Pharmaceuticals Instruments Calibration and Operation PDF Notes

 Pharmaceuticals Instruments Calibration and Operation PDF Notes

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Operation
& Calibration of Melting Point Apparatus

OPERATION

·        
Powder the sample
as fine as possible.

·        
Dry it at a
temperature if specified in the individual monograph.

·        
Heat the bath
until the temperature is about 10° below the expected melting temperature.

·        
Continue the
heating and note the temperature at which the column of the
sample collapse definitely against the side of the tube at any point,
when melting may be considered to have begun and note also the temperature
at which the sample becomes liquid throughout as seen by the formation of
a definite meniscus.

·        
Record the
reading of Melting point.

·        
After observation
remove the capillaries/ sample column from oil bath.

·        
After completing
the test switch off the instrument properly.

CLEANING

·        
Clean the
instrument with Clean Cloth

·        
Remove the Silicon Oil
frequently

PRECAUTION:

·        
Keep the
instrument clean and Dust free.

·        
Do not go to the over
limit of temperature as prescribed.

·        
All the knobs are
very delicate and should be handled carefully.

CALIBRATION PROCEDURE

·        
Melting point
apparatus is calibrated by taking the melting points of vanillin, acetanilide, sulphanilamide
and caffeine anhydrous.

·        
Using the
standard calibrated thermometer the temperatures at which the materials
start melting, & at which they completely melt, are read and recorded.

·        
The melting
points of these materials should fall within the given limits; then the
instrument is calibrated.

S. No.

SUBSTANCE

LIMITS

 1.

 Vanillin

 81 to 83
Deg. C

2.

Acetanilide

114 to 116 Deg.
C

3.

Sulphanilamide

164.5 to 
166.5 Deg. C

4.

Anhydrous
Caffeine

234 to 237
Deg.C

FREQUENCY of CALIBRATION: 

·        
Quarterly or
after any maintenance work.

·        
Record the data
on APPENDIX I Format                

Calibration of EC & TDS analyzer

PROCEDURE

PRE- OPERATION INSTRUCTIONS

·        
Ensure various containers (Polythene and / or glassware
etc.,) are free from alkalinity, especially when they are steam cleaned. 

·        
Conductivity cells selected is appropriate for
the conductance range of interest having its platinum chloride black coating in
good condition. Soaked in distilled water for a least 2 hours before use.
Thoroughly clean.

NOTE:

§  A new
conductivity cell should be soaked in distilled water for at least 24 hours.

§  Thermo-probe
and / or thermometers are / is in good working condition or handy.

§  Keep the
samples to be tested handy in clean proper containers.

·        
Ensure the containers used for the solutions /
samples are ABSOLUTELY CLEAN AND DRY.The quantity of solutions / samples are
sufficient enough to ensure immersion of sensor part of the cells /
probes. 

·        
Ensure 90-260 V AC, 50-60 Hz, single-phase
powder is available in 3 contact 5A AC powder outlet. Cartridge fuse of 3A
F-250 V AC is in its place in the instrument.

Calibration of pH Meter

PROCEDURE

Calibration of an indicator
reference electrode combination is undertaken by the standardization of
the pH meter with different buffer solutions and measuring the response by a
immersing the combination electrode in different pH buffer solutions.

STANDARD REQUIRED FOR
CALIBRATION / VERIFICATION
:

·        
Prepare the buffer solutions of pH
4.0, 7.0 and 9.2 respectively using standard buffer capsules E.
Merck make.

·        
Switch on the instrument and allow
it to stabilize for 20 to 30 minutes.

·        
Under the stipulated operating
conditions measure the pH of the different buffer solutions allowing the
reading to stabilizes before reading the pH.

·        
Adjust the pH meter according to
the pH observed.

Calibration of MIcropipettes

REQUIREMENTS

·        
Purified Water

·        
Calibrated digital Balance

·        
Micropipette with tips

·        
Micropipette Manual

PROCEDURE

Performance
Verification of a Micropipette

·        
Micropipette performance is
evaluated using Gravimetric method. In Gravimetric Method desired volume
of purified water is placed or dispensed from micropipette & checked in
term of weight on calibrated digital balance. The desired volume is termed as
Nominal volume.

·        
Calculation is performed to
convert the weight of water to volume or vice –versa.

·        
If the volume measuring device is
properly operated, then the nominal volume will be identical to the volume as
calculated from the weight of the water.

Calibration Procedure

·        
Label 5 centrifuge tubes, Weigh
each of
them.                                                         

·        
Use Micropipette of any size
whichever is required for calibration, pipette the lower volume of that
micropipette & aspired into the centrifuge tubes with the single tip.

·        
Weigh each of the five tubes. By
subtraction, determine the weight of water in each tube.

·        
A calculation is performed to
convert the weight of water to volume or vice –versa.

          W

V    = 
 ————–

          D

            Where       W
= weight in mg

                              V
=   Nominal Volume in micro litre

                              D   =   density
of purified water in mg/micro litre

·        
Calculate the standard deviation
(repeatability) of micropipette based on these five weights.

·        
Determine the closeness of average
weight of purified water is to the expected Weight. Example If we are
using 5µL micropipette, the average weight to be 5 mg.

·        
Determine the accuracy of the
micropipette by percentage error.

·        
Repeat all the above steps with
two mid ranges of the micropipette.

·        
Repeat all the above steps with
single upper range of the micropipette

CLASS RESULTS: MICROPIPETTORS

Sr.No

Volume

Precision
(expressed as Standard deviation)

Accuracy
(relative per cent)

Measured Accuracy as per Instrument
Specification

Measured CV   %

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 Warning & Precautions    
                     
                     
                     
                     
                     

·        
Never drop a
micropipette.                                                                                            

·        
Never rotate the volume adjuster of and the
micropipette beyond the upper or lower range of the Instrument. 
                     
                     
                     
                     
                     
  

·        
Never pass a micropipette through a
flame.   

·        
Never use a micropipette without a tip thus allowing
liquid to contaminate the shaft assembly.

·        
Never lay a filled micropipette on its side thus
allowing liquid to contaminate the shaft assembly.

·        
Never immerse the barrel of a micropipette in liquid,
only immerse the tip.

·        
Never allow the plunger to snap up when liquid is
being aspirated.

·        
Store micropipette set to their highest volume. This
releases pressure on the spring inside the micropipette

Calibration
of Weight Box

PROCEDURE

VISUAL EXAMINATION

·        
Check and clean the weights under
calibration for the quality and finish of surfaces and edges, freedom from
foreign /extraneous matter and the closeness of the screw knob. 

·        
Ensure that no weight is magnetic
in nature.

·        
Fractional weights should not be
punctured and should be identifiable.

ANALYTICAL BALANCE:

·        
Ensure that the analytical balance
used for weighing should be levelled at zero levelling and is supported on a
vibration free rigid table base.

·        
Ensure that a consistent ambient
or fixed room temperature is maintained.

·        
There should be no movement of
air, like running of ceiling fan etc.

·        
Ensure cleanliness of the
analytical balance.

CALIBRATION

·        
Weigh one by one the standard set
of weights and the weights under calibration.

·        
The standard set of weights must
have traceability to a national accredited body.

·        
Repeat the weighing of each set of
weights at least two or three times to ensure greater accuracy and
reproducibility.

·        
Ensure the precision and accuracy
of weighing. 

REPORTING

Assign
the actual mass value for each weight under calibration with respect to the
weight of the reference set of weights.

Calibration of Single Pan Balance

 

PRECAUTION BEFORE CALIBRATION:

·        
Ensure
that the analytical balance used for weighing should be levelled at zero
levelling and is supported on a vibration free rigid table base.

·        
Ensure
that a consistent ambient or fixed room temperature is maintained. There
should be no movement of air or air from ceiling fan etc.

·        
Ensure
cleanliness of the analytical balance.

APPARATUS REQUIRED FOR CALIBRATION:

·        
Calibrated
weight box traceable to either NPL or NABL accredited laboratory.

