CMY 383

EXPERIMENT 5: Direct Determination of Ascorbic Acid in a Commercial Fruit Juice

ALISSA KRIEL 11123002 BSc Physics Practical Initiated on 28 February 2013

The use of Analytical Voltammetry to determine the Vitamin C content of a Commercial Fruit Juice by means of Differential Pulse Polarography in Conjunction with Standard addition

Abstract:
In this quantitative polarographic chemical analysis, the content of Vitamin C within a commercial juice sample which is claimed to contain 48mg/100mL is investigated by means of both Calibration Plot Differential Pulse Polarography and Standard addition Differential Pulse Polarography. The result from the calibration curve method correlated well with the 48mg/100mL claim by yielding a Vitamin C concentration of 46mg/100mL , whilst the standard addition curve was not entirely linear, although it yielded a concentration of 39mg/100mL. Furthermore the instrument parameters for a DPP and DCTAST were investigated by considering the effects they had upon a recorded Polarogram of the same solution.

Method:
The preparation of the cell solution and instrument settings as described in the CMY 383 laboratory manual was followed without significant deviation. The Instrument settings were as follows, unless stated otherwise as from figure 6- figure 9 And then only singular changes at a time were made.

Results:

1. Firstly, DP Polarograms were recorded for a 0.020 L background electrolyte (0.1M CH3COO- buffer) and consecutive 50l standard 1g/L ascorbic acid additions up to a total of 200l Standard AA solution present. These polarograms are represented on a single graph in Figure 1:

Except for the slight unexpected pre-peak response on the green (DP Polarogram after 100l std Vit C addition), the Polarograms all peak at the same Potential : Setting the appropriate Peak Potential Ep. From these peak heights, a calibration curve was constructed ply plotting the various consecutive standard solution concentrations versus their recorded Peak heights (Ip):

Calculation of Vit C content in 200L juice using recorded Ip which is gained further on in figure 3, (Ip= 1.97 x10-7 A), =4.623 ppm

2. Secondly a standard addition technique where 3 consecutive 50L 1g/L Vit C soln was added to 200L of Juice sample within the 0.02L BE solution yielded the following DPP Polarograms in Figure 3:

Once again, the consecutive peaks all share the same Peak Potential, E p=0.0682V. BE was very successfully purged as continuum is exceptionally flat. From This data a Standard Addition Curve was generated by Matching concentration to Peak Height:

3. Table 1: Data obtained from the Calibration plot polarogram collection Volume Std Vit C soln Added (L) *and 0.02L BE 50 100 150 200 200l Juice (No Std soln) Concentration of Vit C in Polarographic Cell (ppm) Peak Potential Ep (V) Peak Height Ip (A) Area under Peak

2.49 4.98 7.44 9.90

0.0642 0.0642 0.0642 0.0679

9.48 x 10-8 2.06 x 10-7 3.17 x 10-7 4.28x 10-7

5.02 x 10-9 1.07 x 10-8 1.67 x 10-8 2.25 x 10-8

4.62

0.0682

1.97 x 10-7

9.99 x 10-9

b) Since the Vit C concentration in the 200L juice sample was interpolated as 4.623 ppm = 4.623 mg/L (also by substitution into the linear trend line equation) ,

It follows: The Vit C Concentration per 100mL of Juice is 4.623 mg/L /0.1mL =46.2 mg/100mL of Juice

4. Table 2: Data obtained from standard addition polarograms Concentration of Vit C in Polarographic Cell (ppm) FROM STANDARDS 0 Peak Potential Ep (V) Peak Height Ip (A)

Volume of soln added (L) *0.02L BE soln

200L Juice Solution Juice soln + 50L Std Juice soln + 100L Std Juice soln + 150 Std

0.0682

1.97 X 10-7 3.4 X 10-7 4.47 X 10-7 5.58 X 10-7

0.00

0.0682

2.47

0.0682

4.95

0.0682

From figure 4 it is clear by the equation of the trendline fitted onto the standard addition data points that the Vit C concentration in the sample (200L) will equal to 3.93 ppm = 3.93 mg/L.

