In the Laboratory

Carbohydrate Analysis: W
Can We Control the Ripening of Bananas?
S. Todd Deal,* Catherine E. Farmer, and Paul F. Cerpovicz
Department of Chemistry, Georgia Southern University, Statesboro, GA 30460; *[email protected]

Background Table 1. Student Data on Carbohydrate Composition of

Bananas
Laboratory exercises involving carbohydrates tend to Stage of Ripeness Starcha Reducing Sugarsa
be based on classic qualitative analyses employing Benedict’s (Appearance) (% of Total Sample) (% of Total Sample)
reagent, Fehling’s solution, polarimetry, etc. (1). Recent Green with some yellow 12.9 4.0
exceptions are the synthetic experiments published by Lowray
Yellow (ripe) 6.2 4.4
(2) and Norris (3) in this Journal. While the latter are excel-
lent examples of nontraditional carbohydrate investigations, Yellow with black spots
2.0 11.6
(overripe)
we wanted to develop an introductory-level experiment that
focused on quantitative carbohydrate analysis and maintained aBased on wet weight of original sample. Note that the data in ref 5

the theme of our laboratory program, which is the utilization are based on dry matter.
of common foodstuffs to investigate biochemical principles
relevant to nutrition.1 The result is an experiment with a
threefold purpose: (i) determination of the carbohydrate fructose and 3,5-dinitrosalicyclic acid. The reaction results
composition of a banana, (ii) analysis of the changes in in the formation of a reddish brown reduction product, 3-
carbohydrate composition as the banana ripens, and (iii) amino-5-nitrosalicyclic acid, from the colorless 3,5-dintro-
investigation of the control of the ripening process. salicyclic acid. The inclusion of potassium sodium tartrate
In a 1986 study, Englyst and Cummings demonstrated (Rochelle’s salt) as a component of the reagent was a later
that while the total carbohydrate content of a banana remains modification designed to limit the solubility of oxygen in the
relatively constant (~28% of dry matter) during ripening, the reaction mixture, since dissolved oxygen can interfere with
free sugar content increases sharply while the starch content the oxidation of glucose.
shows a similar decrease (4). Using this study as our basis, we To begin the analysis, the students homogenize a 1-g
developed a carbohydrate analysis experiment entitled sample of banana in cold distilled water using a hand-held
“Ripening Bananas: What’s Happening and Can We Control glass homogenizer. The resulting suspension is centrifuged
It?” In it, students determine the carbohydrate composition and the pellet is isolated, dried, and weighed to assess the
of bananas at various stages of ripeness. Using these data as a starch content of the banana. (The dry pellet is composed mostly
benchmark, they then test various storage methods to ascertain of starch.) Then, a sample of the supernatant is analyzed using
which method best preserves the desirable qualities (taste, color, the DNS assay to determine the banana’s reducing sugar
etc.) of bananas for the longest period. content. Each pair of students completes duplicate analyses
of starch and reducing sugar content and records the averages
Description of the Experiment of the analyses on the chalkboard or on a pre-prepared class
data sheet. Using the collective class data, the students can
Determining the Carbohydrate Content of Bananas observe the differences in the carbohydrate composition of
In the first part of the experiment, each pair of students bananas at various stages of ripeness. Table 1 shows some
analyzes the starch and free reducing sugar (glucose and representative student data.
fructose) content of a green, ripe, or overripe banana. For the
free reducing sugar analysis, we chose the 3,5-dinitrosalicyclic Determining the Best Method for Storing Bananas
acid–potassium sodium tartrate (DNS) assay originally de- At the beginning of the first laboratory period, each pair
veloped by Sumner for reducing sugars (5). We decided to of students is given a yellow-green banana (in addition to
analyze only for reducing sugars because, as demonstrated by their green, ripe, or overripe banana) taken from the same
Englyst et al. (4), the change in free sugar content of a banana bunch and told to store this banana (see below). During the
as it ripens is due mainly to an increase in the amount of following laboratory period, the students analyze their stored
reducing sugars. Additionally, the chosen method has been banana to determine the best method of storage to control
shown to be an accurate quantitative measure of reducing the ripening process.
sugar content in solutions containing both unhydrolyzed To design the “storage” part of the experiment, we turned
starch and free reducing sugars (6 ). to popular culture. Many readers will recall the jingle that
The DNS assay is a colorimetric assay based upon the was used long ago by one of the major banana producers,
redox reaction between a reducing sugar such as glucose or which implored consumers not to put their bananas in the
refrigerator. Why shouldn’t we store bananas in the refrig-
erator, we wondered? Thus, the refrigerator became one of
†
Initially presented at the 52nd Southeast/56th Southwest the test storage methods. Next, we noted that it is nearly
Combined Regional Meeting of the American Chemical Society, impossible to walk into the produce section of any grocery
Dec 6–8, 2000, New Orleans, LA. store without seeing a display of white plastic or wooden “ba-

