Flame Photometer – Standard Operating Procedure (SOP).pptx

This presentation on “Flame Photometer – Standard Operating Procedure (SOP)” provides a complete and practical understanding of one of the most widely used analytical instruments in biomedical, pharmaceutical, agricultural and environmental laboratories. It explains the principle, parts, working mechanism, step-by-step SOP, safety precautions, applications, advantages and limitations of Flame Photometer CL-378.

1. Flame Photometer (Standard operating procedure) Presenter: Dr. Debendra Suman (JR3) Dept. of Pharmacology & Therapeutics King George’s Medical University, Lucknow, UP, India-226003 Email id: [email protected]

2. Contents Introduction Specification of Flame Photometer CL 378 Parts of the Flame Photometer Principle Standard operating procedure Safety Precautions Advantages Disadvantages Applications Summary

3. Specific learning objectives By the end of this teaching learning session, my co-learners will be able to: Describe the parts of Flame photometer Explain principle of Flame photometer Explain the standard operating procedure Enumerate the application of Flame photometer Enumerate the advantages and disadvantages of flame photometer

4. Introduction It is an instrument that determines the concentration of alkali and alkaline earth metal ions based on the measurement of intensity of light emitted, when a metal is introduced into the flame Intensity of the flame tells about amount of the element present in the sample Wavelength of the colour gives information about the element Dr. Debendra Suman 1 16-05-2025

5. Specification of Flame Photometer CL 378 Used for detecting and measuring the concentration of certain metal ions, particularly Sodium (Na) , Potassium (K) , Lithium (Li) , Calcium (Ca) , and Barium (Ba) SENSITIVITY The minimum detectable concentration of each ion Na, K, Li : 0.5 ppm (parts per million) — very sensitive Ca : 15 ppm — less sensitive Ba : 50 ppm — least sensitive among the listed element 16-05-2025 Dr. Debendra Suman 2

6. RANGE This indicates the measurable concentration range Sodium (Na) : 0–100 ppm or 0–200 mEq Potassium (K) : 0–100 ppm or 0–100 mEq Lithium (Li) : 0–100 ppm or 0–10 mEq Calcium (Ca) : 15–100 ppm or 0–5 mEq Barium (Ba) : 50–1000 ppm 16-05-2025 Dr. Debendra Suman 3

7. Instrument ID: KG/PH/MBL/07 COMPRESSOR BUFFER TANK MAIN UNIT SOP 16-05-2025 Dr. Debendra Suman 4

8. MAIN UNIT : Flame Photometer CL 378 COMPRESSOR : It provides necessary air for the flame BUFFER TANK : P art of the air compressor system Act as a reservoir that stores compressed air, This air is then used to generate the flame H elps to maintain a consistent air pressure, ensuring a controlled flame for accurate analysis 16-05-2025 Dr. Debendra Suman 5

9. 16-05-2025 Dr. Debendra Suman 6 Parts of the Flame Photometer

10. CHIMNEY FLAME VEIW PORT SOLUTION SUCTION PORT STAGE TO KEEP BEAKER DISPLAY AND KEYBOARD AIR FLOW CONTROLLER GAS FLOW CONTROLLER AIR PRESSURE GAUGE FRONT VEIW 16-05-2025 Dr. Debendra Suman 7 LIGHT INDICATORS

11. BACK VEIW SWITCH AC POWER CABLE PORT GAS INLET AIR INLET 16-05-2025 Dr. Debendra Suman 8

12. LEFT SIDE VEIW PRINTER PORT SERIAL PORT FOR COMPUTER 16-05-2025 Dr. Debendra Suman 9

13. RIGHT SIDE VEIW CHIMNEY STAGE TO KEEP BEAKER 16-05-2025 Dr. Debendra Suman 10

14. INSIDE VEIW Burner Nebulizer Mixing Chamber Lens Filter Photodetector ( SILICON PHOTODIODE ) 16-05-2025 Dr. Debendra Suman 11

15. 1. Source of flame Burner is a source of flame and it can be maintained at a constant temperature Fuel: Natural gas (LPG) Oxidant: Air Temperature: 1900 C° 2. Nebulizer Used to send homogenous solution into the flame at a balance rate 3. Optical system Convex mirror transmits the light emitted from the atoms and focus the emission to the lens The convex lens helps to focuses the light on a filter 16-05-2025 Dr. Debendra Suman 12

16. 4. Photodetector It detect the emitted light and measures the intensity of the emitted light by the flame Here the emitted light is converted into the electrical signal 16-05-2025 Dr. Debendra Suman 13

17. Principle of Flame Photometer The compounds of the alkali and alkaline earth metals dissociates into atoms when introduced to high temperature Flame Some of these atoms further get excited to even higher levels, But these atoms are not stable at higher levels Hence the atoms emits radiations when returning back to the ground state These radiation lies in visible region of the spectrum 16-05-2025 Dr. Debendra Suman 14

18. M + X - MX MX M + X M* M Solution Gas Solid Residue Excited Atom Gas Ground State Solvent evaporates Solid vaporises Atomises vaporises Thermally excited Return to ground state by emitting its own characteristic wavelength 1 2 3 4 5 16-05-2025 Dr. Debendra Suman 15

19. The five steps describe the transformation of the sample into measurable light- Desolvation : Involves drying a sample Vaporization : The metal particles are dehydrated, also lead to evaporation Atomisation : Separation of all atoms Excitation : Atoms jumps to the higher energy state Emission : Higher energy state is unstable, comes back to lower energy state to gain stability. This atoms emits radiation 16-05-2025 Dr. Debendra Suman 16

