Ion Chromatography Station 2

Determining the presence, and concentration of sugar

carbohydrates in unknown samples using both qualitative, and
quantitative analytical techniques.

Experiment 1: Qualitative Analysis of Carbohydrates using Ion

Chromatography
Experiment 2: Quantitative Analysis of Sugars in juices and soft
drinks using a calibration curve method

Submitted By: Chad Adams

Date(s) performed: 20/01/2015, 27/01/2015
Date Submitted: 03/02/2015

I, Chad Adams certify that the data enclosed within this report
was performed solely by me on the dates disclosed above. I
understand the concepts of academic honesty, and certify that no
portion of this lab has been copied without proper credit being
given to the original author. I further understand that if any
portion of this lab has been copied and not properly credited that
the penalty for this is a mark of zero on the entire lab and an
academic alert being issued.

Introduction:
Many of the consumer beverages available on the market nowadays
contain some form of sugar. Often times, this is un-natural sugar, which is
mainly composed of various synthetic version of fructose, sucrose, and
glucose. These carbohydrates are so prevalent in todays soft drinks which
are extremely popular among the young of our population, as well as the old
that they are actually beginning to notice the correlation between health,
and sugar carbohydrate consumption. Due to this sudden surge of sugar
related health risks, the food industry follows strict guidelines for how much
sugar is allowed in their beverages, as well as what types of sugar can be
added such as to manage, and monitor the effects that sugar is having on
the general public. To monitor the amount of sugar that is in their created
beverages, companies can utilize the Ion Chromatography, due to its
incredible ability to separate ionic compounds in solutions. In this laboratory
experiment, the Ion Chromatographer will be used to demonstrate the
qualitative, and quantitative capabilities of Ion Chromatography when
utilized by the food industry.
The instrument that will be used in this laboratory experiment is the
Dionex Ion Chromatographer. This instrument utilizes Ion Chromatography.
This process involves the separation of ions based on their affinity to an
exchange column. Retained analytes are measured for their concentration
area, which is converted into usable data by a detector, which is then routed
through a readout device, such as a desktop computer, where it is then
utilized by a laboratory experimenter to determine the content of a
carbohydrate in an unknown sample usually by use of a calibration curve.
Many additional components can be added to the Ion Chromatography
system to allow it to be utilized for very specific applications.
In this experiment, the ion chromatography is being used to perform
qualitative, and quantitative analysis of sugar carbohydrates in unknown
samples. To do this, two experiments will be enacted, one to showcase the
qualitative aspects of Ion Chromatography, and one to demonstrate the
quantitative aspects. In the first experiment, 5, 50 ppm stock solutions, will
be created of the following sugar carbohydrates: Sucrose, Glucose, Fructose,
Lactose, and Sorbitol. A mix of the 5 sugars will also be made, and labelled

to determine the various peaks in a mixture environment. Using the data

found in lab, an unknown Coca Cola sample will be analyzed to determine
what sugars are present. For this experiment, only qualitative data is being
analyzed. In the second experiment, 100 ppm stock solutions of both
Glucose, and Sorbitol are being created. From there, 4 working standards
(10, 25, 50, 100ppm) are being created for both compounds, and are being
analyzed using the IC. Data recovered will then be formulated into calibration
curves, which will be used to determine the amount of each compound in
unknown Coca Cola, and Apple Juice Samples.

Observations, Data, and Procedure:

The procedure was followed as stated in the Spectroscopy Laboratory
Manual Fall 2014 edition pages 105-106.
Please refer to the lab book sheet attached with this report for detailed
and observational notes.
All safety guidelines, and protocols were followed.

Experiment 1: Qualitative Analysis of Carbohydrates using

Ion Chromatography
Table 1: Analysis of various sugar carbohydrates, a mixture of these
carbohydrates, and an unknown Coca Cola sample
Retent Concentr
Sampl ion
ation
e
Time
Area
Gluco
11135013
se
2.95
9
Lacto
se
Fruct
ose
Sucro
se
Sorbit
ol
Mixtu
re

5.00
3.25

6.20
1.75

67455893
10849044
9

64568174
11435310
4

Observa
tions
Large
Peak
Lower
Conc,
Smaller
peak
Large
Peak
Lower
Conc,
Smaller
peak
Large
Peak

Individual Samples
The concentrations of these
samples are expected to be
around the same, but, it would
seem that as the retention time
increases, the concentration area
decreases, as the lower RT have
similar areas, as do the higher RT.

