave concentration (uM) concentration (mg/L) toxicity Kangen Water 18.5 1.03313Toxic PSU Tap Water 16.9167 0.944713Toxic Gresham Tap Water 19.9167 1.11225Toxic Sandy Well Water 19.5 1.08898Toxic 1.) UV-visible spectrophotometry showed excellent results in comparison to the atomic absorption spectrometer. All the water samples tested with the UV-vis spectrophotometer showed iron concentrations within the allowed levels. This makes sense because the water quality is regulated by the city and levels higher than 0.3 ppm would not be allowed. The atomic absorption failed in comparison giving false toxic absorbance values that were due to poor iron standard preparation resulting in an unuseable calibration curve.

The overall limits of detection of all of the samples were either right at the limit of detection or slightly above it, thus there was no need to perform a statistical analysis to find a concentration beyond the limit of detection, which could have been outside the calibration curves range, respectively. The UV-visible spectrophotometer was excellent for measuring the iron concentrations and was relatively easier to use than the Shimadzu atomic absorption/atomic emission spectrophotometer (AA/AE). However, if the standards were prepared properly then the AA/AE may be just as precise because it gave such trendy results for the calcium and magnesium as well as for the sodium, which is next in the discussion. Atomic Emission Then determination of the concentration of sodium was performed in the exact same manner as was done for the iron, calcium, and magnesium with the burner orientation in the parallel position, except no sodium hollow-cathode lamp was involved in atomic emission for sodium. The parallel burner position showed a dynamic range that resulted in a quadratic fit of the sodium standard data. The concentrations from the water samples themselves may be calculated from this burner orientation, but due to lack of linearity the determined concentrations from this direct calibration may result in concentrations that are different from the true sodium concentrations by orders of magnitude.
Data table 8. Sodium concentrations in their respective water samples. The Atomic Emission spectrometer was in the parallel burner orientation and the standards for sodium were run. The resulting data was tabulated below along with the data for the water samples. Volume Added Concentration uM Absorbance 0 0 0.0003

0.0002 0 0.132 0.1316 0.1299 0.2214 0.221 0.2187 0.2739 0.2722 0.2729 0.3115 0.3134 0.317 PSU 0.3144 PSU 0.3131 PSU 0.3137 Gresham 0.2868 Gresham 0.2826 Gresham 0.2824 Sandy -1.0005 Sandy -1.0005 Sandy -1.0005 Kangen n/a 0.3886 Kangen n/a 0.3781 Kangen n/a 0.3733 1.) Sodium standards were measured the with Atomic Emission burner orientation in the parallel position and the absorbance values were recorded at the respective analytical concentrations. Then the water samples were run in the parallel position and their data was recorded with their respective standard data. Fig 3. Standard sodium calibration curve for atomic emission with the burner in the parallel orientation.

0 0 0.5 0.5 0.5 1 1 1 1.5 1.5 1.5 2 2 2

0 0 87 87 87 174 174 174 260 260 260 346 346 346 335.2 330.358 332.546 265.836 258.226 257.873 -413.419 -413.419 -413.419

Parallel NaCl 0.4 0.3 0.2 0.1 0 0 50 100 150 200 250 300 350

y = -2E-06x 2 + 0.0016x + 0.0028 R2 = 0.9984

Series1 Series2 400 Poly. (Series1)

1.) Parallel orientation produced false readings due to the lack of linearity despite the values of the samples ambiguously falling on the curve. The samples themselves are not in the correct oxidation state to be analyzed by the atomic absorption spectrometer.

The sodium calibration with the atomic absorption burner in the parallel orientation resulted in a dynamic range that was fit with a quadratic equation. All of the samples fell on this curve. The main reason for a dynamic range rather than a linear range was because the sodium ions were self-absorbing. [2] This makes sense because at higher and higher concentrations of sodium atoms the relative amount of energy remains the same that was supplied from the flame. If the energy supplied to vaporize the sample did not change, but the concentration went up that means more of the emitting 3p->3s resonance transitions gets absorbed by the neighboring ground state atoms and dissipated as heat instead of being emitted as electromagnetic radiation. This phenomenon directly produced the effect seen in figure three. Thus, anytime this dynamic response is seen for atomic emission it is because of self-absorption of the emitting photons from the sodium atoms in the sample themselves.
Data table 9. Sodium concentrations in their respective water samples. The atomic emission spectrometer was in the perpendicular burner orientation and the standards for sodium were run. The resulting data was tabulated below along with the data for the water samples.

