Republic of the Philippines

Tabaco National High School

Tabaco City

Parametric Evaluation of Biriran (Averrhoa carambola linn.) Fruit Wine

Fermented in Varying Conditions

Researchers:

John Emmanuel O. Brotamante Jr.

John Marc Carlet

Cyrus Alexander Vizon

Ian Arvin Bellen

Research Adviser:

Mr. Ronaldo Reyes

Table of Contents

Contents Page

Abstract 1

I. Introduction 2

Background of the Study 2

Objectives 3

Significance of the Study 4

Scope and Limitations 4

Related Studies and Literature 5

II. Materials and Methods 8

Materials 8

Procedure 8

Diagram of the Procedure 10

Documentation 11

III. Results and Discussions 16

Table 1: pH of Wines Fermented in different conditions 16

Table 2- Ethanol content (% by vol.) of wines fermented in 17

different conditions

Table 3. Titratable Acidity of Wine Fermented in Different Conditions 18

Conclusion 19

Recommendation 19

References 20

Appendixes 21
 1

Abstract:

Wine making is a very promising industry. That may be a possible business in the

future. The quality and content of the wines produced can be improved using proper

technology. But there are certain wine parameters that are tested when making wines

such as pH, ethanol content, titratable acidity, total soluble solids and volatile acidity.

This study only handled the pH, ethanol content and titratable acidity.

The highest value of ethanol was 4.93 from the unfiltered colored glass set up while

the lowest was using the filtered plastic which is 2.55. The other values were 4.91, 4.34,

4.76 and 4.43 to the unfiltered colorless glass, unfiltered plastic, filtered colorless glass

and filtered colorless glass respectively. The pH values for each fermented set ups are

between the given range of 3-4. The titratable acidity resulting from the experiment are

within the specific range of 0.6-0.9.

Variations in the Ethanol Content, pH and titratable acidity are from the different

conditions of fermenting; use of plastic, colored and uncolored bottle; filtered and

unfiltered set up. The results of the ethanol content did not reach the PNS/FDA 30:2010

given value for fermentation of wine because it was for the fermentation of at least 1

month while the experiment was only for 6 days. Each of the result gathered was

obtained using the proper equipments and procedure. Therefore, considering the different

variations in the wine parameters from the standard, the effect of the filtration and the

different container used to the stability of the wine parameters further studies are needed

to prove the effectivity of the filtration and the different containers used in the wine

parameters.
 2

INTRODUCTION

Background of the Study

Wine making is a challenging and difficult activity due to it requires scientific

knowledge of the fields of biology and chemistry. The process of making tropical fruit

wines was already standardized by the National Standards/Food and Drug Administration

31:2010- Recommend code of practice for the processing and handling tropical fruit

wines. According to PNS/FDA 31:2010, “ The need for more standards for our Philippine

ethnic foods transpired during the round table discussion on the processed food exports.

The development of the standard aims to set the high standard of the product, provide

guide for the assurance of its quality and safety, harmonize export requirements, and

prepare the products to be more competitive in the world market.

The wine parameter that will be monitored are the pH, ethanol content and titratable

acidity. The specific values for the parameters are given by the Philippine National

Standards/ Food and Drug Administration 30:2010 - Tropical Fruit Wines Specifications.

The alcohol content shall be composed mostly of ethyl alcohol and shall not be less than

7% (v/v) and not more than 24% (v/v). The methanol content shall be in accordance to

the provisions of BFAD M.C No. 13 s. 1989: Methanol may be present in alcoholic

beverages provided that it shall be derived from the natural alcohol fermentation process

and not added. The pH of the finished shall be derived from the natural alcohol

fermentation process and not added. The pH of the finished product shall not be less than

3.0 and not more than 4.0 The total acidity must shall not be less than 0.6% and not more

than 0.9%. The soluble solids of the finished product shall not be less than 8.0% m/m, as

determined by a refractometer at 20 °e, uncorrected for acidity and read as °Brix on the
 3

International Sucrose Scales. The volatile acidity (expressed as acetic acid) shall not

exceed 0.14g/100mL. The researches would like to evaluate the pH, ethanol content and

titratable acidity of Biriran (Averrhoa carambola linn.) Wine fermented in different

conditions should comply with the PNS/FDA 31:2010. This research will provide the

protocol for the most favorable conditions in the production of Biriran (Averrhoa

carambola linn.) Wine. The interest to produce Biriran (Averrhoa carambola linn.) Wine

was conceptualized because of the availability and abundance in the backyard of each of

the researchers and wine making is a promising industry if the proper machines for the

production of the wine will be established. Therefore, there is a need to evaluate the wine

parameters in the Biriran wine produced at different conditions to look or the best

technology for a possible business endeavor.

