Cokie Gaich

CH321/Akinbo
Project Report
Freshly Squeezed Apple and Orange Juices vs. Bottled or Canned Apple and Orange
Juices: Acidity and Sugar Content

Background
The acidity of a compound can be defined in two ways. For Brønsted acids, a compound
will act as a hydrogen donor and can be expressed by the acid dissociation constant. Lewis acids
are related to the association constants of Lewis-adducts, meaning when a Lewis acid reacts with
a Lewis base, the base donates a pair of electrons to the acid to form the adduct (product of direct
addition of molecules) (4). The difference between total acidity and titratable acidity is that total
acidity is a measure of the complete acid content while titratable acidity is the measure of the
amount of acid present in a solution (3).

Titratable acidity and dental erosion:

Traditionally it’s been understood that pH can be an accurate indicator of erosive
potential for a food or drink, however baseline pH values only provide a measure of initial
hydrogen concentration and not the presence of undissociated acid. Titrable acidity is presently
more widely accepted as an accurate measurement of acid content in a drink and could be a
better tool for predicting their erosive potential. There have been many reports that link tooth
decay to consumption of juices. In theses cases, diluting the juices reduced their titratable acidity
in proportion to the dilution ration. The results of one report that tested soft drinks and juice
showed that the drinks were still highly acidic even after dilution, however the fruit juice has a
greater buffering effect (1). In another report fruit juices, soft drinks, and sports drinks were
tested for their pH, and all fell in the range of 3.1 – 3.6, however fruit juices had a titratable
acidity of about 2-3 higher than the other drinks (2). Though acidity can be a guide to erosive
potential of a drink, the acid type and concentration are proven to be more important when
determining the amount of damage even if the pH values are similar between the drinks.
Even though high acidity levels are interpreted as being the most harmful for dental
health, the consumption of them stimulates salivary flow making it possible that more of the
acidic drink could be cleared or neutralized more rapidly, spending less time in contact with
teeth. This being said, the clearance from the mouth depends on the drink’s ability to adhere to
the teeth, highly dependable on the drinks viscosity. Though we can compare the erosive
potentials of drinks, we cannot determine the degree in which these drinks can damage teeth
because this depends on how an individual consumes the drink (1). Such as drinking a soft drink
through a straw, the pH decrease in plaque pH was less than a mouth rinse of the same soft drink
(2).

Organic vs non-organic:
There have been several studies that have shown that organic tomatoes have a higher
content of biologically active compounds when compared to conventional tomatoes. A reason for
this is mainly nitrogen fertilization, since in organic farms only natural fertilizers, such as
compost and manure, can be used. It is known that when plants grow in soil that is abundant in
soluble nitrogen, they grow quicker however the amount of biocompounds is lower. When
studies were done comparing the organic and non organic tomatoes, they were compared using
dry matter (tomato juice was dried under certain conditions and were weighed for weight loss),
vitamin C content and the comparison of other biocompounds. The dry matter for both types of
 Gaich 2

tomato juices were very similar, however the fruits themselves had different content of dry
matter. The organic tomato fruit had a high dry matter content and the juice contained less water
compared to the non-organic tomatoes. However the non-organic tomato juice had a high
vitamin C content (19.30 mg/100g fw) compared to the organic juice (16.80/100g fw). An
important factor for levels of vitamin C in plants is if the plant is gown in low nitrogen
availability condition, there will be an increase of vitamin. The pH of the juices help to maintain
the vitamin C content, if the juice is maintained at a pH of 2 it stabilized the vitamin C content
(5).

Acidity and sugar content:

For many soft drinks the low pH and high titratable acidity can lead of enamel erosion,
and the sugar content in these drinks are metabolized by plaque microorganisms to create organic
acids that demineralize the enamel (6). In one study the sugar content (sucrose + fructose +
glucose) was measure in fruit juices, soft drinks, and sports drinks. It was found that fruit juices
and soft drinks have 9.3-9.8% sugar in one serving, while sports drinks contain 4.4% (2). The pH
of a beverage did not give the titratable acidity or the erosion potential, therefore in another study
it was shown that fruit juices had a higher initial pH than soft drinks but required more sodium
hydroxide to raise the pH. This study concluded that all sugars have about the same potential for
acid production in plaque compared to sucrose, however soft drinks cause a more pronounced
drop in pH than fruit juice in the first few minutes (6).

Acidity and taste:

A known characteristic of fruit juice is the tart taste that’s caused by the different
amounts of acids in the different fruit juices. Generally, the acid concentration is directly
proportional to the tartness, however the degree of correlation is not high in natural strength
citrus juice (7).