·        
Ensure
that the calibrated set of weights is clean and free from any dust particles
and any type of deformity.

CALIBRATION PROCEDURE:

·        
Weigh
one by one the standard calibrated set of weights on the balance and record the
weights as indicated on the balance.

·        
Repeat
the weighing of each set of weights at least two to three times to ensure
greater accuracy and reproducibility.

·        
Ensure
the precision and accuracy of weighing. 

REPORTING

·        
From
the assigned weights of the standard set of weights correlate the accuracy of
the weights observed on the balance under calibration.

·        
Make
a table for the observed weight on balance of the standard weights, and the
actual calibrated weight of the set of weights.

·        
The
average of the weights shall be reported same as digits displayed on the
balance screen.

ACCEPTANCE CRITERIA

The balance (to be calibrated)
should display the weights in the tolerance limits as given in the table below
against the standard weights used:

S.No.

Denomination Metric

Tolerance Limits(g)*

1

100
g

0.0005

2

50
g

0.0003

3

20
g

0.0025

4

10
g

0.0002

5

5
g

0.0015

6

2
g

0.0012

7

1
g

0.0001

8

500
mg

0.00008

9

200
mg

0.00006

10

100
mg

0.00005

11

50
mg

0.00004

12

20mg

0.00003

13

10
mg

0.00025

14

5
mg

0.00002

15

2
mg

0.00002

16

1
mg

0.00002

 With reference to
OIML – International Recommendation R111.

Calibration
of High Performance Liquid Chromatography
(Quaternary & Binary)

PROCEDURE

1 MATERIAL REQUIRED

·        
Instrument under calibration (Quaternary & Binary
HPLC)   

·        
Water (HPLC Grade)

·        
Acetone (HPLC Grade)

·        
Caffeine (Working Standard)

·        
Calibrated Digital Thermometer

·        
Naphthalene

·        
Calibrated Glassware

·        
Weighing Balance

2 CALIBRATION

2.1. CALIBRATION
OF PUMP

2.1.1 PRESSURE
PULSATIONS TEST

·        
The Pressure Pulsations Test is to
verify that the output flow fluctuations are within specification (2% peak to
peak of the running pressure).

·        
Connect the pump output to a
restrictor coil. Set the pump flow rate to 1 ml/min. Pressure will be 900-1200
psi of backpressure.

·        
Start the pump and let the
pressure stabilize.

·        
Observe the minimum and maximum
pressure during the 1 min interval.

·        
Determine the minimum and maximum
pressures observed as well as the delta, or difference between observed values.
Determine the mid-point pressure (MP) between the maximum and the minimum, this
is the mid-point pressure used to compute 2 % peak-to-peak error band.

Acceptance criteria: Difference
between the minimum and the maximum pressure should be less than 2% of the mid-point
pressure. 

2.1.1 FLOW
ACCURACY TEST

This test is used to prove that
the flow rate selected on the pump is delivered in the correct time.

2.2 CHECKING FLOW AT 1 ML/MIN (LOW FLOW RATE)

·        
Connect the backpressure regulator
to the output of the pump. The backpressure as read on the pump will be between
900-1200 psi.

·        
Zero the scale of the weighing
balance. Place a 50 ml beaker on a scale and note the reading of the empty
beaker

(W1).

·        
Start the pump and fill the lines
with water.

·        
Set the pump flow at 1 ml/min.

·        
When pressure stabilizes, direct
the flow of water into the beaker and simultaneously start the stopwatch.

·        
Let the pump run for 5
min.    

·        
Remove the beaker after 5 min and
weigh the beaker containing water (W2).

·        
Subtract the final weight with the
initial weight of the beaker (W2-W1) & divide the weight of the water
by 5 & with the water density to get the volume delivered per minute.

2.3 CHECKING FLOW AT
3 ML/MIN (HIGH FLOW RATE) 

·        
Connect the backpressure regulator
to the output of the pump. The backpressure as read on the pump will be
between 900-1200 psi.

·        
Zero the scale of the weighing
balance. Place a 50 ml beaker on a scale and note the reading of the empty
beaker (W1).

·        
Start the pump and fill the lines
with water.

·        
Set the pump flow at 3 ml/min.

·        
When pressure stabilizes, direct
the flow of water into the beaker and simultaneously start the stopwatch.

·        
Let the pump run for 5 min.

·        
Remove the beaker after 5 min and
weigh the beaker containing water (W2).

·        
Subtract the final weight with the
initial weight of the beaker (W2-W1) & divide the weight of the water by 15
& with the water density to get the volume delivered per minute.

Acceptance Criteria: The volume of
water should be ±0.5 ml

3 COMPOSITION
ACCURACY TEST

The Composition Accuracy Test
proves that the proportioning valves are operating correctly. This is
accomplished by varying the amounts of solvent entering the pump through the
proportioning valves. Using acetone solution, which has absorbance at 265 nm as
one solvent, and water, which does not show any absorbance at 265 nm.

·        
Fill reservoirs A and C with
water.

·        
In a 500 ml graduated cylinder,
accurately pipette 1.0 ml of acetone into 250 ml of HPLC- grade water. Dilute to
500ml with HPLC grade water. This is the 0.2% Acetone Solution.

·        
Place acetone solution into
reservoir B and D.

·        
Degas all mobile phases for 5 min.

·        
Purge all reservoirs with 30 ml of
solvent.

·        
Bypass the column.

·        
Set the detector wavelength to 265
nm.

·        
Pump the acetone solution for 30
min through channel B and 30 min through channel D if applicable. Check the absorbance
level is 0.5-0.7 AU.   

·        
Ensure the pump display reads at
least 1000 200 psi of backpressure at 5 ml/min.

·        
Generate and run the method in
table 1 and 2, zeroing the detector at the end of step zero.

Table 1: For Binary and
quaternary (Reservoirs A and B)

STEP

 TIME            

FLOW

%
A              

%
B              

CURV 

AU

0

5.0

5.0

100

0

0.0

 

1

1.0

5.0

100

0

0.0

 

2

3.0

5.0

0

100

0.0

AU1=

3

3.0

5.0

10

90

0.0

AU2=

4

3.0

5.0

49

51

0.0

AU3=

5

3.0

5.0

51

49

0.0

AU4=

6

3.0

5.0

90

10

0.0

AU5=

 

Table 2: For Quaternary
only (Reservoirs C and D)

STEP

TIME

FLOW

%A

%B

CURV

AU

0

5.0

5.0

100

0

0.0

 

1

1.0

5.0

100

0

0.0

 

2

3.0

5.0

0

100

0.0

AU6=

3

3.0

5.0

10

90

0.0

AU7=

4

3.0

5.0

49

51

0.0

AU8=

5

3.0

5.0

51

49

0.0

AU9=

6

3.0

5.0

90

10

0.0

AU10=

 

Compute the actual compositions (C).

% B in A (Binary and Quaternary)

% D in C (Quaternary only)

C (90) = AU2/AU1  100%

C (90) = AU2/AU6  100%

C (51) = AU2/AU1  100%

C (51) = AU2/AU6  100%

C (49) = AU2/AU1  100%

C (49) = AU2/AU6  100%

C (10) = AU2/AU1  100%

C (10) = AU2/AU6  100%

Acceptance Criteria: The actual
composition value should be within 1.0 % of the target %. 

4CALIBRATION
OF DETECTOR

4.1 CAFFEINE
WAVELENGTH VERIFICATION

·        
Dissolve 10 mg of caffeine with
1000 ml of water. Dissolve and mix completely.

·        
Place the 1-liter of dilute
caffeine mixture on an available reservoir of the pump.

·        
In another reservoir place a
bottle of HPLC water. Using the HPLC water, purge the pump lines using
prime purge valve. Connect outlet of pump directly to inlet of the UV/VIS
flow cell.

·        
Set the UV/VIS Detector wavelength
to 300 nm.

·        
Start the pump flow rate at
1ml/min with water and pump for 5 minutes.

·        
Auto zero the detector with water
in the flow cell.

·        
Set the wavelength to 205 nm.

