Performing Calibration of Volumetric Flask in Quality Control Department
For Volumetric Flask
This is applicable up to 100 ml volumetric flask.
Pre-Lab work before Calibration
Clean the glassware & make it free from grease before carrying out calibration.
Clean the wet outer surface with tissue paper.
Before calibration keep all glassware at room temp for 1 hour.
The glassware must not be wetted above the meniscus, if this happens, wipe it with tissue
paper.
Always kept your eye at the same level, while aligning the meniscus.
On receipt of glassware from the supplier, enter the following
details into the register:
a. Date of receipt
b. Description
c. Serial No.
d. Quantity received
e. Supplier Name
f. Challan No.
g. Qty. Approved/ Serial No.
h. Qty. Rejected/ Serial No.
i. Remark
Procedure
1. First take the empty weight of dry volumetric flask and record the weight.
2. Fill the volumetric flask with distilled water up to the mark using funnel.
3. Wipe the outside of volumetric flask with dry paper towel and weigh it, as provided
below in observations.
4. Perform the process in triplicate and record average readings.
5. Calculate the difference between labelled volume and average of observed volume of
glassware as per formula.
6. Calculate the volume by taking the correction factor 0.99602 g (1ml of purified water at
25-degree Celsius equals 0.99602 grams) and record the readings.
Observations I:
Weight of pre-weighed dry, empty, volumetric flask, in g (W1): __________________
Weight of volumetric flask and water, in g (W2): ____________________
Weight of water in gram (W3) = (W2-W1): ________________________
Observed Volume of Glassware: W2-W1/ D = ______________________ml
Observations II:
Weight of pre-weighed dry, empty, volumetric flask, in g (W1): __________________
Weight of volumetric flask and water, in g (W2): ____________________
Weight of water in gram (W3) = (W2-W1): ________________________
Observed Volume of Glassware: W2-W1/ D = ______________________ml
Observation III:
Weight of pre-weighed dry, empty, volumetric flask, in g (W1): __________________
Weight of volumetric flask and water, in g (W2): ____________________
Weight of water in gram (W3) = (W2-W1): ________________________
Observed Volume of Glassware: W2-W1/ D = ______________________ml
Annexure
Calibration of Glassware
Glassware Tolerance
Basics of Cleaning Validation
Cleaning of pharmaceutical instruments after a product has been manufactured is critical to
ensure that the subsequent products to be manufactured are not contaminated. The removal
of the residues of the previous products from manufacturing equipment is known as
cleaning. If the whole process of cleaning has been documented, it is referred to as cleaning
method validation.
Cleaning validation is proof that the cleaning process is effective to remove all residues of
the product that was manufactured, cleaning agents those were used during the cleaning
process and prevents micro-organisms from developing. This process is done as a
requirement of regulatory authorities.
Manufacturing companies should not do it for the sole reason of satisfying the regulatory
authorities but they should put it in mind that it is required to ensure that the patients are
safe.
Requirements for a cleaning validation process
Protocol
First, you must create a protocol. When preparing a protocol, some of the factors that should
be considered are the quality of the water, the detergent to be used, the rinsing period and
the system's size. The protocol should contain the objective of the whole process, the scope
of the protocol, responsibilities of the departments, the procedure of cleaning, acceptance
criteria and cleaning method validation report.
Personnel
The people conducting the process should be trained before they start the process of cleaning
method validation. They must have knowledge of cleaning procedure, standard operating
procedure and validation protocol.
Parts of the Equipment to Clean
There are some parts of the equipment that come into contact with the product during
manufacturing. This place should be labeled contact parts while those that do not come into
contact with the product are labeled non-contact parts. When cleaning, contact parts of the
equipment should be cleaned properly. A lot of care should be taken for cleaning of the
place those are difficult to clean. However, for non-contacts take care that these residues
during cleaning do not move to these places. Consideration should still be given to the design
of the equipment as this influence how it will be cleaned and the time it takes to clean.
Determine the Detergent Used
A good detergent should be easily removed during the cleaning process by rinsing.
Detergents which have residues that are hard to remove usually are discouraged. There
before choosing any cleanser, a manufacturer must know its composition. The manufacturer
should also define the limits of the detergent residue that are acceptable.
Prevent Microorganisms
It’s also a requirement that the validation process does not support the growth of microbes.
In determining if the validation process has supported microbial growth, the storage of the
equipment before cleaning and after cleaning is often considered to decide whether they
support microbial growth. Make sure that after cleaning the equipment is dry. Store it in a
dry place. This is important as any other sterilization procedure that might be applied to the
equipment will more likely achieve the required standard.
Sampling process
Samples are needed to determine the level of residues present in the equipment. There are
two types of sampling used in the validation process. Rinse sampling and direct sampling.
Direct sampling is used to collect samples for areas that are hard to clean. With Rinse
sampling, you can get a sample of a place that is inaccessible or for a large surface area.
Using the two methods is highly recommended.
Calculating the Acceptance Criteria
A cleaning process is determined before the process begins. An appropriate method is
determined by creating a matrix of the product's attributes, and the equipment is used. The
method chosen should be sensitive enough to detect any residuals on the equipment. The
accepted method should also detect an acceptable limit of the contaminants and residues.
