Calibration of FTIR, Purpose:
Calibration of FTIR, The purpose of this SOP is to describe the operation, calibration and cleaning of FTIR Spectrophotometer (Shimadzu, IR Affinity-1) used in the quality control laboratory at XX Pharmaceuticals Limited.
Scope
This procedure is applicable for FTIR Spectrophotometer (Shimadzu, IR Affinity-1), installed in the quality control laboratory of XX Pharmaceuticals Limited.
Definitions / Abbreviation:
Standard Operating Procedure (SOP): Standard Operating Procedure. A written authorized procedure, which gives instructions for performing operations.
[][]QC: Quality Control.
[][]FTIR: Fourier Transform Infrared.
Responsibilities
[][]Executive/ Sr. Executive, QC
[][]To ensure that this procedure is followed.
[][]To maintain the records properly as per SOP
Manager, Quality Control
[][]To ensure that this procedure is kept up to date.
[][]To confirm that the SOP is technically sound and reflects the required working practices.
[][]To arrange training on the SOP to all concerned personnel and to ensure implementation of the SOP after training.
[][]Schedule calibration of the instrument at the defined intervals.
Head of Quality Assurance
[][]Approval of the SOP.
[][]To ensure the overall implementation of the SOP.
Procedure: Calibration of FTIR
General Procedure:
[][]Ensure that the instrument is clean and calibrated.
[][]Connect the instrument properly with the power supply.
[][]Switch “ON” the main switch, power switch of the instrument situated at the backside panel, printer and computer.
[][]Allow the instrument initialize for at least 1 hour.
Disk preparation for solid sample:
[][]Mix 5 to 10 mg of sample with 200 to 400 mg of Potassium Bromide (KBr) [IR Grade].
[][]Grind the mixture to very fine powder with agate mortar.
[][]Pour the mixture into the barrel and carefully insert the plunger with its polished face downward.
[][]Maintain the hydraulic pressure 50 KN to 60 KN of the barrel for 1-2 minutes.
[][]Release the pressure and take out the disk from the barrel.
[][]For sample of liquid material using sealed liquid cell:
[][]Inject the liquid sample (pure or in solution) in sealed liquid cell.
[][]Plug to close both the slots of sealed liquid cell.
[][]For sample of liquid material which converted to solid (Nujol preparation) and measurement using NaCl demountable liquid cell:
[][]Put the cell window on the cell window holder and place the spacer of appropriate thickness on the cell window.
[][]Take 3 to 5 mg of sample with 2 to 3 drop of Nujol in a mortar and mix properly.
[][]Insert sample inside the spacer with a spatula. Put another cell window on the spacer and fit the metal plate on it.
[][]Fasten those plates with the four screws. Do not fix them with excessive force. It is enough to fasten them tightly by hand.
Operation Procedure (FTIR): calibration of FTIR
[][]Double click on “IRsolution” software icon.
[][]Press Measure
[][]Select FTIR Parameters in parameter window as follows:
[][]Data :
[][]Measurement Mode: % Transmitance ,
[][]Resolution: 2
[][]Apodization: Happ-Genzel
[][]No of Scans: 10
[][]Range (cm-1): Min: 400 cm-1 Max: 4000 cm-1
[][]Instrument:
[][]Beam: Internal
[][]Detector: Standard
[][]Mirror speed: 2.8
[][]More:
[][]Mode: Power
[][]Remove sample, if any, from the sample compartment.
[][]Click on Measurement then “Initialize” then “Yes”.
[][]After successful initialization, click BKG then “OK”. Wait till window on the screen is displayed as last Bkg.
[][]After ‘Bkg scan’, put the sample disk in the sample chamber, with the help of sample holder and close the chamber.
[][]Enter the Sample name, Batch No., Sample ID. in comment box and enter the data file location in Data file box.
[][]Click Sample to measure sample. Wait till the actual sample IR scan is displayed.
[][]Click on the spectrum window. Select Calculate peak table. Type threshold: 60.00.
[][]Click on Calc, OK
[][]Compare the Spectrum in computer screen. View the IR scan of standard, click on Manipulation 2 then “Purity” then “Yes”.
