PHARMACEUTICAL AND BIOLOGICAL EVALUATIONS

October 2015; vol. 2 (Issue 5): 204-207.

www.onlinepbe.com ISSN 2394-0859

Research Article

Quantitative determination of ethanol in arishta by using UV-

visible spectrophotometer
S. F. Sayyad1*, S. R. Chaudhari1, B. P. Panda2
1
Amrutvahni College of Pharmacy, Sangamner, Maharashtra, India - 422 608
2
Pharmaceutical Biotechnology Laboratory, Faculty of Pharmacy, Jamia Hamdard, New Delhi, India- 110 062

*For correspondence
Sadik Ali F. Sayyad,
Assistant Professor,
Department of Pharmaceutics,
Amrutvahni College of
Pharmacy,
Amrutnagar, P.O.-Sangamner
(S.K.), Tal. Sangamner,
Dist. Ahmednagar,
Maharashtra, India– 422 608.
Email:
sadik_sayyad@rediffmail.com
Received: 16 September 2015
Accepted: 27 September 2015
ABSTRACT
Objective: The present work aimed to develop a simple and rapid
technique for determination of ethanol in arishta formulations using UV-
visible spectrophotometer and its comparison with conventional method
involving specific gravity.
Methods: Separation of ethanol from arishta formulations carried out by
solvent extraction using TBP (tri-n-butyl phosphate). The resultant
mixture quantitatively analysed for ethanol concentration by dichromate
oxidation method. Oxidation of ethanol imparts blue colour to the liquid
which is easily detected at 595 nm.
Results: Dichromate oxidation method have lesser deviations in results of
replicates as compare to specific gravity technique. Evaluated arjunarishta,
draksharishta and kutajarishta have ethanol content in the range 5.69 -
8.31, 7.65 - 10.26 and 5.63 - 7.89% v/v respectively.
Conclusions: Quantitative determination of ethanol in fermented
ayurvedic arishta formulation is possible after solvent extraction by
dichromate oxidation technique with consistency and lower volume
requirement.
Keywords: Arishta, Ethanol concentration, Solvent extraction, Dichromate
oxidation

Introduction herbal preparations can be determined by

Arishtas are widely used and unique ayurvedic
products with higher stability and palatability.
These are alcoholic medicaments prepared by
fermentation of herbal decoctions with addition
of sugars.1 The alcohol generation facilitates
extraction of active constituents present in plant
drugs. Self generated alcohol also support
preservation and have better therapeutic
properties.2 Total alcohol content of fermented
specific gravity of distillate which involves
distillation of formulation and then measurement
of specific gravity3. This method is laborious,
time consuming and also have less accuracy.
Other techniques used are electric ebulliometer4
and gas chromatography5. These have
involvement of costlier equipments.
The present work illustrates the simple and rapid
method for determination of ethanol in arishta
formulations. Arishtas are brownish liquid due

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Sayyad SF. et al. Pharmaceutical and Biological Evaluations 2015; vol. 2 (5): 204-207.
to presence of tannins and phenolic compounds
in plants. Hence this method employees solvent
extraction of ethanol from arishta followed by
measurement of alcohol by spectrophotometer
using acid dichromate solution. Determination
of ethanol based on oxidation of ethanol by
reacting with excess of acidic potassium
dichoromate solution.
3C2H5OH + 8H2SO4 + 2K2Cr2O7
3C2H5OH + 2Cr2(SO4)3 + 11H2O + K2SO4
When ethanol is present in an aqueous solution,
chromium ions oxidize ethanol, and these ions
are reduced from the +6 oxidation state to +3,
changing the color from orange to green.6
This study aimed to develop a convenient
technique for measurement of alcohol in arishta
and comparison of it with quantitative
determination by specific gravity of distillate.
Materials and Methods
Materials
Traditionally prepared arjunarishta,
draksharishta and kutajarishta has been
selected for quantitative determination of
alcohol content. Three different brands of
selected formulations have also been
procured from market and used in the
present study. TBP (tri-n-butyl phosphate)
has been purchased from Loba Chemie,
Mumbai, India.
Methods
Determination of ethanol by dichromate
oxidation method
Solvent extraction
TBP (tri-n-butyl phosphate) has been
selected as a solvent for extraction of
ethanol from arishta. It has density 0.9727,
distribution coefficient for ethanol in water
is 0.576 and able to show intense colour
©Pharmaceutical and Biological Evaluations
development for alcohol during dichromate
oxidation.6 The 2.5 ml of arishta sample
diluted with equal fraction of distilled water
and mixed with 5 ml TBP by vigorous
vortexing for 15 minutes. Then these tubes
were kept aside for phase separation and
upper layer used for dichromate oxidation.
Standard solutions of ethanol were prepared
by diluting specific amount of absolute
alcohol using distilled water and processed
similarly.
Preparation of dichromate reagent
The dichromate reagent required for study
was prepared by dissolving 40 g potassium
dichromate in approximately 200 ml of
distilled water. Then 270 ml concentrated
H2SO4 added cautiously, the resultant
solution cooled and volume is adjusted to
500 ml by adding sufficient volume of
distilled water.
Dichromate oxidation and
spectrophotometric analysis
3 ml of TBP layer transferred to new tube
and mixed with 3 ml of dichromate reagent
by shaking at 150 rpm for 10 minutes.
Afterward lower layer separated and
subjected to measurement of absorbance at
595 nm using spectrophotometer
(Shimadzu-1800).
Ethanol measurement on the basis of
specific gravity
Not less than 25 ml of preparation being
examined was transferred to the distillation
apparatus and its temperature is noted. It
was diluted with equal volume of water and
distilled. Distillate about 2 ml less than the
total volume collected. The distillate is clear
or not more than slightly cloudy, and does
not contain more than traces of volatile
substances other than alcohol and water.
Water was added to measure exactly same
volume of original test liquid and adjusted to
205
Sayyad SF. et al. Pharmaceutical and Biological Evaluations 2015; vol. 2 (5): 204-207.

