SOP Title: Operation, Cleaning and Calibration of Fourier Transform Infrared Spectrophotometer
- Objective:
To lay down the procedure for Operation, Cleaning and Calibration of Fourier Transform Infrared Spectrophotometer.
- Scope:
This SOP is applicable for Operation, Cleaning and Calibration of Fourier Transform Infrared Spectrophotometer located in Quality Control Laboratory.
- Responsibility:
- Quality Control Department: To prepare and review the SOP. To follow the procedures laid down for Operation, Cleaning and Calibration of Fourier Transform Infrared Spectrophotometer as per this SOP.
- Engineering Department: To carry out preventive maintenance as per schedule and procedure defined.
- Quality Assurance Department: To review and approve the SOP and Annexures.
- Accountability:
Head Quality Control Department, Head Quality Assurance Department
- Procedure:
| 5.1 | Safety Precautions/Instructions: | ||||||||||||||||||
| 5.1.1 | Ensure all the electrical connections are properly earthed. Ensure the power plug of FTIR is fixed in the socket properly. | ||||||||||||||||||
| 5.1.2 | Wear personal protective equipments such as hand gloves, nose mask during operation of instrument. | ||||||||||||||||||
| 5.1.3 | Ensure that the ON/OFF switch is working properly by switching “ON” FTIR. | ||||||||||||||||||
| 5.2 | Precautions: | ||||||||||||||||||
| 5.2.1 | Do not put off the instrument mains, it should be kept on continuously for 24 hours. In case of a power failure turn the instrument to power ‘OFF’ position.
Accessories required for IR must be stored in a dry box or desiccators. |
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| 5.2.2 | When the power is resorted put the instrument ‘ON’ using switch on the right hand side of the instrument allow an equilibration time of 2 hours before using the instrument. | ||||||||||||||||||
| 5.2.3 | Check the temperature and relative humidity. The temperature should be between15 to 30°C and humidity should not be more than 60 %. | ||||||||||||||||||
| 5.2.4 | Use IR Grade solvents and Potassium bromide. | ||||||||||||||||||
| 5.2.5 | Interference of CO2 can be minimized by avoiding crowding near IR Instrument. | ||||||||||||||||||
| 5.3 | Operational Procedure: | ||||||||||||||||||
| 5.3.1 | Check the validity of Calibration before starting any operations. | ||||||||||||||||||
| 5.3.2 | To switch on the instrument, connect the plug to 230 Volts stabilized power supply. | ||||||||||||||||||
| 5.3.3 | Put ‘ON’ the MAINS switch, the “orange” light on the extreme front side of the instrument will glow. This indicates that the dehumidifier is on and working. | ||||||||||||||||||
| 5.3.4 | Turn the power ON of FTIR by pressing the button on extreme front right side of the instrument. It will show green light glowing on left side of button. | ||||||||||||||||||
| 5.3.5 | Turn the power on the computer, it will show green light of computer glowing and monitor will go to initialization mode and runs window program and stops at start window. | ||||||||||||||||||
| 5.3.6 | Turn on the printer, it will indicate green light. | ||||||||||||||||||
| 5.3.7 | Double click on ‘IR SOLUTION’ on the desktop. It will load IR solution on PC and will open the IR SOLUTION window. | ||||||||||||||||||
| 5.3.8 | The window comprises of the following items:
Title bar: The title bar displays the name of the software and selected function tabs. Menu bar: The menu bar displays the menu names like File, Edit, Measurement, AIM, Graph, Admin, Manipulation1, Manipulation 2, Search, Quant, Macro, Print, 3Dim, Window, Environment, Help. |
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| 5.3.9 | Tool bar: Displays commonly used commands as icons. Instead of selecting the command from a menu, click the desired icon for faster and more convenient operation.