·        
Start pump at 1 ml/min with dilute
caffeine solution. After 5 minutes check that absorbance reading is
between 0.9 and 1.3 AU.

·        
Record the observed wavelength
maximum. Record this value in the instrument performance.

CHANNEL PARAMETERS

Run
Time                  :   23.00min

Sampling
Rate          :    2.0000
pts/s

DETECTOR PARAMETERS

Step

Time

Wavelength

Auto zero  

Step

Time 

Wavelength

Auto zero

1

1.0

300

NO

11

1.0

269

NO

2

4.0

200

NO

12

1.0

270

NO

3

1.0

201

NO

13

1.0

271

NO

4

1.0

202

NO

14

1.0

272

NO

5

1.0

203

NO

15

1.0

273

NO

6

1.0

204

NO

16

1.0

274

NO

7

1.0

205

NO

17

1.0

275

NO

8

1.0

206

NO

18

1.0

276

NO

9

1.0

207

NO

19

1.0

277

NO

10

1.0

208

NO

20

1.0

278

NO

PUMP PARAMETERS

STEP

TIME

FLOW

A

B

C

D

CURVE

0

1.0

3.00

100.0

0.0

0.0

100.0

0.0

1

23.0

3.00

0.0

0.0

0.0

100.0

0.0

Acceptance Criteria: The
wavelength maximum at 205 nm and/or 273 nm should be in the range of 1
nm.

5.  CALIBRATION OF
COLUMN OVEN

5.1 TEMPERATURE
ACCURACY

·        
Measure oven temperature accuracy
at three points in the normal operating range of 5°C above ambient to
100°C.

·        
Remove the oven door cover.
Carefully place the probe of the thermometer in the column holder bracket so
the tip of the probe is in the air stream of the oven.

·        
Ensure the oven is installed
properly and power switched on at the Column Oven. Select a Set
temperature of 20C for the pelteir oven or 30C via the oven’s keypad for
the heat only oven.

·        
Monitor the reported oven
temperature (on the oven’s display) vs. the temperature record on the thermometer.

·        
Allow the system to reach the set
point and to stabilize for 15- 30 minutes.

·        
Record the temperature
displayed on the digital thermometer.

·        
Repeat step (c) for a Set
Temperature of 40C for pelteir oven or 50C for the Heat only oven.

·        
Perform the Temperature Stability
Test.

·        
Repeat step(c) for a Set
temperature of 60C for the pelteir oven or 70C for the Heat only oven.

·        
Acceptance Criteria: The value
should be within ±1°C.

5.2 TEMPERATURE
STABILITY

·        
Measure oven stability at 40°C.

·        
With the set temperature set to
40C and stabilized, take six further readings of the Digital Thermometer at
fifteen-minute intervals. Record the temperature range. The range of the
results should be less 1C.          

·        
Acceptance Criteria: The
value should be Max (reading) – Min (reading) <1°C.

6 CALIBRATION OF
AUTOSAMPLER:
The initial system set up conditions:

Wavelength

260 nm

Flow Rate

1.0 ml/min

Mobile Phase

Methanol: Water (80:20
v/v)

Flush Solvent

80% Methanol/Water

Column

C18

Dilution Medium

Mobile Phase

6.1 REPEATABILITY
(PRECISION)

·        
Repeatability will look at the
standard deviation of the area of the Naphthalene peak, the next to last peak
in the chromatogram to check Repeatability.

·        
The Naphthalene peak from each
data file will be averaged and the standard deviation determined.

·        
From the standard deviation the
RSD is calculated.

·        
The purpose of Repeatability is to
verify that the Autosampler can repeatedly inject the same volume of sample
numerous times.

·        
Inject 6 injections of 10 L
each. This test also checks that the Autosampler can accurately inject sample
volumes from multiple positions on the tray.

ACCEPTANCE CRITERIA:

The standard deviation should be
less than or equal to 0.5 %.

6.1.1 LINEARITY:

·        
Linearity will check that the
Autosampler can accurately inject varied amounts of sample and that the
injected volumes are correct.

·        
We do this by injecting multiple
samples of different injection volumes.

·        
The injection volumes are 4 injections
of 5 L from vial at position 1, 6 injections of 10 L from vial 10,
and finally 4 injections of 20 L from vial 100.

·        
There should be linear
relationship between sample volume and area under the peak.

·        
Acceptance Criteria: The R-squared
value should be of 0.999 or greater

6.1.2 CARRYOVER

·        
Carryover is the amount of
previous sample retained by the system and therefore has a presence in the
current sample.

·        
Looking at
the peak of Naphthalene and at the mobile phase blank
checks carryover.

·        
There should be very little if any
of the Naphthalene in the mobile phase blank at the retention time of the
Naphthalene peak.

·        
The specification for this test is
less than or equal to 0.02%.

           Area
of blank

%
Carryover = ————————- X 100

              Area
of sample

Acceptance Criteria: The % carryover should not exceed
0.02%.

Calibration of HPLC Isocratic

Procedure 

Calibration
of Pump

·        
Switch on the instrument.

·        
Using the standard operating
conditions start the operations of the instrument using water as mobile phase.

·        
Set the flow rate of the pump at a
required set point i.e. 1.5ml/minute or 2ml/minute.

·        
Ensure that there are no air
bubbles in the Teflon tube (waste tubing).

·        
Allow the liquid to flow through
the pump and collect the same in a calibrated measuring cylinder,
simultaneously starting the stop watch to note down the time.

·        
Record the volume collected in 2
minutes.

·        
Take atleast 2 to 3 readings for
better accuracy and reproducibility.

·        
Set the next flow rate and record
the volume collected.

·        
Correlate the readings observed
with the set point and make adjustments, if necessary.

Calibration
of Detector

·        
Attach a C-18 column to the
instrument.

·        
Set the UV Detector at a
wavelength of 254nm.

·        
Use a mobile phase of methyl
cyanide and water, (CH3CN : H2O) in the ratio of 80 : 20.

·        
Adjust the flow rate at a set
point 1ml/minute.

·        
Inject 2ml of the test mixture
containing 0.5mg/ml Naphthalene and 0.5mg/liter Anthracene at ambient
temperature and record the graph.

·        
Also inject 2ml of the individual
solutions mentioned above under the same conditions and record the graphs.

·        
The retention time of Naphthalene
is 4.5minutes and Anthracene has a retention time of 6.9minutes.

·        
Take at least 2 to 3 readings for
better accuracy and reproducibility.

·        
Correlate the readings observed
with the set points and make adjustments, if necessary.

Calibration of Viscometer (oscillatory)

PROCEDURE

·        
Switch
on the instrument.

·        
Using
the standard operating conditions start the operations of the instrument.

·        
Select
the proper spindle number i.e. L1, L2, L3, L4 depending
upon the range of viscosity to be determined and attach to the viscometer.

·        
Ensure
that the spindles are cleaned before and after use.

·        
Select
the proper speed in RPM depending upon the range of viscosity to be determined.

·        
Control
the temperature of the sample at which the viscosity is to be determined.

·        
Immerse
the spindle in the sample container up to the set point mark. The spindle should not
touch the container in any way.

·        
Start
the instrument at the selected speed and record the viscosity in cps units.

·        
Change
the speed of the spindle and record the viscosity in cps units.

·        
The
viscosity should not differ with change in the speed of the spindle.

·        
Correlate
the observed values of viscosity with the stated values of the reference
standard and make adjustments, if necessary.

Calibration
of UV/VIS Spectrophotometer

PROCEDURE

1 MATERIALS
REQUIRED

·        
Instrument to be used

·        
Potassium dichromate

·        
Sulphuric acid

·        
Potassium chloride

·        
Toluene

·        
Hexane

2 CONTROL OF ABSORBANCE

·        
Prepare a solution of potassium
dichromate solution, which has been previously dried to constant weight at 130oC
using 57.0 to 63.0mg in sufficient 0.005M Sulphuric acid to make up to volume
of 1000ml.

