Food Science and Technology 6(1): 20-27, 2018 http://www.hrpub.

org
DOI: 10.13189/fst.2018.060103

Authenticity Testing of Coconut Toddy Samples (Palm

Wine) Using Sulfated Ash Content
Gowri Ramanah*, Samantha Wimalasena, Vajira Senadipathi, Eranthi Seniviratne,
Kulali Suntharalingam

Government Analyst’s Department, Sri Lanka

Copyright©2018 by authors, all rights reserved. Authors agree that this article remains permanently open access under
the terms of the Creative Commons Attribution License 4.0 International License

Abstract Coconut toddy (palm wine) is an unopened inflorescence of the coconut palm, is the major
effervescence beverage tapped from young inflorescence raw material used in the manufacture of alcoholic
of coconut tree (Cocos nucifera). The sap contains various beverages in Sri Lanka. The sap is a brownish clear liquid,
nutrients, and it undergoes natural fermentative changes containing 14 to18 percent (w/v) sugars mostly in the form
when it is stored. Toddy is a traditional alcoholic drink inof sucrose. [1]
Sri Lanka. In Sri Lanka artificial fermented liquor is made Toddy is a sweetish, milky white, vigorously
out from the fermentation of sugar solution. There have effervescent alcoholic beverage consumed as mildly
been several complaints with respect to adulteration of alcoholic beverage similar to beer which is collected from
toddy by artificial fermented liquor. Ash values are used topalm tree. [2]
find out quality, authenticity and purity of substances and During ancient period, people drank palm wine in early
these values are important quantitative standards. Hence morning in empty stomach before breakfast for health
this study mainly focused on identifying authenticity of benefits.[2] It is made from the fermented sap of tropical
toddy using the sulfated ash content. Genuine toddy plants of palmate family, such as the coconut palm (Cocos
samples were collected from different areas and artificial nucifera), oil palm (Elaeis guineesis), date palm, nipa palm,
fermented liquor samples were made in the laboratory kithul palm and raffia palm (Raphia hookeri).[2]
using sugar solution (16%) and inoculum of genuine toddy The coconut sap oozing out of the treated inflorescence
samples. Different ratios of genuine toddy and artificial is traditionally collected in earthenware pots and allowed to
fermented liquor mixtures were prepared. Sulfated ash ferment in open pots for varying lengths of time. During
content of genuine toddy samples, artificial fermented this period, microorganisms from the atmosphere
liquor samples and different ratios of genuine and artificial
contaminate, multiply in the sugary liquid, and finally
fermented liquor were determined by using muffle furnace. transform the raw material into various products, mainly
Linear regression was obtained with sulfated ash content vs alcohol. The resulting solution containing about 7 percent
different ratios of genuine toddy samples. Sulfated ash (v/v) ethyl alcohol is either consumed directly as toddy or
content of genuine toddy varied from 0.44 to 0.54 distilled to obtain the sprit, arrack. [1]
gram/100ml. Distilleries in Sri Lanka usually obtain toddy from
contractors in Sri Lanka and they are paid on the content of
Keywords Genuine Toddy, Artificial Toddy, Sulfated
pure toddy. There have been several complaints with
Ash, Fermentation
respect to adulterated toddy which is artificially made
fermented liquor prepared from sugar solution to the
natural toddy.
Therefore, the problem of adulterated toddy has to be
1. Introduction solved and necessary steps have to be taken to prevent such
substandard toddy entering the market. There have been
Coconut Palm (Cocos nucifera) is an important member many research studies about toddy, but none of these
of the monocotyledons. This tree mainly grows in tropical studies focused on adulteration of artificial fermented
coastal areas in Sri Lanka. Coconut is one of the major liquor prepared from sugar solution.
plantation crops in Sri Lanka which accounts for Many researchers have studied the chemical or
approximately 12% of all agricultural produce in Sri Lanka. microbiological compounds of fresh inflorescence coconut
The coconut sap, an exudate obtained by “tapping” the sap (FICS) and natural fermented coconut inflorescence
 Food Science and Technology 6(1): 20-27, 2018 21