Tree view window: The ‘Tree view window’ displays scan parameters, peak tables, data history etc. saved together with data. Function tabs: The function tabs changes the operation menus such as Measure, View, Manipulation, Search, Quant, AIM, Print form, Macro. “Status” window: displays user name, the work history, error messages, etc. Work space: The work space displays spectra and various data. |
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| 5.3.10 | Check that the instrument is on measure mode. Click on ‘measurement’ key then Click on “INITIALISE”. | ||||||||||||||||||
| 5.3.11 | Window showing previous background data will get displayed. It will ask whether to remove the data. Click on Yes. The instrument will get initialized. | ||||||||||||||||||
| 5.3.12 | After initialization 2 icons in green colour will appear on the right side of the window which is interface and mirror. It shows that the instrument is ready to use. | ||||||||||||||||||
| 5.3.13 | Below these parameters like More, Files, Advanced, Data, Instrument is displayed. | ||||||||||||||||||
| 5.3.14 | For routine analysis click on ‘Data’ and set the parameters as follows:
Measuring Mode: % T Apodization: HAPP- Genzel No of scans: 10 Resolution: 4.0 Range (cm-1): Minimum = 400 and Maximum = 2000 |
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| Blank and Sample disc preparation with KBr Press: | |||||||||||||||||||
| 5.3.15 | For operation of KBr Press refer SOP ‘Operation, Cleaning and Calibration of KBr Press’. | ||||||||||||||||||
| 5.3.16 | Check and ensure that the KBr Press is clean and suitable for use. If not clean it with soft cloth duster. | ||||||||||||||||||
| 5.3.17 | Take approx 1 to 3 mg of the sample and 100 to 300 mg of potassium bromide in clean and dried granite mortar and pestle. | ||||||||||||||||||
| 5.3.18 | Grind the potassium bromide with the sample in order to mix it thoroughly. | ||||||||||||||||||
| 5.3.19 | Assemble the dye by inserting the barrel into the base; insert the shiny stainless steel disc into the barrel with the polished side up. | ||||||||||||||||||
| 5.3.20 | Transfer the powder from the mortar into the barrel with the help of clean and dry stainless steel spatula or butter paper in such a way that the surface of the powder in the barrel is levelled and uniformly spread over the barrel. Now insert the plunger slowly and carefully. | ||||||||||||||||||
| 5.3.21 | Place the dye assembly in the centre of the KBr press. Rotate the wheel on top of the KBr press clockwise until it holds the dye assembly tightly. | ||||||||||||||||||
| 5.3.22 | Ensure that the dye assembly is tightly placed between the top and bottom of the KBr press. | ||||||||||||||||||
| 5.3.23 | Close the valve present at the side of the pressure gauge in the clockwise direction and increase the pressure by moving the piston to about 10 tones and wait for about 2 minutes. | ||||||||||||||||||
| 5.3.24 | Release the pressure gently by opening the same valve in the anticlockwise direction. | ||||||||||||||||||
| 5.3.25 | Now rotate the wheel in anticlockwise direction and remove the die assembly carefully. | ||||||||||||||||||
| 5.3.26 | Remove the Disc by separating the barrel from the base. Gently tap the plunger on the inverted side so that the steel pellet and KBr disc can be removed from the top. Check that the pellet prepared is transparent and uniform. | ||||||||||||||||||
| 5.3.27 | For preparation of blank KBr disc follow the same procedure without the sample. | ||||||||||||||||||
| 5.3.28 | The KBr disc/pellet is now ready to be scanned. | ||||||||||||||||||
| Scanning of KBr disc. | |||||||||||||||||||
| 5.3.29 | Transfer the blank disc into the sample holder with the help of butter paper. Put the sample holder into the sample compartment of the instrument. | ||||||||||||||||||
| 5.3.30 | On the left hand side of the screen window showing the dialog box as comment and Data file will be displayed. For measuring the background scan enter ‘BKG’ in comment as well as in data file. | ||||||||||||||||||
| 5.3.31 |
Click on “BKG”, then the dialog box will appear as: |