·        
Determine the absorbance of the
prepared potassium dichromate at different wavelengths of 235nm, 257nm, 313nm
and 350nm.

Based on the absorbance obtained,
at the particular wavelengths, determine the value of A (1%, 1cm) at each
wavelength. The IP-1996 gives the following tolerances:

S. No.

Wavelength (nm)

A (1%, 1cm)

Maximum Tolerance

1.

235

124.5

122.9 to 126.2

2.

257

144.5

142.8 to 146.2

3.

313

48.6

47.0 to 50.3

4.

350

107.3

105.6 to 109.0

 3 LIMIT OF STRAY LIGHT:

·        
Stray light is detected at 200 nm
with 1.2 % w/v solution of potassium chloride at a path length of 1 cm.

·        
The absorbance of 1.2 % w/v
solution of potassium chloride should be greater than 2.0 when compared
with water as reference liquid.

4 RESOLUTION POWER:

·        
Prepare a solution of 0.02 % v/v
of Toluene in hexane.

·        
Take the absorbance at 269 nm and
266 nm.

·        
The ratio of absorbance at the
maximum (269 nm) to that at the minimum (266 nm) is not less than 1.5.

Calibration
of Ultra Low Temperature Freezer

REQUIREMENTS

·        
Calibrated Digital Temperature
(-80 to 199.9 C)

·        
Instrument Instruction
Manual 

 

PROCEDURE

1 Calibration Procedure       
                     
                     
                     
                  

·        
Set -80 0 C
temperature at which temperature calibration of Ultra –Low
Temperature    freezer has to be done.

·        
Use the set key & press it on
the control panel of the Ultra –Low Temperature to memorise the temperature.

·        
Press the numerical Value Shift
Key & digital shift key to set -80 C temperature.

·        
Wait for the set temperature to
flash on control panel display screen.

·        
Wait for few minutes to maintain
the temperature in Ultra –Low Temperature freezer.

·        
Open the door latch & insert
the PT-100 sensor of calibrated digital thermometer (-80 to 199.9 0  C)

·        
Record the temperature observed on
Ultra-Low Temperature freezer & calibrated digital thermometer.

·        
Record the temperature at set
temperature at different time interval first reading at zero hr, Second reading
at the interval of 2 hrs & third reading after 2 hrs of second reading.

·        
Zero hr will be defined as the
time of starting the calibration of Ultra Low Temperature Freezer.

2Calibration Record

Sr.No

Calibration Time Intervals

Temperature UUC

( 0  C
)

Temperature
Reference

0  C )

Correction to UUC

In (  0  C
) if required

     1

  0 hr

           -80 0  C

 

 

     2

  0-  2 hr

           -80 0  C

 

 

     3

  0- 4  hr

           -80 0  C

 

 

Calibration
of Turbidity Meter

PROCEDURE

·        
Switch on the instrument.

·        
Using the standard operating conditions
start the operations of the instrument.

·        
Set the instrument at zero.

·        
Pour the standard turbidity
solution in the glass tube / cuvette and measure the turbidity.

·        
Ensure that the glass tube /
cuvette is cleaned/washed before and after use.

·        
Make the measurements of turbidity
in each range of the instrument or in different ranges of the Instrument.

·        
The turbidity shall not differ
from the stated values of the reference standards.

·        
Correlate the observed values of
turbidity with the specified/stated values of the reference standard and make
adjustments, if necessary.

 

 

 

Calibration of Refractometer

 PROCEDURE

·        
The Refractometer is calibrated
with different liquids or glass test piece of known refractive index.

·        
Ensure that the surface of the
prisms is cleaned each time either with water or acetone before using for the determination
of the refractive index.

·        
Put in a few drops of the
reference liquid of known refractive index in between the two prisms.

·        
Circulate the water from a
separate thermostatic bath within the prism box using the inlet and outlet
tubes provided.

·        
A thermometer to measure the
temperature is provided along with the instrument. By continuously circulating
the water a fixed desired temperature can be obtained.

·        
Make the achromatic critical line
coincide with the index line with the field of view of the telescope.

·        
Keeping the telescope in the
position take the reading of the scale against the index mark.

·        
Record the observed refractive
index reading from the scale and correlate the same with the reference reading.

·        
Adjustment can be made by setting
the scale reading to correct value of refractive index to that fixed
temperature by operating the pin provided over the cone of the telescope till
the critical line moves and coincides with the cross line.

Calibration of Polarimeter

PROCEDURE

·        
Ensure that the instrument is
clean and free from dust particles.

·        
Ensure that the polarimeter tube
is cleaned each time before and after use.

·        
Switch on the instrument and allow
sufficient time to achieve maximum brightness of the sodium lamp.

·        
Ensure that the settings are done
in the zero-zero position.

·        
Standard Solution Required For
Calibration: Prepare reference of known specific
rotation using Sucrose AR.

CALIBRATION:

·        
Open the polarimeter tube
compartment and place the polarimeter tube, which is filled with the standard
solution of sucrose in such a way so as to avoid air bubbles.

·        
The specific rotation observations
are done by viewing through the telescope when the half dark and half-light
portion of the circle are in equilibrium.

·        
The half dark and half-light is
matched with a knob rotating it in the clockwise and anti-clockwise directions.
It is also defined as dextro (+) or levo (-) rotating respectively.

·        
A blank determination is also performed
and corrections if any are applied.

·        
Record the corrected observations.

CALCULATION:

Specific optical rotation
=       (Corrected observed specific
rotation at a specified temp)

     
                     
            (Length of the polarimeter tube in
dm) x Conc. of Sucrose soln. %w/w)

Correlate the observations with the specified values of the
various concentrations of sucrose solutions.

 

Calibration of Photofluorimeter

PROCEDURE

·        
On the instrument 15 minutes
before starting the calibration.

·        
Make dilutions using fluoroscene
in 0.1 M NaOH (1ppm, 2ppm, 3ppm and 4 ppm).

·        
Use 0.1M NaOH as blank and set the
instrument on “0 “with blank knob.

·        
Set the instrument at ‘20’ with
1ppm solution.

·        
Take the readings at 1, 2, 3 and 4
ppm without touching any of the knobs.

·        
Standard readings for above
mentioned concentrations: 

S. No.

Concentration(ppm)

Observed Intensity(Units)

1

1 ppm

21

2

2 ppm

40

3

3 ppm

60

4

4 ppm

79

Calibration
of Muffle Furnace

PROCEDURE

·        
Switch on the instrument.

·        
Ensure that the temperature
indicator/controller is in working condition i.e. the set temperature should be
maintained / retained properly.

·        
Ensure that the door of laboratory
oven is properly closed and does not loosen during operation.

·        
Ascertain and set the
points/points at which calibration is required.

·        
Switch on the laboratory oven and
allow it to reach the set point temperature.

·        
Apparatus Required For
Calibration: Platinum resistance thermometer with sensor and digital
temperature indicator.

CALIBRATION

·        
Insert the sensor of the
reference/calibrator sensor (PRT) in the laboratory oven as near as possible
to the sensing tip or sensor of the temperature controller/indicator of the
laboratory oven.

·        
When the temperature is stable
note down the temperature reading as indicated by on the laboratory oven and
the reference indicator simultaneously.

·        
Take at least three to four
readings for better accuracy and reproducibility.

·        
Set the next point and record the
temperature readings accordingly.

·        
Correlate the readings observed
with the reference standards and apply corrections,
if   necessary.

Calibration
of Laboratory Oven

PROCEDURE

·        
Switch on the instrument.

·        
Ensure that the temperature
indicator/controller is in working condition i.e. the set temperature should be
maintained / retained properly.

·        
Ensure that the door of laboratory
oven is properly closed and does not loosen during operation.

·        
Ascertain and set the
points/points at which calibration is required.

·        
Switch on the laboratory oven and
allow it to reach the set point temperature.

·        
Apparatus Required For Calibration:
Platinum resistance thermometer with sensor and digital temperature
indicator.

