Qualification of GC & FTIR

introduction
qualification
FTIR
GC

1. QUALIFICATION OF GC & FTIR Pharmaceutical Validation Mehul H Jain M. Pharmacy 1 st Sem Pharmaceutical Analysis

2. INTRODUCTION VALIDATION: It is a act of demonstrating and documenting that the process operates effectively and reproducibly to produce a product meeting its predetermined specifications and quality attributes. The aim of validation is to show that the critical steps are under control and lead continuously to the desirable quality. CALIBRATION: Calibration is a comparison between measurements – one of known magnitude or correctness made or set with one device and another measurement made in as similar a way as possible with a second device.

3. QUALIFICATION: Qualification is an act or process to assure something complies with some conditions, standard or specific requirements.

4. Design Qualification: Documented evidence which shows that the plant design agrees with the design specifications of the customer. Installation Qualification : Written evidence is given that all parts of equipment are installed according to the equipment supplier’s and purchase specifications . Operational qualification : Documented evidence which shows that all parts of the plant and equipment work within their specifications and process parameters. Performance Qualification : Provides documented evidence that all parts of a plant and other processes produce products of specified quality under conditions of normal production for a longer period of time.

5. FTIR Fourier Transform Infrared Spectroscopy ( FTIR ) is a technique which is used to obtain an infrared spectrum of absorption, emission, photoconductivity or Raman scattering of a solid, liquid or gas. An FTIR spectrometer simultaneously collects spectral data in a wide spectral range. This confers a significant advantage over a dispersive spectrometer which measures intensity over a narrow range of wavelengths at a time.

6. CALIBRATION WAVE NUMBER PRECISION: This is performed for substances with well known peak wave number(s) position such as carbon dioxide, water vapor, polystyrene ,ammonia. Test is performed to know whether the exact peak wave numbers are shown at that time of validation. Thus the result is obtained from the difference between the peak wave numbers position for a substance with a well known peak wave numbers and the values indicated by the system. 0% TRANSMITTANCE: A sample which do not allow the transmission of light is measured in order to investigate the 0% transmittance. This test thus can be used to find out error caused by stray light and secondary emission spectra.

7. 100% TRANSMITTANCE: This is investigated by performing analysis with out a sample. By performing analysis with out the sample 100% transmittance can be investigated. LINEARITY OF CURVE: A calibration curve for the % transmittance and the concentration is created and the linearity of the inspected. REPRODUCIBILITY: A stable sample is measured twice with in a short period and confirmed whether the variation in the measurement values such as wave numbers and transmittance are obtained.

8. VALIDATION To perform FTIR validation and to confirm that it is operating properly, diverse IR inspection was performed by measuring the spectra of polystyrene film. INSTALLATION OF VALIDATION PROGRAM: Software validates the Shimadzu Fourier transform infrared spectrophotometer IRPrestige-21/IRAffinity-1/FTIR-8000 series by the IR solution software to control them and data processing. The program is automatically installed when the IR solution is installed, therefore the Validation program does not need to be separately installed. PERFORMANCE QUALIFICATION: POWER SPECTRUM: Power spectrum gives the plot of portion of signal's power(energy per unit time) falling with in the given frequency bins. This test estimates the intensity of power spectrum at a specified wave numbers. When the measured intensity is equal to or larger than the criterion value, the test is passed.

9. ACCEPTANCE CRITERIA: Wave number (cm-1) Standard Value FTIR-8400S. 4600 10% or min of max 4000 25% or min of max 3000 50% or min of max Power max value 50.0 700 10% or min of max 500 2% or min of max 403 0.5% or min of max 351 0.01% or min of max

10. RESOLUTION: The resolution is checked by recording the spectrum of polystyrene film of approximately 35µm in thickness. The difference between percentage transmittance at the absorption maximum A at 2870 cm -1 and absorption minima B at 2849.5 cm -1 must be greater than 18. The difference between percentage transmittance at the absorption maximum C at 1589 cm -1 and absorption minima D at 1583 cm -1 must be greater than 12. WAVE NUMBER ACCURACY: The wave number scale is usually calibrated by the use of several characteristic wave numbers of a polystyrene film. 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 The software then judges whether the values are within the allowable range.

