Introduction

Vinegar is a French word where vin means wine .It is a liquid that has 5 acetic acid in
water .Vinegar is derived from various sugary and starchy material .Vinegar is define has “a
liquid carrying sugars, formed by fermentation . That is double fermentation suitable for human
consumption; first fermentation is alcoholic and than fermentation is acetous fermentation.”

Vinegar can be produced from different raw materials .Substrates used are wine ,malted barley,
fruits mustard, cider and alcohol, etc. Vinegar is primarily , widely and traditionally used as
preservatives in food industries .It is corresive with sour taste and pungent smell. Vinegar is
acetic acid solution with 4 to 6% dilution used directly as flavouring agent, food preservative as
cleaning agent as ingredient in salad dressing and marinade

Despite this widespread occurrence , the Sumerians were the first civilization to record the use
of vinegar as condiment .In actuality ,vinegar have exited for nearly 5000years .Babylon vinegar
enhanced to evoke the splendor of 5000BC with fruits ,honey ,malt ,etc.Hippocrates and the
ancient testament both documented the therapeutic benefits of vinegar .A claim made by
Albucases in 1100 that vinegar distillation needed to done over low fire .Chemist Stahl
discovered that aspartic acid was the sour principle of vinegar in the first half of 18 th century . In
1778, Durande produced glacial acetic acid with increased concentration. In 1814, Berzelius
conducted the first thorough analysis of acetic acid . In 1955,Schulzenbach development a rapid
method for producing vinegar from alcohol. Joslyn studied Hromatka evolved a method called
submerged fermentation in 1955.

Apple cider is a pale medium coloured vinegar prepared from cider or apple must .Mother of
vinegar is present in unpasteurized or organic apple cider vinegar .Apple cider is made from the
juice of crushed apples that has been fermented .Pectin, vitamin B1 , vitamin B2 , biotin , folic
acid , niacin , pantothenic acid and vitamin C are likely to be present , as they are in apple
juice .Ingestion of the acetic acid in vinegar provide danger of probable harm t soft tissues of the
mouth, throat and stomach ,according to preliminary studies. Natural vinegar contain all
carbohydrate, organic acid (acetic acid , lactic, malic, citric, succinic and tartaric), alcohol ,
amino acids and peptides , vitamin , salts and polyphenolic substance ( gallic acid, catechin ,
caffeic , ferulic acid) have all been found in natural vinegar .A variety of vinegar are available
on the market . Traditional vinegar is made from regional cuisine is accordance with long
standing traditions . Antimicrobial properties , prevention of inflammation and hypertension ,
lower serum cholesterol , reduction systolic blood pressure , enhanced calcium absorption and
retention and decrease the glycemic index of carbohydrate food for people with and without
diabetes Apple cider vinegar is created by fermenting especially n unfiltered and organic
varieties . Honey can be used to enhance the flavour of apple cider vinegar drinks without adding
extra calories

Acetic acid bacteria play a major role in acetous fermentation and should be kept in good growth
status especially under alcoholic -acidic conditions . During alcoholic fermentation ,quantities of
substrates are consumed by yeast which may causes nutrients in alcoholic mash advisable for
acetous fermentation by acetic acid bacteria . In this case , mash some nutrients essential for
acetous fermentation by acetic acid bacteria but which are lacking in alcoholic mash should be
added .Actually , additional nutrients are supplied before acetous in some kinds of submerged
vinegars production like cider vinegar and spirit vinegar.

Transforming alcohol into acetic acid coupled with oxidation respiration is a well known caracter
for most species of acetic acid bacteria, which plays major role in energy generation for
bacterium during vinegar fermentation. Generally , acetic acid bacteria cannot completely
oxidise acetic acid into carbon dioxide and water until alcohol in medium is
exhausted .Therefore , alcohol and acetic acid cannot serve as the major carbon source for
cytoskeleton precursor formation during acetous fermentation . Acetic acid bacteria may utilize
other nutrients from raw material like glucose as the resource of cytoskeleton precursor
formation. For cider vinegar fermentation , as previous study has show , the total residual content
o carbon sources (containing sucrose .fructose and glucose) in the cider was enough for acetic
acid bacteria growth based on yield of biomass versus glucose and the yield of acetic acid versus
biomass.