CALIBRATION:

·        
Insert the sensor of the
reference/calibrator sensor (PRT) in the laboratory oven as near as possible to
the sensing tip or sensor of the temperature controller/indicator of the
laboratory oven.

·        
When the temperature is stable
note down the temperature reading as indicated by on the laboratory oven and
the reference indicator simultaneously.

·        
Take at least three to four
readings for better accuracy and reproducibility.

·        
Set the next point and record the
temperature readings accordingly.

·        
Correlate the readings observed
with the reference standards and apply corrections, if necessary.

Calibration
of Karl Fischer Apparatus

PROCEDURE

1 APPARATUS REQUIRED FOR
CALIBRATION:

·        
Dried Beakers, which have been
previously rinsed with acetone. 

·        
Calibrated Analytical Balance

2 DETERMINATION OF VOLUME:

·        
Clean the measurement/dispensing
pipette available with the respective set of the equipment.

·        
Fill the measuring pipette with
water and adjust to the zero point.

·        
Dispense the known volumes of
water at random or from the set point of the graduation marks of the pipette.

·        
Repeat the dispensing of water at
different set points and weigh the beaker again and again.

·        
From the different weights
obtained calculate the volume dispensed and co-relate with the graduation of
the pipette and assign the value.

3 CALCULATION:

                                                            Weight
of the water dispensed (gm)

Volume of the pipette (ml), V=
————————————————–

                                                            Density
of Water at 27oC (gm / ml)

Calibration
of Infrared Spectrophotometer

PROCEDURE

·        
Infra-red spectrophotometer is a
technique used for identifying the organic compounds.

·        
Control the humidity of the
environment within the range of 40 to 50RH at 25oC.

·        
Switch on the instrument and
observe that it initializes at 4018  1cm-1. 

1 Standard Required For
Calibration
:

Standard Polystyrene Film

2 Calibration:

Using the stipulated operating
conditions run in the infra-red spectra of the reference polystyrene film
supplied with the instrument.  

All the significant or principal
peaks in the spectrum obtained should be at the same wavelength as that of the
standard reference graph and should correspond to within 0.5% of the wave
number scale. The relative sizes of the bands should be concordant in the two
spectra.

3 FREQUENCY

At every 3 months

Calibration
of Inductively Coupled Plasma Mass Spectrometer

REQUIREMENTS

·        
Argon Gas Cylinders

·        
Elan 9000 Hardware Guide

·        
Elan Inductively Coupled Plasma
Mass Spectroscopy safety manual

·        
Tuning Solution

PROCEDURE

·        
Precaution while start-up the
Inductivel Coupled Plasma Mass Spectrometer

·        
Argon gas cylinder should not be
used below 100 psi in term of gas quantity

·        
Release gas pressure should be
between 60-80 psi.

·        
Switch on the blower &
recirculater.

·        
Before the ignition of the plasma,
vacuum pressure should be 10-6 torr.

·        
Open Software.     
     

·        
Open “Instrument “———Start
Plasma

·        
Leave the plasma for 30 minutes or
more (minimum 25 minutes).

·        
Open —-“ Workspace”
———-“Daily Performance .wrk”

·        
Place the tuning solution.

·        
Analyse the above solution as
“Sample “.

·        
Do  Daily Performance
Check

§ 
Mg intensity > 40,000 cps/10
ppb

§ 
In Intensity > 250 , 000 cps/10
ppb

§ 
U intensity > 200,000 cps/10
ppb

§ 
CeO/Ce ;Ba++/Ba < 3
%

§ 
Background 220 <2 cps

·        
If above test is not OK then
optimize. Optimization is required only when the value are highly disturbed .Do
optimization while referring to the manual. Following parameters must be
optimized. 

§ 
Nebulizer Gas Flow (NEB)

§ 
Auxiliary Gas Flow

§ 
Plasma Gas Flow 

§ 
Lens Voltage

§ 
Inductively Coupled Plasma Radio-frequency
(Rf) power 

§ 
Analog Stage voltage

§ 
Pulse Stage voltage 

§ 
Quadropole Rod Offset (QRD) 

§ 
Cell Path Voltage (CPV) 

·        
Click on get Analyte list Icon
& select Analyte from Analyte Icon i.e.Mg 

·        
Select optimization criteria &
select maximum intensity.

·        
Click on optimize icon. 

·        
Repeat the step for above
mentioned parameters individually i.e. Nebuliser, Gas Flow, Auxiliary Gas Flow,
and Plasma Gas flow

REFERENCE

·        
Working Instructions (Inductively
Coupled Plasma Mass Spectrometer); Issue date: Jan 2006

Calibration
of Gas Chromatograph (GC)

PROCEDURE:

MATERIALS REQUIRED:

·        
Instrument to be used

·        
Hydrogen Cylinder

·        
Air Cylinder

·        
Calibrated Temperature Probe

·        
Ethanol

·        
Glassware

FLOW VERIFICATION:

·        
Using the standard operating
conditions start the carrier gas flow through at a set point.

·        
Allow the gas to pass through the
gas flow meter supplied with the instrument and record the gas flow rate.

·        
Similarly allow the hydrogen gas
and air gas to pass through at a set point flow rate of 30ml/minute and
300ml/minute respectively. The gases are allowed to pass through the gas flow
meter and the flow rates recorded.

·        
Correlate the observed gas flow
rates with the set point flow rates and make adjustments, if necessary. 

TEMPERATURE VERIFICATION:

Insert a calibrated temperature
probe into the oven near the oven sensor. Allow all zones to stabilize and to
become ready. Record the actual temperature probe value for oven temperature.

Detector/Auto sampler
Reproducibility

1 Repeatability
(Precision)

Repeatability will look at the standard deviation of 6 injections
of the same concentration of the 0.5% solution of ethanol. From the standard
deviation the RSD is calculated. If the standard deviation is less than or
equal to 2.0 % the test passes. The purpose of Repeatability is to verify that
the Auto sampler can repeatedly inject the same volume of sample numerous
times. Inject 6 injections of 1L each.

2 LINEARITY

Linearity will check that the Auto
sampler can accurately inject different concentration of ethanol. We do this by
injecting multiple samples of different concentrations. The injection volume is
1L from 0.40%, 0.45%, 0.50%, 0.55% and 0.6 % w/v solution of ethanol. There
should be linear be a linear relationship between sample concentration and area
under the peak.

Calibration
of BOD Incubator

PROCEDURE

·        
Switch on the instrument.

·        
Ensure that the temperature
indicator / controller is in working condition i.e. the set temperature should
be maintained/retained properly.

·        
Ensure that the door of incubator
is properly closed and does not loosen during operation.

·        
Ascertain and set the
points/points at which calibration is required.

·        
Switch on the incubator and allow
it to reach the set point temperature.

·        
Apparatus Required For
Calibration: Platinum resistance thermometer with sensor and digital
temperature indicator.

CALIBRATION:

·        
Insert the sensor of the
reference/calibrator sensor (PRT) in the BOD incubator as near as possible to
the sensing tip or sensor of the temperature controller/indicator of the BOD
incubator.

·        
When the temperature is stable
note down the temperature reading as indicated by the BOD incubator and the
reference indicator simultaneously.

·        
Set the next point and record the
temperature readings accordingly.

·        
Take at least three to four readings
for better accuracy and reproducibility.

·        
Correlate the readings observed
with the reference standards and apply corrections, if necessary. 

Calibration
of Humidity Chamber

PROCEDURE

·        
Switch on the instrument.

·        
Ensure that the temperature
indicator/controller is in working condition i.e. the set temperature
should be maintained/retained properly.

·        
Ensure that the door of incubator
is properly closed and does not loosen during operation.

·        
Ascertain and set the
points/points at which calibration is required.

·        
Switch on the humidity chamber and
allow it to reach the set point temperature.

·        
Apparatus Required: Reference
thermometer with sensor and digital temperature indicator.

CALIBRATION:

·        
Insert the sensor of the
reference/calibrator sensor (PRT) in the humidity chamber as near as possible
to the sensing tip or sensor of the temperature controller/indicator of
the humidity chamber.

