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Apple Wine PDF
Apple Wine PDF
Introduction to Biotechnology:
Prepared by:
AMADO, Rosenn B.
DANDAMUN, Benbellah Ali Y.
EBREO, Mencyn Michelle Kellie
GONZALES, Belinda Jean A
IGAYA, Deanne May G.
JUAGCO, Cris-Anne III
LOPEZ, Charmaine A.
TORRES, Angela Jane R.
BSCHE-IV
Submitted to:
Engr. Ronald Allan Co
ACKNOWLEDGEMENT
Fermentation of Apple Juice for the Determination of Wine and Ethanol Concentrations Page 2
Chapter I: INTRODUCTION
Fermentation of Apple Juice for the Determination of Wine and Ethanol Concentrations Page 3
When yeast respires anaerobically, only a small amount of energy is
released from the sugar and so most of the sugar stays in the alcohol. When the
alcohol concentration reaches about 15% of alcohol by volume, the yeast cells
die. Ethanol concentration can be based by determining the density
measurement of the ethanol.
Fermentation of Apple Juice for the Determination of Wine and Ethanol Concentrations Page 4
OBJECTIVES
The objective of this study is to determine the wine and ethanol concentrations
using bio-chemical reaction and stabilization process.
Specific Objectives:
Fermentation of Apple Juice for the Determination of Wine and Ethanol Concentrations Page 5
Chapter II: Methodology
Materials Needed
Fermentation of Apple Juice for the Determination of Wine and Ethanol Concentrations Page 6
Procedure
Preparation:
1. Bottles obtained were of the same size and brand. Labels were removed.
2. Bottles and covers were cleaned with soap and water.
3. After cleaning, it was sterilized with hot water.
4. Bottles and covers dried by wiping the excess water and turned the bottle
down on a clean cloth for the condensate will run down and be
absorbed by the cloth.
5. Bottles were paired, making up 14 pairs. 7 for the first batch. Another 7 for
the second batch.
6. While it is still drying, covers were being bore with the use of cutter.
7. 2 straws were connected together and locked by scotch tape. And to be
inserted on the bottle caps.
8. One end of the straw must be cut to make it shorter (for the Wine Mixture
bottle). While, the other end of the straw must be long to have contact
with the water.
9. Bottles were labeled from 1 to 7, on both batches.
1. For basis, beaker was put on the analytical balance and set into zero.
2. 300 ml water were poured, and weighed. Record the data (mass, volume,
density, average mass, average volume, average density)
3. For the next basis: beaker was put on the analytical balance and set into
zero.
Fermentation of Apple Juice for the Determination of Wine and Ethanol Concentrations Page 7
4. 150 ml apple juice were poured, plus one pinch of yeast and weighed.
Record the data (mass, volume, density, average mass, average volume,
average density)
5. After the preparation, bottles has to be placed in a dark room and shake
free place.
Actual Set-up:
1. Everyday, take a 3-trial of obtaining twenty (20 ml) for each bottle.
(Everyday there should be four bottles to take samples from. 2 from the
first batch (Wine mixture and Ethanol), and another 2 from the second
batch (Wine mixture and Ethanol)
2. For every trial, It should be put on a dry vial, or any glass bottle to weigh
the sample.
3. Record the data. (mass, volume, density, average mass, average volume,
average density).