11. The program labels the results “PASS” if all the peak numbers are within the range. In case of dispersive spectrophotometer the permissible level of frequency at 1601.2 cm -1 and at 1028.3 cm -1 should be with in +/- 2.0 cm -1 . WAVE NUMBER REPRODUCIBILITY: This program specifies three points to measure the peak wave numbers. Then it obtains the actual peak wave numbers at each point by measuring the polystyrene film twice. It should satisfy 5 cm -1 around 3000 cm -1 of polystyrene absorption wave number, 1 cm -1 around 1000 cm -1 . The software determines whether the differences between each of two measurements are within the allowable range and it labels the result PASS if they are with in the range. EP 4.0 doesn’t include this inspection.

12. TRANSMITTANCE REPRODUCIBILITY: This program specifies peak wave number at three points and the transmittance at each point is measured is twice. The transmittance reproducibility should satisfy 0.5%T when the several points of polystyrene absorption from 3000 cm -1 to 1000 cm -1 are measured twice. Then it is determined whether the differences between the two data are within the allowable range and it labels the result PASS if they are with in the range. AS PER ASTM E1421-94 LEVEL ZERO: This soft ware complies describes with in the description in the ASTM (American Society for Testing and Materials). The FTIR abnormalities or large changes over short term and long term is assessed by these tests. The three parameters checked by this program are: Energy spectrum test. 100% line test. Polystyrene test.

13. ENERGY SPECTRUM TEST: Power spectra obtained in the inspection are compared with reference data and the spectra are checked for changes over long periods. ONE HUNDRED PERCENT LINE TEST: 100% T line spectra are calculated for power spectra & are measured continuously in inspection and the spectra are checked for changes over short periods. POLYSTYRENE TEST: Evaluation is performed using differences between spectra obtained for polystyrene film in inspection and the stored reference data. When the differences are within the standard, "pass” results. All of the above furnished data should be represented in Validation Report.

14. GAS CHROMATOGRAPHY LEVEL-I : SELECTION OF INSTRUMENTS & SUPPLIERS: At level I of the qualification of a GC equipment (selection of instruments and suppliers). It is recommended to select a manufacture of GC that can satisfy the needs of the laboratory and works under ISO 9001 certification. LEVEL-II OF EQUIPMENT QUALIFICATION: INSTALLATION & RELEASE FOR USE: It is recommended to check all requirements set during the selection of the instrument, and calibration should be performed before putting into service by an accredited external service supplier. (or) Internally by appropriately qualified personnel, using certified reference buffers according to an approved procedure.

15. LEVEL-III: PERIODIC & MOTIVATED INSTRUMENT: Instrument Mode Parameter to be checked Typical Tolerance Limit 1. Inlet System i ). Injector leak test. Pressure drop ≤ 1.5 pKa within 5mins. ii). Pressure flow accuracy & stability. Covered by over all test-1. iii). Repeatability of injection (overall test-1) In split mode RSD ≤ 3.0 % In split less mode RSD ≤ 3.0 % iv). Injector temp. accuracy & stability. Covered by overall test-2. v). Carry over (overall test-3). ≤ 0.2 % 2. Oven i ). Repeatability of oven temp. characteristics. Covered by overall test-2. 3. FID Detector i ). Linearity (overall test-3) r 2 ≥ 0.999 ii). Constant detector response Covered by overall test-1 or 2.

16. INLET SYSTEM: INJECTOR LEAK TEST METHOD: The leak test is carried out according to the procedure laid down in the instrument manual or by the built in automatic leak check procedure of the instrument. Otherwise use the test described below: Disconnect the column from the injector and close the injector outlet with a sealed cap. Close the septum purge and the bypass. Adjust the flow and pressure controller to the maximal possible value of the pressure gauge. Adjust the flow controller to zero. Read the pressure after 1 minute and record the value. Record the pressure after 5 minutes. LIMITS: Pressure drop ≤ 15 kPa within 5 minutes.

17. INLET PRESSURE FLOW ACCURACY AND STABILITY: A direct measurement of these parameters was not deemed practical or necessary, but the optimal conditions of flow/pressure can be verified by the overall test 1. LIMITS: Refer to overall test 1. REPEATABILITY OF INJECTION: The verification of this parameter is covered by the overall test 1. This test is to be performed in both split and split less mode. LIMITS: Refer to overall test 1. INJECTOR TEMPERATURE ACCURACY AND STABILITY: Due to the fact that the temperature cannot be reliably measured without opening and modifying the system and due to the difficulties of introducing a probe inside this module, the verification of this parameter is considered to be covered by the overall test 2. LIMITS: Refer to overall test 2.

