CHAPTER 4

ANTICIPATED RESULTS, INTERPRETATION, AND IMPLICATION

For this study, the primary objective is to fabricate an acetic acid generator for efficient
batch production of Acetic acid from fermentation of reject pineapples and mangoes. For higher
acetic acid concentration yield, parameters such as pH, temperature and aeration rate in the
generator are manipulated. The % Acetic acid per day as well as highest acetification efficiency
will also be determined. The calculated results will be presented in graphs and tabulated.

4.1.1 Objective No. 1: To produce high alcohol content wine from alcoholic fermentation
of reject fruit waste

Two of the key major parameters that needs to be observed in alcoholic fermentation is
the pH and temperature to attain maximal ethanol production in the fermentation. 3 batches of
the same mass ratio of the reject pineapple and mango mixture will be treated with the same
amount of yeast (Saccharomyces cerevisiae). This batch fermentations will be optimized with
different degree of pH, temperature and time period. The inoculated fruit juice will be carried out
in different flasks (small scale) and will be incubated at different pH (4, 5, and 6) and at
temperatures 20oC, 25oC, and 30oC for 3 days. Results will be tabulated below.

Table 1.

o
Batch Brix Temperature, oC pH Alcohol
Content
4
1 20 5
6
4
2 25 5
6
4
3 30 5
6
Statistical analysis will also be employed in order to determine if there are any significant
differences in the alcohol concentration when varied at different pH and temperature parameters.
Analysis of Variance (ANOVA) using F-test will be used.

4.1.2 Objective No. 2: To produce more Acetobacter Aceti by controlling the temperature
and pH in bamboo segments.

pH plays an important role in the production of bacteria, one must able to control it to
maximize yield. For this study, Acetobacter Aceti will be used for the fermentation of mangoes
and pineapple peelings. Bamboo shavings, a lignocellulosic material will serve as the packed bed
for the immobilization of the bacteria. The packing material will be made more porous by size
reduction
Behavior of Actobacter was studied in the study of Dirisu (2017) entitled as "pH Effect
and pH Changes during Biocellulose Production by Gluconacetobacter xylinus in Moringa oleifera
Tea-Sugar Medium", a pH of 5 was found to give the maximum yield.

Figure X. Mean Biocellulose yield at different pH in Moringa tea-sugar medium

pH will be strictly monitored by a pH meter. Acetic acid production alters the pH and so,
two reagents might be used. One is hydrochloric acid and the other, sodium hydroxide. Both
concentrated at 1 N. Basically, the results will tell what pH was suitable for the system.

4.1.3 Objective No. 3: To determine percent yield of acetic acid from the acetic acid
generator.
Acetic acid produced from the acetic acid generator will be estimated by titration of the
supernatant broth against a standardized 1N NaOH with a phenolphthalein indicator. The
balanced equation for the neutralization reaction is as follows:

NaOH + CH3COOH → CH3COONa + H2O

The grams of Acetic acid neutralized by the NaOH is calculated upon reaching the end point
where a slight pink color appears. Where it can be obtained through stoichiometry by:

1 𝑚𝑜𝑙𝑒 𝑁𝑎𝑂𝐻 60 𝑔 𝐶𝐻3𝐶𝑂𝑂𝐻

𝑔 𝐶𝐻3 𝐶𝑂𝑂𝐻 = 𝑀𝑜𝑙𝑎𝑟𝑖𝑡𝑦 𝑁𝑎𝑂𝐻 (1 𝑚𝑜𝑙𝑒 𝐶𝐻3𝐶𝑂𝑂𝐻) ( 1 𝑚𝑜𝑙𝑒
) (𝑣𝑜𝑙. 𝑁𝑎𝑂𝐻 𝐿𝑖𝑡𝑒𝑟𝑠) Equation X

Calculating for percent yield of acetic acid from the vinegar obtained from the generator, is then
computed by:

𝑚𝑎𝑠𝑠 𝑜𝑓 𝑎𝑐𝑒𝑡𝑖𝑐 𝑎𝑐𝑖𝑑

% 𝐴𝑐𝑒𝑡𝑖𝑐 𝑎𝑐𝑖𝑑 = 𝑚𝑎𝑠𝑠 𝑜𝑓 𝑣𝑖𝑛𝑒𝑔𝑎𝑟
𝑥100 Equation X

Batch production of acetic acid will be carried in the generator spanning 5 days for 3 trials. The
results will be plotted as %Acetic acid for every run vs the number of days as shown below:

Trial Number
6
5
Acetic acid (%)