Hence, the Vit C concentration per 100mL of Juice is found to be 39.3 mg/100mL.

*Please see figure 4 where the graphical determination was used to mark linear trend line intercept with the x-axis which marks the sample concentration.

Investigation of instrumental parameters on recorded Polarograms:

Following are 5 figures wherein one specific parameter was changed (depending on whether a DCTAST Polarogram was to be recorded or a DP Polarogram). An analyses of what effect takes place and why follows each.

4.00E-07 3.50E-07 3.00E-07

Figure 5: DCTAST wave Polarograms of a solution using different Voltage Step Times
DCTAST Polarogram obtained using 0.8 s Voltage Step time for 200ul Juice Solution + 150ul Std Vit C Soln DCTAST Polarogram of Same Soln with Voltage step time = 1.5s

Current (A)

2.50E-07 2.00E-07 1.50E-07 1.00E-07 5.00E-08

0.00E+00 -0.2 -0.1 0

U(V) 0.1

0.2

0.3

There is a slight increase in the Height (Id) as the Voltage step time has been increased. This may have been caused by the fact that voltage steps are now made farther apart, and since the halfwave potential is also marked a bit farther toward the positive potentials, it reinforces that the change in step time caused the DC wave to reach its equivalent points later on.

4.00E-07 3.50E-07 3.00E-07 2.50E-07 2.00E-07 1.50E-07 1.00E-07 5.00E-08 0.00E+00

Figure 6: DCTAST wave Polarograms with a change in Voltage Step Value

DCTAST Polarogram obtained using juice + Std solution Voltage step = 0.004V DCTAST Polarogram obtained using same Soln Voltage Step=0.012V

-0.2 -0.15 -0.1 -0.05

0.05

0.1

0.15

0.2

0.25

There is a slight decrease in height when the Voltage step was increased by a factor 2. The diffusion current may not even have been affected by the Voltage step being altered since the change seems relatively small and the two Polarograms seem indistinguishable.

There is a significant decrease in peak height as the Pulse amplitude is halved. In fact, the height (Ip) seems to also have halved, creating the possibility that the resultant peak height is directly proportional to the Pulse amplitude implemented.

The aforementioned direct proportionality between peak height and applied Pulse amplitude does not seem to carry through successfully here since the increase in peak height (Ip) is not the expected value exactly, although there is still the general trend that with larger Pulse amplitudes, larger peak heights are recorded.

When the pulse time is increased, the peak height is decreased by an amount. This may be due to the fact that pulse times are now extended and the pulse applied over a longer period of time. Thus the amplitude will not reach as high a value as before thus not such high peaks as the measurement is elapsed/completed.

Discussion
The Juice Producer claims that 48 mg/100mL of Vitiman C is present in the juice. The results obtained here, both by calibration plot method of DP Polarography and a standard addition method, are somewhat lower one being ~46mg/100mL and the standard addition method yielding ~39 mg/100mL . Firstly, we note that both methods also yielded significantly different results. It is also considered that in the Standard addition technique, we assume that the response is linear (Skoog et al, 2004, 210). The trendline shows some deviation and was also set to intersect the y-axis at the Ip (Current peak) of the 200L Juice sample. This affects the credibility of the ultimate intercept with the x-axis- and hence the obtained concentration of Vitamin C in the juice sample. If the Calibration curve which was obtained as inherently linear, is to be considered as more credible, then the difference between obtained concentration and manufacturers claimed concentration is minimal. 2mg/100mL. This also reinforces the ability of the Polarogrophy analytical method to accurately quantitatively analyse the organic content within a sample matrix.

References:
Skoog et al, 2004, Fundamentals of Analytical Chemistry, 8th Edition, Brooks/Cole, 210-212

Kealey D. And Haines P.J. , 2005, Instant Notes in Analytical Chemistry,2nd Edition , BIOS Scientific Publishers Ltd., 46.

J. N. Miller and J. C. Miller, Statistics and chemometrics for analytical chemistry,2000, 4th Edition, Prentice Hall