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Pharmaceuticals Instruments Calibration and Operation PDF Notes

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.

V = W/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 %.

4. CALIBRATION 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 130^oC 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 ⁰C)
Instrument Instruction Manual

PROCEDURE

1 Calibration Procedure

Set -80⁰C 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 ⁰C 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 ⁰C)
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.

2. Calibration Record

Sr.No	Calibration Time Intervals	Temperature UUC (⁰C )	Temperature Reference ( ⁰C )	Correction to UUC In ( ⁰ C ) if required
1	0 hr	-80 ⁰C
2	0- 2 hr	-80 ⁰C
3	0- 4 hr	-80 ⁰C

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:

Volume of the pipette (ml), V= Weight of the water dispensed (gm) / Density of Water at 27^oC (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 50% RH at 25^oC.
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^-6torr.
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)

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 37^oC.
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:

Volume of the pipette V=Weight of the water dispensed / Density of Water at 27^oC

Density of Water at 27^oC 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.