There are many non food application of vinegar is its use as disinfectant , which makes it good
house hold cleaner .wiping and spraying vinegar on wood panels is one of technique used for
prevention of mold growth in furniture which is a problem in tropical climate. The advantage of
using vinegar rather than synthetic chemicals are as vinegar is an inexpensive product ,functional
for cleaning and disinfecting , and non toxic
Review of literature

Abhishek Kumar singh (2020) evaluated the ,different raw material ,different substrate for
production of vinegar . Vinegar is important role in food preservation in food industries. It is
used as flavouring agent, for pickling and beverages and is also contain medicinal properties .
Qualities of vinegar are depending on the efficiency of microorganisms. There different method
for vinegar production from Orleans method to generator method to submerged fermentation

Sunil jayant kulkari he obtained exclusively through biotechnological process such as double
fermentation, alcoholic and acetic fermentation of liquids or other substances of agriculture
origin . There are various types of vinegars obtained from various sources such as wine fruit and
berry , cider , alcohol . Vinegar is used as a food additive and also it acts as preservative against
food spoilage . Various investigators have carried out investigations on vinegar production from
various raw material .

Bernadette M
Material and methodology

 Ripe apple
 Distilled water
 Yeast(Saccharomyces cerevisiae)
 Mother culture
 Fermentation jar
 pH meter
 0.1 N NaOH
 Phenolphthalein indicator
 Nutrient agar plate

Methodology

Production of Apple cider vinegar

 Collection of Apples: Apples should be firm and ripe when purchased. Avoid green apple
as the have a bland flavor when they are not fully developed.
 Washing : The apples must washed after they mellowed to remove leaves , twigs,
insects ,spray residues, and hazardous bacteria. They are dumped out of the bins into a
scrubber . This cleanses and rinsed each apple , removing the majority of chemical
residues.
 Grinding : Because apple are hard fruit, slicing is required to obtain juice . This is
accomplished by high speed rotary blades to reduce the fruit to a pulp. It is done to
extract the maximum amount of juice from the apples.
 The apple and apple juice both poured in the glass jar ,sugar was added . On the other
hand yeast was activated in warm water and little amount sugar . Activated yeast was
added in jar containing apple and apple juice
 1)Fermentation : As apple juice has two phase of fermentation this one is the alcoholic
fermentation . In this yeast strain perform traditional alcoholic fermentation of
carbohydrates into ethanol This fermentation for kept for 21 days (3 weeks) temperature
at room temperature in dark condition. It was covered with muslin cloth so that CO2 can
get out
 2)Fermentation :After the alcoholic fermentation , all the solid of apple were separated
from the liquid , the fermented apple juice was filtered multiple time and it was poured in
clean sterile jar
 Introducing of starter culture:
1. Preparation of starter culture : 50ml of apple juice was inoculate with unfiltered
and unpasteurized apple cider vinegar which contain acetic acid producing
bacteria it was kept fermentation for 30 days
2. After 30 days it was filtered and it was as inoculated in the main fermentation jar
 After the inoculation of starter culture it was loosely tighten in this phase there is ethanol
is converted to vinegar by acetic acid producing bacteria. This fermentation take about
21days days .The optimum condition are maintain such as temperature warm temperature
, it was kept in dark condition
 After fermentation some test were performed
 Initial and final pH was checked with help of pH meter
 Sugar concentration was checked with the help of hand refractometer
 Estimation of acetic acid was done by titration against 0.1N NaOH
In this 25 ml of apple cider vinegar was pipette out and was diluted in 250ml
volumetric flask with distilled water , in burette 0.1N NaOH was filled and in
flask 25 ml diluted apple cider vinegar was pipette out with indicator .Three
reading were taken C.B.R.taken.
 Antimicrobial active was checked against E. Coli.
In this spp. Of E. Coli suspension was prepared in saline water. The suspension
was spread on Nutrient agar plate with spread plate technique. The hole were
 dwell with cork borer .The apple cider vinegar was filled in hole and it was
incubated for 24hrs at 37dregre in incubator
 For the confirmation of acetic acid
The apple cider vinegar was added with 1N AgNO3 and observe the changes
occurred.
 Test for flavonids
2ml of apple cider vinegar was taken in test tube ,few drop of dilute 10%NaOH
solution were added to it . Then dilute HCl was added to the solution and yellow
colour formed .
 Test for Alkaloids –
Wagner’s reagent 3-5 drops in 2 ml of sample in test tubes and brown reddish
precipitate was obtained
Observation and Result:
Sample Before fermentation After fermentation
In
1 4.0 2.5
bottle :

 Smell is floral and apple

 Colour is slight brown
 Transparency is cloudy and opaque

Result of test performed:

 pH
I

 Sugar concentration

Sample Before fermentation After fermentation

1 21% 3.5

 Estimation of acetic acid in the apple cider vinegar

Sample 1 2 3 C.B.R.(x) ml
 Final level 24.5 24.5 24.9
Initial level 00.0 00.0 00.0 24.5
Difference 24.5 24.5 24.9

Estimation of percentage of acetic acid in apple cider vinegar .