·        
When the temperature is stable
note down the temperature reading as indicated by on the humidity chamber and
the reference indicator simultaneously.

·        
Take at least three to four
readings for better accuracy and reproducibility.

·        
Set the next point and record the
temperature readings accordingly.

·        
Correlate the readings observed
with the reference standards and apply corrections, if necessary.                                                      
                     
                   
      

Calibration of Burette

Materials and Equipment Used:

·        
A weighing
balance that is calibrated with traceable weights and regularly maintained.

·        
A calibrated
thermometer.

·        
A weighing
vessel.

·        
Distilled water.

·        
Acetone.

·        
4% Acetic acid
solution

·        
Soap solution

·        
Burette
stand 

·        
Worksheets or
forms to record the raw data.

Procedure: 

The determination of capacity of
burette is based on weighing water and calculating mass and volume based on the
density at the appropriate temperature.

·        
Clean the burette
thoroughly with soap solution and water to remove any adhering dirt or grease.
Rinse the volumetric flask with acetic acid solution and then wash with water till
free from all acid.

·        
Dry the burette
by rinsing with acetone and clamp it vertically on a burette stand.

·        
Fill the burette
with distilled water at ambient temperature sufficiently above the zero
mark. 

·        
Place a glass
vessel below the jet of the burette and open the stopcock fully.

·        
Stop the outflow
when the meniscus is slightly above the zero graduation line, ensuring that no
air bubble is entrapped within the burette.     

·        
Open the stopcock
slowly and set the meniscus at the zero graduation line. Remove any drop
adhering to the jet by bringing a glass vessel into contact with the jet.

·        
Setting of the
meniscus at any graduation line shall be performed as follows – 

§ 
Set meniscus so
that the plane of the top edge of the graduation line is horizontally tangential
to the lowest point of the meniscus, the line of sight being in the same plane.

§ 
In order to
minimize possible errors the same method of setting shall be used for both zero
and other readings.

·        
Take a tared
weighing vessel (W1), place it below the jet of the burette and open the
stopcock fully to allow free descent of water meniscus, taking care that jet
does not come into contact with the wall of the receiving vessel during the
delivery period.

·        
Stop the outflow
when the meniscus reaches a few ml above the graduation line up to which
accuracy is to be determined. Allow a ‘waiting time’ of 30 seconds and then
make the final setting of meniscus by controlling the outflow and gradually
stopping it when the meniscus touches the graduation line.

·        
After the setting
of meniscus at a particular graduation line is done, add any drop, adhering to
the jet, to the delivered volume by bringing the inside of the receiving vessel
into contact with the jet and weigh the vesel (W2).

·        
Note the
temperature of water and calculate the volume of water delivered by the burette
at the reference temperature of 270C.

 

Calculation: Calculate the volume of water delivered by the
burette using the following formula – 

V= W2 – W1 / D

Where:

V    = Volume of the
burette, Cm3 (ml)

W1 = Weight of empty weighing vessel.

W2 = Weight of empty weighing vessel
& distilled water.

D    = Density of water at
270C (i.e. 0.9965 g /cc)

 

Reporting: Record all the observations in the worksheets
or forms. Calculate the correct volume of water delivered by the burette and,
the amount by which this differs from the indicated volume, is the volumetric
error.                   
   

Calibration
of Measuring Cylinder

Materials and Equipment Used:

·        
A weighing
balance that is calibrated with traceable weights and regularly maintained.

·        
A calibrated
thermometer.

·        
A weighing
vessel.

·        
Distilled water.

·        
Acetone.

·        
4% Acetic acid
solution

·        
Worksheets or
forms to record the raw data. 

Procedure: 

The determination of capacity of
volumetric flask is based on weighing water and calculating mass and volume
based on the density at the appropriate temperature.

Clean the
measuring cylinder with soap solution and water to remove any adhering dirt or
grease. Rinse the measuring cylinder with acetic acid solution and then wash
with water till free from all acid.

·        
Dry the measuring
cylinder by rinsing with acetone.

·        
Weigh a cleaned
and dried measuring cylinder (W1).

·        
Fill the
measuring cylinder with distilled water at ambient temperature to a few ml
below the maximum graduation line.

·        
Then hold the
volumetric flask in a vertical position, and setting of the meniscus at any
graduation line shall be performed as follows – 

§ 
Add more
distilled water gradually to set the meniscus to the graduation line so that
the plane of the top edge of the graduation line is horizontally tangential to
the lowest point of the meniscus, the line of sight being in the same plane.

§ 
In order to
minimize possible errors the same method of setting shall be used for maximum
mark or other readings.     

·        
Again weigh the
measuring cylinder containing distilled water (W2).

·        
Note the
temperature of water and calculate the volume of water in the measuring
cylinder at the reference temperature of 270C.

 

Calculation: Calculate the volume of water filled in
measuring cylinder by using the following formula – 

V= W2 – W1 / D

Where:

V   = Volume of the
measuring cylinder, Cm3 (ml)

W1 = Weight of empty measuring
cylinder.

W2 = Weight of empty measuring
cylinder & distilled water.

 D = Density of water at
270C (i.e. 0.9965 g /cc)

 

Reporting: Record all the observations in the
worksheets or forms. Calculate the correct volume of water contained in
measuring cylinder and, the amount by which this differs from the indicated
volume, is the volumetric error. 

Calibration
of Volumetric Flask
 

Materials and Equipment Used:

·        
A weighing
balance that is calibrated with traceable weights and regularly maintained.

·        
A calibrated
thermometer.

·        
A weighing
vessel.

·        
Distilled water.

·        
Acetone.

·        
4% Acetic acid
solution

·        
Worksheets or
forms to record the raw data.

Procedure: 

 

The determination of capacity of
volumetric flask is based on weighing water and calculating mass and volume
based on the density at the appropriate temperature.

 

·        
Clean the
volumetric flask with soap solution and water to remove any adhering dirt or
grease. Rinse the volumetric flask with acetic acid solution and then wash with
water till free from all acid.

·        
Dry the
volumetric flask by rinsing with acetone.

·        
Weigh a cleaned
and dried volumetric flask (W1).

·        
Fill the
volumetric flask with distilled water at ambient temperature to a few ml below
the graduation line.

·        
Then hold the
volumetric flask in a vertical position, and by adding more water, set the
meniscus to the graduation line so that the plane of the top edge of the
graduation line is horizontally tangential to the lowest point of the meniscus,
the line of sight being in the same plane.

·        
Again weigh the
volumetric flask containing distilled water (W2).

·        
Note the
temperature of water and calculate the volume of water in the volumetric flask
at the reference temperature of 270C.

Calculation: Calculate the volume of water filled in
volumetric flask by using the following formula –

V= W2 – W1 / D

Where:

V = Volume of the volumetric flask,
Cm3 (ml)

W1 = Weight of empty volumetric flask.

W2 = Weight of empty volumetric flask
& distilled water.

D = Density of water at 270C (i.e.
0.9965 g /cc)

 

Reporting: Record all the observations in the worksheets
or forms. Calculate the correct volume of water contained in volumetric flask
and, the amount by which this differs from the indicated volume, is the
volumetric error.

Calibration
of Pipette, Graduated

Materials and Equipment Used:

 

·        
A weighing
balance that is calibrated with traceable weights and regularly maintained.

·        
A calibrated
thermometer.

·        
A weighing
vessel.

·        
Distilled water.

·        
Acetone.

·        
4% Acetic acid
solution

·        
Worksheets or
forms to record the raw data.

Procedure: 

The determination of capacity of
pipettes is based on weighing water and calculating mass and volume based on
the density at the appropriate temperature.

·       Clean the pipette
with soap solution and water to remove any adhering dirt or grease. Rinse the
pipette with acetic acid solution and then wash with water till free from all
acid.

·        
Dry the pipette
by rinsing with acetone.

·        
Weigh a cleaned
and dried weighing vessel (W1).

·        
Fill the pipette
with distilled water at ambient temperature sufficiently above the 
 zero mark.