[][]Again view the IR scan of sample, click on Manipulation 2 “Purity” “Calc” “OK”.
[][]After comparison with standard and sample then click file and open file as required. Then click Window “Join visible” “Calc” “OK”.
[][]Purity index should be within 0 .9500 – 1.0000.
[][]A disc is rejected if visual examination shows lack of uniform transparency or when transmittance at about 2000 cm-1 (5 µm) in the absence of a specific absorption band is less than 60 % without compensation, unless otherwise prescribed in individual monograph.
[][]For printing of spectrum, click File “Print preview”. Select respective necessary template and then print.
[][]Close the parameter window and the Software window.
[][]Shutdown PC, switch off Printer.
[][]Power OFF Spectrophotometer.
[][]Take out the liquid cell from the instrument.
[][]Discard the disk /Nujol / liquid sample.
[][]Clean the holder and liquid cell with dry and clean tissue paper.
[][]In case of sealed liquid cell mop and wash with IPA or absolute Ethanol and in case of demountable liquid cell mop and wash with Chloroform or Carbon Tetrachloride.
[][]Keep them in specified desiccator.
[][]Rub mortar, pestle, barrel & plunger with tissue paper to make them free from dust.
[][]Mop them with a tissue paper soaked in absolute ethanol.
[][]Keep them in a desiccator for next operation.
[][]Record all analysis in operation log book as per Annexure-II.
Operation Procedure (ATR):
Running a sample:
[][]Turn on the computer and crick on the respective software on the desktop.
[][]Set the MIRacle/ MIRacle A Base Unit to the instrument.
[][]Before initialization, set FTIR Parameters in parameter window as follows:
[][]Data :
[][]Measurement Mode: % Transmitance ,
[][]Resolution:2
[][]Apodization: Happ-Genzel
[][]No of Scans: 10
[][]Range (cm-1): Min : 700 cm-1 Max : 4000 cm-1
[][]Click on Measurement then “Initialize” then “Yes”. Click “cancel” option for auto adjustment.
[][]Click on Measurement then “Auto adjustment (Fine)” then “Yes”.
[][]After autoadjustment, click BKG then “OK”. Wait till window on the screen is displayed as last Bkg.
[][]Set the clamp to the base. Place sample on the ZnSe crystal- Liquids -place a drop of sample on the ZnSe crystal. Place a small neat solid sample on the ZnSe crystal with a spatula. Adjust the knob of the ATR until the pressure reads 12.
[][]Make sure that no acid sample (pH below 5), strong alkaline sample (pH more than 9) or liquid which give damage to PTFE should not be place on ZnSe prism.
[][]Enter the Sample name, Batch No., Sample ID. in comment box and enter the data file location in Data file box. Click Sample to measure sample. Wait till the actual sample ATR scan is displayed.
Working up the Data:
[][]Compare the Spectrum in computer screen. View the ATR scan of standard, click on Manipulation 2 then “Purity” then “Yes”.
[][]Again view the ATR scan of sample, click on Manipulation 2 “Purity” “Calc” “OK”.
[][]After comparison with standard and sample then click file and open file as required. Then click Window “Join visible” “Calc” “OK”.
[][]For printing of spectrum, click File “Print preview”. Select respective necessary template and then print
Calibration:
[][]Calibrate the FTIR Spectrophotometer at 6 month’s frequency either by following procedure or as per supplier’s protocol:
[][]Power ON the Spectrometer. Wait for at least 1 hour. Power ON PC & Printer.
[][]Double click on “IRsolution” software icon. Press Measure
[][]Select FTIR Parameters in parameter window as follows:
[][]Data :
[][]Measurement Mode: % Transmitance ,
[][]Resolution : 2
[][]Apodization : Happ-Genzel
[][]No of Scans : 10
[][]Range (cm-1) : Min : 400 cm-1 Max : 4000 cm-1
[][]Instrument :
[][]Beam : Internal
[][]Detector : Standard
[][]Mirror speed : 2.8
[][]More :
[][]Mode : Power
[][]Remove sample, if any, from the sample compartment.