We have 1ml 0.1 N NaOH solution =0.006 of oxalic acid

1ml 0.096 N NaOH solution=0.096× 0.006/0.1

=0.00576 g of acetic acid

X ml of 0.1 N NaOH solution =0.00576×24.5

= 0.14112

Now, 25ml diluted solution =0.14112 of acetic acid

Therefore, 250ml diluted solution=0.14112×10

=1.4112 g of acetic acid

Ask for given volume of vinegar , it is 25 ml

The 25ml of given solution =1.4112 g of acetic acid

Therefore 100ml of given solution=1.4112×100/25

=5.648 g of acetic acid

Percentage of acetic acid is the sample is 5.64%

 Antimicrobial activity of apple cider vinegar

Sample E coli zone (mm)

Apple cider vinegar 0.6 mm
  Test for flavonids
From this test it conclude that flavonid are present in the test sample

Fig: Antimicrobial activity of

apple cider vinegar against
E. Coli

 Test for Alkaloids

Fig: test for flavonids
 due to reddish brown precipitate it conclude that alkaloids are present in this

sample
Fig: Test for Alkaloids
Confirmatory test
The apple cider vinegar was added with 1N Silver Nitrate and observes the change , white
precipitation is observed it confirm the acetic acid in the sample

Fig 4: confirmatory test for

acetic acid in sample
 Conclusion
 Vinegar is the product as the result of oxidation of alcohol in a fermentation sugar
containing apples.
 The addition of starter culture speed of the fermentation process of apple cider
vinegar .Naturally fermentation process takes to 8to 6 weeks but addition of starter
culture speed up the fermentation process which completes in 2 to 4 weeks
 Additionally , the activity of discovered compounds can be docked with target
receptors to find the compound’s affinity to the target site
 Apple cider was also obtaining higher antioxidant activity
 As it has antimicrobial activity and it is in expensive and non-toxic .The high level
capacity of acetic acid in killing microorganisms
Reference

1. Abigail C ,Leeder(2016) The social network :Deciphering fungal language ,

Natural review microbiology(vol.9 Issue 6)
2. Vinod K., Rakesh S.,(2018)Optimization of a process for preparation of
base wine for cider vinegar production, Proceedings of national academy of
sieves, India section B : Biological science volume 89,page 1007 -
1016(2019)
3. Zhenjiang Q., Die D ., (2016) Improve fermented quality of cider vinegar
via rational nutrient feeling strategy, Food chemistry Volume 224, Q June
2017, Page 312-319
4. Swati M., Sindh R.,(2017) Antimicrobial efficacy of apple Enterococcus
faecalis and candida albicans :An in vitro study , Journal of advanced
pharmacy education and research Apr-Jan 7(2)137 -141,
5. Simon H., Shimelis A.,(2015) Vinegar production technology -An
overview ., Beverages and food world November
6. Ankul S . ,Anuragh S ,(2023) GC- MS based metabolite profiling ,
antioxidant and antiurolithiatic properties of apple cider vinegar, Future
Science OA , vol. 9,no.4
7. Mumtahinul K.,Bushra M.,(2022), A perfect Detoxifying Drink- Apple cider
vinegar,Just Agriculture(e- ISSN : 2582 -8223) ,Vol. 2 Issue-7, article-001
8. Bernadette M., Shaily A. Nadita N., (2019),Isolation of acetic Acid Bacteria
and Preparation of starter culture for apple cider vinegar fermentation, Vol.9
no.6
9. Asma S.(2017),Antimicrobial activity of apple cider vinegar,Mapana journal
of science ,Vol. 16 , no.2, 11-15
10.Judy G. , Vimala A , Manukandan M.,(2017), Authenticating apple cider
vinegar ‘s home remedy claims :antifungal ,antiviral properties and
cytotoxicity aspect,Vol33 , Issue6 ,page 906 -910
11. Hatice O., Buse K. (2022),The antioxidant and antibacterial properties of
chitosan encapsulated with the bee pollen and apple cider vinegar, Journal of
Biomaterial science ,Vol. 33, Issue8 ,page 995-1011