INVESTIGATING SCIENCE

SCIENTIFIC REPORT

Alark Sharma | Investigating Science | 26/03/21

Aim: To observe and compare the effect of applying many solutions of different pH to
determine which solution is the best preventative of enzymatic browning.
Hypothesis: If the pH of the solution the apple slices is being soaked in higher, the lower
the rate of enzymatic browning of the apples.

Materials and Equipment:

 Apple 3x (Pink Lady variety)

 Milk
 Baking soda
 Vinegar
 Lemon Juice
 Tap water
 Paper plates 2x
 Paper towel
 pH meter
 Tongs
 Stopwatch 2x
 Nitrile gloves (FDA and HACCP certified)
 Permanent marker (soft, felt-tipped)
 Labels (Avery Handy, rectangle 34 x 76mm)
 Beaker 5x
 Kitchen knife
 Chopping board
 Distilled Water
 Camera/Phone
 SRM scale

Independent Variable: Use of different solutions/treatments

Dependent Variable: Degree of enzymatic browning of each apple slice

Controlled Variable: Untreated apple slice (no coating)

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METHOD:

1. Collect all materials and equipment you will require for the experiment
2. Prepare the solutions and respectively pour them in each beaker at 150ml. Note
that you only need a small quantity of each, enough to fully submerge the apple
slice in the beaker.
3. Record the pH of each respective solution before moving forward with the
experiment using a pH meter (clean the pH meter every time after attaining
readings for each solution using distilled water)
4. Ensure you have nitrile gloves worn (which is the preferred material of choice for
handling foods in a safe, contaminate-free and additive-free manner).
5. Place an apple onto the chopping board with the stem facing up
6. Using a kitchen knife, make a cut ½-inch to the right or left of the stem and cut all
of the way down.
7. Cut the two halves into 6 equally sized apple slices (untreated control, milk,
baking soda, vinegar, lemon juice, tap water) using proper cutting technique.
8. Set one untreated apple slice at room temperature (this is your control)
9. Ensure to label using a permanent marker of each slice and treatment on 2 paper
plates with 3 slices on each.
10. Using tongs, dip an apple slice into the beaker filled with lemon juice for 30
seconds (with your stopwatch correctly zeroed).
11. Repeat step 10 for each of the differing treatments for the apple slices (make sure
to rinse and clean tongs after every use to avoid cross-contamination).
12. Place each of the different treatments to their corresponding labels on the paper
plates
13. Set a timer on your phone/device and monitor the oxidation progress of each slice
for 10 minutes for 3 intervals
14. Observe, photograph, and record your qualitative data after each consecutive
interval. Use a SRM scale/colour chart to determine the degree of browning of
each apple slice.

SRM scale/colour chart from:

http://www.popsci.com/science /article/2012-
12/beersci-how-beer-gets-its-color

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RESULTS:
Table 1. Enzymatic browning of apple slices in various pH treatments over time based on
SRM scale/colour chart

Trial 1
Solution/Treatmen pH value Amount of Amount of Amount of
t browning after browning after browning after
10 minutes 20 minutes 30 minutes
Untreated control Not applicable 6 7 8

Lemon Juice 2.3 1 1 2

Baking Soda 6.4 4 5 6
Vinegar 2.6 2 3 5
Milk 6.6 2 3 4
Tap water 7.1 2 3 4

Table 2. Enzymatic browning of apple slices in various pH treatments over time based on
SRM scale/colour chart

Trial 2
Solution/Treatment pH value Amount of Amount of Amount of
browning after browning after browning after
10 minutes 20 minutes 30 minutes

Untreated control Not applicable 4 5 7

Lemon Juice 2.2 1 2 2
Baking soda 6.4 3 4 5
Vinegar 2.6 2 3 4
Milk 6.7 4 5 5
Tap water 7.0 2 3 3

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 Figure 1(Trial 1): Clustered Column Chart

Figure 2 (Trial 2): Clustered Column Graph

DISCUSSION:
Throughout the experiment, the shade of brown in which apple slices developed into was
recorded, photographed and compared with each other over the two trials. Most focally, the