Methods and Materials Commented [CG1]: Make in paragraph form and make
into past tense
Preparing 500mL 0.1 sodium hydroxide (NaOH) from 50% (w/w) NaOH
1. Measure 4g of 50% (w/w) NaOH into a 500mL volumetric flask (KIMAX volumetric
flask) using an analytical balance
2. Add water to the mark

Standardization of the sodium hydroxide solution

1. Weigh 0.25g of potassium hydrogen phthalate (KHP) into a beaker or Erlenmeyer flask
2. Dissolve it with about 20mL of deionized water
3. Add three drops of phenolphthalein
4. Obtain weight of beaker or flask plus content (calls this A)
5. Place the beaker or flask on a stirrer and add stir bar into it. Stir gently
6. Insert rinsed pH probe (Accumet® AB15 pH Meter) into solution
7. Pour NaOH solution into burette (50mL KIMAX burette) and note initial volume. Call
this Vi
8. Titrate content of beaker or flask with NaOH from burette by adding 0.5mL at a time and
recording the pH every time
 Gaich 3

9. When the titration gets closer to the end point (when the colors stays longer) add NaOH
drop-wise. Record the pH and volume added
10. At the end point:
a. Read the burette volume again (call this Vf)
b. Remover the stir bar and weigh beaker/flask with content (call this B)
c. Put stirrer back into the beaker/flask
d. Titrate further by adding 1mL NaOH and obtain the pH
e. Repeat step 10d four more times
11. Repeat steps 1-10 four more times
12. Calculate the average concentration of NaOH and its precision based on:
a. Volume of NaOH used to reach the end point (Vep = Vf – Vi)
b. Mass of NaOH used (Me = B – A)
c. Volume of NaOH at equivalence (Ve) determined from the titration plot obtained
by plotting the volume (x-axis) vs pH (y-axis)

Method Validation for Determination of Acidity and Sugar Content:

1. Prepare 100mL of 1.0 x 10-2 g/mL spike solution of the acid that is predominant in the
juice (e.g. citric acid for orange juice; malic acid for apple juice)
2. Carry out triplicate titration of unspiked fruit juice using the 0.1M NaOH(aq). For each
replicate use 5 mL of juice + 5 mL of deionized water. Calculate the acidity for each.
a. End point of the titration is when the pH reaches 8.2 or above after one drop
3. Carry out replicate titration of fruit juice spiked at three levels (2 mL, 5 mL, 7 mL).
Calculate the acidity for each replicate
a. End point of titration is when the pH reaches 8.2 or above after one drop
4. Calculate accuracy:
Acidity of spike juice-acidityof unspiked juice
Accuracy = ( )´100
Expected acidity of spike

The Actual Thing:

Three SunPacific California Navel oranges were weighed in triplicate and their average were
calculate. The oranges were peeled. The weight of the juice collector cup was weighed in
triplicate and the average weight calculated. A Black & Decker JE2200B vegetable juice
extractor was used to extract the juice from the oranges into the collection cup and the cup was
subsequently measured. The juice volume, in mL, per gram of fruit, the volume of juice

Results and Discussion:

Standardization of NaOH
KHP KHP Vf Vi Vep Ve
B (g) A (g) Me (g)
wt (g) moles (mL) (mL) (mL) (mL)
Replicate
0.25 0.00124 46.4 35.5 10.9 100.1715 89.658 10.5135
1
Replicate
0.251 0.001229 31.5 19 12.5 101.438 89.68 11.758
2
Replicate
0.25 0.00124 27 16.5 10.5 100.785 90.225 10.56
3
Replicate 0.251 0.001229 19 6.4 12.6 102.734 90.185 12.549
 Gaich 4

4
Average 0.001227 11.625 11.34513

Concentration calculations:
Moles (NaOH) = mass (NaOH) / Molecular weight (NaOH)
= 11.3345113g / 39.997 gmol-1 = 0.28338 moles NaOH
Concentration = moles / volume = 0.28338 moles/ 11.625 mL
= 0.0243771214 moles/mL *(1000mL/1L) =

From graphs:

Method Validation:

Acidity of Orange Juice:

Average
acidity
Spike Mass of Expected
Volume of 0.1M Acidity of acid solution of acid
Volume acid in acidity
NaOH used (mL) from titration solution
(mL) spike (g) (g/mL)
from
titration
1 2 3 1 2 3
0 0 0 4.57 5.03 4.4 8.7799 9.664 8.453 8.966
2 0.0202 22.876 8.1 9.4 8.6 15.562 18.059 16.522 16.714
5 0.0505 57.191 17.3 16.4 17.2 33.237 31.508 33.045 32.596
7 0.0707 80.067 20.6 20.7 19.6 39.577 39.769 37.656 39.000

