Atomic absorption spectroscopy is an analytical technique used for the determination of elements, and at times compounds, based on the

absorption of radiation by unexcited gaseous free atoms. The determinations of metals in a variety of real-life samples with flame atomic absorption spectroscopy (FAAS) have been reported in this Journal. Those studies include the determination of zinc in human hair (1), lead in paint chips and gasoline (2, 3), iron in multivitamin tablets (4), iron and manganese in dry pet food (5), calcium in fruit juice and analgesics (6, 7), and calcium in foodstuffs (8). Although the last reference (8) provided results on the determination of calcium in foodstuffs, the procedure was designed for a nonscience major course. The quantity of calcium present in foodstuffs was determined using the standard calibration method, which does not compensate for determinate errors caused by the matrix effect when analyzing real-life samples. Also, the experiment used wet-ashing with concentrated nitric acid, where some samples had to be filtered and the wet-ashing repeated. The experiment described here is designed for science majors who have completed two semesters of general chemistry and are enrolled in the Quantitative Methods of Analysis course. This experiment uses the two increment standard addition method in conjunction with FAAS to determine the quantity of calcium in cereal. It eliminates the need for wetashing with concentrated nitric acid and minimizes the generation of hazardous waste by dry-ashing the sample in a silica crucible using a high temperature muffle furnace. This experiment (i) introduces the students to the principles of atomic absorption spectroscopy, (ii) gives the students handson experience in operating and obtaining measurements using a modern FAAS instrument, (iii) gives experience in dry-ashing sample preparation, (iv) gives the opportunity to work with and understand the principle of the two increment standard addition approach for the analysis of real-life samples, and (v) illustrates the importance of FAAS in the analysis of samples relevant to their daily lives. This experiment was successfully used in three different laboratory sections of the Quantitative Methods of Analysis course in the fall 2001 term. The data presented in this article were obtained by 36 students majoring in biochemistry, biology, chemistry, Earth science, or environmental science, the majority of whom are required to take the course. The experiment is normally completed in six hours of laboratory time.

Procedure Sample Preparation A 5-g sample of the cereal to be analyzed is crushed into a fine powder using a mortar and pestle. Samples, 0.5-g, of the crushed cereal are placed in three weighed glazed silica crucibles and weighed to the nearest 0.0001 g. The samples are then placed in a muffle furnace maintained at 600 _C. An empty crucible is also placed in the furnace to serve as a

blank. After two hours, the four crucibles are removed, cooled, and the residue is treated with 6 M HCl. The residue readily dissolves and filtration of the resulting solution is not necessary. The content of each crucible is transferred to 100-mL volumetric flasks and diluted to volume with distilled water. The two increment standard addition technique is performed by using two 5.00-mL aliquots from each sample. A single addition of 20.00 mL of the standard calcium solution is added to one of the aliquots and both are diluted to volume using 50-mL flasks. This treatment generates three sets of unspiked and spiked samples to be used for the FAAS measurements. A blank solution is also prepared from the empty crucible and measured.

Atomic Absorption Measurement and ppm Calculation of Calcium in Cereal A Varian SpectrAA-600 instrument with a calcium hollow cathode lamp and nitrous oxideacetylene flame is used. Because of its higher temperature, the nitrous oxideacetylene flame eliminates the need for the addition of a releasing agent, such as lanthanum or strontium ion, to the solutions. A releasing agent is added to eliminate chemical interferences owing to the formation of refractory compounds when air acetylene flame is used in the analysis of calcium. Absorbance measurements are made using the calcium resonance line at 422.7 nm, 0.5-nm slit width, and 5.0-mA lamp current. Additional experimental details are included in the Supplemental

Material.W The calcium level is calculated as,

where Csam and Cstd are the concentrations in g mL_1 of the sample and standard solutions, respectively, and Asam and Asam+std are the corrected absorbance values of the unspiked and spiked solutions, respectively. These values are obtained by subtracting the absorbance of the blank solution from their respective absorbance measurements. Vstd is the number of mL of the standard solution added and Vsam is the number of mL of the cereal sample solution. Equations 1 and 2 allow for the determination of Csam and the ppm Ca in the cereal sample. Hazards Flame-resistant leather or glass-fiber gloves and jumbo (18 in.) crucible tongs must be used when inserting and removing the silica crucibles from the muffle furnace. Hydrochloric acid, HCl, is corrosive; preparation of dilute solutions from concentrated HCl must be done in the fume hood. Acid-resistant gloves and laboratory coats are recommended in addition to mandatory safety goggles. Owing to potentiall fire or explosion hazards when using nitrous oxide as the oxidant, the instrument manufacturer safety practices and recommendations should be followed carefully. Results and Discussion The data obtained by the students for the determination of calcium in two brands of cereal, Kix and Cheerios, from three different laboratory sections in Fall 2001 are shown in Table 1. Each student worked independently and was responsible for making her or his standard solutions, sample preparation, and measurements using the Varian SpectrAA-600 instrument. Each value represents the average of three separate determinations. For the Kix results, the RSD of the student triplicate determinations ranged from 0.31 to 16.50% with the majority (78%) having values 5%. For the Cheerios results, the RSD for the student triplicate determinations ranged from 0.59 to 11.64% with the majority (67%) having values between 0.59 and 5.25%. Based on the averages of the students results and a value of 1200 mg for the recommended dietary allowance (RDA) for calcium for the 1924 year age group (the pertinent age group for the students; ref 9), one 30-g serving of Kix and Cheerios provides 17.0% and 13.1% of the RDA, respectively. Typical absorbance values for blanks and Kix and Cheerios samples are shown in Tables 2 and 3. Each sample was measured three

times and the average was used in the calculation in eq 1. Although this experiment is designed for the two increment standard addition approach, it may be easily adapted to the multiple-increment standard addition method. A standard addition graph for a Cheerios sample, with additions ranging from 0 to 20 mL of the standard calcium solution to 5.00-mL aliquots of the sample followed by dilution to 50.0 mL, is shown in Figure 1. The quantity of calcium in this sample, Csam, was found to be 4500 ppm. This value, which is comparable to those obtained with the two increment approach, was calculated using the following equation in conjunction with eq 2,

where Cstd is the concentration of the standard solution added, Vsam is the number of mL of the cereal sample solution, m is the slope, and b is the intercept of the standard addition graph.

Conclusion The implementation of this laboratory experiment in the Quantitative Methods of Analysis course was successful. The analysis of cereal products was of interest to the students since these are foods they regularly consume. The results for the determination of calcium in two cereal brands obtained by 36 students from three different laboratory sections showed very good precision with 72% having RSD 5%. We plan to use this approach in the future for the determination of other metals such as iron, zinc, and copper in cereal products.
W Supplemental

Material Instructions for the students and notes for the instructor are available in this issue of JCE Online. Literature Cited
1. Pomeroy, R. K.; Drikitis, N.; Koga, Y. J. Chem. Educ. 1975, 52, 544. 2. Markow, P. G. J. Chem. Educ. 1996, 73, 178. 3. Bye, R. J. Chem. Educ. 1987, 64, 188. 4. Pinnel, R. P.; Zanella, A. W. J. Chem. Educ. 1981, 58, 444. 5. Paschal, D. C. J. Chem. Educ. 1976, 53, 165. 6. Strohl, A. N. J. Chem. Educ. 1985, 57, 1322. 7. Quigley, M. N. J. Chem. Educ. 1994, 71, 800. 8. Kostecka, K. S. J. Chem. Educ. 2000, 77, 1321. 9. Snyder, C. H. The Extraordinary Chemistry of Ordinary Things, 3rd ed.; Wiley: New York, 1998; p 498.