Perpindicular NaCl

Abs 0 -0.0003 0 -0.0007 0 -0.0006 87 0.1244 87 0.1254 87 0.1258 174 0.2546 174 0.2539 174 0.2515 260 0.3675 260 0.368 260 0.3691 346 0.4903 346 0.4904 346 0.4889 Gresham 174.571 0.2463 Gresham 166.929 0.2356 Gresham 163.786 0.2312 PSU Tap 174.429 0.2461 PSU Tap 173.714 0.2451 PSU Tap 174.571 0.2463 Kangen 212.143 0.2989 Kangen 213.357 0.3006 Kangen 208.643 0.294 Sandy -715.571 -0.9999 Sandy -715.571 -0.9999 Sandy -715.571 -0.9999 1.) Sodium standards were measured with the atomic emission burner orientation in the perpendicular position and the absorbance values were recorded at their respective analytical concentrations. Then the water samples were run in the perpendicular position and their data was recorded with the respective standard data.

uM 0 0 0 0.5 0.5 0.5 1 1 1 1.5 1.5 1.5 2 2 2

Fig 4. Standard sodium calibration curve for atomic emission with the burner in the perpendicular orientation.

Perpindicular NaCl 0.6 0.4 0.2 0 -0.2 0 50 100 150 200 250 300 350 400 y = 0.0014x + 0.0019 R2 = 0.9997 Series1 Series2 Linear (Series1)

concentration (uM)

1.) Perpendicular orientation produced excellent readings due to the lack of dynamic response that was seen in figure three. The samples follow a linear response that was determined from direct calibration. The R squared value of 0.999 makes this calibration curve reliable.

The sodium calibration curve in the perpendicular position produced a linear range that did not appear to be affected by self-absorption from the samples themselves. The linearity was a good indicator that the standard concentrations were exactly proportional to the absorbencies that were detected from the atomic emission data. All of the data from data table nine was plotted in figure four. Since the response was linear this direct calibration was used to determine the actual concentrations of sodium in the water samples. Sodium matches the trend that was seen for the atomic absorption part of the experiment, which was Kangen had the highest relative concentration of sodium, then PSU, Gresham and lastly Sandy with the lowest sodium concentration.
Data table 10. Standard iron calibration curve error analysis for atomic emission.

slope intercept Parameter 0.001415 0.001877 Std Dev 6.53E-06 0.001386 R^2 0.999723 0.003094Std Dev (y) 1.) This gives the error of the slope and intercept from the standard calibration curve, along with the R squared value that tells the precision of the standards preparation

The limit of detection was reached for sodium with atomic emission requiring that some sort of value be found beyond the initial portion of the linear response calibration curve. Thus, data table ten above provided the error analysis of the slope, and intercept. When performing error analysis the limitation laid within the largest standard deviation in whichever variable it occurred. In this case the highest standard deviation was from the intercept for the direct calibration of the sodium standards. The standard deviation was 0.001386 abs in the y-intercept. The y-intercept itself had a value of 0.001877 abs. This means the value of the Sandy water sample was 0.001386 abs in the y-intercept. This gave values of either 0.003263 abs, or 0.000491 abs. Using the equation in figure four gave a concentration of 0.973571 uM, or 1.00643 uM. That means the Sandy water sample could be ~ 1 uM sodium present. Considering that filtered water always has some concentration of analytes that makes the most sense. The other concentrations are 1 uM as well.
Data table 11. Sodium concentrations in their respective water samples. The atomic emission spectrometer was in the perpendicular burner orientation and the standards for sodium were run with potassium chloride ionization suppressant. The resulting data was tabulated below along with the data for the water samples. Perpindicular with Ion Suppresant uM Abs 0 0 -0.0007 0 0 -0.0011 0 0 -0.0011 0.5 87 0.1113 0.5 87 0.1121 0.5 87 0.1128 1 174 0.2137 1 174 0.2131 1 174 0.213 1.5 260 0.3053 1.5 260 0.3099 1.5 260 0.3085 2 346 0.3901 2 346 0.3933 2 346 0.3919 2.5 432 0.4708 2.5 432 0.4813 2.5 432 0.4762 PSU 159.909 0.1882 PSU 157 0.185 PSU 158.182 0.1863 Gresham 142.364 0.1689 Gresham 142.273 0.1688 Gresham 144.182 0.1709 Kengan 198.182 0.2303 Kengan 199.636 0.2319 Kengan 198.727 0.2309 Sandy -919.182 -0.9988 Sandy -919.182 -0.9988 Sandy -919.182 -0.9988 1.) Sodium standards with KCl ion suppressant were measured with the atomic emission burner orientation in the perpendicular position and the absorbance values were recorded at their respective analytical concentrations. Then the water samples with KCl were run in the perpendicular position as well and their data was recorded with the respective standard data.