Spike
Standard deviation of
Volume Accuracy of Method Precision of method (RSD)
Acidity found
(mL)
0 0.6262 0 6.984311866
2 1.2598 33.87 7.53728566
5 0.9477 41.32 2.907396579
7 1.1686 37.51 2.996434744

From the tables above the average acidity of orange found from titration increases with the
increase of spike volume in the juice sample. Though for all levels of spiked and unspiked
samples the average acidity found is lower than the expected acidity, with the average acidity of
the 7mL spiked solution being half of the expected value. The accuracy of this method for all
spiked solutions are below 50% and the precision of the method is low therefore this method for
orange juice is not accurate or precise. This could have been due to the systematic error from the
pH meter.

Acidity of Apple Juice

 Gaich 5

Average
acidity
Spike Mass of Expected
Volume of 0.1M Acidity of acid solution of acid
Volume acid in acidity
NaOH used (mL) from titration solution
(mL) spike (g) (g/mL)
from
titration
1 2 3 1 2 3
0 0 0 2.7 3.2 5.5 3.6204 4.291 7.3745 5.095
2 0.02 14.963 8.7 10.4 15.1 11.666 13.945 20.2478 15.286
5 0.05 37.407 9.91 9.49 20.11 13.288 12.725 26.966 17.660
7 0.07 52.369 13.5 15 26.1 18.102 20.114 34.998 24.404

Spike
Standard deviation of
Volume Accuracy of Method Precision of method (RSD)
Acidity found
(mL)
0 2.002 0 39.298
2 4.445 66.667 29.080
5 8.064 71.147 45.664
7 9.229 79.121 37.816

From the tables above the acidity of the apple juice increased with the volume of spike added to
the solution. All of the found acidities are lower than the expected values calculated, but the
acidity found for the 2mL spiked solution is within a 1g/mL difference and the acidity found for
the 7mL spiked solution is about half of the expected acidity. The accuracy of this method for
apple juice is above 50% but all below 80%, therefore it is more accurate than the orange juice.
The precision of the data is higher than that for the orange juice making this method more
precise for the apple juice. Once again this could be due to the differences in the pH meters used
for each type of juice.

Percent Sugar in Orange Juice:

Refractive % Sugar of
Sample Brix Accuracy
Index spike
water 1.333 0
0% 1.34 4.75 4.5
2% 1.344 7.5 7 125
5% 1.3475 10 9.1875 234.375
10% 1.364 20 19.5 750
Average 10.046875
St Dev 6.586637159
%RSD 65.55906348

Percent Sugar in Apple Juice:

Sample Refractive Index Brix % Sugar of spike Accuracy

water 1.333 0.00
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0% 1.351 12.0 11.375

2% 1.353 13.5 12.625 62.5
5% 1.357 15.75 15.125 75
10% 1.3655 21.0 20.4375 90.625
Average 16.0625
St Dev 3.989732917
%RSD 24.83880415

1.368
y = 0.1469x + 1.3504
1.366 R² = 0.9903
1.364
1.362
Refraction Index

1.36
1.358
1.356
1.354
1.352
1.35
1.348
0% 2% 4% 6% 8% 10% 12%
%Sucrose

The Brix values indicate the sugar content in a solution with 1 Brix being 1g of sucrose in 100g
of solution, thus representing the strength of that solution. We used the water sample as a blank
to calibrate and ensure that the instruments are working correctly. As the percent of spiked
sucrose solution increased the refractive index and brix increased as well. The accuracy of this
method for apple juice is accurate especially as the percent of sucrose increases. The correlation
coefficient is close to 1 therefore there is a strong correlation between percent sucrose and
refraction index, this graph also shows that there is a linear relationship between percent sucrose
and refraction index which can allow us to find either using the linear equation above.

Average Orange Fruit weight

Fruit 1 Fruit 2 Fruit 3
Weight 1 304.711 310.383 323.915
Weight 2 304.698 310.304 323.897
Weight 3 304.675 310.247 323.863
Average 304.6946667 310.3113333 323.8916667
Overall Average 312.9658889
Standard Deviation 9.869964597

Juice Content per Orange Fruit:

 Gaich 7

Fruit 1 Fruit 2 Fruit 3

Collection cup (g) 62.124 62.124 62.124

Cup + juice(g) 214.116 197.662 233.664
Volume of juice
144 122 160
(mL)
juice (g) 151.992 135.538 171.54
ml/g fruit 0.472604268 0.393153543 0.493992332
ml/ fruit 144 122 160
g juice/fruit 151.992 135.538 171.54

There should be a correlation between the average weight of each orange and the volume of juice
produced. However fruit 2 does not follow this trend, as its mass is greater than fruit 1’s but fruit
2 produced less juice out of all of the oranges.