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lemon juice consistently outperformed each of the different treatments in both trials, leading to
apple slices exhibiting little to no browning. It was noticed that milk and baking soda, the
development of enzymatic browning was gradual and increased by similar amounts after each
interval, due to their more basic pH values. Surprisingly, the performance for tap water (7.0 pH)
was effective in retarding discoloration of the apple slices, where it seemed that tap water
impeded the exposure of oxygen and the ability for the enzymes to catalyze the first step in the
oxidation process. In contrast, vinegar appeared to show relatively low shades of brown in the
first interval (10 minutes) as denoted by Figure 2, but eventually experienced a dramatic increase
in pigmentation even though it is closer to Lemon juice in terms of pH value. Lemon juice has a
pH level of 2 which makes it extremely acidic. It can be inferred, that the PPO enzymes are
inhibited by acids as well, so the more acidic the liquid, the more effective it is in slowing the
oxidation process, and cut off the enzyme’s access to oxygen. It can be deduced that Lemon Juice
is considered an anti-browning gent due to its properties as an acid because it is full of ascorbic
acid (Vitamin C) and it has a low (acidic) pH level. Ascorbic acid works because oxygen will
react with it before it reacts with the polyphenol oxidase enzyme in the fruit, acting as
conservative. Despite being as low as lemon juice in terms of pH value, vinegar did very poorly
due to its properties as an acid. The acetic acid in vinegar is a not an antioxidant, unlike Lemon
Juice which thereby hindered its ability to restrict oxygen coming in contact with the fruit tissues.
This means that the pH level of the liquids plays a large role in the slowing of the oxidation
process, but there are always some exceptions, such as vinegar.

Sources of error identified during the experiment included, the distinguishability of each liquid.
When the apple slices were dipped into each liquid, the dripping liquid from the apple slice wiped
of the permanent marker labels for each treatment which resulted in misconception or error of
where to put the apple slice on the two paper plates. This error can be remedied with proper use
of labels and not using permanent marker on the plates to write down where each slice is to be
placed. In addition, another source of error experienced was accurately measuring the different
shades of brown on the apple slices, as the SRM chart had many different shades of brown which
were all extremely similar to each other. This affected the ability to differentiate which shade of
brown each slice had.

In future experimentation improvements would include being wise to expand to vegetables as

well as fruits. This can compare the different levels of polyphenol oxidases in both fruits and
vegetables and not just apples, creating a better understanding of the distribution of polyphenol
oxidases in nature. Expanding to many different liquids with distinctly different pH levels will
also create a more accurate depiction of the effect that pH levels play on the oxidation of produce.
In addition, another improvement can be the type of knife used to cut the apple. The type of knife
that was used also affected the oxidation process of the fruits. Stain-less steel metal knives or
kitchen knives accelerate the process because they are usually made up of a mixture of iron and
copper which react with the polyphenol oxidase enzymes. An improvement to this would be
using a ceramic knife which would have kept the oxidation process to be completely natural and
would not have sped up the oxidation process unnaturally.  

Lastly, another main source of uncertainty was the moisture in the room during the experiment.
The experiment was conducted in one day for each of the two trials, with certain weather
conditions. On the day of experimentation, it was rainy. The rain created a slightly more humid

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environment in the room that the experiment was conducted in, as the warmth and moisture of the
rain entered through the open windows and open doors. Polyphenol oxidases need a sufficient
supply of water to be properly activated, so on the results attained on that specific day could vary
if the experiment was conducted on a dryer day, which could possibly impact the validity and
reliability of the experiment, since the amount of water in air would’ve been greater to fully
sustain the enzymes as opposed to a dryer day. Thus, to improve the reliability of the results
attained, it would be better to conduct trial-to-trial experimentation on dryer and wetter days to
reduce any alterations to our experiment.

CONCLUSION:
In conclusion, my results were able to confirm my hypothesis. In my hypothesis, I predicted that
if the pH of the solution the apple slices is being soaked in higher, the lower the rate of enzymatic
browning of the apples. Based on my results, I was able to see that lemon juice outperformed and
delayed enzymatic browning the most, thus being the best preventative for enzymatic browning.
As there are many factors that affect the rates of enzymatic browning; such as concentrations of
both active polyphenol oxidase and phenolic compounds present, the pH, and the temperature, I
cannot confidently confirm the results of my data. For future experimentation, it would be more
accurate if we expanded on the variety liquids on the pH scale, fruits composed of different
phenolic amounts, and attaining results in different weather conditions.

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