Fig 5. . Standard sodium with KCl ion suppressant calibration curve for atomic emission with the burner in the perpendicular orientation.

Perpindicular NaCl 0.6 0.5 0.4 0.3 0.2 0.1 0 -0.1 0 y = 0.0011x + 0.0123 R2 = 0.9966 Series1 Series2 Linear (Series1) 100 200 300 400 500

1.) The perpendicular burner position produced a dynamic range when the sodium with potassium chloride ion suppressant was added. All of the samples absorbances are within the linear response range of the calibration curve.

The perpendicular burner orientation of sodium with potassium chloride ion suppressant produced lower concentrations of sodium than sodium without potassium chloride. The absorbance values for the Sandy water samples also showed a direct relation between data table eleven and nine. The absorbance values were -0.999 abs for both sets of calibrations. That means adding potassium chloride in those samples had no effect. Essentially, no sodium was present in the Sandy well water to begin with and the potassium chloride had nothing with which to interact. Thus, the other samples did have sodium to interact with and figure five showed the same self-absorption curve that was noticed when the burner was in the parallel position. However, the effect of an added salt to the flame resulted in a lower temperature non-homogeneous flame and at lower temperatures resonance lines are broadened with a dip in the center. [2] The detector measures the signal being sent directly from the sodium itself. The resulting non-homogeneous temperature results in a lower detection limit for the sodium. This was because the detector takes measurements over an interval of time. When the temperature was homogeneous the emitted photons of the excited state had a continuous signal, but when the flame was non-homogeneous the detector measured less emitted photons over the same interval of time. That resulted in a dynamic response exactly like the self-absorption. Thus, flame temperature was not hot enough to handle the excess potassium chloride being delivered to the burner. However, the calibration curve for the sodium with potassium chloride can still be fit with a linear curve, but it was not chosen as the actual sodium concentration data. The actual sodium concentration data came from the sodium in the perpendicular orientation without ion suppressant potassium chloride. The error analysis for the sodium standard calibration curve with ion suppressant is below.
Data table 12. Sodium with potassium chloride standard calibration curve with error analysis of atomic emission. slope intercept Parameter 0.001098 0.012282 Std Dev 1.59E-05 0.004174 R^2 0.996643 0.009971Std Dev (y) 1.) This gives the error of the slope and intercept from the standard calibration curve, along with the R squared value that tells the precision of the standards preparation

Iron Spectrophotometry The iron spectrophotometry experiment yielded results at better precision than atomic absorption as was seen when comparing the concentrations of the same samples of water with both the atomic absorption/atomic emission spectrophotometer and the ultra-violet

visible spectrophotometer. The concentrations of iron from the standard and concentrations from the water samples were recorded in data table 13 below. The standard calibration response was linear allowing for determination of the iron concentrations in the water samples, respectively.
Data table 13.Iron Spectrophotometry standard calibration data with corresponding water sample data. Fe Spectrophotometry mL Iron added Concentration (uM) Abs 0 0.00E+00 0 0.00E+00 0 0.00E+00 0.5 5.08E+00 0.5 5.08E+00 0.5 5.08E+00 1 1.02E+01 1 1.02E+01 1 1.02E+01 1.5 1.53E+01 1.5 1.53E+01 1.5 1.53E+01 2 2.03E+01 2 2.03E+01 2 2.03E+01 2.5 2.54E+01 2.5 2.54E+01 2.5 2.54E+01 -3.09E-02 -1.34E-01 -1.34E-01 2.78E-01 2.34E+00 3.47E+00 3.47E+00 2.43E+00 3.47E+00 3.37E+00 3.37E+00 3.784 1.75E-01 -3.09E-02 -3.09E-02 Concentration (ppm) 0 0 0 0.07 0.07 0.08 0.12 0.1 0.11 0.17 0.17 0.17 0.21 0.18 0.2 0.26 0.27 0.26 0.01 0.01 0.01 0.01 0.03 0.05 0.05 0.03 0.05 0.04 0.04 0.05 0.01 0.01 0.01 0.00E+00 0.00E+00 0.00E+00 2.84E-01 2.84E-01 2.84E-01 5.70E-01 5.70E-01 5.70E-01 8.54E-01 8.54E-01 8.54E-01 1.13E+00 1.13E+00 1.13E+00 1.42E+00 1.42E+00 1.42E+00 -1.73E-03 -7.48E-03 -7.48E-03 1.55E-02 1.31E-01 1.94E-01 1.94E-01 1.36E-01 1.94E-01 1.88E-01 1.88E-01 2.11E-01 -9.77E-03 -1.73E-03 -1.73E-03