Juice Acidity in Orange:

Acidity of Acidity of Acidity
Volumeof Volume Volume Average
acid acid of acid
0.1M of 0.1M of 0.1M acidity of
solution solution solution
Juice Brand NaOH NaOH NaOH acid
found found found
used for used for used for solution
from from from
titration titration titration found
titration titration titration
mL ml ml ml 1 2 3
OJ 1 3.85 4.3 4 7.39662 8.26116 7.6848 7.78086
OJ 2 7.3 3.55 3.25 14.02476 6.82026 6.2439 9.02964
OJ 3 5.35 5.45 4.9 10.27842 10.47054 9.41388 10.05428
Tropicana 4.2 5.05 4.7 10.5666 9.70206 9.02964 9.7661
Simply Orange 5.5 5.35 4.95 10.5666 10.27842 9.50994 10.11832

The average acidity of the natural orange juices have a wide range this could be due to the
ripeness of the oranges as it was just purely the juice with no modified content. The average
acidity of the bottle juices is similar to the acidity of fruit juice 3 falling on opposites sides of the
acidity value.

Standard Accuracy Precision of

Juice Brand deviation of of Method
acidity found method (RSD
mL
OJ 1 0.440202221 5.657500858
OJ 2 4.335489081 48.01397488
OJ 3 0.562860235 5.598215233
Tropicana 0.770478651 7.88931765
 Gaich 8

Simply
0.546220282 5.398329781
Orange
For all the juices except for that of fruit 2 the standard deviation between the acidity found is less
than 1. Fruit juice 2 has a much higher standard deviation of 4.33 this is due to the first titration
of the juice where 14 mL of NaOH was need to reach the end point.

Sugar Content in Orange Juices:

RI Brix
OJ 1 1.3495 11.25
OJ 2 1.3435 7.25
OJ 3 1.345 8.25
Tropicana 1.344 7.5
Simply
1.344 7.5
Orange
Water 1.3325 0

Deionized water was used during this test as a control variable and to compare the amount of
sugar in juices to water. The bottle fruit juices, Tropicana and Simply Orange, have exactly the
same RI and Brix value suggesting that fruit juice companies must reach a standard for all of
their oranges juices before they can be sold. These values are the same ones as the values given
for a 2% sucrose solution in the method validation, indicating that companies may strive to reach
a similar sweetness in their orange juice to 2% sucrose solution. The three naturally squeezed
oranges have similar RI values but have a wide range for their Brix values (7.25 - 11.25). A
reason for this range in Brix values could be due to the ripeness of the fruits when they are
picked. Since they are naturally squeezed no extra sugar is added or the sugar content isn’t
modified to meet a standard, this is a similar concept to why the sweetness of fruits vary from
individual fruits.
 Gaich 9

References
(1) Cairns, A.; Watson, M.; Creanor, S.; Foye, R. The Ph And Titratable Acidity Of A Range Of
Diluting Drinks And Their Potential Effect On Dental Erosion. Journal of Dentistry
2002, 30, 313-317.
(2) Birkhed, D. Sugar Content, Acidity And Effect On Plaque Ph Of Fruit Juices, Fruit Drinks,
Carbonated Beverages And Sport Drinks. Caries Research 1984, 18, 120-127.
(3) pH, Total Acidy, And Titratable Acidity (TA) Defined; 1st ed.; M&M Wine Grape Company,
LLC, 2013; p. 1.
(4) Definition of acidity - Chemistry Dictionary
http://www.chemicool.com/definition/acidity.html (accessed Jan 31, 2017).
(5) Hallmann, E.; Lipowski, J.; Marszałek, K.; Rembiałkowska, E. The Seasonal Variation In
Bioactive Compounds Content In Juice From Organic And Non-Organic Tomatoes. Plant
Foods for Human Nutrition 2013, 68, 171-176.
(6) Saeed, S.Al-Tinawi, M. Evaluation Of Acidity And Total Sugar Content Of Children′S
Popular Beverages And Their Effect On Plaque Ph. Journal of Indian Society of
Pedodontics and Preventive Dentistry 2010, 28, 189.
(7) Kilburn, R. The Taste Of Citrus Juice; 1st ed.; 1958; pp. 251-254.