analytical calibration

CALIBRATION MANAGEMENTCALIBRATION MANAGEMENTBUSINESS PERSPECTIVEBUSINESS PERSPECTIVE

WHAT IS CALIBRATION:WHAT IS CALIBRATION: Calibration is the process of determining the absolute values corresponding to the Calibration is the process of determining the absolute values corresponding to the

graduations on an arbitrary scale on an instrument.graduations on an arbitrary scale on an instrument. It is a set of operations that establish relationship between values indicated by an instrument It is a set of operations that establish relationship between values indicated by an instrument

or system for measuring, recording and controlling and the corresponding known values of a or system for measuring, recording and controlling and the corresponding known values of a reference standard.reference standard.

It is a demonstration that an instrument produces results within specified limits when It is a demonstration that an instrument produces results within specified limits when compared to those produced by a reference standard or a standard which is traceable to a compared to those produced by a reference standard or a standard which is traceable to a national or international standard.national or international standard.

WHY CALIBRATE:WHY CALIBRATE: Because: All equipment degrades slowly over time. The electronic equipment is no exception. Because: All equipment degrades slowly over time. The electronic equipment is no exception.

As they age they drift from their specifications. Rough handling of the equipment adversely As they age they drift from their specifications. Rough handling of the equipment adversely affects calibration. Calibration is required also because different people handle equipment affects calibration. Calibration is required also because different people handle equipment differently.differently.

Laboratory output is analytical data. This data is used in sentencing batches of drugs for Laboratory output is analytical data. This data is used in sentencing batches of drugs for patients use. Hence, the data generated in the laboratory should be reliable, precise and patients use. Hence, the data generated in the laboratory should be reliable, precise and accurate.accurate.

The Initial stage of pharmaceutical manufacturing starts with laboratory testing of input The Initial stage of pharmaceutical manufacturing starts with laboratory testing of input materials both, raw and packaging materials. Several in process control tests form an integral materials both, raw and packaging materials. Several in process control tests form an integral part of pharmaceutical manufacturing. The final product is sentenced based on the laboratory part of pharmaceutical manufacturing. The final product is sentenced based on the laboratory testing.testing.

Accuracy of testing and testing equipments if not established make the whole exercise of Accuracy of testing and testing equipments if not established make the whole exercise of testing meaningless. Erroneous data results in wrong interpretations adverse to the business. testing meaningless. Erroneous data results in wrong interpretations adverse to the business. In legal or other disputes testing and test equipment accuracy becomes very critical.In legal or other disputes testing and test equipment accuracy becomes very critical.

Reporting unreliable and inconsistent data leads to: reanalysis, reworks, batch recalls, Reporting unreliable and inconsistent data leads to: reanalysis, reworks, batch recalls, complaints, regulatory punitive actions, poor credibility, and financial loss.complaints, regulatory punitive actions, poor credibility, and financial loss.

WHAT TO CALIBRATE:WHAT TO CALIBRATE: Any instrument / equipment used for controlling, monitoring, measuring, testing, Any instrument / equipment used for controlling, monitoring, measuring, testing,

calibrating.calibrating. All analytical instruments and equipments having an impact on product quality; gauges All analytical instruments and equipments having an impact on product quality; gauges

and recorders used in production; flow meters, measuring equipment and devices; tanks and recorders used in production; flow meters, measuring equipment and devices; tanks and storage vessels.and storage vessels.

Analytical balances. Spectrophotometers, chromatographs, pH meters, polarimeter, Analytical balances. Spectrophotometers, chromatographs, pH meters, polarimeter, particle counter.particle counter.

Thermometers; other temperature measuring devices; temperature recorders; Thermometers; other temperature measuring devices; temperature recorders; disintegration / dissolution apparatus; measuring equipment like Vernier Calipers; disintegration / dissolution apparatus; measuring equipment like Vernier Calipers; volumetric glass ware like burettes, pipettes, measuring cylinders.volumetric glass ware like burettes, pipettes, measuring cylinders.

WHO CALIBRATES:WHO CALIBRATES: Company staff – skilled and trained; external engineers technicians – skilled and trained; Company staff – skilled and trained; external engineers technicians – skilled and trained;

external laboratory or external calibration agency; government agencies.external laboratory or external calibration agency; government agencies.

CALIBRATION PROCESS:CALIBRATION PROCESS: Subject the equipment to comparison; confirm that it already meets the specifications; Subject the equipment to comparison; confirm that it already meets the specifications;

bring it up to specifications; make adjustments during calibration to fall within certain bring it up to specifications; make adjustments during calibration to fall within certain tolerances.tolerances.

Tolerances need to be small, acceptable deviations from the equipment's specified Tolerances need to be small, acceptable deviations from the equipment's specified accuracy, as suggested by the manufacturer of the equipment or published literature.accuracy, as suggested by the manufacturer of the equipment or published literature.