JChemEd.chem.wisc.edu • Vol. 79 No. 4 April 2002 • Journal of Chemical Education 479

 In the Laboratory

nana hangers” with their attendant claim to keep bananas obviously the best storage method to slow the ripening process,
fresher, longer. The “banana hanger” became another test but many consumers would refuse to eat a refrigerated banana
storage method. Then, one of us (PFC) recalled the instructions because of its appearance. The brown paper bag method yields
of his wise grandmother to “always store bananas in a brown the most appealing fruit, but the outward appearance belies
paper bag.” So, the brown paper bag became yet another test the almost overripe flesh that lies beneath the skin.
storage method. Finally, we threw in the “conventional wisdom”
method of fruit storage and chose the lab drawer as a cool, Summary
dark place to serve as the final test storage method.
In this portion of the experiment, the students are assigned While we utilize this experiment in our Nutritional
one of the test storage methods and instructed to store their Biochemistry course, we believe that it is broadly applicable.
bananas for the week. During the following lab period, they We feel confident that this experiment is appropriate for
repeat the analyses from the first week, taking samples from courses such as introductory chemistry and organic chemistry
their stored banana. We also ask them to record the appearance in which the basic principles of carbohydrate analysis are
of the skin of the banana when they remove it from storage. introduced. Techniques taught in this experiment include
The students enjoy this second part of the lab exercise, and construction and use of standard curves, spectrophotometric
the collective results are always eagerly anticipated. analysis, and centrifugation. Additionally, interpreting the
results of the experiment requires both critical-thinking and
Hazards decision-making skills.

3,5-Dinitrosalicylic acid is toxic and an irritant. Potas- W

Supplemental Material
sium sodium tartrate is an irritant. Always wear gloves and
appropriate eye protection when handling these compounds. Notes for the instructor and a handout for students with
detailed instructions, a data sheet, and questions are available in
Discussion this issue of JCE Online.

As any trained chemist might expect, the refrigerated Note

banana “ripens” the least during the week of storage. Typically,
students find that the composition of the refrigerated banana 1. This is the fourth report in a series on the development of
has changed very little, whereas the composition of each of our nutritional biochemistry laboratory program. For the first three
the other three bananas is similar to that of the ripe banana parts see ref 7.
from the first part of the experiment. After all the data are
collected, we pose the question, “Which of the storage methods Literature Cited
would you recommend to consumers?” While this seems like
a straightforward question, it turns out to be far more complex 1. See for example: Williamson, K. L. Macroscale and Microscale
than it first appears. Organic Experiments; Heath: Lexington, MA, 1989; pp 499–506.
Since most of our students are nutrition majors, they Boyer, R. F. Modern Experimental Biochemistry, 2nd ed.;
understand that food is more than taste and smell. We’ve often Benjamin/Cummings: Redwood City, CA, 1993; pp 347–362.
overheard them in class discussing mouth feel, consistency, 2. Callam, C. S.; Lowray, T. L. J. Chem. Educ. 2001, 78, 73–74.
and appearance as important facets of food and eating. It is the 3. Norris, P.; Freeze, S.; Gabriel, C. J. J. Chem. Educ. 2001, 78,
last characteristic—appearance—that presents the dilemma. 75–76.
At the end of a week, a banana stored in the refrigerator is 4. Englyst, H. N.; Cummings, J. H. Am. J. Clin. Nutr. 1986,
not a pretty sight! Typically, a refrigerated banana has a 44, 42–50.
mottled green-brown color that makes it appear almost 5. Sumner, J. B. J. Biol. Chem. 1921, 47, 5–9.
bronze—not at all an appetizing appearance. The banana 6. Bernfeld, P. Methods Enzymol. 1955, 1, 149–158.
stored in the brown paper bag, on the other hand, has a rich, 7. Deal, S. T.; Pope, S. R. J. Chem. Educ. 1996, 73, 547. Pope, S. R.;
bright yellow color with very few brown spots, while the other Tolleson, T. D.; Williams, R. J.; Underhill, R. D.; Deal, S. T.
two are yellow but have many patches of brown. Thus, the J. Chem. Educ. 1998, 65, 761. Mullis, T. C.; Winge, J. T.;
students are forced to make a tough decision. Refrigeration is Deal, S. T. J. Chem. Educ. 1999, 76, 1711.

480 Journal of Chemical Education • Vol. 79 No. 4 April 2002 • JChemEd.chem.wisc.edu