18. INJECTOR CARRY OVER: After having injected the solutions for the linearity test of the FID detector, in increasing order, inject the blank and measure the peaks that correspond to the major peaks (= analytes ) in the linearity solutions. The verification of this parameter is covered by the overall test 3. LIMITS: Refer to overall test 3. OVEN: REPEATABILITY OF OVEN TEMPERATURE CHARACTERISTICS: Due to the fact that the temperature cannot be reliably measured without opening and modifying the system conditions and that even when introducing a probe inside the oven, its location would not reflect the real temperature conditions at all points, the verification of this parameter is covered by the overall tests 2A and 2B. LIMITS: Refer to overall test 2.

19. FID DETECTOR: FID DETECTOR LINEARITY: Increasing amounts of analyte are injected and a linear response should be obtained. The verification of this parameter is covered by the overall test 3. LIMITS: Refer to overall test 3. CONSTANT FID DETECTOR RESPONSE: The proper and reproducible functioning of the FID can be demonstrated by checking the peak areas obtained from a pre- defined standard solution. The verification of this parameter is covered by the overall test 1 or 2. LIMITS: Refer to overall test 1 or 2.

20. OVERALL TEST-1 The overall test 1 covers the following parameters: Pressure/flow accuracy and stability in the inlet system: Retention time repeatability Repeatability of injection: peak area precision In split mode In split less mode The test may be combined with overall test 3. SPLIT MODE: Test solution: 1-octanol in n-hexane 1% (v/v). Settings: Column: SPB-1 (30m x 0.32mm ID x 0.25µm film) Carrier gas: Helium Velocity: 25cm/sec Split: 1:100

21. Injection: 1µl Injector temperature: 220°C Oven temperature: 100°C isotherm Detector temperature: 300°C Runtime: 8 min Retention time of 1-octanol: about 5 mins . SPLIT LESS MODE: Stock solution: 1-octanol in n-hexane 1% (v/v) Test solution: Dilute 10 ml of the stock solution with n- hexane to 100 ml (corresponds to 1µl/ml of 1-octanol in n- hexane). Settings: Column: SPB-1, 30m, 0.32mm ID, 0.25µm film Carrier: Helium Velocity: 30cm/sec Split less injection: purge valve closed during 2 min Injection: 0.2µl of the test solution

22. Injector Temperature: 220°C Oven Temperature: Initial 60°C for 4 min, 15°C/min. up to 135°C, final time 1min. Detector temperature: 300°C Runtime: 9.5 min Retention time of 1-octanol: about 8 mins . METHOD: Carry out 6 consecutive injections of the test solution and calculate the RSD of the different peak areas and retention times. LIMITS: Retention time repeatability: The RSD of the retention times should be ≤ 2.0% Peak area precision (split and split less mode): The RSD of the peak areas should be ≤ 3.0%

23. OVERALL TEST 2 The overall test 2 covers the following parameters: Injector, oven and detector temperature accuracy and stability: retention time repeatability Two alternative tests are proposed. OVERALL TEST 2A: TEST SOLUTION: 0.035 ml 1-octanol 0.035 ml 2-octanone 0.035 ml 2,6-dimethylanilin 0.035 ml n- tridecane 0.035 ml n- tetradecane 35 mg n- eicosane Dissolved in 50 ml Dichloromethane

24. SETTINGS: Column: SPB-1 (30m x 0.32mm ID x 0.25µm film) Carrier gas: Helium Velocity: 25 cm/s Split: 1:100 Injection volume: 1 µl Injector temperature: 220°C Detector: FID Detector temperature: 300°C Gradient program: 60°C (4 min), 5°C/min, 270°C (3 min) METHOD: Inject the solution twice and calculate the relative retention times in relation to n- eicosane (RRT = 1). The following table shows the approximately expected relative retention times.

25. LIMITS: The RSD of each RRT from two consecutive injections should be ≤ 1.0% OVERALL TEST 2B: TEST SOLUTION : 1.0% (W/W) n- Nonane and Hexadecane in Tetradecane . SETTINGS: Column: Ultra-1 (25m x 0.32mm ID x 0.52µm film) Injection volume: 1 µl Solvent: Tetradecane Oven temperature: 110°C Gradient program: 110°C, 20°C/min, 180°C (final time: 3.5min) Detector temperature: 250°C