COST OF CALIBRATION:COST OF CALIBRATION: Machines need to be calibrated regularly for proper functioning. A qualified and Machines need to be calibrated regularly for proper functioning. A qualified and

trained engineer or mechanic is required for calibration.trained engineer or mechanic is required for calibration. Time taken to test and calibrate one unit could take hours depending on how Time taken to test and calibrate one unit could take hours depending on how

complex or sophisticated it is. complex or sophisticated it is. If a problem has been found the machine should not be used first until a If a problem has been found the machine should not be used first until a

replacement part has arrived. For minor cases a simple adjustment may suffice to replacement part has arrived. For minor cases a simple adjustment may suffice to make it work again make it work again

IN-HOUSE CALIBRAITON LAB:IN-HOUSE CALIBRAITON LAB: The cost of calibration includes setting up a fully equipped in-house calibration lab. The cost of calibration includes setting up a fully equipped in-house calibration lab.

Things need to be ensured are; Space requirements, availability of references Things need to be ensured are; Space requirements, availability of references standards with traceability to national standards, maintenance of standards, trained standards with traceability to national standards, maintenance of standards, trained staff and their retention, documentation, and calibration process audits.staff and their retention, documentation, and calibration process audits.

EXTERNAL CALIBRATION LAB OR AGENCY:EXTERNAL CALIBRATION LAB OR AGENCY: If not in-house calibration lab, an external reputed lab or agency has to be If not in-house calibration lab, an external reputed lab or agency has to be

identified.identified. Their accreditation, their infrastructure and capabilities, timely support, due Their accreditation, their infrastructure and capabilities, timely support, due

diligence and or auditing, suitability of calibration certificates, and charges per diligence and or auditing, suitability of calibration certificates, and charges per calibration need to be looked into.calibration need to be looked into.


FREQUENCY:FREQUENCY: Derive own frequency of calibration based on full justifications duly recorded and Derive own frequency of calibration based on full justifications duly recorded and

approved. Criticality of equipment – direct impact, indirect impact or no impact; approved. Criticality of equipment – direct impact, indirect impact or no impact; and past history and present performance trend of the equipment need to be and past history and present performance trend of the equipment need to be looked into.looked into.

Use risk based approach. Frequency may change depending on operation and use Use risk based approach. Frequency may change depending on operation and use of instrument / equipment.of instrument / equipment.

BENEFITS OF CALIBRATION:BENEFITS OF CALIBRATION: The benefits of calibration are: Reliable measurements; assured product quality The benefits of calibration are: Reliable measurements; assured product quality

and patient safely; financial benefits through reduced or no rework, reprocessing, and patient safely; financial benefits through reduced or no rework, reprocessing, retesting, and recalls; no regulatory observations and penal actions; reduced retesting, and recalls; no regulatory observations and penal actions; reduced customer complaints; and motivated staff.customer complaints; and motivated staff.


DEALING WITH CALIBRATION DEALING WITH CALIBRATION FAILURESFAILURES

CALIBRATION: DefinitionsCALIBRATION: Definitions Calibration is a comparison between measurements. One of known magnitude or Calibration is a comparison between measurements. One of known magnitude or

correctness made or set with one device and another measurement made in a correctness made or set with one device and another measurement made in a similar way with a second device.similar way with a second device.

The device with the known or assigned correctness is called the standard. The The device with the known or assigned correctness is called the standard. The second device is the unit under test (UUT) or the test instrument (TI).second device is the unit under test (UUT) or the test instrument (TI).

ELEMENTARY REQUIREMENTS OF CALIBRATION:ELEMENTARY REQUIREMENTS OF CALIBRATION: Calibration must be traceable to a Standard (National / International / Regional); at Calibration must be traceable to a Standard (National / International / Regional); at

a defined frequency – as per schedule / programme; as per defined procedure – a defined frequency – as per schedule / programme; as per defined procedure – specific directions and limits for accuracy and precision; recorded – log books, specific directions and limits for accuracy and precision; recorded – log books, tags, reports, certificates; and performed by suitably qualified and trained tags, reports, certificates; and performed by suitably qualified and trained personnel. And finally, compendia / specification requirements.personnel. And finally, compendia / specification requirements.

COMMONLY FOUND INSPECTION COMMONLY FOUND INSPECTION OBSERVATIONSOBSERVATIONS

Failure to maintain equipment at appropriate intervals to prevent malfunctions Failure to maintain equipment at appropriate intervals to prevent malfunctions or contamination.or contamination.

Calibration stickers are illegible and damaged making ascertaining calibration Calibration stickers are illegible and damaged making ascertaining calibration status difficult.status difficult.

Calibration trending is not done for critical instruments, and the responsibilities Calibration trending is not done for critical instruments, and the responsibilities of staff carrying out calibration are not well defined.of staff carrying out calibration are not well defined.

There are no controls on vendor calibration procedures, original calibration There are no controls on vendor calibration procedures, original calibration data is not available for review; no robust calibration programme in place; no data is not available for review; no robust calibration programme in place; no SOP for calibration; routine calibration, inspection, and checking equipment is SOP for calibration; routine calibration, inspection, and checking equipment is not performed according to a written program record of training on calibration not performed according to a written program record of training on calibration is not available.is not available.