Objectives:

The main objective of the study is to evaluate the wine parameters of Biriran wine

produced in different conditions and look for the protocol of Biriran wine making.

Specific objectives:

A. To ferment wine in different conditions

B. To determine the wine parameters pH, ethanol content and titratable

acidity after fermentation

C. To determine the quality of the fermented wines using PNS/FDA 30:2010

and PNS/FDA 31:2010

D. To formulate the suggested procedure for biriran wine.

 4

Significance of the Study:

This study is highly significant due to the fact that tropical fruit wine is a healthy

beverage and in can help in solving stomach food problems. Farmers with Biriran tree at

their backyard will have a use for it since making wine from Biriran is a simple process

with the aid of the right materials. This study also makes a use out of the Biriran that is

not used a lot. This study will increase the chance for a possible business endeavor for the

researchers.

Scope and Limitations:

This study is limited only to the use of Biriran fruit to produce a tropical fruit wine. It

will also measure the different wine parameters limited only to; pH, ethanol content,

titratable acidity. This will not observe the volatile acidity and total soluble solids. The

only different conditions to be observed are the kind of container for fermenting juice and

the filtration of the fermented juice. The length of fermenting the extract is not tested

during the experimental process of this research. Same amount of the extract will be

observed as well.
 5

Related Literature and Studies:

Tropical Wines are made from tropical fruits such as pineapples, mangoes, guavas,

star fruits etc. Tropical fruits are mostly found on tropical countries which is found

between the tropic of Cancer and tropic of Capricorn. These fruits are pretty well known

in the world due to its uniqueness and flavor that is why it was made to wine. The

advantages of making tropical fruit wines is forming a wine that has a lot of healthy

benefits. Wine making is the process of producing wine, using the selected raw materials

(any fruit). Grape is the most usual fruit used in making wine due to the fact that it has a

lot of juice and has a great taste. But other fruits are used for a different taste. In making

wines, there are parameters to be measured; pH, ethanol content, titratable acidity, total

soluble solids and volatile acidity. There are standards to be measured as well, especially

when using tropical native fruits. Balimbing is commonly known as biriran. Is an edible

fruit which, though known to many, has not been fully utilized despite its good economic

potentials (Brion 2015). Balimbing is described as a small tree reaching up to 6-9 meters

in height. It is characterized by its dark-green colored leaflets and reddish purple flowers

clustered in leaf axils. The fruit turns greenish yellow to yellow when ripe and has a

sweet to sour taste. It has an oblong to ellipsoidal shape with a translucent, soft, and waxy

skin. About 6-9 centimeters long, it presents itself as a five-cornered fruit with its five

prominent longitudinal, angular wings. When cut across, it resembles a star hence the

name starfruit (Coronel). It can be processed to be a fruit juice, jam, jelly and can also be

eaten fresh and raw. Consumption may provide the body some essential vitamins and

minerals it needs including calcium, iron, thiamine, riboflavin, niacin and ascorbic acid.

Too much of anything is bad so consuming a lot of this may result to an upset of a
 6

balanced diet or may even pose threats to human health.