·        
Then hold the
pipette in a vertical position, and adjust the falling meniscus at any
graduation line shall be performed as follows –

§ 
Set the meniscus
so that the plane of the top edge of the graduation line is horizontally
tangential to the lowest point of the meniscus, the line of sight being in the
same plane.

§ 
Remove any drop
adhering to the jet of the pipette by bringing the surface of a glass vessel
into contact with the tip of the jet.

·        
Deliver the water
thus measured into cleaned tared vessel slightly inclined so that the tip of
the jet   is in contact with the inside of the vessel, but without
movement of the one against the other throughout the delivery and waiting
periods.

·        
Stop the delivery
of distilled water when the meniscus reaches a few ml above the graduation line
up to which accuracy is to be determined. Make the final setting of meniscus by
controlling the delivery and gradually stopping it when the meniscus touches
the graduation line.

·        
After the setting
of meniscus at a particular graduation line is done, add any drop, adhering to
the jet, to the delivered volume by bringing the inside of the receiving vessel
into contact with the jet. 

·        
Again weigh the
vessel containing distilled water (W2).

·        
Note the
temperature of water and calculate the volume of water delivered by the pipette
at the reference temperature of 270C.

Calculation: Calculate the volume of water delivered by
pipette using the following formula

V= W2 – W1 / D

Where:

V    = Volume of the
pipette, Cm3 (ml)

W1 = Weight of empty weighing vessel.

W2 = Weight of empty weighing vessel
& distilled water.

D   = Density of water at
270C (i.e. 0.9965 g /cc)

Reporting: Record all the observations in the worksheets
or forms. Calculate the correct volume of water delivered by the pipette and,
the amount by which this differs from the indicated volume, is the volumetric
error. 

Calibration
of Pipette, Single Mark

Materials and Equipment Used:

·        
A weighing
balance that is calibrated with traceable weights and regularly maintained.

·        
A calibrated
thermometer.

·        
A weighing
vessel.

·        
Distilled water.

·        
Acetone.

·        
4% Acetic acid
solution

·        
Worksheets or
forms to record the raw data.

 

Procedure: 

The determination of capacity of
pipettes is based on weighing water and calculating mass and volume based on
the density at the appropriate temperature.

Clean the pipette
with soap solution and water to remove any adhering dirt or grease. Rinse the
pipette with acetic acid solution and then wash with water till free from all
acid.

·        Dry the pipette
by rinsing with acetone.

·        
Weigh a cleaned
and dried weighing vessel (W1).

·        
Fill the pipette
with distilled water at ambient temperature to a few ml above the graduation
line.

·        
Then hold the
pipette in a vertical position, and adjust the falling meniscus to the graduation
line as given below –

§ 
Set the meniscus
so that the plane of the top edge of the graduation line is horizontally
tangential to the lowest point of the meniscus, the line of sight being in the
same plane.

§ 
Remove any drop
adhering to the jet of the pipette by bringing the surface of a glass vessel
into contact with the tip of the jet.

·        
Deliver the water
thus measured into cleaned tared vessel slightly inclined so that the tip of
the jet   is in contact with the inside of the vessel, but without
movement of the one against the other throughout the delivery and waiting
periods.

·        
Allow the pipette
to empty till the meniscus comes to rest in the jet.

·        
Again weigh the
vessel containing distilled water (W2).

·        
Note the
temperature of water and calculate the volume of water delivered by the pipette
at the reference temperature of 270C.

 

Calculation: Calculate the volume of water delivered by
pipette using the following formula – 

V= W2 – W1 / D

Where:

V    = Volume of the
pipette, Cm3 (ml)

W1 = Weight of empty weighing vessel.

W2 = Weight of empty weighing vessel
& distilled water.

D    = Density of water at
270C (i.e. 0.9965 g /cc)

Reporting: Record all the observations in the worksheets
or forms. Calculate the correct volume of water delivered by the pipette and,
the amount by which this differs from the indicated volume, is the volumetric
error. 

Operation of Microscope

PROCEDURE

OPERATION

Keep the Microscope
in Position.

·        
Switch on the main
switch of microscope so that bulb will glow.

·        
Adjust the low power
objective, by rotating the revolving nosepiece in position.

·        
Adjust the focus of
light with the help of mirror and condenser.

·        
Now place the
objective (high power or oil immersion whichever is required, use, place one drop of immersion oil on the slide before
placing it on the stage and then lower the O.I objective just to touch the oil
drop.

·        
Make the field
clearer with the help of fine adjustment and then observe the slide.

·        
Slide can be moved
in left, right, upper and lower direction to observe the complete smear area
with the help of slide stage screw and coarse adjustment.

·        
After completion of
observation, remove the slides, clean the objective lens and condenser with
cotton soaked in xylene. Lower down the condenser and place low power objective
in position.

CLEANING

Clean the apparatus externally with clean Tissue Paper.

Clean the unit surface with wiping cloth supplied with the
unit before and after use.

Clean the
all-using slide with distilled water.

PRECAUTION

Keep the instrument
clean and dust free.

·        
Do not allow the oil
to harden on the lens.

·        
Calibrate the lens
twice in year.

 

Calibration of Antibiotic Zone Reader

PROCEDURE

·  
Ensure that the instrument is clean and free
from any dust particles.

·  
Switch on the instrument. 

·  
Circle of zone reader is set on the edge of
bored medium coincidence.

·  
Place a reference circular glass microscopic
slide and measure the accurately the zone reading.

·  
Correlate the observed scale readings with
reference microscopic slide.

·  
Apply corrections, if necessary.

Operation of Digital Colony Counter

PROCEDURE

OPERATION

·        
Ensure that
instrument is clean and free from dust.

·        
Ensure that the
magnifying lens is fixed on its appropriate plate.

·        
Ensure that the
market is connected to either right or left side of the Instrument.

·        
Switch on the mains.

·        
Switch on the power
button. The internal light will be flashed ‘ON’ and the Digital screen will
show ‘0000’ figure. Again pressing this button will turn The Power ‘OFF’.

·        
Now press the plate
containing colonies to be counted.

·        
Now, Mark each and
every colony one after the other by pressing the tip of the marker.

·        
The number of
colonies will automatically get displayed on the screen can be set to 0000 by
pressing the RESET button.

·        
When the counting is
over, the screen can be set to 0000 by pressing the ‘manual’ Button.

·        
The counting can be
done manually by pressing the MANUAL button.

·        
Section In charge
witch off the instrument and put under cover when not required.

 

CLEANING

·        
Clean the apparatus externally with clean Tissue Paper

·        
Clean the unit surface with wiping cloth supplied with the unit before
and after use.

 

PRECAUTION

·        
Keep the instrument
Clean.

·        
Calibrate the
instrument at defined time interval.

Operation of Incubator

PROCEDURE

OPERATION

·        
Switch on the main
power supply.

·        
Switch on the
instrument switch.

·        
While heater is on
the green light will be on.

·        
When the incubator
attains temperature green light turns off.

·        
Read the temperature
inside by the calibrated thermometer and record.

·        
Open the door and
keep the objects inside.

·        
After the required
time of incubation put off the instrument switch.

·        
Put off the main
power supply.

CLEANING

·        
Clean the apparatus externally with clean Tissue Paper                 

·        
Lean the unit surface with wiping cloth supplied with the unit before and
after use.

 

PRECAUTION

·        
Keep the Instrument
Dust free and clean

·        
Check the
Temperature three or four time in a day.

·        
Do the surface
sterilization with 70 % IPA once in a week.

 

 

OPERATION OF MAGNETIC STIRRER HOT PLATE

PROCEDURE

OPERATION

·        
Switch “ON” the
machine after connection it with power supply.

·        
Set the temperature
setting by Knob.

·        
Kept the flask or
beaker to be heated on it.

·        
Switch “off” the
power supply when the operation is completed.

CLEANING

·        
Clean the instrument
with Clean Cloth.

Precaution:

·        
Keep the instrument
clean and Dust free.

·        
Never do the Over
Heat to the beaker etc. kept on Hot Plate.