New working standard are standardized against old working standards; weights New working standard are standardized against old working standards; weights used for calibration of balances are stored in uncontrolled manner; change used for calibration of balances are stored in uncontrolled manner; change control SOP does not mention changes related calibration; calibration status control SOP does not mention changes related calibration; calibration status label does not have all the required details; and instruments which are label does not have all the required details; and instruments which are calibrated prior to use are not labelled “Calibrate Prior To Use”calibrated prior to use are not labelled “Calibrate Prior To Use”

CALIBRATIONCALIBRATIONREGULATORY PERSPECTIVEREGULATORY PERSPECTIVE

Calibration:Calibration: Calibration is required to ensure monitoring data and that the reports are Calibration is required to ensure monitoring data and that the reports are

accurate; to ensure continued process control; and for safety of drug product accurate; to ensure continued process control; and for safety of drug product and process employed in producing “drug substances” and “drug products”.and process employed in producing “drug substances” and “drug products”.

Definition:Definition: Schedule M or L1 or any other guidelines issued by drug regulatory agencies in Schedule M or L1 or any other guidelines issued by drug regulatory agencies in

India are yet to define “calibrate”.India are yet to define “calibrate”. Regulatory officers during their inspections expect calibration compliance as Regulatory officers during their inspections expect calibration compliance as

they feel fine. (Mostly balances in stores are insisted to have to calibration they feel fine. (Mostly balances in stores are insisted to have to calibration status).status).

DOCUMENTATION REQUIRED:DOCUMENTATION REQUIRED: SOPsSOPs

For operationFor operation For calibrationFor calibration

Calibration RecordCalibration Record The results of calibration carried out for each instrument shall be reported accurately, The results of calibration carried out for each instrument shall be reported accurately,

clearly and unambiguously in a pre-designed calibration record format for every clearly and unambiguously in a pre-designed calibration record format for every instrument.instrument.

Maintenance RecordMaintenance Record To achieve performance consistency of instrument proper maintenance of every To achieve performance consistency of instrument proper maintenance of every

instrument should be maintained per the predetermined maintenance schedule. This instrument should be maintained per the predetermined maintenance schedule. This should be properly documented. Historical Data on “Out Of Calibration” should be should be properly documented. Historical Data on “Out Of Calibration” should be documented for each instrument along with investigation report.documented for each instrument along with investigation report.

CALIBRATION OF GLASSWARE & CALIBRATION OF GLASSWARE & ANALYTICAL EQUIPMENT USED FOR ANALYTICAL EQUIPMENT USED FOR

PHARMACOPOEIAL ANALYSISPHARMACOPOEIAL ANALYSIS

CALIBRATION OF GLASSWARESCALIBRATION OF GLASSWARES

There are two grades of apparatus Class A and Class B. Glass apparatus There are two grades of apparatus Class A and Class B. Glass apparatus may be employed in routine work after calibration. Class A is intended for may be employed in routine work after calibration. Class A is intended for use in work of the highest accuracy.use in work of the highest accuracy.

The tolerance on capacity for volumetric flasks, pipettes and burettes as The tolerance on capacity for volumetric flasks, pipettes and burettes as laid down in the relevant Indian Standards given an page A-8, table – 2 in laid down in the relevant Indian Standards given an page A-8, table – 2 in IP 1996.IP 1996.

Volumetric glassware is normally calibrated a 270C as per Volumetric glassware is normally calibrated a 270C as per pharmacopoeial requirement, otherwise calibrate at room temperature by pharmacopoeial requirement, otherwise calibrate at room temperature by measuring temperature of water and applying the correction factor.measuring temperature of water and applying the correction factor.

Calibration of volumetric flasks: (Capacity – 5, 10, 50, 100, 250, 1000ml)Calibration of volumetric flasks: (Capacity – 5, 10, 50, 100, 250, 1000ml) Clean the volumetric flask with chromic acid solution. Clean the volumetric flask Clean the volumetric flask with chromic acid solution. Clean the volumetric flask

with distill water. Dry at 70°C temperature. Do not use cloth or tissue paper for with distill water. Dry at 70°C temperature. Do not use cloth or tissue paper for drying.drying.

First weigh empty flask and then fill up with double distill water up to mark and First weigh empty flask and then fill up with double distill water up to mark and again weigh it at required temperature. Ensure to consider lower miniscus all again weigh it at required temperature. Ensure to consider lower miniscus all the time. Find the difference between the two weights, and convert it into the time. Find the difference between the two weights, and convert it into volume and apply the correction factor.volume and apply the correction factor.

Volumetric Flasks: IS 915: 1975Volumetric Flasks: IS 915: 1975

Nominal CapacityNominal Capacity 55 1010 2525 2020 100100 250250 500500 10001000

Tolerance + ml (Class A)Tolerance + ml (Class A) 0.020.02 0.020.02 0.030.03 0.040.04 0.060.06 0.10.1 0.150.15 0.20.2

Tolerance + ml (Class B)Tolerance + ml (Class B) 0.040.04 0.040.04 0.060.06 0.080.08 0.150.15 0.20.2 0.30.3 0.80.8

One Mark pipettes: IS 1117: 1975One Mark pipettes: IS 1117: 1975

Nominal CapacityNominal Capacity 11 22 55 1010 2020 2525 5050 100100

Tolerance + ml (Class A)Tolerance + ml (Class A) 0.010.01 0.010.01 0.020.02 0.020.02 0.030.03 0.00.033