Wine making in the Bicol Region is a very promising industry. The quality and

packaging of the wines produced can be improved using proper technology. Six tropical

fruit wine manufacturers, two lambanog distillers, one nipa wine distiller and five tuba

makers were identified, profiled and sampled. Characterization of the wine parameters

pH, volatile acidity (VA), titratable acidity (TA), ethanol content (EC), methanol content

(MC), total soluble solids (TSS) and total plate count (TPC) of the wine samples was

made and the values were compared with the standards set by the Philippine National

Standard (PNS) for tropical fruit wines, distilled wines and tuba. The pH was taken by a

glass electrode, gas chromatography for EC and MC, titration for TA as tartaric acid and

VA, refractometry for TSS and disc injection method for TPC. The mean pH was 3.78 ±

0.28 for tropical fruit wines, 3.55± 0.29 for distilled wines and 3.87 ± 0.11 for tuba. The

mean TA was 0.52 ± 0.31% for tropical fruit wines, 0.50 ± 0.45% for distilled wines and

0.31 ± 0.003% for tuba. The mean VA was 0.25 ± 0.15% by vol. for tropical fruit wines,

0.29 ± 0.14% by vol. for distilled wines and 0.12 ± 0.005% by vol. for tuba. The mean

EC was 12.44 ± 3.34% by vol. for tropical fruit wines, 23.43 ± 6.08% by vol. for distilled

wines and 3.80 ± 0.07% by vol. for tuba. The mean TSS was 9.34 ± 3.33 oBrix for

tropical fruit wines, 8.25 ± 1.72 oBrix for distilled wines and 13.8 ± 0.007 oBrix for tuba.

There was no detection of methanol in the MC determination for all the wine samples.

Analysis of the result of the wine parameters showed deviation from the standard in the

VA, TA and TSS and no deviation in the pH, EC and MC. There was no detection of

methanol and bacteria in the TPC for all the wine samples. One-way ANOVA and

Scheffe’s test showed statistically significant difference among the VA, TA and TSS and
 7

no significant difference in the pH, EC, MC and TPC.Variations in the EC, TSS, TA and

VA in the tropical fruit wines and distilled wines came from the deviation of the

production procedure and packaging of the wine samples. The proposal for each wine

manufacturer was formulated based from the findings of the study and a wine catalog

featured the wine parameters, description and information of the manufacturer. Therefore,

considering the variations in the wine parameters from the standard and the effect of the

container and temperature to the stability of the wine parameters, further studies are

required on the compliance of the wine manufacturers to the proposal made for the

improvement of the quality and packaging of the wines.

 8

Materials and Methods

Materials:

15 Kg of Balimbing

5L distilled water

2 750mL colored wine bottle

2 750mL transparent wine bottle

2 800mL plastic bottle

Thermometer

Juicer

2 stainless container

Strainer

Bread yeast

2 plastic container

7 specific gravity bottle

Plastic cups

6 Calibrated Droppers

Procedure:

Raw materials (Balimbing) were gathered from one of the researcher’s backyard. The

obtained material were cleaned using tap water. The gathered balimbing were sliced to

remove the hard parts; bones and the edges of the fruit. The balimbing (Averrhoa

carambola linn.)were inserted in the juicer to extract all the juice. The collected juice was

divided into two containers; one for the filtered and one for the unfiltered. The juices were

introduced to the process of pasteurization to avoid public health hazards involving

 9

pathogenic microorganisms at a maximum heat of 63˚. The extracts were cooled to less than

40˚ in a short period of time to avoid the inactivation of inoculum (facilitates the

fermentation of the extract).The extracts were formed using 2 cups of pure extract + 1 cup

of bagasse + 1 cup of red sugar .The extracts were transferred different containers; one

colored, one transparent, one plastic bottle for the filtered extract; and one colored, one

transparent, one plastic bottle for the unfiltered extract. The containers were filled up to 3/4

of its maximum capacity. The bottled extracts were kept at a drum which temperature is

between 19˚ and 28˚. The fermentation lasts for one week to get the desired amount of

alcohol. The extracts were separately filtered after the fermentation. The filtered fermented

wine was pasteurized for 63˚ in 15 minutes to inactivate spoilage microorganism. The

pasteurized fermented wine was cooled. Each sanitized bottle were filled not reaching 1

inch from the cap to avoid oxidation. The wine parameters are tested using different

methods; for pH using glass electrode method, ethanol content using Annex G of PNS/FDA