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| 5.3.32 | Click on OK. The background measurement will be started. The progress in measurement and the number of scans will be displayed on the status bar at the left hand corner of the screen. | ||||||||||||||||||
| 5.3.33 | After the measurement is over the blank spectrum will be directly saved in the ‘VIEW’ window and the ‘VIEW’ window will automatically open on the screen. | ||||||||||||||||||
| 5.3.34 | Remove the sample holder containing blank disc from the sample holder. Put the sample disc into the sample holder and insert it back into the sample compartment. | ||||||||||||||||||
| 5.3.35 | In the comment window enter the information related to spectra such as sample name, Batch no., A. R. No. and Container no. (For individual identification test) or composite sample. | ||||||||||||||||||
| 5.3.36 | In the ‘Data file’ box, type the file name on which the spectrum is to be saved. The IR solution saves all the measured spectra to the hard disc as soon as measurements are finished. | ||||||||||||||||||
| 5.3.37 | Click the [….] button at the right end of the data file to select the destination. Set and enter the destination (hard disk and folder) and the file name here. All the files get saved as “smf “files. | ||||||||||||||||||
| 5.3.38 | Put the check mark on ‘Auto increment’ option provided on the right side of the data file name to automatically update the file name by putting a number at the end of the file name in every measurement. | ||||||||||||||||||
| 5.3.39 | Put the check mark on ‘View background’ to display the measured power spectrum for confirmation when the background measurement is finished. | ||||||||||||||||||
| 5.3.40 | Then click on sample. The sample measurement will be started. | ||||||||||||||||||
| 5.3.41 | The progress in measurement and the number of scans will be displayed on the status bar at the left hand corner of the screen. | ||||||||||||||||||
| 5.3.42 |
During measurement, measurement can be paused and restarted by clicking on “STOP” button in the measurement window. Then the dialog box will open as:
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| 5.3.43 | To stop measurement and display the spectrum: Click the [Stop] button. | ||||||||||||||||||
| 5.3.44 | To stop measurement and abandon data acquired: Click the [Abort] button. | ||||||||||||||||||
| 5.3.45 | To continue measurement: Click the [Cancel] button. | ||||||||||||||||||
| 5.3.46 | After the measurement is over the spectrum will be directly saved in the ‘VIEW’ and the ‘VIEW’ window will automatically open on the screen. | ||||||||||||||||||
| 5.3.47 | ‘VIEW’ window will show measured spectrum in the upper and lower windows. | ||||||||||||||||||
| 5.3.48 | The upper window is called the “Overview” window and the lower window is called “Zooming” window. | ||||||||||||||||||
| 5.3.49 | For overlaying the two spectrums one of sample and other of standard, go to “Window” click on “Join visible” and for splitting the two spectra’s again go to “Window” and click on “Split”. | ||||||||||||||||||
| 5.3.50 | To display previous spectra from the hard disk, click on ‘File’ on the menu bar and select ‘open’. Then the dialog box will open. | ||||||||||||||||||
| 5.3.51 | On the dialog box select a drive and folder in which a spectrum to be open is saved and then select the file name of the spectrum and click ‘OPEN’. | ||||||||||||||||||
| 5.3.52 | To open multi spectra at one time, use the [Shift] or [Ctrl] key. | ||||||||||||||||||
| For Creation of Print Report Template: | |||||||||||||||||||
| 5.3.53 | Go to print form option. Click on template .It will show 4 options i.e. Body, Header, Footer, Sizes. | ||||||||||||||||||
| 5.3.54 | Click on each option & select / type the parameters, which are required on report, with the help of the window showing the parameters like date, time, file name etc. | ||||||||||||||||||
| 5.3.55 | Then save the template. For printing of the report first select the template and then give the print command to the computer. | ||||||||||||||||||
| 5.3.56 | For giving the identification of the spectrum annotations can be given. For that go to ‘Graph’ then click on’ Annotations’. | ||||||||||||||||||
