Voltammetric Determination of -Tocopherol in Vitamin E Supplements Cabotaje, Princess Rosery Ibale, Jonathan Rey A.

Institute of Chemistry, College of Science, University of the Philippines, Diliman, Quezon City Date Performed: February 14, 2014 Date Submitted: February 26, 2014 Abstract This experiment aims to study the actual analytical and biological amount of -tocopherol in Mayla Natural Vitamin E 600 IU using voltammetry, a simple and affordable electroanalytical technique. Using cyclic voltammetry, the electrochemical behavior of the analyte was observed using ethanol:toluene:sulfuric acid solution (300.0 mL, 150.0 mL, 3.0 mL, respectively) as supporting electrolyte (SE) solution. The quantitative analysis employed differential pulse voltammetry using the same SE solution and a 1.016 g/L -tocopherol standard. The latter analysis also incorporated the use of standard addition technique for the calculations. It was found that the analyte undergoes an irreversible redox reaction and exhibits an anodic potential peak, 0.591 V, with an observed current of -4.42 A. From a 0.3314 g capsule, the solution (50.0 mL) was prepared from 0.3244 g (484 IU) extracted liquid. The -tocopherol present in the sample was 7.82 mg/mL in solution and about 391 mg or 583 IU. This data was +18.6% different from the expected 484 IU, assuming that all extracted liquid was -tocopherol, while -2.77%error was calculated analyzed against label value (theoretical). These difference may have been due to preparation errors or random errors accumulated from electroderelated interferences. I. Introduction Voltammetry is an electroanalytical method that employs of variable potential from a working electrode which generates a current measured as a potential function from which quantitative analysis would later be employed further. It is one of several electro chemical techniques of interest in analyses due to their inherent simplicity, availability of equipment, and affordability. In voltammetry, the qualitative parameter is taken to be the cathodic or anodic potential (Epc and Epa, respectively), depending on the analytes electrochemical behavior, while the quantitative parameters are the current peaks (ipc and ipa), given by the Randles-Sevcik equation (1). Ip = 2.686x105

popularly used by the pharmaceutical industry as an aid against free radical oxidation of polyunsaturated fatty acids1 and some other particular diseases and disorders.[1]

Figure 1. 2,5,7,8-tetramethyl-2-(4,8,12trimethyl-tridecyl)-6-chromanol or tocopherol.[1] In particular, cyclic voltammetry will be used for the characterization of the electrochemical behavior of the analyte. Differential pulse voltammetry would be used for the sample analysis aided by standard addition analytical technique. II. Methodology Before the analysis, the solutions prepared initially were the 1.016 g/L -tocopherol standard solution, the sample solution, and the ethanol:toluene:H2SO4 solution (300.0 mL, 150.0 mL, 3.0 mL, respectively). The sample solution was prepared by obtaining 0.3244 g liquid from a 0.3314 g Mayla Natural Vitamin E 600 IU capsule. The sample solution was prepared to 50.0 mL volume.[2] Other than preparing the solutions to be used, the system was also prepared by cleaning the working

AC

(1)

, where n = no. of e- involved in the half-reaction, A = electrode area (cm2) D = diffusion coefficient (cm2/s) C = analyte concentration (mol/cm3) V = scan rate (V/s) In this experiment, -tocopherol (Figure 1) will be quantified from extractions from Myra E vitamin supplement capsules using voltammetry. The analyte, 2,5,7,8-tetramethyl-2-(4,8,12trimethyltridecyl)-6-chromanol, is widely used in the industry as an active and stabilizing agent against oxidation of other components. It

electrode (glassy carbon) by polishing it on a mat with Al2O3 slurry in a figure-8 pattern. The cleaning was done by wiping the electrode evenly on the mat 100 times. Then, all of the electrodes were washed with deionized water and wiped dry before finally attaching to the equipment via alligator clips. The analyte behavior was first identified, i.e. whether or not the subject reaction was reversible or not and whether it was primarily oxidation or reduction. This was done by employing cyclic voltammetry. First, 20.0 mL of the supporting electrolyte (SE) solution was added into the electrochemical cell, deaerated with nitrogen gas for 60 seconds, and was analyzed while maintaining the N2(g) atmosphere now outside and above the solution. The parameter values used for the program was that in Table 1. Table 1. Cyclic Voltammetry parameter input. Initial -100 mV Final -100 mV Ramp Rate 10 mV/s Width 200 ms Upper 1000 mV Limits Lower -100 mV Cycles # of cycles 1 Range 1000 mV Speed 10 kHz Scan Period 100 ms Rest Time 2s Range 20 A After obtaining the voltammogram of the SE solution, the -tocopherol standard (100 L) was pipetted into it, then the new voltammogram was obtained, superimposing the graphs of the two to obtain a resulting cyclic voltammogram. For the sample analysis, differential pulse voltammetry was used. The same SE solution was added (20.0 mL) and deaerated with N2(g) and read before analysis while maintaining the N2(g) atmosphere during analysis. The new parameter for the differential pulse voltammetry shown in Table 2 was inputted. Table 2. Differential Pulse Voltammetry parameter input. Initial 0 mV Ramp Final 1000 mV