Kangen Kangen Kangen Kangen PSU Tapwater PSU Tapwater PSU Tapwater PSU Tapwater Gresham Tapwater Gresham Tapwater Gresham Tapwater Gresham Tapwater Sandy Well Sandy Well Sandy Well Iron

slope intercept Parameter 0.009695394 0.01 Std Dev 0.000296139 0 R^2 0.985292317 0.01STD Dev (y) 1.) The iron standard calibration curve from UV-visible spectrophotometry. The concentrations of iron present in the standards and in the water samples are recorded in uM and ppm units. The error analysis for the calibration data is at the bottom. The error in the slope was the highest of the parameters measured. This time Kangen and Sandy water samples had the least amount iron present. Gresham had the highest concentration of iron. Fig 6. Iron spectrophotometry calibration curve in uM units.
Fe Spectrophotometry Calibration Curve

y= 0.0097x +0.01 3 1 0.3 0.2 0.1 0 -5.00E+ 00 0.00E+ 00 -0.1 5.00E+ 00 1 .00E+ 01 1 .50E+ 01 2.00E+ 01 2.50E+ 01 3.00E+ 01 R2 = 0.9853 Fe Spectrophotometry Samples Linear (Fe Spectrophotometry)

c oncentration ( uM)

1.) The iron standard solutions were plotted with the water samples. The water samples were all within the linear response range of the calibration curve. The error of the derived equation was recorded at the bottom of data table 13.

Fig 7. Iron spectrophotometry calibration curve in ppm units.

Fe Spectrophotometry Calibration Curve y = 737x +0.01 2 0.1 1 0.3 0.2 0.1 0 -2.00E-01 0.00E+ 00 -0.1 2.00E-01 4.00E-01 6.00E-01 8.00E-01 1 .00E+ 00 1 .20E+ 00 1 .40E+ 00 1 .60E+ 00 R2 = 0.9856 Fe calibration Samples Linear (Fe calibration )

conc entration ( ppm)

1.) The iron standard solutions were plotted with the water samples. The water samples were all within the linear response range of the calibration curve. The error of this plotted was found in figure six, essentially this figure is that of figure six except for the change to ppm notation, respectively.

Figures six and seven above showed the linearity of the iron spectrophotometric standard calibration curves. One calibration curve was plotted with units of micromolar and the other plotted with units of parts-per-million notation in order to relate the iron concentrations determined from the UV-visible spectrophototmeter with the federally regulated limit of 0.3 ppm (mg/L) of iron. Data table 13 shows the error analysis in the calibration curve in figure six above. The relative error applies to both figure six and seven because they are both plotted from the same data found in data table thirteen, of course. The water samples in data table thirteen fall right on the very edge of the curve in figure six. All the water samples had a standard deviation in the slope of 0.0003 abs that corresponded to an error of virtually nothing. That means the calculation was reasonable and was accepted because the Kangen and Sandy water samples were both filtered with soft water filters prior to testing. Since replicate measurements were made in data table 13, all of the replicate standard deviations were recorded along with the average concentrations for each water sample in data table 14 below. The standard deviation of the iron spectrophotometry showed no greater deviation than 0.63 uM in iron concentration determinations. That means the average concentration recorded in the final row of table 14 below was the respective Std Dev value in the third column. As was seen from analyzing table 14 the standard deviation in the spectrophotometry was small compared to the atomic absorption method of iron concentration determination.
Data table 14. Iron spectrophotometry average concentration of iron in the water samples. Fe Spectrophotometry concentration (uM) Std Dev in samples (uM) Ave conc (uM) Kangen -3.09E-02 0.2 Kangen -1.34E-01 Kangen -1.34E-01 Kangen 2.78E-01 PSU Tapwater 2.34E+00 0.63 PSU Tapwater 3.47E+00 PSU Tapwater 3.47E+00 PSU Tapwater 2.43E+00 Gresham Tapwater 3.47E+00 0.2 Gresham Tapwater 3.37E+00