| 5.3.57 | Type the batch number of sample or standard and then click on “OK”. The name will appear on the spectrum. | ||||||||||||||||||
| 5.3.58 | Update the “Instrument/Equipment Usage Log Book” as per SOP “Calibration of Instruments and Equipments”. | ||||||||||||||||||
| 5.4 | Cleaning Procedure: | ||||||||||||||||||
| 5.4.1 | Clean the sample compartment with soft nylon brush before and after use. | ||||||||||||||||||
| 5.5 | Calibration Procedure: | ||||||||||||||||||
| Frequency: Quarterly or after carrying out any major maintenance work. | |||||||||||||||||||
| 5.5.1 | Generate the A. R. No. on the “Instrument/Equipment Calibration Register” as per SOP and give requisition to QA Department for calibration formats of instruments on the due date of calibration. | ||||||||||||||||||
| 5.5.2 | For issuance of calibration report refer SOP “Control of documents, Preparation, Approval, Issuance and Maintenance. | ||||||||||||||||||
| 5.5.3 | Perform the calibration of instrument as per Annexure and procedure mentioned below. | ||||||||||||||||||
| 5.5.4 | Calibration of FTIR is performed for below parameters:
1. Control of Resolution Performance 2. Verification of the Wave number scale 3. Reproducibility of the wave number |
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| 5.5.5 | First follow the procedure from point 5.3.1 to 5.3.13 and then proceed. | ||||||||||||||||||
| 5.5.6 | For calibration click on ‘Data’ and set the parameters as follows: | ||||||||||||||||||
| 5.5.7 | Maximize “IR SOLUTION” window. Set the parameters as follows:
Measuring Mode : % T Apodization : HAPP- Genzel No. of scans : 45 Resolution : 2.0 Range : Minimum = 4700, Maximum = 400 |
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| 5.5.8 | On the left hand side of the screen window showing the dialog box as comment and Data file will be displayed. | ||||||||||||||||||
| 5.5.9 | For measuring the background scan enter ‘BKG’ in comment as well as in data file. Click on “BKG”, then the dialog box will appear as:
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| 5.5.10 | Click on OK. The background measurement will be started. The progress in measurement and the number of scans will be displayed on the status bar at the left hand corner of the screen. | ||||||||||||||||||
| 5.5.11 | After the measurement is over the blank spectrum will be directly saved in the ‘VIEW’ window and the ‘VIEW’ window will automatically open on the screen. | ||||||||||||||||||
| 5.5.12 | After completing background scan, click on the pull down menu of [Measurement] on the menu bar and select [JP Validation]. | ||||||||||||||||||
| 5.5.13 | Select ‘Instrument Model” and then click OK. | ||||||||||||||||||
| 5.5.14 | Now again window [A] will open, click on [Settings], then window [B] will open, click on [Language]. | ||||||||||||||||||
| 5.5.15 | Select English language and click OK. | ||||||||||||||||||
| 5.5.16 | Again window [A] will open, click on [Settings], then window [B] will open, click on [Change-Beam] and then the following window will open.
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| 5.5.17 | Select Change-Beam Unusing and click OK. | ||||||||||||||||||
| 5.5.18 | Change temperature, relative humidity. | ||||||||||||||||||
| 5.5.19 | After changing the parameters, click OK. | ||||||||||||||||||
| 5.5.20 | Following window will open.
Lab control Macro Language
Remove sample from sample compartment.
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| 5.5.21 | Remove if any sample is present in the sample compartment and click on OK. | ||||||||||||||||||
| 5.5.22 | Wait till the background scan gets complete. | ||||||||||||||||||
| 5.5.23 | After the completion of scanning, following window will open.
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| 5.5.24 | Insert polystyrene film into sample compartment. Click on OK. | ||||||||||||||||||
| 5.5.25 | Wait till the sample gets scanned. | ||||||||||||||||||
| 5.5.26 | After the completion of scanning, the results will be automatically printed out. | ||||||||||||||||||
| 5.5.27 | The following window will be displayed on the screen.
