PHARMA DEVILS
QUALITY CONTROL DEPARTMENT
GENERAL TESTING PROCEDURE
Title: Determination of Water
SOP No.: Department: QC
Effective Date: Review Date:
Revision No.:
Supersede SOP No.:
1.0 OBJECTIVE:
1.1 To lay down a procedure for Determination of Water.
2.0 SCOPE:
2.1 It is applicable for the estimation of Raw material.
3.0 RESPONSIBILITY:
3.1 Analyst / Officer / Executive follow the procedure.
3.2 Head-QC are responsible for effective implementation of this SOP.
4.0 REFERENCE:
4.1 BP (Ph. Eur. Method)
5.0 DEFINITION:NA
6.0 PROCEDURE: Use Method IA unless otherwise directed
6.1 Method I: Semi-micro Determination of Water-The semi-micro determination of water is based
upon the quantitative reaction of water with sulfur dioxide and iodine in, a suitable anhydrous
medium in the presence of a base with sufficient buffering capacity.
Apparatus: The apparatus consists of a titration vessel with:
2 identical platinum electrodes.
Tight inlets for introduction of solvent and titrant.
An inlet for introduction of air via a desiccant.
A sample inlet fitted with a stopper or, for liquids, a septum.
Inlet systems for introduction of dry nitrogen or for aspiration of solvents may also be fitted. The
titration is carried out according to the instrument supplier's instructions. Care is taken throughout
the determination to avoid exposure of reagents and solvents to atmospheric moisture. The end-
point is determined using 2 identical indicator electrodes connected to an electrical source that
PHARMA DEVILS
QUALITY CONTROL DEPARTMENT
GENERAL TESTING PROCEDURE
Title: Determination of Water
SOP No.: Department: QC
Effective Date: Review Date:
Revision No.:
Supersede SOP No.:
maintains between the electrodes either aconstant current (2.2.65. Voltametric titration) or a
constant voltage (2.2.19. Amperometric titration). Where direct titration is used (method A),
addition of titrant causes either a decrease in voltage where constant current is maintained or an
increase in current where constant voltage is maintained, until the end-point is reached.
Instruments with automatic end-point detection are commonly used. Instrument qualification is
carried out according to established quality system procedures, for example using a suitable
certified reference material (sodium aminosalicylate dihydrate for equipmentqualification CRS
may be used).
Standardisation: To the titration vessel, add methanol, dried if necessary, or the solvent
recommended by the supplier of the titrant. Where applicable for the apparatus used, eliminate
residual water from the measurement cell or carry out a pre-titration. introduce a suitable amount
of water in an appropriate form(water R or a certified reference material) and carry out the
titration, stirring for the necessary time. The water equivalent is not less than 80 per cent of that
indicated by the supplier Standardise the titrant before the first use and at suitable intervals
thereafter.
Unless otherwise prescribed, use Method A.
METHOD A: Introduce into the titration vessel methanol, or the solvent indicated in the
monograph or recommended by the supplier of the titrant. Where applicable for the apparatus
used, eliminate residual water from the measurement cell or carry out a pre-titration. Introduce the
substance to be examined rapidly and carry out the titration, stirring for the necessary
extraction time.
METHOD B: Introduce into the titration vessel methanol, or the solvent indicated in the
monograph or recommended by the supplier of the titrant. Where applicable for the. apparatus
used, eliminate residual water from the measurement cell or carry out a pre-titration. Introduce the
substance to be examined rapidly and in a suitable state of division. Add an accuratelymeasured
volume of the titrant, sufficient to give an excess of about 1 mL or the prescribed volume. Allow
to stand protected from light for 1 min or the prescribed time, with stirring. Titrate the excess of
reagent using methanol or the prescribed solvent, containing an accurately known quantity of
water.
PHARMA DEVILS
QUALITY CONTROL DEPARTMENT
GENERAL TESTING PROCEDURE
Title: Determination of Water
SOP No.: Department: QC
Effective Date: Review Date:
Revision No.:
Supersede SOP No.:
Suitability: The accuracy of the determination with the chosen titrant must be verified for each
combination of substance, titrant and solvent to be examined. The following procedure, given as
an example, is suitable for samples containing 2.5-25 mg of water.