( ) Where:
%
Fermentation of Apple Juice for the Determination of Wine and Ethanol Concentrations Page 8
Cwine = Concentration of wine (g/ml) ρEtOH= density of pure ethanol
(0.8059 g/ml)ρwine = density of wine
@t=0, initial value
Table of Data
DATA AND RESULTS: Apple Wine
Basis:
Fermentation of Apple Juice for the Determination of Wine and Ethanol Concentrations Page 9
Trial 1 20 20.9 1.045
Trial 2 20 20.9 1.045
Trial 3 20 20.6 1.03
Average 20 20.8 1.04
Fermentation of Apple Juice for the Determination of Wine and Ethanol Concentrations Page 10
Day 1 Volume (ml) Mass (g) Density (g/ml)
Trial 1 20 21.4 1.07
Trial 2 20 21.9 1.095
Trial 3 20 21.9 1.095
Average 20 21.7333 1.0867
Fermentation of Apple Juice for the Determination of Wine and Ethanol Concentrations Page 12
Day 4 Volume (ml) Mass (g) Density (g/ml)
Trial 1 20 20.6 1.03
Trial 2 20 20.6 1.03
Trial 3 20 20.4 1.02
Average 20 20.5333 1.0267
Fermentation of Apple Juice for the Determination of Wine and Ethanol Concentrations Page 13
Average 20 21.7333 1.0867
Concentration Data
Batch Time ρ(EtOH) ρ x C EtOH C Wine
(Wine)
1 0 1 1.2 0 0 1.2
1 Day 1 1.0533 1.0867 0.28749 0.23169 0.855011
1 Day 2 1.0367 1.0367 0.41436 0.33393 0.702766
1 Day 3 1.04 1.0733 0.32149 0.25909 0.81421
1 Day 4 1.0317 1.0367 0.41436 0.33393 0.702766
1 Day 5 1.0333 1.07 0.32987 0.26584 0.804161
1 Day 6 1.03 1.0517 0.3763 0.30326 0.748439
1 Day 7 1.045 1.0583 0.35955 0.28976 0.768536
Fermentation of Apple Juice for the Determination of Wine and Ethanol Concentrations Page 15
2 Day 0 1 1.2 0 0 1.2
2 Day 1 1.0583 1.0867 0.28749 0.23169 0.8550114
2 Day 2 1.0467 1.0283 0.43568 0.35111 0.6771885
2 Day 3 1.0283 1.045 0.3933 0.31696 0.7280386
2 Day 4 1.0267 1.0633 0.34687 0.27954 0.7837605
2 Day 5 1.0483 1.0717 0.32555 0.26236 0.8093377
2 Day 6 1.0517 1.0583 0.35955 0.28976 0.7685359
2 Day 7 1.06 1.0717 0.32555 0.26236 0.8093377
Fermentation of Apple Juice for the Determination of Wine and Ethanol Concentrations Page 16
1.4
1.2
1
Figure 1.1 Line graph
0.8
C EtOH obtained from data
0.6 on Batch 1 (Ethanol
C Wine
and Wine
0.4 Concentration)
0.2
0
0 2 4 6 8 10
1.4
1.2
0.8
C EtOH Figure 1.2 Line
0.6 C Wine graph obtained from
data on Batch 2
0.4
Fermentation
0.2 of Apple Juice for the Determination of Wine and Ethanol Concentrations Page 17
0
0 2 4 6 8 10
(Ethanol and Wine Concentration)
Time Xa
Day 0 0
Day 1 0.28749
Day 2 0.414362
Day 3 0.321492
Day 4 0.414362
Day 5 0.329866
Day 6 0.3763
Day 7 0.359553
Zero Order Reaction (1)
0.5
y = 0.0333x + 0.1629
0.45 R² = 0.3713
0.4
0.35
0.3
0.25 Series1
Fermentation of Apple Juice for the Determination of Wine and Ethanol Concentrations Page 18
Time Xa 1/Xa2
Day 0 0 0
First Order Reaction (1) Day 1 0.28749 12.0991
Day 2 0.414362 5.824257
0 Day 3 0.321492 9.675193
0 2 4 6 8 Day 4 0.414362 5.824257
-0.2 y = 0.018x - 1.1081 Day 5 0.329866 9.190225
R² = 0.0817 Day 6 0.3763 7.062046
-0.4
Day 7 0.359553 7.735229
-0.6 Series1
-1
-1.2
-1.4
Time Xa 1/Xa
Day 0 0 0
Day 1 0.28749 3.478376
Day 2 0.414362 2.41335
Day 3 0.321492 3.110497
Day 4 0.414362 2.41335
Day 5 0.329866 3.031538
Day 6 0.3763 2.657451
Second Order Reaction (1)
Day 7 0.359553 2.781228
2.5
2 Series1
Linear (Series1)
1.5
0.5
Fermentation of Apple Juice for the Determination of Wine and Ethanol Concentrations Page 19
0
0 2 4 6 8 10
Third Order Reaction (1)
14
y = -0.3965x + 9.7873
12 R² = 0.1412
10
8
Series1
6 Linear (Series1)
0
0 2 4 6 8 10
Time Xa 1/Xa^2
Day 0 0 0
Day 1 0.28749 -1.24657
Day 2 0.414362 -0.88102
Day 3 0.321492 -1.13478
Day 4 0.414362 -0.88102
Day 5 0.329866 -1.10907
Day 6 0.3763 -0.97737
Day 7 0.359553 -1.02289
Fermentation of Apple Juice for the Determination of Wine and Ethanol Concentrations Page 20
Zero Order Reaction (2)
0.5
y = 0.0269x + 0.1881
0.45 R² = 0.2464
0.4 Time Xa
0.35 Day 0 0
0.3 Day 1 0.28749
0.25 Series1 Day 2 0.435676
0.2 Linear (Series1) Day 3 0.393301
0.15 Day 4 0.346866
Day 5 0.325552
0.1
Day 6 0.359553
0.05
Day 7 0.325552
0
0 2 4 6 8 10
0
Time Xa LN Xa
0 2 4 6 8 10 y = -0.0071x - 1.0195
-0.2 R² = 0.0128 Day 0 0 0
Day 1 0.28749 -1.24657
-0.4 Day 2 0.435676 -0.83086
Day 3 0.393301 -0.93318