Table 1: Ethanol content of different arishta formulations.

Ethanol content (% v/v)

Formulation
By dichromate oxidation By specific gravity
Arjunarishta A-I 6.65  0.16 5.71  0.36
A-II 5.69  0.30 4.85  0.39
A-III 8.31  0.16 7.64  0.35
Draksharishta D-I 7.65  0.17 8.29  0.47
D-II 9.58  0.16 8.62  0.39
D-III 10.26  0.24 8.81  0.76
Kutajarishta K-I 5.63  0.29 4.61  0.56
K-II 7.89  0.29 7.08  0.46
K-III 6.74  0.25 5.75  0.32
Where, n=3

temperature noted before. Specific gravity of
this liquid was determined at 25oC and
alcohol content analyzed on the basis of
alcoholometric table given in United States
Pharmacopoeia.7
bring the ethanol concentration in linearity
range. Dichromate oxidation method is suitable
for determination of alcohol concentration upto
16% v/v with limit of detection (LOD) 0.32%
v/v and limit of quantification (LOQ) 0.98% v/v.

Results and Discussion

Arishta consist sugar and nitrogen rich
constituents making it susceptible to microbial
attack but production of ethanol improves its
stability by preventing microbial spoilage.
Alcohol also facilitates extraction of some key
bioactive molecules from plant material
contributing for the activity of formulation. But
higher concentration may leads to toxicity,
hence monitoring of alcohol concentration is
vital parameter for quality control of arishta
formulations. In current study evaluated three Figure 1: Comparison of ethanol content of
different brands of arjunarishta, draksharishta arishta formulations determined by
and kutujarishta shown wide variation in ethanol dicromate oxidation and specific gravity
content (Figure 1). Standard deviations of method.
obtained results indicate dichromate oxidation
method is more consistent in replicates whereas Conclusions
a specific gravity technique is less reliable due
to variations (Table 1). As compare to specific Dichromate oxidation is a consistent and reliable
gravity technique determination of ethanol by method for quantitative determination of ethanol
dichromate oxidation method require very less in fermented ayurvedic formulation arishta.
sample. Standard curve plotted by dichromate Alcohol measurement is feasible at lower
oxidation method have correlation coefficient sample volume making it ideal for small
0.996 with equation of line Y = 0.218X + 0.022. research scale batches.
Appropriate dilution of test sample required to

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Sayyad SF. et al. Pharmaceutical and Biological Evaluations 2015; vol. 2 (5): 204-207.
Funding: No funding sources
Conflict of interest: None declared
References
1. Ayurvedic Formulary of India. First edition,
Government of India, Ministry of health and
family welfare, Department of Indian system
of medicine and homeopathy, New Delhi,
2000; part 1: pp. 33.
2. Sekar S, Mariappan S. Traditionally
fermented biomedicines, arishtas and asavas
from Ayurveda. Ind J Trad Knowl.
2008:7(4):548-56.
3. Swain MR, Kar S, Sahoo AK, Ray RC.
Ethanol fermentation of mahula (Madhuca
latifolia L.) flowers using free and
immobilized yeast Saccharomyces
cerevisiae. Microbiol Res. 2007;162:93-8.
4. Weerasooriya WMB, Liyange JA, Pandya
SS. Quantitative parameters of different
©Pharmaceutical and Biological Evaluations
brands of asava and arishta used in
ayurvedic medicine: an assessment. Indian J
Pharmacol. 2008;38(5):365.
5. Kroes BH, van den Berg AJJ, Abeysekara
AM, de Silva KTD, Labadie RP.
Fermentation in traditional medicine: the
impact of Woodfordia fruticosa flowers on
the immunomodulatory activity, and the
alcohol and sugar contents of Nimba arishta.
J Ethannopharmacol. 1993;40(2):117-25.
6. Seo HB, Kim HJ, Lee OK, Ha JH, Lee HY,
Jung KH. Measurement of ethanol
concentration using solvent extraction and
dichromate oxidation and its application to
bioethanol production process. J Ind
Microbiol Biotechnol. 2009;36(2):285-92.
7. USP 32 NF 27. The official compendia of
standards, The United States Pharmacopeial
Convention, Rockville, MD, USA.
2009;1:224-5,954.
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