A proper cleaning method validation will enhance the process of the company’s equipment
cleaning and will free the company from facing legal actions for not performing it. Therefore,
every company where a pharmaceuticals or whatsoever industries it operates in must always
observe this process.
Performing the Calibration of Glass Pipette in Quality Control Department
For Glass Pipette
This is applicable up to 20 ml glass pipette.
Pre-Lab work before Calibration
Clean the glassware & make it free from grease before carrying out calibration.
Clean the wet outer surface with tissue paper.
Before calibration keep all glassware at room temp for 1 hour.
The glassware must not be wetted above the meniscus, if this happens, wipe it with tissue
paper.
Always kept your eye at the same level, while aligning the meniscus.
On receipt of glassware from the supplier, enter the following
details into the register:
a. Date of receipt
b. Description
c. Serial No.
d. Quantity received
e. Supplier Name
f. Challan No.
g. Qty. Approved/ Serial No.
h. Qty. Rejected/ Serial No.
i. Remark
Procedure
1. Fill the pipette with distilled water up to the mark using bulb sucker.
2. Wipe the outside of pipette with dry paper towel and transfer the water in a dry pre-
weighed beaker and weigh it, as provided below in observations.
3. Perform the process in triplicate and record average readings.
4. Calculate the difference between labelled volume and average of observed volume of
glassware as per formula.
5. Calculate the volume by taking the correction factor 0.99602 g (1ml of purified water at
25-degree Celsius equals 0.99602 grams) and record the readings.
Observations I:
Weight of pre-weighed dry, empty beaker, in g (W1): __________________
Weight of beaker and water, in g (W2): ____________________
Weight of water in gram (W3) = (W2-W1): ________________________
Observed Volume of Glassware: W2-W1/ D = ______________________ml
Observations II:
Weight of pre-weighed dry, empty beaker, in g (W1): __________________
Weight of beaker and water, in g (W2): ____________________
Weight of water in gram (W3) = (W2-W1): ________________________
Observed Volume of Glassware: W2-W1/ D = ______________________ml
Observation III:
Weight of pre-weighed dry, empty beaker, in g (W1): __________________
Weight of beaker and water, in g (W2): ____________________
Weight of water in gram (W3) = (W2-W1): ________________________
Observed Volume of Glassware: W2-W1/ D = ______________________ml
Annexure
Calibration of Glassware
Glassware Tolerance
Performing Calibration of Glass Burette in Quality Control Department
For Glass Burette
This is applicable up to 50 ml glass burette.
Pre-Lab work before Calibration
Clean the glassware & make it free from grease before carrying out calibration.
Clean the wet outer surface with tissue paper.
Before calibration keep all glassware at room temp for 1 hour.
The glassware must not be wetted above the meniscus, if this happens, wipe it with tissue
paper.
Always kept your eye at the same level, while aligning the meniscus.
On receipt of glassware from the supplier, enter the following
details into the register:
a. Date of receipt
b. Description
c. Serial No.
d. Quantity received
e. Supplier Name
f. Challan No.
g. Qty. Approved/ Serial No.
h. Qty. Rejected/ Serial No.
i. Remark
Procedure
1. Fill the burette with distilled water up to the mark using funnel.
2. Wipe the outside of glass burette with dry paper towel and transfer the water in a dry pre-
weighed beaker and weigh it, as provided below in observations.
3. Perform the process in triplicate and record average readings.
4. Calculate the difference between labelled volume and average of observed volume of
glassware as per formula.
5. Calculate the volume by taking the correction factor 0.99602 g (1ml of purified water at
25-degree Celsius equals 0.99602 grams) and record the readings.
Observations I:
Weight of pre-weighed dry, empty beaker, in g (W1): __________________
Weight of beaker and water, in g (W2): ____________________
Weight of water in gram (W3) = (W2-W1): ________________________
Observed Volume of Glassware: W2-W1/ D = ______________________ml
Observations II:
Weight of pre-weighed dry, empty beaker, in g (W1): __________________
Weight of beaker and water, in g (W2): ____________________
Weight of water in gram (W3) = (W2-W1): ________________________
Observed Volume of Glassware: W2-W1/ D = ______________________ml
Observation III:
Weight of pre-weighed dry, empty beaker, in g (W1): __________________
Weight of beaker and water, in g (W2): ____________________
Weight of water in gram (W3) = (W2-W1): ________________________
Observed Volume of Glassware: W2-W1/ D = ______________________ml
Annexure
Calibration of Glassware
Glassware Tolerance
Performing the Validation of Autoclave
(Standard operating procedure to validate the steam sterilizer using biological indicators having spores of Bacillus
stearothermophilus)
PROCEDURE
Validation of autoclave is done once in three months using the Bioindicator System.
The bioindicator system consists of ampoules containing spores of Bacillus
stearothermophilus.
These ampoules are stored under refrigeration when not in use.
On the day of validation remove three ampoules from the refrigerator and allow them to come
to room temperature.