0.040.04 0.060.06

Tolerance + ml (Class B)Tolerance + ml (Class B) 0.020.02 0.020.02 0.030.03 0.040.04 0.050.05 0.00.066

0.080.08 0.120.12

Graduated pipettes: IS 4162:1967Graduated pipettes: IS 4162:1967

Nominal CapacityNominal Capacity 11 22 55 1010 2525 ------ ------ ------

Subdivision mlSubdivision ml 0.010.01 0.20.2 0.050.05 0.100.10 0.020.02 ------ ------ ------

Tolerance + mlTolerance + ml

Tolerance + ml (Class A)Tolerance + ml (Class A) 0.000.0066

0.010.01 0.030.03 0.050.05 0.010.01 ------ ------ ------

Tolerance + ml (Class B)Tolerance + ml (Class B) 0.020.02 0.060.06 0.100.10 0.100.10 0.150.15 ------ ------ ------

Burettes: IS 1997:1967Burettes: IS 1997:1967

Nominal CapacityNominal Capacity 11 2525 5050 100100 ------ ------ ------ ------

Subdivision mlSubdivision ml 0.050.05 0.050.05 0.10.1 0.10.1 ------ ------ ------ ------

Tolerance + ml (Class A)Tolerance + ml (Class A) 0.010.01 0.010.01 0.050.05 0.010.01 ------ ------ ------ ------

Tolerance + ml (Class B)Tolerance + ml (Class B) 0.020.02 0.060.06 0.10.1 0.20.2 ------ ------ ------ ------

Volumetric Glassware IPVolumetric Glassware IPCALIBRATION OF GLASSWARESCALIBRATION OF GLASSWARES

CALIBRATION OF ANALYTICAL CALIBRATION OF ANALYTICAL INSTRUMENTS / EQUIPMENTINSTRUMENTS / EQUIPMENT

Weighing on an Analytical Balance:Weighing on an Analytical Balance: Weighing is a frequent step in analytical procedures and balance is an essential Weighing is a frequent step in analytical procedures and balance is an essential

piece of laboratory equipment in most analyses. In spite of this, weighing is a piece of laboratory equipment in most analyses. In spite of this, weighing is a common sourced of error that can be difficult to detect in the final analytical common sourced of error that can be difficult to detect in the final analytical results. Unless balance is checked before each weighing operation is performed, results. Unless balance is checked before each weighing operation is performed, errors can easily occur, resulting in faulty analytical data.errors can easily occur, resulting in faulty analytical data.

Balance Environment:Balance Environment: Balance should be kept in suitable location with sufficiently low level of Balance should be kept in suitable location with sufficiently low level of

Vibration and Air current.Vibration and Air current. The surrounding area should be clean and tidy.The surrounding area should be clean and tidy. When the balance location is changed it must be allowed to adjust to the When the balance location is changed it must be allowed to adjust to the

temperature, its new environment and be re-calibrated.temperature, its new environment and be re-calibrated. The balance users should check the balance: Environment, calibration, and The balance users should check the balance: Environment, calibration, and

balance uncertainties.balance uncertainties. Do not assume that the balance has been left in the proper operating condition Do not assume that the balance has been left in the proper operating condition

by the previous user.by the previous user. Balance Calibration:Balance Calibration:

Turn on the balance and allow to equilibrate for at least 1 hour before Turn on the balance and allow to equilibrate for at least 1 hour before proceeding with calibration. (Microbalances may require up to 24 hours to proceeding with calibration. (Microbalances may require up to 24 hours to reach equilibrium).reach equilibrium).

Re-calibrate balance, if the balance power has gone and resumed. Balance may Re-calibrate balance, if the balance power has gone and resumed. Balance may not give correct weighing until it has been calibrated.not give correct weighing until it has been calibrated.

Electronic analytical balance has an internal calibration system based on the Electronic analytical balance has an internal calibration system based on the applied load.applied load.

Calibration applies for the current ambient temperature.Calibration applies for the current ambient temperature. Balance should be calibrated daily prior to first weighing of the day. The Balance should be calibrated daily prior to first weighing of the day. The

calibration should typically include 2-3 weights in the operating range.calibration should typically include 2-3 weights in the operating range. Calibration of the full operating range should be carried out every quarter.Calibration of the full operating range should be carried out every quarter. Weight box used for calibration should be certified and have traceability to Weight box used for calibration should be certified and have traceability to

National Standard Laboratory (NPL) or equivalent.National Standard Laboratory (NPL) or equivalent.


Calibration parameters:Calibration parameters: Accuracy: Check accuracy using different standard weights. Tolerance: ± 0.1 % of Accuracy: Check accuracy using different standard weights. Tolerance: ± 0.1 % of

the actual weight.the actual weight. Linearity: Check 6 different weights. Plot a graph of actual weights against observed Linearity: Check 6 different weights. Plot a graph of actual weights against observed

weights. It should be linear.weights. It should be linear. Precision: Check 6 times weights of similar weight (100mg). Calculate % RSD for Precision: Check 6 times weights of similar weight (100mg). Calculate % RSD for

observed weight. Tolerance: NMT 0.5%observed weight. Tolerance: NMT 0.5% Corner Load Test: Weigh 100mg in four corner and middle of the pan of the balance. Corner Load Test: Weigh 100mg in four corner and middle of the pan of the balance.