4
3
2
1
0
1 3 4 5
Days of batch acetic acid production in generator

Figure X. % Acetic acid per day of acetic acid production

4.1.4 Objective No. 4: To determine the acetification efficiency of the generator

Acetification efficiency measures how efficient the generator will be. The acetification
efficiency will be calculated and expressed by calculating the theoretical yield and total
concentration. The production of acetic acid will be observe and compared with the theoretical
yield which will be calculated from the ethanol consumption and from the stoichiometry of the
conversion reaction of ethanol to acetic acid:

C2H5OH + O2 → CH5COOH + H2O Equation X

The total concentration (or Gesammte Konzentration), is largely used in the vinegar
industry, expressing fermentation efficiency as the sum of the ethanol (% v/v) and acetic acid (%
w/v) concentrations at the beginning and at the end of the fermentation (Ilha, et.al 2000). This
relation may be applied when calculating the conversion efficiency of ethanol to acetic acid: GK
yield = (final GK / initial GK) x 100 (Leonel, et.al, 2015). In the absence of losses through
evaporation, over oxidation and conversion to biomass, the GK value should remain constant
throughout. Efficiency, expressed as GK yield, reaches up to 90%. Typical GK values are up to
10 (Adams & Twiddy, 1987).

Figure X. The behavior of GK over a period of 24 hours for three sets of aerations

Ideally the expected result is likely the same from the figure above. That over time, the
GK value decreases due to the evaporation of volatile acids present in the solution. There will be
slight variation of GK values over time due to expected evaporation of volatile acids, the longer
fermentation time and strong aeration the more effects are noticeable. This is unchanging by what
was shown from the study of Ilha et al., 2000 shown in Figure 1.

After obtaining the results statistical analysis will be employed in this objective using the
ANOVA and F-test. For this study there is only one dependent variable, acetification rate while
there are two independent variables, aeration rate and recirculation rate. Results will be then
compared whether to accept or reject the null hypothesis. The null hypothesis considered is that
all the means are equal, with an alternative hypothesis that at least one mean is equal. For the
F-statistics if it is greater than the F-critical value, this will imply that null hypothesis must be
rejected; this will be the basis that there is a significant difference among the GK yield. As
anticipated, there is more evaporation losses with the stronger the aeration then the one with the
highest efficiency is the smallest aeration rate while the lowest efficiency is the highest aeration.

Minimal foaming is another factor that affects the efficiency it is likely to occur during the
acetification process. This will occur on the mouth of the showerhead where the feed hose is
attached (Gallardo et al., 2018) This is due to the solutions containing biomaterials that
accompanies the fermentation process. Foaming can be affected by the gas introduction, medium
composition, cell growth, metabolite formation, surface-active substance formation and indirectly,
vessel geometry (Taticek et al., 1991; Vardar-Sukan, 1992).

4.2 Potential implications

This section should join the data analysis and possible outcomes to the theory and
questions that you have raised. It will be a good place to summarize the significance of the
work, i.e. summarize how the results would add to the knowledge base of the research area,
especially in the context of the questions and hypotheses that were posed in Sections
1.2 and 1.3 in Chapter 1. In addition, provide the broader potential applications of the research
findings. Additional notes on captions for Figures and Tables:
• If your figure or table is essentially the same as or based on another author’s, but you
recreated or adapted it, it is standard to include the words “Adapted from” or “After” followed by
the author’s name and a citation at the end of the caption.
• Always cite the figure or table if it—or its data—came from a source, using the same
citation style that you have used throughout the paper. The most logical place for the citation to
appear is at the end of the caption.
4.3 Plans for disseminating the findings of this research

The purpose of the research is to fabricate a low-cost, efficient Acetic acid generator capable
of batch production, with the produced Acetic acid from reject pineapple and mangoes enough to
compare with commercial vinegar in the market. The team also aims to determine the parameters
to yield the highest acetic acid concentration to impart new knowledge and improve the current
Acetic Acid generator process used in the industry.
Vinegar had a variety of uses ranging from the industry to the medical sect or our very own
homes. From this, potential stakeholders would include manufacturers in the vinegar industry and
researchers in vinegar-making and those who are generally interested in the subject matter of
acetic acid production and equipment design.
When do you disseminate? As with all aspects of your study, it is important that your
dissemination activities are an integral part of your research implementation plan. This means
that you need to plan your dissemination at the outset. Having identified exactly what it is your
research will be disseminating, you need to give some thought to the timing of particular
dissemination activities. You will need to consider each of your target audiences, and the level
of dissemination required and begin to plan the timing.
The media of which the information will be disseminated will be in the forms of poster
presentations and its publication in journals ADD PLS

How will you disseminate? It is at this stage that you need to begin to match vehicles for
dissemination to your objectives. The following list offers you with some ideas of different types
of dissemination media that you might consider using:
 Digital archiving (i.e.
ProQuest)
 Publications in journals or
books
 Poster presentations
 Oral presentations
 …