[][]Click on Measurement then “Initialize” then “Yes”.
[][]After successful initialization, click BKG then “OK”. Wait till window on the screen is displayed as last Bkg.
[][]After ‘Bkg scan’, place the Polystyrene film in the sample chamber and close the chamber.
[][]Enter the Polystyrene 1 and Wave number verification in comment box and enter the data file location in Data file box.
[][]Click Sample to scan the polystyrene film (0.05 mm thick). Wait till the actual sample IR scan is displayed.
[][]Click on the spectrum window. Select Calculate peak table. Type threshold: 65.00.
[][]Click on Calc OK
[][]For printing of spectrum, click File “Print preview”. Select respective necessary template and then print. Collect Polystyrene spectrum and peak table printed in two pages.
Limit of Wave Number Accuracy
[][]Check the maximum at following wave numbers (cm-1): 3060.0 (±1.5) cm-1, 2849.5 (±1.5) cm-1, 1942.9 (±1.5) cm-1, 1601.2 (±1.0) cm-1, 1583.0 (±1.0) cm-1, 1154.5 (±1.0) cm-1, 1028.3 (±1.0) cm-1.
[][]Resolution of apparatus
[][]Resolution due to transmittance:
[][]Record the Spectrum of a polystyrene film of approximately 35 µm thickness using Measuring Mode: % T.
[][]Measure the percentage transmittance at the transmission maximum at 2870 cm-1 (3.48 µm) and that at the transmission minimum at 2849.5 cm-1 (3.51 µm).
[][]Measure the percentage transmittance at the transmission maximum at 1589 cm-1 (6.29 µm) and that at the transmission minimum at 1583 cm-1 (6.32 µm).
[][]Record the results in calibration information sheet as per Annexure-I.
Acceptance criteria:
[][]The difference between the % transmittance at the transmission maximum at 2870 cm-1 (3.48 µm) and that at the transmission minimum at 2849.5 cm-1 (3.51 µm) must be greater than 18.
[][]The difference between the percentage transmittance at the transmission maximum at 1589 cm-1 (6.29 µm) and that at the transmission minimum at 1583 cm-1 (6.32 µm) must be greater than 10.
Resolution due to Absorbance:
[][]Record the Spectrum of a polystyrene film of approximately 35 µm thickness using Measuring Mode: Abs.
[][]Measure the absorbance at the absorption minimum at 2870 cm-1 and the absorption maximum at 2849.5 cm-1.
[][]Measure the absorbance at the absorption minimum at 1589 cm-1 and the absorption maximum at 1583 cm-1.
[][]Record the results in calibration information sheet as per Annexure-I.
Acceptance criteria:
[][]The difference between the absorbance at the absorption minimum at 2870 cm-1 and the absorption maximum at 2849.5 cm-1 is greater than 0.33.
[][]The difference between the absorbance at the absorption minimum at 1589 cm-1 and the absorption maximum at 1583 cm-1 is greater than 0.08.
[][]If the calibration is not within the specified standards, check the cleanliness of polystyrene film again and repeat the calibration.
Wavelength Repeatability Test:
[][]Repeat the Spectrum of the polystyrene film and Superimposed one after another of the absorption bands at 2849.5 cm–1 (3.51 µm), 1601.2 cm–1 (6.25 µm), 1028.3 cm –1 (9.72 µm).
[][]Acceptance Criteria: Compare the difference within 0.5% of the wave number scale and Purity index should be within 0.9500 – 1.0000
[][]Affix the label “CALIBRATED “on the equipment if all results are satisfactory. If the equipment found out of calibration limit affix “NOT FIT FOR USE” label and inform supplier.
Cleaning procedure:
[][]Clean the instrument with clean dry cloth daily.
[][]Remove your sample from the ATR, by using a cloth that is damped with acetone.
[][]Clean the plunger in the same way, if it made contact with sample.
[][]Never squirt (spray) acetone directly on the platform.
Annexure: Calibration of FTIR
Annexure-I: Calibration Information Sheet for FTIR Spectrophotometer
Annexure-II: Operation Logbook for FTIR Spectrophotometer