Mixture
Peaks decrease in height as the
RT decreases. All peaks present.

Unknown Sample

Sample from Experiment 2 was

used. It would seem that Glucose,
and Fructose are the major
components of the high sugar
content of Coca Cola. This makes
sense, although one would expect
Sucrose to be present. There will
be no Lactose due to the fact that

Sorbit
ol
Gluco
se
Fruct
ose
Lacto
se
Sucro
se
Unkn
own
(used
from
Exp.
2)
Coca
Cola
Sorbit
ol
Gluco
se
Fruct
ose
Lacto
se
Sucro
se

1.80

Tallest
6970033 Peak

3.00

5776796
Peak size
5323656 decrease
s as RT
4381669 increases
Smallest
3879724 Peak

3.45
5.15
6.35

N/A

N/A

3.00

14100656

Only
containe
3.40 15884392 d high
concs of
N/A
N/A
Glucose,
and
N/A
N/A
Fructose

Experiment 2: Quantitative Analysis of Sugars in juices

and soft drinks using a calibration curve method
Table 2: Quantitative Analysis of Sorbitol present in Apple Juice, and Glucose
present in Coca Cola
Samp
le
(ppm
)
Gluco
se
10.06
25.20
50.40

Glucose
Retention
Time

3.00
3.00
3.00

Concentratio
n Area

Observati
ons

Removed
1557732 as outlier
6308225 N/A
13898055 N/A

10.06ppm standard was removed

as an outlier due to the fact that
it was completely out of range,
and did not correspond to other
data points. Retention Time
remained constant. 3 data points
were used, using the 25, 50, and
100
ppm solutions.
Sorbitol
51.60ppm
was omitted
Unknownstandard
Sample
from data due to the fact that the
sorbitol
peak formed
tangent,
Both samples
were inarange
of
which
threw off the
data.
Area/RT
their respective
data
points.
remain relatively constant.

100.8
0
Sorbi
tol
10.32
25.80

51.60
103.2
0
Apple
Juice
Sorbi
tol
Coca
Cola
Gluco
se

3.00

27539909 N/A

1.80
1.80

3063731 N/A
7058792 N/A

1.75

9115317

Tangent
formed (see
Printouts) removed as
outlier

1.75

30840790 N/A

1.85

20196638 In range

3.00

10100856 In range

Figure 1: Calibration Curve of Glucose

Glucose Content Calibration Curve

f(x) = 279389.43x - 512702
R = 1

Glucose
Calibration Curve
Equation: y =
279389x 512702

Figure 2: Calibration Curve of Sorbitol

Sorbitol Content Calibration Curve

f(x) = 301708.14x - 356888.38
R = 1

Sorbitol
Calibration Curve
Equation: y =
301708x 356888

Calculations:

Stock Solution Preparation

5, 50ppm Stock Sugar solutions are to be created. To do this, we weigh a
50mg sample of a sugar, and transfer it to a 100mL volumetric.
Glucose
50mg (Actual weight: 50.4mg) 100mL volumetric flask
Ppm = mg/L = 50.4mg/.1L = 504ppm
On average, the solutions were all around 500ppm.

Glucose Unknown Determination

For the determination of Glucose to be possible, we must have a calibration
curve. If we refer to Figure 1, we can see that our equation is:
Y = 279389x 512702
x = y + 512702/279389 = 10100856 + 512702/279389 = 37.99ppm
Since we took a 1mL sample of Coca Cola into a 1000mL volumetric flask, we
assumed a dilution factor of 1000x.
To account for this, we simply multiply the determined concentration by
1000.
x = 37.99ppm x 1000 = 37990ppm
It would seem that our unknown Coca Cola sample contains 37990ppm of Glucose.