Pulse Scan Period Rest Time Range Speed Range

Rate Width Width Height 50 ms 2s 2000 mV 10 kHz 20 A

10 mV/s 200 ms 100 ms 10 mV

Afterwards, the sample solution (500 L) was added followed subsequently by a reading. The standard solution was also added at 500 L increments to the SE solution then followed by a voltammogram reading. The analysis was continued until 2500 L of the standard solution was added. After all the data was gathered, the anodic peak values were determined and quantitative analysis as well as qualitative analysis was followed through. III. Results and Discussion In the cyclic voltammetry, the potential obtained was 0.591 V, an anodic potential peak. This implies that the analyte undergoes oxidation, and since there were no cathodic potential peaks observed, the analyte half reaction was an irreversible redox reaction. The sample analysis was then expected to yield anodic values of negative potential with an increasing magnitude. The corrected data, i.e. subtracting the blank signal, was tabulated in Appendix A. When plotted the anodic peaks were plotted against the -tocopherol concentration Figure 1 was obtained with a linear equation of y = -4.8809x-0.9306 and a linearity coefficient of R2 = 0.9964.
0 0 -0.5 -1 -1.5 -2 Std concentration (mg/mL) 0.05 0.1 0.15

Figure 1. Anodic peaks of -tocopherol standard as a function of concentration.

Anodic Peak (corrected, A)

Extrapolating to y = 0 A, a concentration for the blank was obtained, 0.190661558 mg/mL which corresponds for the solution of sample and blank solutions only. Solving for the original concentration, the 50.0 mL sample solution was 7.82 mg/mL. The original 0.3314 g capsule of Mayla Natural Vitamin E 600 IU that yielded 0.3244 g liquid contains about 391 mg -tocopherol. The amount of -tocopherol can be expressed in its biological quantity, 583 IU.[3] Against the label IU, the difference of values was 2.77%(%error). Assuming that all of the liquid obtained from the sample capsule was -tocopherol, the value difference would be 18.6%(%difference), i.e. against 484 IU. Since, the generic label retains that the paraphernalia contains Vitamin E only, the latter value must be considered for error analysis of the procedure. The error was a positive error which may be attributed to the presence of anodic electrolytes that may have added to the peak signals resulting to the difference by the end of the analysis. These may have been due to procedural errors during solution preparations and even during experimental run, e.g. from a contaminated electrochemical cell. It may also be that the electrodes have not been properly cleansed or that calibration was not optimal due to contamination of the main electrolyte in the cell via the frits. Other errors related to the electrode may also be pointed out. IV. Conclusion

V. Reference [1] Ziyatdinova, G.K., E.R. Giniyatova, H.C. Budkinov (29 Jun 2011). Voltammetric determination of tocopherol in the presence of surfactants. A.M. Butlerov Institute of Chemistru, Kazan (Volga Region) Federal University, ul. Kremlevskaya 18, Kazan, 420008 Russia. (ISSN 1061-9348, Journal of Analytical Chemistry, 2012, Vol. 67, No. 5, pp. 467473. Pleiades Publishing, Ltd., 2012) [2] Ballesteros, J. (2008). Voltammetric determination of -tocopherol in vitamin E supplements. Institute of Chemistry, University of the Philippines-Diliman. [3] Office of Dietary Supplements (5 June 2013). Vitamin E-Health Professional Fact Sheet. Office of Dietary Supplements, National Institute of Health, US Department of Health & Human Services|Online Resource (HHS.gov). http://ods.od.nih.gov/ factsheets/VitaminE-HealthProfessional/ (viewed 25 Feb 2014) [4] Institute of Chemistry (2013). Chemistry 101.2 Intermediate Chemistry Laboratory II-Laboratory Manual. 2013 Ed. Institute of Chemistry, University of the Philippines, Diliman Quezon City, Philippines. [5] Skoog, D., D. West, Holler, F.J. Crouch, S.R. (2004). Fundamentals of Analytical Chemistry. 8th Ed. Books/Cole, Singapore. Ch. 23, pp. 666-668. VI. Appendix

Appendix A Table 3. Data for DP Voltammetry Ip Ip Added Solution (raw,A) (corrected,A) Blank (20.00 mL SE) -0.61 0 500 L sample solution -1.55 -0.94 500 L Std solution -1.64 -1.03 1000 L Std solution -1.77 -1.16 1500 L Std solution -1.89 -1.28 2000 L Std solution -1.99 -1.38 2500 L Std solution -2.07 -1.46 Appendix B Calculation for Sample Concentration

The analyte, -tocopherol or Vitamin E, was found to be an oxidized species that undergoes a irreversible redox reaction as the cyclic voltammetry exhibits. The differential pulse voltammetry analysis for the sample analysis showed that the sample contained about 391 mg or 583 IU -tocopherol. This was 18.6% from the calculated IU of the total sample obtained, 484 IU, with the assumption of being 100% -tocopherol constitution. Several errors were pointed out which includes introduction of unknown electrolyte species and random electrode-related errors. This experiment may be improved by careful preparation of solutions and set-up and the employment of multiple trials to improve precision of data and mitigation of random errors.

(For blank + sample solution concentration (20.5 mL)) y = -4.8809x 0.9306 = 0 x= = - 0.190661558 mg/mL sample (For actual sample concentration) Concn = = 7.817123891 mg/mL

Appendix C Calculation for International Units (IU)[2] IU = mg x = 390.8561946 mg x = 583.3674546 IU Appendix D Calculation for %Error %Error =
| |

x 100%

= x 100% = 2.772090896%error Appendix E Calculation for %Difference %Diff = =

| |

x 100%

x 100%

= 18.58248697%difference