Calculate % RSD for observed weight. Tolerance: NMT 0.5%Calculate % RSD for observed weight. Tolerance: NMT 0.5% Balance Uncertainties:Balance Uncertainties:

Drift is one of the common errors. It is easiest to reduce or eliminate. Check for the Drift is one of the common errors. It is easiest to reduce or eliminate. Check for the following causes:following causes:

Balance door is open.Balance door is open. Temperature of the balance and material to be weighed are not the same.Temperature of the balance and material to be weighed are not the same. Sample is losing or gaining weight.Sample is losing or gaining weight. Balance moved, but not allowed to equilibrate to its surroundings or re-calibrated.Balance moved, but not allowed to equilibrate to its surroundings or re-calibrated. Air currents are present in the laboratory.Air currents are present in the laboratory. Temperature in the laboratory varies.Temperature in the laboratory varies. Balance is not properly leveled.Balance is not properly leveled. Laboratory operations cause vibration.Laboratory operations cause vibration. Hysteresis of the mechanical part occurs during weighing. (Hysteresis I the balances caused by Hysteresis of the mechanical part occurs during weighing. (Hysteresis I the balances caused by

excessive stretching of the strings and it is primarily due to accidental dropping of object onto excessive stretching of the strings and it is primarily due to accidental dropping of object onto the pan).the pan).

Karl Fischer TitratorKarl Fischer Titrator Calibration parameterCalibration parameter

Factor: Calculate the factor for 6 different weights of di-sodium tirtrate or known quantity of Factor: Calculate the factor for 6 different weights of di-sodium tirtrate or known quantity of water in methanol. Calculate % RSD for factor. Tolerance: NMT 1 %water in methanol. Calculate % RSD for factor. Tolerance: NMT 1 %


Auto Titrator:Auto Titrator: Burette Calibration:Burette Calibration:

Set burette 5ml and 20ml. Measure the volume of water by using measuring cylinder for 3 Set burette 5ml and 20ml. Measure the volume of water by using measuring cylinder for 3 different setting of volumes. Tolerance: ± 0.05ml.different setting of volumes. Tolerance: ± 0.05ml.

Auto Titrator Calibration:Auto Titrator Calibration: Perform 6 sets of assay of known material. Calculate correlation coefficient between the Perform 6 sets of assay of known material. Calculate correlation coefficient between the

weights taken and ml of the volumetric solution required for assay. Calculate % RSD for six weights taken and ml of the volumetric solution required for assay. Calculate % RSD for six different assays. Tolerance: NMT 1%.different assays. Tolerance: NMT 1%.

pH Meter:pH Meter: #pH (as predetermined schedule). Adjust pH meter with Buffer pH 7.0 solution and #pH (as predetermined schedule). Adjust pH meter with Buffer pH 7.0 solution and

pH4.0 solution and check with pH 10.0 solution. Tolerance of check solution is ± 0.05.pH4.0 solution and check with pH 10.0 solution. Tolerance of check solution is ± 0.05. Refractometer:Refractometer:

Refractive Index: Check the refractive index of water, Carbon Tetrachloride and Refractive Index: Check the refractive index of water, Carbon Tetrachloride and Toluene. (Use certified reference materials). They are 1.3325 (at 25°C), 1.4603 and Toluene. (Use certified reference materials). They are 1.3325 (at 25°C), 1.4603 and 1.4969 (both at 20°C) respectively. (When the white light is used, the refractometer is 1.4969 (both at 20°C) respectively. (When the white light is used, the refractometer is provided with a compensating system. The thermometer is graduated at intervals of provided with a compensating system. The thermometer is graduated at intervals of 0.5°C or less).0.5°C or less).

UV-VIS Spectrophotometer:UV-VIS Spectrophotometer: Calibration of Wavelength:Calibration of Wavelength:

Check it with Homium filter or Holmium Perchlorate solution. Tolerance: UV Range: +/- 1nm; Check it with Homium filter or Holmium Perchlorate solution. Tolerance: UV Range: +/- 1nm; VIS Range: +/- 3nm.VIS Range: +/- 3nm.

Calibration of Absorbance:Calibration of Absorbance: Check A (1%, 1cm) value of Potassium dichromate solution (60mcg/ml in 0.01 N Sulphuric Check A (1%, 1cm) value of Potassium dichromate solution (60mcg/ml in 0.01 N Sulphuric

Acid) at different wavelengths.Acid) at different wavelengths. Stray Light:Stray Light:

Measure the absorbance and transmittance of 1.2% (w/v) Potassium Chloride solution at 200 Measure the absorbance and transmittance of 1.2% (w/v) Potassium Chloride solution at 200 nm. Abs: NLT 2.0; Trans: NMT 1.0%nm. Abs: NLT 2.0; Trans: NMT 1.0%

