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Carbohydrate Analysis
Karasawa, Lindsey
lindseykarasawa@gmail.com
California State University, Long Beach (CSULB)
Department of Family and Consumer Sciences
1250 Bellflower Boulevard, Long Beach CA 90840
ABSTRACT
A series of tests were performed to analyze the composition of carbohydrates in a variety
of solutions. First, the sucrose concentrations of 5 beverages were measured by using a digital
refractometer. The beverage with the highest amount of sucrose was found to be the white grape
juice with a Brix of 14.1. With the same beverages, the sugar content was also determined by
using a hydrometer. This test also expressed the white grape juice to have the highest sugar
content at 16.0 g/100 mL of liquid. In the final experiment, Gatorade and coconut water were
both examined using Anthrone reagent and a UV Spectrometer to determine the sugar content.
Surprisingly, the sugar contents listed on the nutrition labels of the beverages did not match up
with those calculated. Using the equipment found in the lab, the legitimacy of Nutrition labels
was challenged. Evidently, these labels should not be seen as a firm measurement of the contents
of food or beverages.
Key Words: carbohydrate, concentration, beverage, sugar, anthrone
INTRODUCTION
Carbohydrates are the building blocks of most structures and food substances we come
into counter with. They are composed of carbon, hydrogen, and oxygen. These molecules take on
several different forms, including sugars such as sucrose. In order to construct food labels and
screen for adulteration, the investigation of sugar concentration must be performed upon
production. Multiple methods can be used to analyze sugar composition and concentration in
beverages. Sucrose contcentration is measured in Brix degrees (Bx), the amount of sugar in
grams per 100 mL of solution. This measurement is often used to determine if fruits are ripe
enough for harvesting (Silva and others 2014). Bx can be determined using a variety of tools.
Digital refractometers shine light into a drop of a liquid sample and measure at what degree the
right is refracted. Each degree of refraction indicates 1 Bx. Another tool that can be used is a
hydrometer. A hydrometer is a slim, graduated tube with a weight at the bottom. When placed in
a cylinder of liquid, it is able to measure the % of soluble solids (which in this case would be
sugar) in grams per 100 mL of solution (Son and others 2009). This tool is also used in the
fermentation of wine and alcoholic beverages.
The measurement of light absorbance is also a worthy way to measure sugar
concentration in a liquid. Ultra Violet Spectrometers (UVSpec) shine light rays into a sample of
liquid and measure the amount of light is absorbed by the sample. This measurement allows
professionals to determine the concentration of pigments in a solution (Czegan and other 2012).
A pigmented indicator must be used to identify the sugar concentration of liquids using the
UVSpec. Anthrone reagent is a qualified solution that binds to sugars upon boiling. When a
sugar solution with Anthrone reagent is measured for absorption by the UVSpec, the results
would directly correlate to the concentration of sugars in the solution (Zweens and other 1968).

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MATERIALS AND METHODS


Determination of the Sucrose Content of Selected Beverages using a Digital Refractometer
The digital refractometer was cleaned and calibrated by applying 1 drop of distilled,
deionized water (DDW) to the lens. Any remaining DDW was wiped away using a Kim wipe.
Using a transfer pipette, 1 drop of each sample liquid was transferred to the lens and analyzed,
one at a time as seen in Figure 1. The lens was cleaned in between each sample. The results,
means, SDs, and CVs were recorded in Table 1.
Determination of the Sugar Content of Selected Beverages Using a Hydrometer
The Brix hydrometer was gently inserted into a 250 mL measuring cylinder. Each of the
sample beverages were poured over the hydrometer allowing the device to float as seen in Figure
2. The value at which the meniscus leveled at was recorded in Table 2.
Determination of Total Sugars of Brand-Name Coconut Water/Gatorade by Anthrone
Reagent
Two boiling water baths and one cold water bath were prepared. DDW was used to make
1:10,000 dilutions of Gatorade and coconut water in test tubes. Samples were collected from four
different bottles of each beverage. 100 mg of glucose was combined with 100 mL of DDW in a
beaker to create a stock solution. 5 glucose standards were prepared using the stock solution:
0.02 mg/mL, 0.04 mg/mL, 0.06 mg/mL, 0.08 mg/mL, and 0.10 mg/mL. The final volumes of all
of the standards were 1 mL. The samples were all vortexed and then 4 mL of Anthrone reagent
was added to each tube. The samples were vortexed once more and placed in the boiling water
bath for 10 minutes as see in Figure 3. Upon completion, the tubes were allowed to cool in a
cold-water bath until the liquids reached room temperature. 2 mL from all of the samples of
Gatorade, coconut water, and the standards were transferred into cuvettes. First the blank was
entered into the UVSpec (Orion AquaMate 8000) and the color intensity was determined using
an absorbance of 630 nm. Then, all of the other samples were analyzed in the same fashion. The
color intensity values were recorded in Table 3. A standard curve was created on Excel to
determine the glucose concentrations of the Gatorade and coconut water.
RESULTS AND DISCUSSION
The Bx was determined using both the refractometer and the hydrometer. When the
results were collected from the refractometer, as seen in Figure 1, the mean absorbances were
calculated in Table 1. The beverage with the highest Bx was the white grape juice at 14.1
0.130. All of the values recorded from the refractometer were very precise however not
completely accurate when compared to the respective nutrition labels. For example, the white
grape juice label stated that there should be 40.0 g of sugar in 240 mL of juice. According the
refractometer, in 240 mL there was 33.9g. In contrast, H.S. Son did a study in 2009 where the
Bx of a known wine was recorded as higher using the refractometer than the known Bx. His
study also stated that the hydrometer produced lower results than the known Bx of wines (Son
and others 2009). These statements were not complementary to the findings of this experiment.
When measured with the hydrometer as seen in Figure 2, the white grape juice was, again, the
beverage with the highest recorded Bx at 16 g/100 mL as seen in Table 2. This translates to 38.4
g of sugar in 240 mL, which was still less than the previously stated sugar content of the juice.
At large, the results of the refractometer were similar to those of the hydrometer, but all
of them were slightly lower. There are many factors that could have contributed to these