-0.005225

2.9275

3.4985

Gresham Tapwater Gresham Tapwater Sandy Well Sandy Well Sandy Well

3.37E+00 3.784 1.75E-01 -3.09E-02 -3.09E-02

0.12

0.037733

Atomic Absorption Iron concentration (uM) Std Dev in samples (uM) Ave conc (uM) Kangen water 18.25 2.14 18.5 Kangen water 16.5 Kangen water 20.75 PSU Tap Water 18 1.13 16.9167 PSU Tap Water 15.75 PSU Tap Water 17 Gresham Tap water 25.25 5.51 19.9167 Gresham Tap water 20.25 Gresham Tap water 14.25 Sandy Well water 15 3.91 19.5 Sandy Well water 22 Sandy Well water 21.5 1.) The concentrations from data table 13 were taken and the standard deviation was analyzed for each respective set of replicates taken and the average concentration from those concentrations was recorded above in micromolars. 2.) The direct comparison of the iron spectrophotometry and atomic absorption spectrometry were tabulated in this data table for direct comparison of both methods. The atomic absorption method yield bad results in comparison to the spectrophotometry method.

In directly comparing iron UV-vis spectrophotometry with atomic absorption the average concentrations are the standard deviations from table 14 above for the average concentrations in table 15 below. Still none of the water samples tested with the spectrophotometry method were hazardous to human health and as was seen previously atomic absorption did not give any results for iron.
Data table 15. Direct comparison of spectrophotometry with atomic absorption spectrometry for the amount of iron in water samples. If concentraion greater than 0.3mg/L, toxic. Fe Spectrophotometry Kangen Water PSU Tap Water Gresham Tap Water Sandy Well Water Ave concentration (uM) Concentration ppm (mg/L) toxicity -0.005225 0safe 2.9275 0.16safe 3.4985 0.2safe 0.037733 0safe

Atomic Aborption Fe Ave concentration (uM) concentration ppm (mg/L) toxicity Kangen Water 18.5 1.03Toxic PSU Tap Water 16.9167 0.94Toxic Gresham Tap Water 19.9167 1.11Toxic Sandy Well Water 19.5 1.09Toxic 1.) The spectrophotometry method yielded average concentrations of iron present only the Gresham, and PSU water samples. Kangen and Sandy water samples had very low responses. The concentrations were tabulated in ppm notation. The results showed that all water samples in the table are safe for consumption with the UV-visible spectrophotometry method. 2.) The atomic absorption method yielded average concentrations of iron present in all water samples with all water samples being extremely toxix. The means the atomic absorption method was not as precise as measuring the iron concencetration levels as the spectrophotometry method.

The values of the molar absorptivity coefficient for iron helped to discern if iron was the actual analyte tested. Thus using Beers law the values of the absorptivity coefficient were calculated using the iron standard calibration data. Typically, the y-intercept is taken as the value for the molar absorptivity coefficient, of course. The value calculated below was 11595 (1/M cm).
Data table 16. Iron molar absorptitivity coefficient (1/M cm)

Fe Molar Absorptivity Coefficient Abs coefficient values (1/M cm) Conc (uM) 11466.9 5.08E+00 10849.3 1.02E+01 10405.5 1.53E+01 10292.4 2.03E+01 10138.4 2.54E+01 1.) Tabulated values represent the standard calibration curve data for determining the molar absorptivity coefficient of iron. Essentially the beers law a=bce was used for all the data points on the Fe spectrophotometry calibration curve. The values of e versus c were plotted and the linear regression was performed. The resultant R squared value was 0.89 because of limited calibration points at higher concentrations. The intercept were concentration was zero was taken as the Fe molar absorptivity coefficient. Fig 8. Iron molar absorptitivity Coefficient extrapolation

Fe Molar Absorptivity Coefficient Absorptivity coefficient values at successive conc 12000 11500 11000 10500 10000 9500 0.00E+0 5.00E+0 1.00E+0 1.50E+0 2.00E+0 2.50E+0 3.00E+0 0 0 1 1 1 1 1 Conc (uM)
1.) Standard calibration plot from the concentration(uM) and absorptivity coefficients calculated from the standard calibration data in data table 13 from Beers law of a=bce and plotted in this figure. The linear regression shows a y-intercept were the concentration of iron equals zero to be 11595 (1/M cm), respectively.