Discussion:

Sugar is responsible for the sweetness of consumer products. Due to

human nature, and the occasional depressing occurrence, sugar has become
one of the most popular carbohydrates. With sugar becoming more, and
more popular, scientists are beginning to notice a decline in health in heavy
sugar users. Due to the increased concern of sugar in consumer drinks, we
must know exactly HOW MUCH we are putting into them. Sugar, in the form
of sucrose, glucose, fructose, lactose, and sorbitol, can be both qualitatively,
and quantitatively with relative ease. This laboratory experiment will
demonstrate the Ion Chromatographers ability to do just that.
In the first experiment, the Ion Chromatographer is being used to
qualitatively analyze an unknown Coca Cola sample. To make this possible, 5,
50ppm stock solutions were made in 100mL volumetric flasks, consisting of:
Sucrose, Fructose, Glucose, Lactose, and Sorbitol. These solutions were
analyzed, and their retention times/peak height/concentration area were
analyzed (see Table 1). A mixture of the 5 stock solutions was created, and
the peaks were labelled using the retention times observed in the singular
tests (see Attached Lab Printouts). A scan of an unknown Coca Cola
sample was then qualitatively analyzed, and the peaks were determined
using the single peaks, and the knowledge gained from the mixture. It was
found that Coca Cola contains a majority of glucose, and fructose, with no
chemical content of the remaining sugar compounds. This makes sense, as
the coke cannot contain lactose, or sorbitol, due to the fact that it isnt milk,
and it isnt created with sorbitol. Oddly enough, youd expect sucrose to be
present in coke, due to the fact that it is a sweetener, but, it would seem that
it is not present, and that glucose is responsible for the sweetness of coke.
In the second experiment, the Ion Chromatographer is being used to
quantitatively determine the content of sucrose in Coca Cola, and sorbitol in
Apple Juice. To do this, 100 ppm stock solutions, which were made by
weighing approximately 0.025g of each compound, and diluting them in a
250mL volumetric flask. These stock solutions were then made into working
standards (10, 25, 50, and 100ppm), using 100mL volumetric flasks. Once
the standards were made, and ran, data was recorded (see Table 2, Figure
1, and Figure 2), calibration curves were formulated. The unknown Coca
Cola sample was prepared by diluting 1mL of the sample in a 1000mL
volumetric flask, and produced an area concentration of Glucose of
10100856. The unknown Apple Juice sample was prepared by diluting 1 mL
of the sample in a 100mL volumetric flask, and produced an area
concentration of Sorbitol of 20196638. Using the calibration curves produced
using lab found data, and after accounting for the dilution factors (see

Calculations Determination of Glucose), it was found that Coca Cola

contained 37990 ppm of glucose, while Apple Juice contained 681.20 ppm of
sorbitol. Now, although the value for Coca Cola is so high, it is mostly due to
the extremely high content of sugar in Coca Cola. This is also a friendly
reminder that Coca Cola is extremely bad for you. Apple Juice was found to
contain 681.20 ppm of the sugar sweetener sorbitol, which corresponds the
logic that apple juice is sweet, but not extremely, unbearably sweet. This
sorbitol also gives apple juice its sweet taste. When looking at these results,
it is fair to say that coke achieves its sweet flavour by packing its soft drinks
with the unhealthy sugar glucose, while apple juice does have its fair share
of glucose, a lot of its sweetness is derived from sorbitol.
No experimental error was encountered in this attempt of this
experiment although a few of the samples were out of range. To account for
this, the data points were simply omitted from the calibration curve due to
the fact that only three data points were required.

Conclusion:
In this experiment, the Ion Chromatographer was used to demonstrate
the qualitative, and quantitative analysis abilities of Ion Chromatography. In
experiment 1, Coca Cola was qualitatively analyzed to determine the sugar
that is most present. It was found that Coca Cola contains a majority of
Glucose, and Fructose. In the second experiment, Coca Cola was analyzed for
its Glucose content, and Apple Juice was analyzed for its sorbitol content.
This was made possible by the use of calibration curves. After testing, and
calculation, it was found that Coca Cola contains 37990 ppm of Glucose, and
Apple Juice contains 681.20 ppm of Sorbitol.

References:
Spectroscopy Laboratory Manual Fall 2014 edition, pages 106-107,
accessed 02/02/2015.
-

Nutritional/Human based information came from my own knowledge of the

human body, and the nutrition needed to keep it running