This document outlines experiments to determine the alcohol content of herbal formulations using distillation and chromatography methods. It describes the distillation process which involves transferring a sample to a distillation flask with water and pumice, distilling the mixture, and collecting the distillate to measure specific gravity and calculate ethanol percentage. It also describes determining alcohol content using gas chromatography by preparing ethanol standard solutions, diluting samples, injecting them, and using a calibration graph to obtain concentration from peak areas. The overall aim is to quantify the alcohol present in herbal formulations like Asava and Arista.

1. HERBAL DRUG TECHNOLOGY PRACTICAL
EXPERIMENT NO. 2
Mrs. Poonam N. Chougule
HOD of Pharmacognosy department and
Assistant Professor
AMCP, Peth- Vadgaon.

2. AIM: TO DETERMINE THE ALCOHOL CONTENT OF
ASAVA AND ARISTA
 Requirement- Distillation assembly, ethanol, volumetric
flask, HPLC grade water, measuring cylinder etc.
 Theory- Alcohol content present in Ayurvedic formulations
like Asava and Arista can be determined by using two
methods i.e. distillation method and chromatographic
method. There will be a separate setup called distillation
unit in order to determine the alcohol content.
 Alcohol is lesser in density than water. Based on this factor
content is determined. Chromatography depends on the
principle of the color formation. Each compound gives
different color for reactant solution. Color indicates the
presence of alcohol.

3. PROCEDURE
 Alcohol content apparatus consists of a round-
bottomed flask fitted with a distillation head with a
steam trap and attached to a vertical condenser.
 A tube is fitted to the lower part of the condenser
and carries the distillate in to the lower pail of a
100ml or 250ml volumetric flask. The volumetric
flask is immersed in a beaker containing mixture of
ice and water during the distillation, a disc with a
circular aperture, 6 cm in diameter, is placed under
the distillation flask to reduce the risk of charring of
any dissolved substances.

4. DETERMINATION ALCOHOL CONTENT BY
DISTILLATION AND ANY DISSOLVED SUBSTANCES
 Take 25ml of the given preparation and transfer,
accurately it to the distillation flask. Dilute it with 150ml of
water and to it add a little pumice powder. Distill it and
collect not less than 90ml of the distillate in 200 ml
volumetric flask and then dilute the volume with distilled
water.
 Determine the specific gravity at 25ᵒ C read off
percentage of ethyl alcohol corresponding to the specific
gravity from the alcohol or metric table given in United
States pharmacopeia, if the specific gravity is found to be
between two values the percentage of ethanol should be
obtained by interpolation after circulation of Ethanol
content the report or result to one decimal place.

5. DETERMINATION OF ALCOHOL CONTENT BYE
GAS CHROMATOGRAPHY
 It is a performed by using carbowax 20M stationary
phase Packed into Steel column with internal diameter
of 2 mm nitrogen gas is used as carrier gas, at a flow
rate off 1kg/cm2/minute, a range of standard solutions
of Ethanol is prepared containing 1, 2, 3, 4, 5% v/v of
ethanol using ethanol 99.98% and HPLC grade water
from the standard solution 1 ml is a diluted to 200 ml
with HPLC grade water, then inject 1micro ml of
solution and record the chromatogram. plot the area
against concentration of alcohol to obtain calibration
graph.

6. THANK YOU