Resolution Power:Resolution Power: Check the absorbance of 0.02% v/v solution of Toluene in Hexane (UV grade) at maxima and Check the absorbance of 0.02% v/v solution of Toluene in Hexane (UV grade) at maxima and

minima at about 269 and 266nm. Ratio of 269/266 = NLT 1.5.minima at about 269 and 266nm. Ratio of 269/266 = NLT 1.5. Photometric Accuracy: Measure the absorbance of Potassium Nitrate solution of Photometric Accuracy: Measure the absorbance of Potassium Nitrate solution of

different concentrations (0.355%, 0.710% and 1.065%) at 302nm. Abs: 0.250, 0.500 different concentrations (0.355%, 0.710% and 1.065%) at 302nm. Abs: 0.250, 0.500 and 0.751 respectively. Tolerance: upto + 0.010.and 0.751 respectively. Tolerance: upto + 0.010.


Fourier Transform Infra Red Spectrophotometer (FTIR):Fourier Transform Infra Red Spectrophotometer (FTIR): Verification of wave numbers:Verification of wave numbers:

Check the wave numbers using polystyrene film of thickness 0.05 mm. Tolerance: ±0.3 Wave Check the wave numbers using polystyrene film of thickness 0.05 mm. Tolerance: ±0.3 Wave number.number.

Spectral Resolution: Record the spectrum of polystyrene film. Check the depth from Spectral Resolution: Record the spectrum of polystyrene film. Check the depth from the maximum absorption at about 2851 cmthe maximum absorption at about 2851 cm-1-1 to the minimum 2870 cm to the minimum 2870 cm-1-1. It should be . It should be NLT 18 absorption at about 1583 cmNLT 18 absorption at about 1583 cm-1-1. It should be NLT 12% Transmittance.. It should be NLT 12% Transmittance.

High Performance Liquid Chromatograph (HPLC):High Performance Liquid Chromatograph (HPLC): Flow Rate:Flow Rate:

Set flow rate at 1 ml, 2ml and 2.5ml/min. Measure the volume of water by weight using Set flow rate at 1 ml, 2ml and 2.5ml/min. Measure the volume of water by weight using measuring cylinder for three different settings of flow rate. Tolerance: ±1% of the set flow rate.measuring cylinder for three different settings of flow rate. Tolerance: ±1% of the set flow rate.

System Suitability:System Suitability: Peak Tailing Factor, Column Efficiency, and Capacity Factor.Peak Tailing Factor, Column Efficiency, and Capacity Factor.

Column Oven:Column Oven: Set the column ± oven at 35°C. Place NIST traceable temperature probe in the column oven. Set the column ± oven at 35°C. Place NIST traceable temperature probe in the column oven.

Measure temperature at different temperature settings after stabilizing. Tolerance ± 2°C.Measure temperature at different temperature settings after stabilizing. Tolerance ± 2°C. Sample compartment of Auto Sampler:Sample compartment of Auto Sampler:

Set the column ± oven at 10°C. Place NIST traceable temperature probe in the column oven. Set the column ± oven at 10°C. Place NIST traceable temperature probe in the column oven. Measure temperature at different temperature settings after stabilizing. Tolerance: ±2°C.Measure temperature at different temperature settings after stabilizing. Tolerance: ±2°C.

Polarimeter:Polarimeter: Angle of Optical Rotation:Angle of Optical Rotation:

Check the alignment of sodium lamp compartment. Ensure zero setting. Measure the angle of Check the alignment of sodium lamp compartment. Ensure zero setting. Measure the angle of optical rotation (five times each) of 10%, 20%, 30%, 40%, and 50% solution previously dried optical rotation (five times each) of 10%, 20%, 30%, 40%, and 50% solution previously dried sucrose at 25°C. Tolerance of angle of rotation should be ±0.50.sucrose at 25°C. Tolerance of angle of rotation should be ±0.50.


Standard angle of rotation of different solutions.Standard angle of rotation of different solutions.

ConcentrationConcentration Angle of RotationAngle of Rotation

10 gm / 100ml10 gm / 100ml 13.33013.330

20 gm / 100ml20 gm / 100ml 26.61026.610

30 gm / 100ml30 gm / 100ml 39.86039.860

40 gm / 100ml40 gm / 100ml 53.06053.060

50 gm / 100ml50 gm / 100ml 66.23066.230

Oven / Vacuum Oven:Oven / Vacuum Oven: Temperature:Temperature:

Set temperature at 60°C. Measure the temperature of the oven at five different locations (4 Set temperature at 60°C. Measure the temperature of the oven at five different locations (4 corners and middle ) using liquid paraffin bath and calibrated thermometer. Take average of corners and middle ) using liquid paraffin bath and calibrated thermometer. Take average of these 5 values.these 5 values.

Similarly set the temperature at 105°C and measure the temperature as above. Tolerance: Similarly set the temperature at 105°C and measure the temperature as above. Tolerance: Difference between set and observed temperatures should be ±1 °C.Difference between set and observed temperatures should be ±1 °C.

Temperature mapping should be performed annually to identify be performed annually to Temperature mapping should be performed annually to identify be performed annually to identify weak sopt and decide sensor position.identify weak sopt and decide sensor position.