The water content of the substance to be examined is determined using the reagent/solvent system
chosen. Thereafter in the same titration vessel, sequential known amounts of water, corresponding
to about 50-100 per cent of the amount found in the substance to be examined, are added in an
appropriate form (at least 5 additions) and the water content is determined after each addition.
Calculate the percentage recovery (r) after each addition using the following expression:
W2
r = 100 -------
W1
Where W1 = Amount of water added, in milligrams.
W2 = Amount of water found, in milligrams.
Calculate the mean percentage recovery (̄ r)
The reagent/solvent system is considered to be acceptable if rˉ is between 97.5 per cent and 102.5
per cent.
Calculate the regression line. The x-axis represents the cumulative water added whereas the y-
Axis represents the sum of the initial water content determined for the substance (M) and the
cumulative water determined after each addition.
Calculate the slope (b), the intercept with the y-axis (a) and the intercept of the extrapolated
calibration line with the x-axis (d) Calculate the percentage errors (e1 and e2) using the following
expressions:
PHARMA DEVILS
QUALITY CONTROL DEPARTMENT
GENERAL TESTING PROCEDURE
Title: Determination of Water
SOP No.: Department: QC
Effective Date: Review Date:
Revision No.:
Supersede SOP No.:
a= The y-axis intercept, in milligrams of water;
d= The x-axis intercept, in milligrams of water;
M =Water content of the substance, in milligrams of water
The reagent/solvent system is considered to be acceptable if:
Ie1I and Ie2I are not greater than 2.5 per cent.
b is between 0.975 and 1.025.
6.2 Method II Determination of Water by Distillation: The apparatus (see Figure 2.2.13.-1)
consists of a glass flask (A) connected by a tube CD) to a cylindrical tube (B) fitted with a
graduated receiving tube (E) and reflux condenser (C). The receiving tube (E) is graduated in0.1
mL. The source of heat is preferably an electric heater with rheostat control or an oil bath. The
upper portion of the flask and the connecting tube may be insulated. Method Clean the receiving
tube and the condenser of the apparatus, thoroughly rinse with water, and dry. Introduce 200 mL
of toluene and about 2 mL of water into the dry flask. Distil for 2 h, then allow to cool for about
30 min and read the water volume to the nearest 0.05 ml. Place in the flask a quantity of the
substance, weighed with an accuracy of 1 per cent, expected to give about 2 mL to3 mL of water.
If the substance has a pasty consistency, weigh it in a boat of metal foil. Add a few pieces of
porous material and heat the flask gently for 15 min. When the toluene begins to boil, distil at the
rate of about two drops per second until most of the water has distilled over, then increase the rate
of distillation to about four drops per second. When the water has all distilled over, rinse the inside
of the condenser tube with toluene. Continue the distillation for 5 min, remove the heat, allow the
receiving tube to cool to room temperature and dislodge any droplets of water which adhere to the
walls of the receiving tube. When the water and toluene have completely separated, read the
volume of water and calculate the content present in the substance as milliliters per kilogram,
using the formula
PHARMA DEVILS
QUALITY CONTROL DEPARTMENT
GENERAL TESTING PROCEDURE
Title: Determination of Water
SOP No.: Department: QC
Effective Date: Review Date:
Revision No.:
Supersede SOP No.:
1000(n2-n1)
-----------------
M
m =The mass in grams of the substance to be examined,
nl= The number of milliliters of water obtained in the first distillation,
n2 the total number of milliliters of water obtained in the 2 distillations.
Apparatus for the determination of water by distillation Dimensions in millimeters
6.3 Method III: Coulometric Titration:
Principle: The coulometric titration of water is based upon the quantitative reaction of water with
sulfur dioxide and iodine in an anhydrous medium in the presence of a base with sufficient
buffering capacity. In contrast to the volumetric method described in general chapter 2.5.12.