31:2010 by specific gravity method, titratable acidity using Annex E of PNS/FDA 31:2010

by process of titration.
 10

Data Analysis

Gathering of raw materials

Juice Extraction

Filtered Unfiltered

Pasteurization Pasteurization

Cooling Cooling

Preparation of Juices for Preparation of Juices for

fermentation fermentation

Fermentation Fermentation

Plastic Colored Transparent Plastic

Colored Transparent
Container Container

Filtration Filtration

Pasteurization Pasteurization

Filling of Filling of
Containers Containers

Cooling and Cooling and

Sealing Sealing

Wine Parameters

Ethanol Titratable
pH Acidity
Content

Data Analysis

Figure 1: Schematic Diagram of the Procedure

 11

Figure 2: The Biriran Tree

Figure 3: Washing/Cleaning of Balimbing

Figure 4: Slicing and removing of hard parts

 12

Figure 5: Juice Extraction

Figure 6: Division of extracts to filtered and unfiltered

Figure 7: Filtration of Extract

 13

Figure 8: Pasteurization (63˚)

Figure 9: Filling of Containers for Fermentation (bread yeast added)

Figure 10: Filtration of fermented juice

 14

Figure 11: Separate Pasteurization of Fermented Juice in different conditions

Figure 12: Filling of containers for Testing of Wine Parameters

Figure 13: Sealing of Containers

 15

Figure 14: Titration

Figure 15: Glass Electrode Method

Figure 16: Specific Gravity Bottle filled with juice extract

 16

Results and Discussions

The bottled extracts were fermented for exactly six days before introducing it to

pasteurization at room temperature. The gathered data for each wine parameters are shown in

the tables below. The wine parameters which were experimented upon are pH, ethanol content

and titratable acidity. Each of the data gathered from the different wine parameters are

compared to the acceptable data for each in the Philippine National Standards.

Table 1: pH of Wines Fermented in different conditions

Wine pH Acceptable Value Interpretation

FCdG 3.97 3-4 Acceptable
FCsG 3.98 3-4 Acceptable
FP 3.97 3-4 Acceptable
UCdG 3.91 3-4 Acceptable
UCsG 3.94 3-4 Acceptable
UP 3.92 3-4 Acceptable

The resulting pH of wine that was fermented in different conditions were acceptable,

ranging between 3-4. The pH of wine is important to determine because it will affect the

quality of the product in terms of taste, color, oxidation, chemical stability and other factors.

The general rule of pH in winemaking is the higher the pH reading, the lower the amount of

acidity in wine.

Any pH above 4.0 indicates that the wine will spoil quickly and be chemically unstable.

Lower pH values allow the wine to stay fresher for a longer period and retain its original color

and flavor (Kearney and Bogolawski; n.d).

 17

Table 2- Ethanol content (% by vol.) of wines fermented in different conditions

Wine Ethanol Content Acceptable Value Interpretation

UCsG 4.91 Not less than 7% Not acceptable
(v/v) and not more
than 24% (v/v)
UCdG 4.93 Not less than 7% Not acceptable
(v/v) and not more
than 24% (v/v)
UP 4.34 Not less than 7% Not acceptable
(v/v) and not more
than 24% (v/v)
FCsG 4.76 Not less than 7% Not acceptable
(v/v) and not more
than 24% (v/v)
FCdG 4.43 Not less than 7% Not acceptable
(v/v) and not more
than 24% (v/v)
FP 2.55 Not less than 7% Not acceptable
(v/v) and not more
than 24% (v/v)

The ethanol content value should not be less than 7% and not more than 24% for the

fermentation of at least 1 month. The fermentation only occurred for six days so the

ethanol content (% by v/v) did not reach the range. The results showed that not filtering

wine before fermentation has a higher ethanol content and based from the

experimentation the preferred fermentation set up for Biriran wine is fermentation using

colored glass. The study followed the filling of glass 1nch or more below the cap to avoid

oxidation. The suggested procedure in making wine is with the use of sanitized

equipments. Fermentation should at least be 1 month to have the proper ethanol content.
 18

Table 3. Titratable Acidity of Wine Fermented in Different Conditions

Wine Conditions Acceptable Value Interpretation

TA (%)
UP 0.750% 0.6-0.9 Acceptable
UCsG 0.700% 0.6-0.9 Acceptable
UCdG 0.953% 0.6-0.9 Acceptable
FP 0.795% 0.6-0.9 Acceptable
FCsG 0.550% 0.6-0.9 Acceptable
FCdG 0.688% 0.6-0.9 Acceptable

The titratable acidity of each extract complies to the Philippine National Standard’s

value of 0.6 to 0.9. Titratable acidity is the total amount of hydrogen ions present in the

wine sample with the exception of those bound to alkaline ions. The hydrogen ions can

be either attached to acids or in a form of free ions or anions.