·        
Never use necked
hand to pick up heated objects.

OPERATION OF VACCUME PUMP

PROCEDURE

OPERATION

·        
Switch on main power supply.

·        
Switch on the power switch of instrument.

·        
Attach the pipe between the pump and
filtration assembly.

·        
After completion of filtration detached the
filtration assembly and switch off the power switch of the instrument.

CLEANING

·        
Clean the outer Surface with Clean Cloth.

Precaution:

·        
Always ensure that the inlet port is covered
before turn on the pump.

OPERATION OF WATER BATH

PROCEDURE

OPERATION

·        
Keep the instrument
in the area where Exhaust Fan is provided.

·        
Ensure that
instrument is clean and Dust free.

·        
Check the water
Level in pan provided for the same.

·        
Set the required
temperature with the help of Knob.

·        
Switch
“ON” the Instrument.

·        
Keep the sample in
conical flask on the fume hole at upper surface.

·        
After completion of
test, remove the sample and switch “OFF” the instrument properly.

CLEANING

·        
Clean the outer Surface with Clean Cloth.

·        
Change the water in the bath frequently.

Precaution:

·        
Always use Exhaust
Fan when instrument is under working.

·        
Do not run the
instrument in case of low water level.

·        
While keeping the
sample on fume hole, make sure that sample containing flask is in steady state.

Calibration of Analytical Balance-Electronic

Procedure 

Analytical Balance

·        
Ensure that the
analytical balance used for weighing should be levelled at zero levelling and
is supported on a vibration free rigid table base.

·        
Ensure that a
consistent ambient or fixed room temperature is maintained.

·        
There should be no
movement of air or air blast like ceiling fan etc.

·        
Ensure cleanliness
of the analytical balance.

Calibrated Weight Box 

·        
Ensure that the
calibrated set of weights are clean and free from any dust particles.

·        
The standard set of
weights must have a traceability to national accredited body.

Calibration

·        
Check the balance’s
calibration and ensure its calibration by internal calibration system.

·        
Weigh one by one the
standard calibrated set of weights on the balance and record the weights as
indicated on the balance.

·        
Repeat the weighing
of each set of weights at least two to three times to ensure greater accuracy
and reproducibility.

·        
Ensure the precision
and accuracy of weighing. 

Reporting

·        
From the assigned
weights of the standard set of weights correlate the accuracy of the weights
observed on the balance under calibration.

Calibration of Conductivity Meter

Procedure 

·        
Calibration of the
conductivity meter is undertaken by the standardisation with different
reference solutions of known conductivity and measuring the response on the
instrument.

·        
Prepare the
reference conductivity solutions ranging from 0.01M to 1.00M having a
conductivity range of 1.4 to 111.9milli-mhos/cm. (Ref.: APHA, Ed. 20, 1998,
7.4565gm of KCl. in 100ml will give 1Molar solution).

·        
Switch on the
instrument and allow to stabilize for 20 to 30 minutes.

·        
Under the stipulated
operating conditions measure the conductivity of different reference conductivity
solutions allowing the readings to stabilize before reading the conductivity.

·        
Adjust the
conductivity meter (if possible) according to the conductivity observed.

Calibration of Nephelometer

Procedure 

·        
Switch on the
instrument.

·        
Using the standard
operating conditions start the operations of the instrument.

·        
Set the instrument
at zero.

·        
Pour the standard
turbidity solution in the glass tube/cuvette and measure the turbidity.

·        
Ensure that the
glass tube/ cuvette are cleaned/washed before and after use.

·        
Make the
measurements of turbidity in each range of the instrument or in different
ranges of the instrument.

·        
The turbidity shall
not differ from the stated values of the reference standards.

·        
Correlate the
observed values of turbidity with the specified/stated values of the reference
standard and make adjustments, if necessary.

Calibration of Muffle Furnace

Procedure 

·        
Switch on the
instrument.

·        
Ensure that the
temperature indicator/controller is in working condition i.e. the set temperature
should be maintained/retained properly.

·        
Ensure that the door
of muffle furnace is properly closed and does not loosen during operation.

Calibration

·        
Ascertain and set
the points/points at which calibration is required.

·        
Switch on the muffle
furnace and allow it to reach the set point temperature.

·        
Insert the sensor of
the reference/calibrator sensor (PRT) in the muffle furnace as near as possible
to the sensing tip or sensor of the temperature controller/indicator of the
muffle furnace.

·        
When the temperature
is stable note down the temperature reading as indicated by on the muffle
furnace and the reference indicator simultaneously.

·        
Take at least three
to four readings for better accuracy and reproducibility.

·        
Set the next point
and record the temperature readings accordingly.

·        
Correlate the
readings observed with the reference standards and apply corrections, if
necessary. 

Calibration of Disintegration Test Apparatus

Procedure 

·        
Ensure that the
equipment is clean.

·        
Ensure that the
equipment is free from any operational defects. 

Calibration

·        
Switch on the
equipment and start the motor. 

·        
Switch on the
temperature controller knob and set the temperature at 37oC.

·        
Count the paddle
movement raising and lowering cycles of the assembly (RPM) manually using a
stopwatch.

·        
Verify the counts
with the set point specified count of 30 RPM.

·        
Take at least three
to four readings for better accuracy and reproducibility. 

·        
IP/BP/USP allows a
tolerance of ± 2 RPM on
the specified count of 30 RPM. 

Calibration of Humidity Chamber

Procedure 

·        
Switch on the
instrument.

·        
Ensure that the
temperature indicator/controller is in working condition i.e. the set
temperature should be maintained/retained properly.

·        
Ensure that the door
of incubator is properly closed and does not loosen during operation.

Calibration

·        
Ascertain and set
the points/points at which calibration is required.

·        
Switch on the
humidity chamber and allow it to reach the set point temperature.

·        
Insert the sensor of
the reference/calibrator sensor (PRT) in the humidity chamber as near as
possible to the sensing tip or sensor of the temperature controller/indicator
of the humidity chamber.

·        
When the temperature
is stable note down the temperature reading as indicated by on the humidity
chamber and the reference indicator simultaneously.

·        
Take at least three
to four readings for better accuracy and reproducibility.

·        
Set the next point
and record the temperature readings accordingly.

·        
Correlate the
readings observed with the reference standards and apply corrections, if
necessary. 

Calibration of Auto-titrator Apparatus

Procedure 

Apparatus

·        
Dried Beakers, which
have been previously rinsed with acetone. 

·        
Calibrated
Analytical Balance

Determination of Volume

·        
Clean the
measurement/dispensing pipette available with the respective set of the
equipment.

·        
Fill the measuring
pipette with water and adjust to the zero point.

·        
Dispense the known
volumes of water at random or from the set point of the graduation marks of the
pipette.

·        
Repeat the
dispensing of water at different set points and weigh the beaker again and
again.

·        
From the different
weights obtained calculate the volume dispensed and co-relate with the
graduation of the pipette and assign the value.

Calculation:

                                                                                      
Weight of the water dispensed

                                   
Volume of the pipette V=

                                                                                                      Density
of Water at 27oC

Density of Water at 27oC is
0.9965gm/ml

 

 

Calibration of Dissolution Test Apparatus

Procedure 

·        
Ensure that the
equipment is clean.

·        
Ensure that the
equipment is free from any operational defects. 

Calibration

·        
Ascertain and set
the RPM points at which the calibration is required.

·        
Set the RPM with the
help of RPM control knob.

·        
Switch on the
equipment.

·        
Start the motor with
the speed regulator.

·        
Count the RPM of the
paddle manually using a stop watch. Verify the counts with the set points at
25, 50, 75 and 100 RPM respectively.

·        
Take at least three
to four readings at each set point for better accuracy and reproducibility.

·        
Correlate the actual
readings observed with the set point RPM and apply corrections, if necessary.

·        
IP/BP/USP allows a
tolerance of ± 4 % on the
specified RPM.

Pharmaceuticals Instruments Calibration and Operation PDF Notes 

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