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inconsistencies such as the calibration of the hydrometer or possible impurities in any of the
beverages. The electronic refractometer is more detailed and finely calibrated than the
hydrometer, which can lead to less precise results as well. The hydrometer is used in the
production of alcoholic beverages to indicate when all of the sugars have fermented by
measuring the buoyancy of the hydrometer tube and therefor determining the sugar content of the
liquid (Lorefice and other 2006).
Lastly, the sugar concentrations of coconut water and Gatorade were determined using an
UVSpec. The preparation of this experiment is seen in Figure 3. These instruments shine light
through a sample of liquid and measure the amount of light absorbed. This allows for researchers
to calculate the concentration of a liquid and predict its characteristics. The results collected from
the UVSpec were recorded and made into a graph, as seen in Table 3. A trend line and its
equation were used to estimate the sugar content of the Gatorade (62.9 g/ml) and coconut water
(21.6 g/ml). A correlation coefficient value was also determined in Table 3. This value expresses
how accurately the values observed correlates with the results of the experiment. The results
were very precise, however not accurate when compared to the nutrition labels. The Gatorade
bottle stated that there were 21.0 g of sugar in 355 mL and the coconut water bottle stated that
there were 15.0 g of sugar in 414 mL. These results were so drastically different, that it leads to
the assumption that somewhere in the UVSpec methodology there was an error.
CONCLUSION
Consumers often use food labels to help them determine what foods and beverages they
would like to purchase for their families. However, when the results of these experiments are
considered, it can be determined that the labels are often ill advising and can lead the consumer
astray. It is important for food scientists to carefully analyze the sugar composition of all
beverages and construct food labels with integrity.

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REFERENCES
Czegan DAC, Hoover DK. 2012. UV-Visible spectrometer: versatile instruments across the
chemistry curriculum. J of Chem Educ 89: 304-309. doi: 10.1021/ed200776x
Lorefice S, Malengo A. 2006. Calibration of hydrometers. J of Meas Sci Technol 17:2560-2566.
doi: 10.1088/0957-233/17/10/005
Silva SA, Queiroz DM, Pinto FAC, Santos NT. 2014. Coffee quality and its relationship with
Brix degree and colorimetric information of coffee cherries. J of Precision Agric 15: 543-554.
doi: 10.1007/s11119-014-9352-y
Son HS, Hong YS, Park WM, Yu MA, Lee CH. 2009, A novel approach for estimating sugar
and alcohol concentrations in wines using refractometer and hydrometer. J of Food Sci Food
Chem 74: 106-111. doi: 10.1111/j.1750-3841.2008.01036.x
Zweens J, Bouman PR. 1968. Simple and accurate determination of urinary glucose excretion
with anthrone reagent. J of Diabetologia 4: 278-280.

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APPENDIX

Figure 1: Determination of Sucrose Content with a Digital Refractometer


Photo courtesy of CSULB Food Science Lab

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Figure 2: Determination of Content with a Hydrometer


Photo courtesy of CSULB Food Science Lab

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Figure: 3 Heating of Samples with Anthrone Reagent


Photo courtesy of CSULB Food Science Lab

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