Water: semi-micro determination, iodine is produced electrochemically in the reaction cell by
PHARMA DEVILS
QUALITY CONTROL DEPARTMENT
GENERAL TESTING PROCEDURE
Title: Determination of Water
SOP No.: Department: QC
Effective Date: Review Date:
Revision No.:
Supersede SOP No.:
oxidation of iodide. The iodine produced at the anode reacts immediately with the water and the
sulfur dioxide contained in the reaction cell. The quantity of water in the substance is directly
proportional to the quantity of electricity (in coulombs), corresponding to electric current (in
amperes) multiplied by time (in seconds), used for iodine generation up until the titration end-
point. When all of the water in the reaction cell has been consumed, the end-point is reached and
thus an excess of iodine appears. 1 mole of iodine corresponds to 1 mole of water, an amount of
electricity of 10.71 C corresponds to 1 mg of water. Moisture is eliminated from the reaction cell
by pre-titration, i.e. the electrolyte reagent is titrated to dryness before starting the sample
analysis. Individual determinations can be carried out successively in the same reagent solution,
under the following conditions:
Each component of the test mixture is compatible with the other components.
No other reactions take place.
The volume and the water capacity of the electrolyte reagent are sufficient.
Coulometric titration is intended for the quantitative determination of small quantities of water
(from 10 ug), however a working range of 100 µg to 10 mg of water is recommended for
reproducibility reasons.
Accuracy and precision of the method are predominantly governed by the sample preparation
and the extent to which atmospheric moisture is excluded from the system. Control of the
system must be monitored by measuring the amount of baseline drift.
Apparatus: The apparatus consists of a reaction cell, electrodes and a magnetic stirrer. The
reaction cell consists of a large anode compartment and a smaller cathode compartment.
Depending on the design of the electrode, both compartments Can be separated by a
diaphragm. Each compartment contains a platinum electrode. liquid or solubilized samples are
introduced through a septum, using a syringe. Alternatively, an evaporation technique may be
used in which the sample is heated in an oven and the water is evaporated and carried into the
cell by means of a stream of dry inert gas. The introduction of solid samples into the cell
should.in general.be avoided. However, if it has to be done it is effected through a sealable
port; appropriate precautions must be taken to avoid the introduction of moisture from air,
such as working in a glove box in an atmosphere of dry inert gas. The analytical procedure is
PHARMA DEVILS
QUALITY CONTROL DEPARTMENT
GENERAL TESTING PROCEDURE
Title: Determination of Water
SOP No.: Department: QC
Effective Date: Review Date:
Revision No.:
Supersede SOP No.:
controlled by a suitable electronic device, which also displays the results. Instrument
qualification is carried out according to established quality system procedures, for example
using asuitable certified reference material. Sodium amino salicylatedihydrate for equipment
qualification CRS may be used when proceeding by direct or liquid sample introduction,
whereas amoxicillin trihydrate for performance verification CRS may beused with the
evaporation technique.
Method: Fill the compartments of the reaction cell with electrolyte reagent for the micro
determination of water R according to the manufacturer's instructions and perform the
coulometric pretitration to a stable end-point. Introduce the prescribed quantity of the
substance to be examined into the reaction cell and titrate again to a stable end-point, stirring
for at least 30 s, unless otherwise indicated in the monograph. If an oven is used, the
prescribed quantity of sample is introduced into the oven and heated. After evaporation of the
water from the sample into the reaction cell, the titration is started. Alternatively, the
evaporated moisture is immediately titrated while heating the sample in the oven to avoid loss
of evaporated water already collected in the reagent solution during prolonged heating. Read
the value from the instrument's output and calculate if necessary the percentage or quantity of
water that is present in the substance. When appropriate to the type of sample and the sample
preparation, perform a blank titration.
Verification of Accuracy: At appropriate intervals, such as at least at the beginning and the
end of a series of sample titrations, introduce a defined quantity of water, in the same order of
magnitude as the quantity of water in the sample, using a suitable certified reference material
and perform the coulometric titration. The recovery is within the range of 97.5 per cent to
102.5 per cent for an addition of 1000 ug of H2O and within the range of 90.0 per cent to 110.0
per cent for the addition of 100 ug of H2O.
7.0 Annexures: NA
8.0 Distribution:
8.1 Display copy 1 : Quality Control Lab
PHARMA DEVILS
QUALITY CONTROL DEPARTMENT
GENERAL TESTING PROCEDURE
Title: Determination of Water
SOP No.: Department: QC
Effective Date: Review Date:
Revision No.:
Supersede SOP No.:
9.0 Abbreviation:
GTP : General Test Procedure
QC : Quality Control laboratories
10.0 Revision History:
10.1 Revision history table:
CC
Document Number Brief Description of Change
Number/Date