Dissolution Calibration:Dissolution Calibration: Calibration of dissolution machine includes: Level test, Centering test, Wobble test, Calibration of dissolution machine includes: Level test, Centering test, Wobble test,

Paddle / Basket depth, Dissolution vessel test, Temperature control test, Rotation Paddle / Basket depth, Dissolution vessel test, Temperature control test, Rotation speed accuracy test, and Dissolution suitability test.speed accuracy test, and Dissolution suitability test.

Level test:Level test: Check level of plate holding dissolution vessel using sprit level.Check level of plate holding dissolution vessel using sprit level.

Centering test:Centering test: Put 900ml water. Lower the shaft in dissolution vessel. Centering should be within +/- mm of Put 900ml water. Lower the shaft in dissolution vessel. Centering should be within +/- mm of

vessel center axis. Check using centering gauge.vessel center axis. Check using centering gauge. Wobble test: Wobble test:

No more than 1 mm from center axis.No more than 1 mm from center axis. Paddle / Basket Depth:Paddle / Basket Depth:

Use height gauge. It should be exactly 25mm (+/-mm) from bottom of vessel. Use height gauge. It should be exactly 25mm (+/-mm) from bottom of vessel. Dissolution vessel test: Dissolution vessel test:

Inside surface should free of abnormalities. The lowest point in the vessel must be in line with Inside surface should free of abnormalities. The lowest point in the vessel must be in line with vertical axis of the shaft.vertical axis of the shaft.

Temperature control test: Temperature control test: Set the temperature of water bath at 37°C. Start heater. Allow water bath to attain the set Set the temperature of water bath at 37°C. Start heater. Allow water bath to attain the set

temperature. Measure the temperature using calibrated thermometer.temperature. Measure the temperature using calibrated thermometer. Rotation speed accuracy test: Rotation speed accuracy test:

Set the rotation speed to 50 rpm, 75rpm, 100rpm. It should be within +/- 4 %Set the rotation speed to 50 rpm, 75rpm, 100rpm. It should be within +/- 4 % Dissolution suitability test:Dissolution suitability test:

Prednisone is a disintegrating tablet and it is highly affected by mechanical parameters and Prednisone is a disintegrating tablet and it is highly affected by mechanical parameters and vessels. These tablets are sensitive to centering, vibrations, shaft wobbling, shaft verticality vessels. These tablets are sensitive to centering, vibrations, shaft wobbling, shaft verticality and degassing.and degassing.

Salicylic acid is non-disintegrating tablet. It is affected by poorly de-aerated media, warped or Salicylic acid is non-disintegrating tablet. It is affected by poorly de-aerated media, warped or dirty baskets and unclean or scratched vessel. These tablets are not sensitive to mechanical dirty baskets and unclean or scratched vessel. These tablets are not sensitive to mechanical parameters.parameters.


Sources of errors in Dissolution Calibration:Sources of errors in Dissolution Calibration: De-aerated media.De-aerated media.

Dissolved air can slow down dissolution by creating barrier either by adhering to the tablet Dissolved air can slow down dissolution by creating barrier either by adhering to the tablet surface or basket screen or particles can clig to bubbles on the glass surface of vessel or shaft.surface or basket screen or particles can clig to bubbles on the glass surface of vessel or shaft.

Use properly dried standard:Use properly dried standard: Reference standard used must be dried properly. Solution must be prepared on the day of use. Reference standard used must be dried properly. Solution must be prepared on the day of use.

For dissolving the standard small of alcohol can be used.For dissolving the standard small of alcohol can be used. Vibration interference:Vibration interference:

Vibration can arise from bearing can get worn and cause vibration and wobble. Drive belt Vibration can arise from bearing can get worn and cause vibration and wobble. Drive belt should be checked for wear and dirt. Vessels need to be locked in the place so they are not should be checked for wear and dirt. Vessels need to be locked in the place so they are not moving with the flow of water in bath. External sources might include bench top, sonicators, moving with the flow of water in bath. External sources might include bench top, sonicators, shakers, centrifuges. Tester should not be near hood. Heavy foot traffic and door slamming shakers, centrifuges. Tester should not be near hood. Heavy foot traffic and door slamming should be avoided.should be avoided.

Instrument:Instrument: Basket and Paddle shaft must be straight and true. If paddles are partially coated with teflon, Basket and Paddle shaft must be straight and true. If paddles are partially coated with teflon,

the layer can get peeled off causing flow disturbances. the layer can get peeled off causing flow disturbances. The baskets screen size may change over time due to usage of acidic medium. The attachments The baskets screen size may change over time due to usage of acidic medium. The attachments

may be clip type or with O rings can contribute to variation.may be clip type or with O rings can contribute to variation. Vessels have problems of irregular inner surface which can affect significantly. Vessel may Vessels have problems of irregular inner surface which can affect significantly. Vessel may

develop residue buildup either from oily products or sticky excipients. On prolong use vessel develop residue buildup either from oily products or sticky excipients. On prolong use vessel might develop scratches on inner surface which can affect dissolution. If surfactants are used might develop scratches on inner surface which can affect dissolution. If surfactants are used in concentration more than 0.5% while preparing dissolution medium then more rinsing to in concentration more than 0.5% while preparing dissolution medium then more rinsing to vessel are required as it is difficult to remove residue easily.vessel are required as it is difficult to remove residue easily.