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| 5.5.28 | Close all windows. Remove the polystyrene film from the sample compartment and keep it in a safe place. | ||||||||||||||||||
| 5.5.29 | Enter the details in calibration report as per Annexure. Ensure that the results obtained, comply with the acceptance criteria. | ||||||||||||||||||
| 5.5.30 | Update the “Instrument/Equipment Usage Log Book”, “Annual Calibration Schedule” and “Monthly Calibration Schedule” as per SOP “Calibration of Instruments and Equipments”. | ||||||||||||||||||
| 5.5.31 | Affix the “Calibration Status Label” on the Instrument as per SOP “Calibration of Instruments and Equipments”. | ||||||||||||||||||
| 5.5.32 | If calibration activity is outsourced, the values are to be transcribed in the Annexure, and external party certificate to be attached. | ||||||||||||||||||
| 5.5.33 | If any breakdown occurs, follow procedure as per SOP “Calibration of Instruments and Equipments”. | ||||||||||||||||||
- Definitions / Abbreviations:
- Definitions:
- Breakdown: Any activity leading to operation of instrument/equipment other than the set parameters or unusual sound or vibration observed in the instrument/equipment.
- Calibration: Tests performed to ensure that the instrument/equipment is functioning as per set parameters. These tests are performed as per predefined frequency and also after any major repair or replacement of parts.
- Preventive Maintenance: Maintenance activity performed to ensure that the instrument/equipment will function smoothly and to avoid breakdowns. These activities are performed as per predefined frequency.
- Abbreviations:
| Abbreviation | Expansion |
| SOP | Standard Operating Procedure |
| QC | Quality Control |
| QA | Quality Assurance |
| FTIR | Fourier Transform Infra Red |
| mg | milligram |
| °C | Degree Centigrade |
| % | Percentage |
| BKG | Blank |
| IR | Infra Red |
| CO2 | Carbon dioxide |
| cm-1 | Centimetre inverse |
| Calc | Calculation |
| KBr | Potassium Bromide |
| i.e. | That is |
| A. R. No. | Analytical Reference Number |
Calibration Report
- Control of Resolution Performance:
| Parameters | Observations | Acceptance Criteria |
| The difference between the percentage transmittance at the transmission maximum at 2870 cm-1 and that at the transmission maximum at 2849.5 cm-1. | Should be greater than 18% | |
| The difference between the percentage transmittance at the transmission maximum at 1589 cm-1 and that at the transmission maximum at 1583 cm-1. | Should be greater than 10% |
- Verification of the Wave number scale:
Carry out calibration using standard polystyrene film and verify the following Wave numbers (in cm-1).
| Std. Wavelength | Observed Wavelength (cm-1) | Acceptance Criteria |
| 3060.0 cm-1 | 3059.0 – 3061.0 cm-1 | |
| 2849.5 cm-1 | 2848.5 – 2850.5 cm-1 | |
| 1942.9 cm-1 | 1941.9 – 1943.9 cm-1 | |
| 1601.2 cm-1 | 1600.2 – 1602.2 cm-1 | |
| 1583.0 cm-1 | 1582.0 – 1584.0 cm-1 | |
| 1154.5 cm-1 | 1153.5 – 1155.5 cm-1 | |
| 1028.3 cm-1 | 1027.3 – 1029.3 cm-1 |
3) Reproducibility of the Wave number:
| Wave number (cm-1) | No.1(cm-1) | No.2(cm-1) | Error(cm-1) | Tolerance(cm-1) |
| 2849.5 | ± 5.0 | |||
| 1601.2 | ± 1.0 | |||
| 1028.3 | ± 1.0 |
The results Complies / Does not complies as per acceptance criteria.
Performed By: ________________ Date: ________________ Checked By: ______________ Date: ______________
*Note – Ready to use SOP available in “DOWNLOAD” Section.