Volumetric, Potentiometric, and Flame Atomic Absorption Analysis of Calcinate Tablets

Jonathan Barnett, Joshua Hiatt, Ryan Nguyen, and Matthew Scorpio

ABSTRACT: Calcinate pills were tested for their calcium concentration because consumers believed that the labeled amount of calcium was an overestimation of the actual amount. Prime Eastern Pharmaceuticals states that their Calcinate pills contain 800 mg of calcium in each tablet; our study was to deduce the actual amount of calcium per tablet and see if the company was falsifying their product. To determine the calcium concentration we used three different methods: atomic absorption spectroscopy (AAS), EDTA titration and potentiometry with an ionselective electrode. The amount of calcium per pill found using AAS was 649 mg, using EDTA titration it was 640 mg, and using potentiometry it was 603 mg. From the information collected we concluded that Prime Eastern Pharmaceuticals was falsifying their product because the calcium contained in each pill was lower than the labeled 800 mg.

INTRODUCTION Calcium is the most abundant mineral in the body, and is absolutely necessary to perform several bodily functions. One percent of bodily calcium is used for vascular contractions, muscle function, nerve transmission, and hormonal secretions. The other 99% is in teeth and bones. Calcium is constantly used in bone because bones are continuously being remodeled and calcium is constantly removed and added as the body sees fit. Based on the many functions calcium provides the body, calcium deficiencies can negatively affect many key bodily functions. These negative effects are particularly prevalent in people age 50 and above, as that is the point at which people start to develop osteoporosis (a disease resulting in low bone mineral density, meaning very frail bones that are prone to

fracture). Because calcium deficiency is a large problem in people older than age 50, it is recommended that they consume 1,0001,200 milligrams of calcium every day, a large amount of which comes from calcium tablets. There are 4 popular types of calcium tablets that use either: calcium carbonate, calcium citrate, calcium gluconate or calcium lactate. Calcium carbonate and calcium citrate are the two preferred options because of the higher percent elemental calcium per tablet. Calcium carbonate is the cheaper of the two options. Prime Eastern Pharmaceuticals has been the producer of top-selling Calcinate calcium tablets for years. These dietary supplements claim to contain approximately 800 mg of calcium in every pill. Recently however, Prime Eastern Pharmaceuticals has been taken to court multiple times for reducing

the amount of calcium in their pills without saying anything about it. If the claim is true, there could be huge consequences, as people need a specific amount of calcium per day. The purpose of this study was to determine the concentration of calcium per tablet by using three different methods: atomic absorption spectroscopy, EDTA titration and potentiometry. Results from this study will prove or disprove the claims against Prime Eastern Pharmaceuticals. BACKGROUND Complexometric Titrimetry Titration is a method of measuring the concentration of an analyte by reacting it with a standard reagent. In complexometric titration, complexes are formed between the analyte (metal ion) and a ligand. This can also be done in reverse where the ligand is the analyte and the metal ions are the standard solution. A common ligand is Ethylenediamintetraacetic acid (EDTA) because of its ability to make 6 bonds with metal ions. The two nitrogens have one lone pair of electrons each that can bond with metal ions. In addition there are four carboxylic acid groups that can further interact with metal ions. The high amount of possible bonds with EDTA give metal ions a large relative formation constant (Kf). Calcium metal ions have an approximate Kf of 5.0X1010. Calmagite is used as an indicator ligand reaction, because calmagite has two OH groups that form complexes with the metal ions. The Calmagite-metal complexes are red. Ca2+ binds more preferably with EDTA than Calmagite, and with titration of EDTA the Calmagite-metal

complexes change into EDTA-metal complex and Calmagite which turns blue. Ion-selective Electrode Potentiometry In ion-selective electrode potentiometry, an electrode is used that interacts with positive ions in solution that will affect the charge interactions inside the electrode system. As calcium ions interact with some interface the amount of positive charges increase at the interface. Inside the electrode there is a solution or some sort of charge displacing medium where protons suspended in a membrane near the interface are repelled by the increase in calcium and released into the internal solution where the increase is seen as electron movement measured by an actual electrode submerged in the internal solution. The change is measured as mV and can only be interpreted with standard mV data obtained using standard solutions of known concentrations. Flame Atomic Absorption Analysis Flame atomic absorption spectroscopy (FAAS) works like any other spectroscopic technique, where light is passed through a sample with an initial intensity is attenuated. The final intensity of light picked up by a detector can be used to determine sample concentration, compared to standard attenuated light intensities measured by running standard solutions of standard solutions of known concentrations. In FAAS the sample is aspirated into a heat source that vaporizes the sample. Light is passed through the flame, and the vaporized sample will attenuate the passing light more, relative to the light attenuation of the flame. Using the beer-lambert law the differences in light intensities before and after passing

through the sample can be related to the concentration of analyte in the sample.