sap (NCIS). The chemical composition of the volatiles 2. Material and Methods
from FICS and NCIS which were fermented for 24 h at 30
± 20C; 21 compounds making up more than 98 % of the 2.1. Samples
volatiles in FCIS, and 12 compounds representing more
This Study involved 90 toddy samples in which
than 95 % of the volatiles in NCIS, were characterized[3].
fermentation was arrested by adding salicylic acid,
The distribution of microorganisms and changes of received from excise department for quality control
physical and chemical contents during the natural purpose and courts for legal action.
fermentation of coconut inflorescence sap and 166 isolates Further artificially fermented liquor and 05 mixtures of
of yeasts and 39 isolates of bacteria were identified, while artificial and natural toddy were prepared in the laboratory.
17 species of yeasts belonging to eight genera were  3 samples of genuine toddy directly collected from
recorded [7]. The Chemical composition changes of the tapper in 2014 denoted G1*
post-harvest coconut inflorescence sap (PCIS) during  4 samples of genuine toddy directly collected from
natural fermentation. Ethanol content increased the tapper in 2015 denoted G2*
significantly from 1 to 5 days of fermentation and achieved  74 samples of market toddy received from excise
maximum on the seventh day, but decreased later. After department in 2015 denoted MMT
five days total acidity rose sharply and amino acid contents  8 Samples of toddy received from Courts denoted
decreased [3]. In “Toddy Authentic or adulterated - How CL
do we Know?” it is explained, as how to identify the starch  Artificial Toddy denoted A*
which is not a component of toddy. The samples were  5 Mixtures of toddy and artificial toddy denoted
tested with iodine (I2)/ potassium iodide (KI) reagent and M*
the colour development was compared using analytical
grade starch as the control. The microorganisms have the 2.2. Method of Preparation of Artificial Fermented
potential of producing many chemical compounds in Liquor (Artificial Toddy)
addition of ethanol. Therefore, there is tendency of giving
2.2.a. Preparations of 16% Sugar Solution
various colours such as reddish purple and light blue with
the presence of amylopectin and amylose, respectively. 160 g of sugar was dissolved in 1 l tap water in ambient
Light blue colour of the toddy sample was solely due to the temperature.
presence of the microorganisms introduced from the 2.2.b. Inoculum
environment. The samples were tested microscopically to
750 ml of Suspended toddy i.e. toddy found in the
confirm the results. It can be concluded that an adulterated
bottom part of tappers pot was collected with the help of
toddy sample gives a deep blue colour with I2/KI reagent
excise department.
due to the presence of starch [1]. However there have been
no reports about the adulterated toddy due to presence of 2.2.c. NPK as Nutrient
sugar. 0.5 g of KH2PO4, 0.5 g of NH4CO3
We studied chemical parameters such as alcohol content,
acidity, total solids, sulfated ash, brix value; conductivity 2.2.d. Anti-bacterial Agent
and original extract in 4 fresh coconut sap samples which 0.5 g of Na2S2O5
were allowed to spontaneous fermentation and their
2.2.e. Preparation of Artificial Fermented Liquor
fermentation were arrested in different time interval. While
(Artificial Toddy).
sulfated ash content remains constant, rest of the chemical
parameters were changed with the time during the 70 ml of toddy inoculum, NPK nutrients and
fermentation processes. Results of sulfated ash content Antibacterial agent were added to the 16 % 1000ml Sugar
showed in Appendix 2 solution in 2.5 l conical flask. Flask was plugged with
Hence this study mainly focused on identifying cotton wool and left for fermentation. At the eighth day
authenticity of toddy using the sulfated ash content. Ash maximum alcohol content was obtained. On that day
values are used to find out quality, authenticity and purity different mixtures of natural and artificial toddy were
prepared as shown in table 1.
of substances and these values are important quantitative
standards. It provides a measure of the total amount of Table 1. Mixture % of natural and artificial toddy
minerals within a food. Sulfuric acid is added prior to Sample no Toddy % (G) Artificial toddy % (A)
heating to facilitate the destruction of organic matter and to M1 100 -
fix certain metals as their sulfate salts to prevent M2 75 25
volatilization. When sulfuric acid is used, the resulting M3 50 50
material is known as sulfated ash. M4 25 75
M5 - 100
22 Authenticity Testing of Coconut Toddy Samples (Palm Wine) Using Sulfated Ash Content