y = -63.276x + 11595 R2 = 0.8949 Fe absorptivity coefficents plotted versus concentration Linear (Fe absorptivity coefficents plotted versus concentration)

The complete summary of all the water testing results showed that atomic absorption/ atomic emission spectrophotometry coupled with UV-vis spectrophotometery were complimentary instrumental techniques for analyzing environmental water concerns. After analyzing all the results and discussions the meaning of the results show that AA/AE and UV-visible spectrophotometry both have merits. However in terms of instrumental use the UV-visible spectrophotometer was more practical to work with. The AA/AE machine was difficult to get to work properly and required a thorough troubleshooting knowledge of the machine and all its weaknesses. The cost of running acetylene and the cost of the instrument lamps and other various parts make it undesirable for use in a fast-paced analytical laboratory. Whereas the UV-visible spectrophotometer was easier to use. The only complications were with making sure the cuvette was inserted in a certain repetitive orientation time and again while measuring the samples. The limits of detection were easy to analyze because all of the zero values in the iron spectrophotometry section were located on the external calibration curve or right at the beginning of the calibration curve. This translated into the concentration being just the standard error in the linear regression equation, respectively. Working with the two complimentary instruments allows for further future exploration with these instruments, respectively.

Reference

1.) D, C, Harris,. Quantitative Chemical Analysis 7th ed. W H Freeman NY, 2007. 2.) Skoog, West, Holler, Crouch Fundamental of Analytical Chemistry. 8th Ed. 2004. p856. 3a.) Atkinson, D, PHD,.Webnotes for Instrumental Analysis Lab-Chemisty 427. Determination of Ca, Mg, Fe, and Na by Atomic Spectrophotometry. http://web.pdx.edu/~atkinsdb/teach/427/webnotes.htm. Accessed 07-05-2011. 3b.) Atkinson, D, PHD,.Webnotes for Instrumental Analysis Lab-Chemisty 427. Spectrophotometric Determination of Iron in Drinking Water. http://web.pdx.edu/~atkinsdb/teach/427/webnotes.htm. Accessed 07-05-2011.

Appendix Data table 6/with associated least squares spread sheet.

Least Squares Spreadsheet Highlight cells B24:C26 x Type"=LINEST(C4:C21,B4:B21, TRUE,TRUE) y 0 0 0 51 51 51 112 112 112 153 153 153 204 204 204 255 255 255 -0.0038 -0.0021 -0.0037 0.0131 0.0143 0.0151 0.0399 0.0404 0.0358 0.0658 0.0701 0.0632 0.0961 0.0978 0.0872 0.0972 0.1083 0.1024

For PC, press CTRL+SHIFT+ENTER

m Sm R^2 n= Mean y= Sum(Xi-meanX)^2

0.000446874 1.72869E-05 0.976616726

-0.005660165b 0.002689158Sb 0.006358004Sy

18B28=COUNT(B4:B13) 0.052061111B29=AVERAGE(C4:C13) 135272.5B30=DEVSQ(B4:B13) Each water sample has three replicate abs values. Take the average value and plug into this space. The derived X and Sx will automatically be computedInput 2Input 15.05381611B34=(B24-C16)/B16 B35=(C26/B28)*SQRT((1/B33)+(1/B28)+ 11.486791((B32-B29)^2)/(B24^2*B30))

Measured y= k=Number of replicate measurements of y= Derived x= Sx=

50% confidence interval for x, x+ tSx degrees of freedom=n-2 95% confidence interval for x, x+ tSx degrees of freedom=n-2 Gresham Gresham Kangen Kangen Sandy Sandy PSU PSU 50% level 95% level 50% level 95% level 50% level 95% level 50% level 95% level

t=0.765 t=2.306

17.739+ 0.765*11.45 26.5,9 17.739+2.306*11.45 44.1427,-8.67 16.47+ 0.765*11.47 16.47+ 2.306*11.47 17.37+ 0.765*11.45 17.37+ 2.306*11.45 25.3,7.7 42.92,-10 26.13,8.61 43.77,-9.034

15.05+ 0.765*11.486 23.84,6.26 15.05+ 2.306*11.486 41.54,-11.44