The document provides details on qualification and validation procedures for gas chromatography (GC) and Fourier transform infrared spectroscopy (FTIR) systems used in pharmaceutical analysis. It describes calibration tests for both instruments, including tests to check wave number precision and reproducibility, linearity, and temperature accuracy and stability. Validation tests are also outlined, such as resolution checks and tests to evaluate performance against acceptance criteria over time as specified in guidance documents. The document provides a thorough overview of qualification processes for critical analytical instruments.

1. QUALIFICATION OF GC &
FTIR
Pharmaceutical Validation
Mehul H Jain
M. Pharmacy 1st 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) r2 ≥ 0.999
ii). Constant detector response Covered by overall test-1 or 2.

16. 1. 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.
2. 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. 3. 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.
1. 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%
2. 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