Melting Point Apparatus:Melting Point Apparatus: Melting Point:Melting Point:

Check the melting points of Vanilin, Acetanilide, Sulphanilamide and Caffeine using calibrated thermometer Check the melting points of Vanilin, Acetanilide, Sulphanilamide and Caffeine using calibrated thermometer capable of determining temperature to level of 0.5°C. They are between 81 – 83°C, 115-116°C, 164-166°C capable of determining temperature to level of 0.5°C. They are between 81 – 83°C, 115-116°C, 164-166°C and 234-237°C respectively.and 234-237°C respectively.


Thermometers:(IS4825:1968 and IS 6274:1971)Thermometers:(IS4825:1968 and IS 6274:1971) D.T. Machine Cycles per minute: D.T. Machine Cycles per minute:

Check the number of cycles per minute three times. It should be 30 cycle/min. tolerance: ±2 Check the number of cycles per minute three times. It should be 30 cycle/min. tolerance: ±2 cycles/mincycles/min

Thermostat Temperature:Thermostat Temperature: Set the temperature 25°C and 37°C. Start the instrument. Measure the temperature of water in Set the temperature 25°C and 37°C. Start the instrument. Measure the temperature of water in

beaker at different time intervals such as 15, 30, 45 and 60 minutes using calibrated beaker at different time intervals such as 15, 30, 45 and 60 minutes using calibrated thermometer. Tolerance: ±2°C of set temperature.thermometer. Tolerance: ±2°C of set temperature.

Distance traveled during oscillation: Distance traveled during oscillation: Check the distance traveled during oscillation three times. Take average value. It should be Check the distance traveled during oscillation three times. Take average value. It should be

55mm. Tolerance: ±5mm.55mm. Tolerance: ±5mm. Time Display:Time Display:

Check the time for set 15, 30 and 45 minutes using calibrated clock. It should match with the Check the time for set 15, 30 and 45 minutes using calibrated clock. It should match with the time of calibrated clock.time of calibrated clock.

Historical Background:Historical Background: Instrument Identification for according to EN45001:Instrument Identification for according to EN45001:

Protocol No.Protocol No. Lab/Dept. NameLab/Dept. Name Producer NameProducer Name Catalog NameCatalog Name New/Old/RefurbishedNew/Old/Refurbished Arrival DateArrival Date Place in the labPlace in the lab Calibration / ValidateCalibration / Validate Company NameCompany Name Instrument NameInstrument Name Distributor NameDistributor Name Serial NameSerial Name Purchasing DatePurchasing Date Installing DateInstalling Date Servicing DetailsServicing Details Calibration / Validation FrequencyCalibration / Validation Frequency


Traceability of Standards:Traceability of Standards: The Quality Standard regulation requires that standards used to calibrate equipment The Quality Standard regulation requires that standards used to calibrate equipment

be traceable to the National Institute of Standards and Technology (NIST), or other be traceable to the National Institute of Standards and Technology (NIST), or other recognized national or international standards. Traceability also can achieved through recognized national or international standards. Traceability also can achieved through a contract calibration laboratory which in turn used NIST services.a contract calibration laboratory which in turn used NIST services.

Instrument Calibration Programs:Instrument Calibration Programs: The accuracy of a measurement can only be assured if the measuring device is known The accuracy of a measurement can only be assured if the measuring device is known

to accurate both before and after the measurement is mode.to accurate both before and after the measurement is mode. External Calibration:External Calibration:

Calibrated prior to use; formal testing of accuracy and precision; traceable reference Calibrated prior to use; formal testing of accuracy and precision; traceable reference standards; approved calibration methods; tolerances and full results documented rage standards; approved calibration methods; tolerances and full results documented rage conditions investigated; qualification of calibration laboratory; status label conditions investigated; qualification of calibration laboratory; status label (identification, recalibration date); reports certified; records consolidated and (identification, recalibration date); reports certified; records consolidated and archived.archived.

Internal Calibration – Performance Checks:Internal Calibration – Performance Checks: Internal weekly or as used checks; upscale and downscale, running logs reviewed by Internal weekly or as used checks; upscale and downscale, running logs reviewed by

management; tolerances and results on the record; traceable reference standards; out management; tolerances and results on the record; traceable reference standards; out of range conditions investigated; statement of adjustment or repair.of range conditions investigated; statement of adjustment or repair.““MANUFACTURERS ARE RESPONSIBLE FOR ENSURING THE MANUFACTURERS ARE RESPONSIBLE FOR ENSURING THE

ESTABLISHMENT OF ROUTINE CALIBRATION ON THEIR ESTABLISHMENT OF ROUTINE CALIBRATION ON THEIR TEST EQUIPMENT (VOLUMETRIC & MEASURING TEST EQUIPMENT (VOLUMETRIC & MEASURING

INSTRUMENT). SO IT WILL BE SUITABLE FOR ITS INSTRUMENT). SO IT WILL BE SUITABLE FOR ITS INTENDED USE.”INTENDED USE.”

analytical calibration

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