Figure 1: Flow diagram of a sample run in an atomic absorption spectrometer. EXPERIMENTAL SECTION Safety Goggles and gloves were worn at all times in the lab. Care was taken with glassware, and protective gloves were worn when handling hot glassware. Care was taken when handling acids and bases, particularly with the 6M HCl, as all were harmful upon skin contact. Procedures The experiment as a whole is composed of three separate experiments that each calculate the amount of calcium in the average calcium tablet. For the set of experiments, five Calcinate calcium pills were weighed and then were placed into a mortar and pestle, crushed, and stored. Analysis through EDTA Titration Procedure: 1.0 gram of EDTA salt was weighed and placed into a 250 mL volumetric flask. 200mL of slightly heated water was added to dissolve, and then the flask was filled to the mark. 0.11 grams of Calcinate powder was accurately weighed and placed into a 150 mL beaker. 6M HCl was added drop wise to dissolve the powder

and then 50mL of water was added and was brought to a boil to expel excess CO2. The solution was taken off the hotplate and allowed to cool to room temperature and then 1M NH4OH was added drop wise to bring the pH back up to 6. The solution was then quantitatively transferred to a 250-mL volumetric flask and was filled to the mark. Three 50.00mL aliquot of the powder solution were transferred to three separate 250mL Erlenmeyer flasks and 15mL of preprepared pH 10 buffer, 6 drops of Calmagite, and 4 drops of methyl red were added to each flask and to a fourth blank flask with no powder solution. The blank was blue, so no extra titration was necessary, meaning that the titration was not off. Each solution was then titrated with EDTA, and the volume of EDTA needed was used to calculate the amount of calcium in each tablet.

Analysis through Potentiometry. Procedure: 1.0g of standard CaCO3 was accurately weighed out into a 100 mL beaker. 6M HCl was added drop wise until the standard dissolved. 50 mL of water was added and the beaker was placed on a hot plate to boil out all CO2 in solution. Add 1M NH4OH until the solution has a pH of 6. The solution was then quantitatively transferred to a 100mL volumetric flask and diluted to the mark with deionized water. 10mL of the solution was then transferred to a second 100mL volumetric flask and diluted to the mark with deionized water 0.01M CaCO3 solution. Prepare four separate standard solutions by pouring 10 mL of 1.0M KCl and 1, 2, 5 and 10 mL of the 0.01M CaCO3 into four separate 100 mL volumetric flasks

and dilute to the mark with deionized water. Accurately weigh out 0.2125 grams of Calcinate Powder into a 100mL beaker. Add HCl dropwise until the powder dissolves, and then add 50mL of water and boil to release all excess CO2. Add 1M NH4OH drop wise until the solution has a pH of 6. Quantitatively transfer to a 100 mL volumetric flask and dilute to the mark with HCl. Transfer 2mL of solution and 10 mL of KCL to a 100mL volumetric flask and dilute to the mark. Transfer 60mL of each of the standards and the unknown solution to individual 100mL beakers and take two measurements of the potentials of each solution. Use the standards to create a calibration curve and use that to calculate the mass of calcium per tablet. Analysis through Atomic Absorption Procedure: 0.34 grams of calcium tablet were accurately weighed into a 100mL beaker. 6M HCl was added drop wise until the powder dissolved. 50 mL of special deionized water for Flame Atomic Absorption was added and the excess CO2 was boiled out. The solution was then quantitatively transferred to a 100mL volumetric flask and diluted to the mark with special deionized water. 1mL of that solution was transferred to a second 100mL volumetric flask and was diluted to the mark with water. The solution was then sent off for the absorption to be measured using Atomic Absorption Spectroscopy.

CONCLUSION From the results of the three experiments it can clearly be seen that Prime Eastern Pharmaceuticals has decreased the amount of calcium per tablet without notifying their customers of the change. Despite the conclusiveness of the groups results, there were many ways that the accuracy and the reliability of the experiments could be improved. One factor that contributes to the data not being as reliable as prefered could be traced to the small amount of trials run by the group and the lack of multiple samples that could be tested. The entirety of the pills that the group tested were crushed and mixed together. While this was a good way to average the amount of calcium in the tablets, it does not, however, replace testing on multiple samples from separate batches of the tablets individually. If this experiment were to be done in the future, or in future research where a similar procedure would be implemented, it would be recommended that a larger amount of samples should be used and more trials run. One further way the accuracy and reliability of the results could be improved would be to introduce a fourth test to determine the average amount of calcium in the tablets. A possible fourth test could be done by reacting the CaCO3, in a solution made by crushing the tablets, with HCl and then back titrating the excess HCl in the solution. These methods could greatly increase the reliability and accuracy of the results. Because the resulting amounts of calcium in each tablet were so significantly less than 800 mg, (an average 631mg of calcium per tablet) extra data is not needed to determine that the amount of calcium per pill was reduced.

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