-0.6 Series1 Day 4 0.346866 -0.10582
-0.8 Linear (Series1) Day 5 0.325552 -1.12223
Day 6 0.359553 -1.10289
-1 Day 7 0.325552 -1.12223
-1.2
-1.4
Fermentation of Apple Juice for the Determination of Wine and Ethanol Concentrations Page 21
Second Order Reaction (2)
4
y = 0.01x + 2.8347
3.5 R² = 0.0031
Time Xa 1/Xa
3
Day 0 0 0
2.5 Day 1 0.28749 3.478376
2 Series1 Day 2 0.435676 2.295282
Linear (Series1) Day 3 0.393301 2.542581
1.5
Day 4 0.346866 2.882955
1 Day 5 0.325552 3.071707
Day 6 0.359553 2.781228
0.5
Day 7 0.325552 3.071707
0
0 2 4 6 8 10
14
y = -0.0031x + 8.4052
12 R² = 9E-06 Time Xa 1/Xa^2
Day 0 0 0
10 Day 1 0.28749 12.0991
8
Day 2 0.435676 5.268332
Series1 Day 3 0.393301 6.464716
6 Linear (Series1) Day 4 0.346866 8.311432
Day 5 0.325552 9.435384
4
Day 6 0.359553 7.735229
2 Day 7 0.325552 9.435384
0
0 2 4 6 8 10
Fermentation of Apple Juice for the Determination of Wine and Ethanol Concentrations Page 22
DOCUMENTATION
Fermentation of Apple Juice for the Determination of Wine and Ethanol Concentrations Page 23
Fermentation of Apple Juice for the Determination of Wine and Ethanol Concentrations Page 24
INTERPRETATION OF RESULTS
The results of the experiment depicts how an ideal fermentation process
would occur wherein the growing cells are consuming the substrate, and
producing more cells as time passes. In this case, the concentrations of ethanol
and wine increases and decreases respectively with time.
The stages of yeast growth are noticeable in the graphs. From Day 0 to
Day 2, the yeast undergoes an exponential growth. From Figure 1.1, the ethanol
concentration increases from 0 to 0.333934 and the wine concentration
decreases from 1.2 to 0.702766. From Figure 1.2, the concentration of ethanol
increases from 0 to 0.351111 and the concentration of the wine decreases from
1.2 to 0.677189. Then from Day 2 to Day 5, the deceleration phase is seen. From
Figure 1.1, the concentration of ethanol begins to decrease from Day 2,
0.333934, to Day 3, 0.25909 while the concentration of wine increases from
0.702766 to 0.81421. From Figure 1.2, the concentration of ethanol begins to
decrease from Day 2, 0.351111, to Day 5, .262362 while the concentration of the
wine increases from .728039 to 0.809338. The concentrations of wine and
ethanol begin to become constant as the yeast approaches the stationary
phase.
As for the order of reaction, Zero Order Reaction yields an R2 closer to 1 (In
the first batch R2 is equal to .3713 and in the second batch it’s value is .2464). For
batch 1, the R2 that is farther to 1 is from the First Order Reaction which yields an
R2 of .0817 while for batch 2; the R2 from the Second Order Reaction is farther to
1 which is equal to .0031.
Fermentation of Apple Juice for the Determination of Wine and Ethanol Concentrations Page 25
Chapter IV: CONCLUSION
The fermentation of apple juice is relatively fast because it only takes 1-2
weeks for the juice to ferment.
The lag phase (wherein the yeast matures and acclimates to the
environment and no growth occurs) was not visibly seen in the graph. The cause
of which may have been the variation of time (Day to Day). The rate of reaction
in the exponential growth of the yeast follows first order kinetics. Parameters like
the substrate and waste concentrations have an effect on the kinetics of yeast
growth as seen in the deceleration phase of the yeast. The stationary phase
(wherein no growth occurs) was only partially seen in the graph. Other factors
that affected the fermentation process may have been the stabilization of the
set-up, the thermal conditions, and the lighting in the area. The application of
other kinds of yeast would also vary the fermentation process.
The order that is best fit for the fermentation of apple juice is the Zero
Order Reaction. The zero order reaction yielded a value of R2 that is closer to 1
than the first, second, and third order reactions.
Fermentation of Apple Juice for the Determination of Wine and Ethanol Concentrations Page 26
Chapter V: Recommendation
Fermentation of Apple Juice for the Determination of Wine and Ethanol Concentrations Page 27