Take two beakers and place an ampoule each, in each beaker, to avoid contamination due to
accidental breakage during sterilization.
Place the two beakers, containing an ampoule each, into the autoclave, along with the material
to be autoclaved.
The position of the two ampoules.
Switch on the autoclave and carry out the autoclaving cycle as per the procedure described in
SOP
After autoclaving, remove the ampoules and allow them to cool to room temperature.
Mark the two ampoules and incubate at 60 ± 2° C for 48 hrs.
The third non-sterilized ampoule is incubated along with the sterilized ampules, to serve as a
control.
If sterilization is effective, the contents of the sterilized ampules remains clear, red violet color.
If sterilization is ineffective, the contents of the sterilized ampules turn turbid, yellow in color.
The contents of the control ampoule should also turn yellow.
Record and store the results obtained.
Label the sterilized ampoules and the control ampoule, mentioning the date of validation, and
store in the refrigerator, for reference.
NOTE
Each ampoule of Bioindicator System consists of nutrient broth, sugar, a pH indicator and
spores of Bacillus stearothermophilus.
The thermal resistance of the spores is such that they are totally killed after 15 minutes when
heated in compressed steam at a temperature of 121°C ± 0.5°C.
If sterilization is adequate, the Bacillus stearothermophilus spores get killed and the contents
of the ampoule remain a clear red violet color.
If sterilization is inadequate, the spores survive. Within 24 hours of incubation, the
contents of the ampoule turn turbid due to microbial growth and the color turns yellow due
to the production of acid, as a result of sugar fermentation.
Precautions
Ensure that the ampoules are stored in the refrigerator, when not in use.
Ensure that the ampoules are not used beyond the date of expiry printed in the pack.
Ensure that the ampoules are at room temperature before loading them into the autoclave.
Performing the Calibration of Weighing Balance
PROCEDURE
1. Operate the instrument as per respective Standard Operating Procedure.
2. Switch ‘ON’ the instrument.
3. Switch ‘ON’ the balance.
4. The display will blink with 8.8.8.8.8.8.
5. After few seconds, the display will show 0.00 g.
6. If display is not stable, press the TARE key & wait till the display shows 0.00 g.
For Accuracy
1. Place 5 grams on the weighing pan.
2. Note the weight.
3. Calculate the difference between the weight in certificate and observed weight.
4. Repeat the above steps using 50gm & 100 gm. weights.
5. Record the readings in Annexure I.
For Precision
1. Place 5 grams in the weighing pan.
2. Note the weight.
3. Repeat the above two steps nine times.
4. Record the weight in Annexure-II.
Calculate the Standard deviation.
Calculate the measurement uncertainty using following equation.
Measurement of uncertainty = 2 x SD/ Actual weight
Annexure-I
Annexure-II
Performing Calibration of UV / Visible Spectrophotometer
(Calibration of the UV spectrophotometer including control of absorbance using potassium dichromate solution, resolution
power using toluene in hexane, limit of stray light and wavelength accuracy)
Control of Absorbance
Potassium Dichromate Solution
Dry a quantity of potassium dichromate by heating to constant weight at 130°C. Weigh &
transfer accurately a quantity not less than 57.0 mg & not more than 63.0 mg. Dissolve & dilute
in sufficient 0.005M H2SO4 to produce 1000 ml.
Measure the absorbance of potassium dichromate solution at the wavelengths given below.
Calculate the value of A (1% 1cm) for each wavelength.
A (1% 1cm) = Absorbance X 10000 / Weight of Potassium dichromate in mg.
Acceptance criteria
Wavelengths (nm) A [1% 1cm] Limit
235.0 124.5 122.9 - 126.2
257.0 144.0 142.8 - 145.7
313.0 48.6 47.0 - 50.3
350.0 106.6 105.6 - 108.2
Stringent limits are adopted from BP for acceptance criteria
Resolution power
Record the spectrum of a 0.02% v/v solution of toluene in hexane in the range of 260 nm to 420
nm (before use check the hexane for transmittance, using water as a blank between 260nm to
420nm & use only if transmittance is not less than 97%).
Acceptance criteria
The ratio of the absorbance at the maximum at about 269nm to that at the minimum at about
266nm is not less than 1.5.
Limit of Stray Light
Prepare a 1.2 % w/v solution of potassium chloride in water.
Measure absorbance of the above solution at 198.0, 199.0, 200.0, 201.0,
202.0 nm using water as blank.
Acceptance criteria
Absorbance is greater than 2.
Wavelength accuracy, Resolution &
Baseline flatness (inbuilt test)
Attach printer directly to the instrument
(instead of the computer).
Go to Þ MODE press F3 key i.e. Maintenance.
Press ‘1’.
Press Start/Stop key.
After screen changes, ensure that nothing is kept in the optical path & press the Start/Stop key
again.
Prints will come after all the three tests are over.
Acceptance criteria
For wavelength accuracy: at 656.1 ± 0.3 nm & at 486.0 ± 0.3 nm.
Resolution: 1.0 nm or less.
Baseline flatness: ± 0.002Abs.
Frequency: Once a month.