2.3. Sulfated Ash chromatogram were obtained. Gas chromatograms showed

in Appendix 3
Ten (10) ml of samples were pipetted out in a silica dish,
Five (5) drops of con H2SO4 were added to the samples and
kept for drying on a water bath and heated until they were
carbonized by the burner. They were placed in a muffle
3. Results and Discussion
furnace at 5000C± 250C. Three (3) drops of con H2SO4 was LOD 0.015 g/ml, LOQ 0.023 g/ml, RSD 9.6%
added to the samples again and after drying, they were
placed in the muffle furnace. Ash weight was determined. Table 2. Mean Sulfated ash content of the Genuine/artificial toddy
* This process was carried out three times for each Description % w/v
sample. Sulfated ash content unfermented sap 6
0.48
0.47±0.02 (n=3)
Mean of Sulfated ash content of G1 toddy 2014
2.4. Determination of Limit of Detection (LOD) and (0.46-0.50)
Limit of Quantitation (LOQ) 0.50±0.01(n=12)
Mean of sulfated ash content of G2 toddy 2015
(0.48-0.52)
LOD and LOQ were determined based on the standard 0.13±0.04(n=3)
Mean of sulfated ash content of artificial toddy A
deviation of the blank. Alcohol content 7 % w/v was (0.102-0.183)

prepared using absolute alcohol, was used as blank. The similarity of sulfated ash content of the year 2014
Sulfated ash content of the blank samples was determined and that of the year 2015 shows the consistency in sulfated
ten times. Mean and standard deviation (S) were ash content of coconut toddy. Further similarity of
calculated. literature value of the sulfated ash content of unfermented
LOD = Mean + 3*S sap is more confirmed the consistency in the sulfated ash
content. Sulfated ash content of artificial fermented liquor
LOQ = Mean + 10*S was significantly low. This show the mineral components
are higher in genuine toddy than artificial fermented liquor.
2.5. Calculation of the Amount of Variation Explained
by Linear Regression Equation, and its Table 3. Observed and calculated mean sulfated ash content of mixtures
of genuine and artificial toddy
Significance
Sulfated ash content (% g/ml)
SS Sum of square, RSS residual sum of square, Stdev Genuine toddy % in mixture
observed calculated
Standard deviation SEM standard error of mean, R2
0 0.13 0.12
correlation coefficient of degrees of freedom
25 0.22 0.23
SStotal
= ∑ (Yi − Y) 2
(1) 50 0.33 0.32

Yi is the value of the dependent y observed for Xi 75 0.42 0.43

Y is the mean value of the dependent y observed for n 100 0.54 0.53
number of observations
SS mod
= el ∑ (Ye − Y) 2
(2)
Ye is the value of the dependent y for Xi the expected on
the basis of the linear model
RSS = SS total-SS model (3)
SSmodel
R2 = (4)
SStotal
SSmodel / ( p − 1)
F = (5)
RSS / ( n − p )

n = number of observations
P = number of parameters estimated in the equation
Figure 1. Graph of relation between Genuine toddy (%)in a mixture of
2.6. Volatile Organic Compounds genuine and artificial toddy and their sulfated ash content (% g/ml)