20. Each of alkali and alkaline earth metals has specific wavelength 16-05-2025 Dr. Debendra Suman 17

21. Standard operating procedure Switch On The Compressor (10min) Switch On Main Instrument Adjust Air Control To Read (10 Psi) Release Gas Regulator Adjust Gas Regulator To Get Silent Blue Cone Flame 16-05-2025 Dr. Debendra Suman 18

22. Select Calibration Select Measurement Mode Select Element To Be Analysed Select Curve Fit Select No. Of Standards (3) Enter All Three Standard Value Concentrations One By One 16-05-2025 Dr. Debendra Suman 19

23. Feed DW Feed standard no 1 Feed DW Feed standard no 2 Feed DW Feed standard no 3 Feed DW 16-05-2025 Dr. Debendra Suman 20

24. Save This Standards Reading Select Sample Feed Sample Feed With DW Save Sample Reading Print Standards And Sample Readings 16-05-2025 Dr. Debendra Suman 21

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26. Safety Precautions Wear lab coat, gloves, and safety goggles Handle LPG gas cylinders and glass equipment's with care Ensure proper ventilation in the laboratory Ensure that the gas and air pipes have been connected correctly and there is no gas leaks before igniting Do not leave the flame photometer unattended during operation 16-05-2025 Dr. Debendra Suman 23

27. Do not look down the chimney when igniting or operating the equipment Allow sufficient time for the chimney to cool before handling Make sure a fire extinguisher is located nearby 16-05-2025 Dr. Debendra Suman 24

28. Advantages of flame photometer Method of analysis is very simple and economical Quick, convenient, selective and sensitive analysis Both qualitative and quantitative analysis very low concentration of metals in the sample can be determined Method can be used to estimate elements that are rarely analysed 16-05-2025 Dr. Debendra Suman 25

29. Disadvantages of flame photometer Accurate concentration of a metal ion in the solution cannot be measured Does not provide information about the molecular structure of the metal Only liquid sample may be used Elements such as carbon, hydrogen, and halides cannot be detected due to their non-radiating nature 16-05-2025 Dr. Debendra Suman 26

30. Applications of flame photometer Determination of Bio-Medical Investigations: Na & K in body fluids Na, K and Li electrolytes in Serum Ba in Faecal matter (from Barium meal, Barium swallow etc.) Concentration of Na+ and K+ ions in the blood serum samples Pharmaceuticals: Na, K, Са nutrient solutions for cultivation of Antibiotics Pollution Monitoring & Control : Na, K & Ca in Natural, Spring, River, Sea boiler feed, Rings and Waste Water 16-05-2025 Dr. Debendra Suman 27

31. 4. Food and Agriculture: Na & K content in Soils, Plant materials, Food & Beverages, Ca in Wine. 5. Research Laboratories and Industries: Alkali metal in Pastes, Soda Glass, Ceramic and Chemical Industries, Na, K and Ca in Cement and its raw materials. 16-05-2025 Dr. Debendra Suman 28

32. Summary Definition : It is used to determine the concentration of alkali and alkaline earth metal ions based on the intensity of light emitted when a sample is introduced into a flame. Working Principle : Atoms get excited in the flame and emit light when returning to their ground state. The intensity of this light correlates with the concentration of the element Commonly Detected Elements : Sodium (Na), Potassium (K), Lithium (Li), Calcium (Ca), and Barium (Ba) are typically measured Parts of Instrument : Includes burner, nebulizer, mixing chamber, lens, filter, photodetector, and a display unit, along with air and gas flow controllers 16-05-2025 Dr. Debendra Suman 30

33. 5. Process Steps : De-solvation Vaporization Atomization Excitation Emission 6. Advantages : Simple, quick, and cost-effective Sensitive to low metal concentrations Suitable for both qualitative and quantitative analysis 16-05-2025 Dr. Debendra Suman 31

34. Specific learning objectives achieved By the end of this teaching learning session, my co-learners are able to: Describe the parts of Flame photometer Explain the principle of Flame photometer Explain the standard operating procedure Enumerate the application of Flame photometer Enumerate the advantages and disadvantages of flame photometer 16-05-2025 Dr. Debendra Suman 32

35. References Elico Flame Photometer CL 378 User’s Manual Gandhi K, Sharma N, Gautam PB, Sharma R, Mann B, Pandey V. Atomic absorption spectroscopy and flame photometry. In Advanced analytical techniques in dairy chemistry 2022 Jan 29 (pp. 219-247). New York, NY: Springer US. Ahluwalia VK. Flame Photometry, 2023, p. 505–17. https://doi.org/10.1007/978-3-031-38355-7_35 . Powell FJN. The Determination of Calcium in Biological Fluids by Flame Photometry. Journal of Clinical Pathology. 1953 Nov 1;6(4):286–9. Available from: https://jcp.bmj.com/content/6/4/286 https://youtu.be/vvp0OAtQ2pY?si=MVvcuaxJIgXzZGx6 https://youtu.be/5zFEQgSyfI0?si=urB9L7U4kqoxzDUA 16-05-2025 Dr. Debendra Suman 33

36. THANK YOU 16-05-2025 Dr. Debendra Suman 34

37. Questions What is the principle behind the operation of a flame photometer? Which parts are essential for the functioning of the Flame Photometer CL 378? List the advantages of using a flame photometer in laboratory analysis? What are the applications of flame photometer? 16-05-2025 Dr. Debendra Suman 35