Titratable acidity is an analysis that is useful in different winemaking situations. It

can be used as an indicator of sensory properties. Knowing the titratable acidity of a wine

help a winemaker if a wine is in balance or stylistically correct (enartisvinquiz.com, 290).

Legend: UCsG-Unfiltered Colorless Glass UCdG-Unfiltered Colored Glass

UP- Unfiltered Plastic Bottle FCsG- Filtered Colorless Glass

FCdG- Filtered Colored Glass FP-Filtered Plastic Bottle

 19

Conclusion:

The study about evaluating of wine parameters of biriran wine fermented in different

conditions was conducted to determine whether the different use of container and

filtration will have a variety of results to the different wine parameters. This study

showed that the different conditions of fermenting wine results to a variation in the

results of the wine parameters. Results showed that the titratable acidity and pH of every

extract conformed to the standards.

Recommendations:

The study is suggested to be repeated again for a more exact result. The study may

include the length of fermentation as one of the conditions in fermenting wine and

include all of the wine parameters in the Philippine National Standards.

Acknowledgement:

The researchers would like to extend their profound gratitude to God for the gift of

wisdom and unending guidance for the endeavor. To their research advisers Mr. Ronaldo

Reyes and Prof. Carina B. Orden and to their parents for invaluable support.
 20

References:

PNS/FDA 30:2010 - Tropical Fruit Wines – Specification page 18

PNS/FDA 30:2010 - Tropical Fruit Wines – Specification page 20

PNS/FDA 30:2010 - Tropical Fruit Wines – Specification page 22

PNS/FDA 31:2010 - Recommended Code of Practice for the Processing and Handling of

Tropical Fruit Wines page 14

PNS/FDA 31:2010 - Recommended Code of Practice for the Processing and Handling of

Tropical Fruit Wines page 16

PNS/FDA 31:2010 - Recommended Code of Practice for the Processing and Handling of

Tropical Fruit Wines page 18

Coronel, Roberto E. MD, Important and Underutilized Edible Fruits of the Philippines

Kearney, C. And Bogolawski (n.d) Winemaking and the importance of pH testing. Retrieved

July 15, 2015 from hannainst.com/USA/whitepaper/Winemaking and pH.pdf

 21

Appendixes

A. Computation for Ethanol content

Specific Gravity of Juice for Fermentation

OSG ( Original Specific Gravity ) = 1.037264

Specific Gravity of Fermented Wine

FSG ( Final Specific Gravity )

UCsG = 0.999816

FCsG = 1. 073564

UCdG = 0.999728

FCdG = 1.070992

UP = 1. 004196

FP = 1.056656

% alc by wt = / OSG – FSG / x105

% alc by vol = % alc by wt 1.25

UCsG

% alc by wt = / 1.037264 – 0.999816 / x105 = 3.93204%

% alc by vol = 3.93204 x 1.25 = 4.9125% or 4.91%

UCdG = 4.93

UP = 4.34

FCsG = 4.76

FCdG = 4.43

FP = 2.55
 22

B. Computation of Titratable Acidity

TA as tactaric acid

TA=mL NooH x NNooH x Factor

Volume of Sample

Factor=0.075

Filtered Plastic

TA=1.06 mL x 0.1 N x 0.075 x100 = 0.795%

1 mL

Filtered Colored Glass

TA=0.688%

Filtered Colorless Glass

TA=0.550%

Unfiltered Plastic

TA=0.750%

Unfiltered Colorless Glass

TA=0.700%

Unfiltered Colored Glass

TA=0.953%