25.0 ml of each mixtures were distilled, 0.5 µl samples Plotting the values (table 3) of genuine toddy % in
were injected to the Gas chromatograph (Shimadzu 2010, mixture against sulfated ash content % (w/v) of the
Column RTX 624,30m*0.32mm,1.8µm) and gas mixtures gives linear relations. (Fig 1)
 Food Science and Technology 6(1): 20-27, 2018 23

% sulfated ash = 0.0041 genuine toddy% in mixture + deviation, Genuine toddy sulfated ash content vary from
0.124 (R2 =0.998) 0.44 to 0.54 % w/v. Furthermore sulfated ash content of
The correlation between the genuine toddy % in mixture artificially fermented liquor varies from 0.102 to 0.183.
at ambient temperature and sulfated ash contents shown in Accordingly Samples were classified as genuine (G),
these results indicates the possible influence of mineral adulterated (AD) and artificially fermented liquor (AFM).
components of toddy on its genuine toddy %. Sulfated ash content % w/v genuine toddy % in the sample
and classification are shown in Appendix 1. Out of eighty
Table 4. Analysis of variance explained by a linear regression equation
and its significance (F-test using the F table) nine samples analyzed from excise and courts, forty two
percent (42%) were genuine, fifty seven percent (57%)
Observed sulfated Calculated
Statistical Parameters ash contents Sulfated ash contents were adulterated with artificial fermented liquor and one
Mean 0.328 0.328 percent was artificially fermented liquor.
Std dev 0.164 0.163
SEM 0.06458 0.6452 4. Conclusions
Number of data points 5
Sulfated ash content of coconut toddy varies from 0.44
SS total 0.10428 Df=4 to 0.54 % w/v. There was a correlation between the
SS model 0.10408 Df=1 genuine toddy % in mixture at ambient temperature and
RSS 0.0002 Df=3 sulfated ash contents. Linear relations ship shows by this
equation, % sulfated ash = 0.0041 genuine toddy% in
R2 0.998
mixture + 0.124 (R2 =0.998). Above Linear regression
Fcalculated 1561.2
model can be used to calculate the adulteration of artificial
F critical for 1%
34.12 fermented liquor in market toddy.
significance
Limitation:- This equation can be applied only the toddy
The calculation of the test table showed that the test for which is adulterated by artificially fermented liquor
regression was significant while the F observed value for prepared by sugar solution.
analyte was much higher than the critical value of 34.1, Recommendation:- further research has to be done on
which corresponded to F (1,3,1%).This meant the sulfated metal analysis, that will help in Authenticity identification
ash content was significantly correlated to the genuine of toddy when adulterated other than artificial fermented
toddy % in mixture of artificial and genuine toddy. It can liquor which prepared by sugar solution.
be concluded that the regression equation is highly
significant (p<0.01), and explains r2 =0.998 of the total
variation dataset. Acknowledgements
From the observed sulfated ash content, genuine toddy %
in samples were calculated by using the above equation on Authors would like to thank Excise Commissioner
the samples received from the excise department and courts. General and staff of Excise Department for providing the
From year 2014 and 2015 sulfated ash content and standard genuine toddy samples and assistance of laboratory staff.

Appendix 1
Sulfated ash content % w/v, Genuine toddy % in the sample and classification of samples, G-Genuine, AD- Adulterated, AFM- Artificially fermented
liquor

Sample no Sulfated ash content % w/v Genuine toddy % in sample Classification

1 0.48 86 G
2 0.40 67 AD
3 0.39 64 AD
4 0.40 67 AD
5 0.39 64 AD
6 0.50 91 G
7 0.50 91 G
8 0.50 91 G
9 0.50 91 G
10 0.30 42 AD
11 0.50 91 G
12 0.50 91 G
24 Authenticity Testing of Coconut Toddy Samples (Palm Wine) Using Sulfated Ash Content

13 0.50 91 G
14 0.30 42 AD
15 0.40 67 AD
16 0.40 67 AD
17 0.40 67 AD
18 0.50 91 G
19 0.50 91 G
20 0.50 91 G
21 0.40 67 AD
22 0.45 79 AD
23 0.40 67 AD
24 0.50 91 G
25 0.40 67 AD
26 0.30 42 AD
27 0.40 67 AD
28 0.44 77 AD
29 0.48 86 AD
30 0.50 91 G
31 0.08 -10 AFM
32 0.23 25 AD
33 0.41 69 AD
34 0.33 50 AD
35 0.25 30 AD
36 0.46 81 G
37 0.42 72 AD
38 0.48 86 G
39 0.33 50 AD
40 0.37 60 AD
41 0.55 103 G
42 0.44 77 G
43 0.44 77 G
44 0.40 67 AD
45 0.38 62 AD
46 0.40 67 AD
47 0.38 62 AD
48 0.29 40 AD
49 0.46 81 G
50 0.51 94 G
51 0.48 86 G
52 0.47 84 G
53 0.39 64 AD
54 0.46 81 G
55 0.49 89 G
56 0.37 60 AD
57 0.36 57 AD
58 0.40 67 AD
59 0.48 86 G
60 0.46 81 G
61 0.46 81 G
62 0.46 81 G
63 0.47 84 G
 Food Science and Technology 6(1): 20-27, 2018 25

64 0.48 86 G
65 0.48 86 G
66 0.40 67 AD
67 0.24 28 AD
68 0.45 79 G
69 0.40 67 AD
70 0.34 52 G
71 0.36 57 G
72 0.20 18 AD
73 0.20 18 AD
74 0.32 47 AD
75 0.24 28 AD
76 0.29 40 AD
77 0.39 64 AD
78 0.29 40 AD
79 0.41 60 AD
80 0.42 72 AD
81 0.42 72 AD
82 0.32 47 G
83 0.38 62 G
84 0.27 35 G
85 0.55 104 G
86 0.49 90 G
87 0.49 90 G
88 0.49 90 G
89 0.53 100 G

Appendix 2
Sample Alcohol Acidity Total Solids Sulfated Original
Brix value Conductivity
content (V/V) (W/V) (W/v) ash(W/V) Extract
A1 5.5 0.3 8.15 0.55 10 5.41 17.32
A2 7.6 0.45 4.86 0.51 5.0 5.78 17.26
A3 8.0 0.5 2.26 0.51 3.5 5.9 15.9
B1 6.5 0.5 5.13 0.55 6.5 5.9 16.2
B2 6.8 0.4 5.1 0.55 6.5 5.9 16.4
B3 7.9 0.5 4.35 0.49 5.5 5.37 17.25
C1 6.1 0.52 5.97 0.47 6.5 5.5 16.13
C2 7.1 0.52 4.13 0.47 6.0 5.6 15.67
C3 7.8 0.58 3.34 0.49 4.0 5.59 16.07
D1 6.0 0.66 6.1 0.47 6.5 5.35 15.47
D2 6.9 0.65 4.0 0.46 5.0 5.35 15.09
D3 7.1 0.72 3.3 0.49 4.0 5.38 14.4
1 denoted just draw from tree, 2 denoted after 7 hours fermentation processes, 3 denoted after 12 hours fermentation processes
26 Authenticity Testing of Coconut Toddy Samples (Palm Wine) Using Sulfated Ash Content

Appendix 3: Gas Chromatograms of Toddy

 Food Science and Technology 6(1): 20-27, 2018 27

[5] Borse B B, Rao L J M, Ramalakshmi K, Ragahavan B

(2007) chemical composition of volatiles from coconut sap
(neera) and effect ofprocessing. Food chem. 101(3):877-880
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[4] P. C. Leong, (1952) The nutritive value of Coconut Toddy [11] Sulphated ash content was determined by the method in I.S.I.
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