Q10. What Is a Change control?

Ans: Change Control is a general term describing the process of managing how changes
are introduced into a controlled System. Into validation, this implies how changes to the
validated system are made. Change control is needed to demonstrate to regulatory
authorities that after system modifications, validated systems remain under control after
system changes.

Q11. What Is SOP?

Ans: A Standard Operating Procedure (SOP) is a certain type of document that describes
in a step-by-step outline form how to perform a particular task or operation. To ensure that
tasks are conducted consistently and appropriately, everybody in an organization must
follow the same procedures. Many organizations have a broad range of SOPs that
illustrate how to execute various tasks. In many companies, technicians and operators are
trained in how to follow individual SOPs and their training record specifies SOPs they are
trained on and are authorized to use.
Q12. Tell me about the content of the SOP
Ans) 1. Objective or Purpose/Aim
2.Scope
3. Responsibility
4.Procedure
5. Precautions
6. Annexure
7. Abbreviations
8. Reference
9. Revisions History
Q13. What do you know about stability studies?
Ans: These are necessary for developing pharmaceutical products. The influence of
environmental factors (e.g. light, humidity, temperature, etc.) on active pharmaceutical
ingredients (API) or pharmaceutical ingredients (API) may be evaluated.
Q14. Difference between validation and calibration?
Ans: Validation provides written evidence to ensure that a particular method or operation
continuously develops a product with predetermined requirements and quality credits. It is
performed according to validation protocol.
Calibration denotes that equipment produces the values in specified limits by comparing
the values produced by a standard. It Is done according to the calibration SOP.

Q15.Why we calibrate an instrument on a particular interval?

Ans) because it can be possible for instruments to drift out of accuracy after qualification.
so it needs to requalify the instrument at a specific time interval.

pH Meter Principle
it measures the voltage between the two electrodes one is a glass electrode and the other
is reference electrodes. it displays the result of that voltage that is related to the
corresponding pH value.

Sometimes there if both electrons are present is called the combination electrode and they
are inserted into the solution in which pH is to be tested. the solution of which the ph is to
be tested. these two electrodes are immersed and after immersing these electrodes in a
solution. that H+ ions in the test solution exchange for other positively charged ions present
on the glass ball. so there is an action that takes place between these plus ions of the
solution and H+ions or positively charged ions present on the glass bulb. amplifier detects
the difference in electric potential between the two electrodes. the difference of these
potentials is called the ph unit.
Why Range of pH (1 to14)
pH always measured between (1 to14). The solution having the pH=1 is called an acidic
solution or generally, it is highly acidic in nature and the solution which has the pH=14 is
highly alkaline or it is highly basic solution than the solutions having more H+ ion
concentration. it is highly acidic and the solutions which have more OH– ion is a highly
basic or highly alkaline solution.
Key components or parts of pH instrument
It consists of a probe typically a three-in-one combination. The electrode has a hydrogen
ion-sensitive glass electrode, a reference electrode, and a temperature probe. the
temperature probe is used to ensure any temperature variation is corrected automatically.

At the tip of the probe is a sensitive glass bulb that detects the acidity or basicity of the
solution and at the other end of the probe is a high input electronic meter that measures
and displays the pH.
 First to calibrate the pH meter takes, three color-coded standard buffer solutions of
pH 7.0, 4.01, and 9.21 for calibration. the first buffer used for calibration is always the
neutral buffer with a pH of 7.0, the second should always be near the expected sample pH
either a pH of 4.01 or 9.21.

Bases should be measured with buffers that have a pH of 9.21, while acidic samples
should be measured with buffers that have a pH of 4.01. Place the electrodes in the buffer
solution with a pH of 7.0 and allow the pH reading to stabilise at 7. if the needed pH is
determined by the H+ ion concentration

if the concentration of H+ ions inside the glass membrane electrode and solution of buffer
solution present outside the electrode is the same the pH equals 7. once the standard with
pH 7.0 is calibrated rinse the electrode with distilled water and blood dry with Kim wipes.
In the next step if the expected pH of the sample is acidic then select the buffer solution of
pH 4.01 place the electrodes in the buffer with a pH value of 4.01 and press the calibrate
button. allows the pH reading to stabilize at 4.01. if the concentration of H+ ions inside the
glass membrane electrode is lower as compared to the solution of buffer solution present
outside the electrode then pH will be less than 7.
once the standard with pH 4.01 is also calibrated rinse the electrode with distilled water
and blood dry with Kim wipes in a similar manner you may skip the previous step if the
expected pH of the sample is on the alkaline side and follow this step by using the buffer
solution of pH 9.21.

Place the electrodes in the buffer with a pH value of 9.21 and press the calibrate button.
allows the pH reading to stabilize at 9.21 if required sometimes pH 10 buffer solution is
also used for the same concentration of H+ inside the glass membrane electrode is higher
as compared to the solution of buffer solution present outside the electrode that pH
displayed is more than 7. repeat the rinse process just like previous steps now the pH
meter is calibrated and ready to determine the pH of the test sample.
place the electrodes in the given sample and then press the measure button to leave the
electrodes in your sample until the reading has stabilized this will be the exact pH value of
your sample take the electrodes out rinse them with distilled water and blot dry with Kim
wipes immerse the probe in three molar potassium chloride solution for storage like this
consult your operation manual for optimal storage practices for your specific pH meter.
Application of pH Meter
 It is used in the agriculture industry to determine the pH of soil.
 It’s also used to test the quality of municipal drinking water and swimming pools.
 It is used to measure the pH value of solutions in numerous chemical and
pharmaceutical businesses.
 The pH metre is also used in the food business, particularly for dairy products such
as cheese, curds, and yoghurts.
A pH metre is an electronic equipment that is used to determine the pH of liquids and
semi-solids. The features of this indicator were used to precisely determine the acidity or
alkalinity of diverse substances. Because it provides reliable readings, the pH metre is
more useful than other pH indicators. It can be used to determine the pH of a liquid or a
semi-solid substance.
What is the full form of pH

pH is denoting the potential of hydrogen or the power of hydrogen.

What is pH value or pH scale

The pH scale is used to determine how acidic or basic an aqueous solution. Lower pH
values indicate solutions that are more acidic, whereas higher pH values indicate solutions
that are more basic or alkaline in nature. The pH scale is logarithmic and indicates the
concentration of hydrogen ions in a solution in the inverse order At 25 degrees Celsius, a
lower pH indicates a higher concentration of hydrogen ions. with the pH less than 7 are
acidic and solutions with the pH greater than 7 are basic.
What is the equation of pH

The negative logarithm of hydrogen ion activity is used to calculate pH? formula is pH = -
log10(aH+).
what are pH indicators

pH indicators may be used to measure pH by making use of the fact that their color
changes with pH visual comparison of the color of a test solution with a standard color
chart provide a means to measure pH accurate to the nearest whole number more precise
measurements are possible if the color is measured spectrophotometrically using a
colorimeter or The universal indication in a spectrophotometer is a blend of indicators that
produces a continuous colour change from around pH 2 to pH 10. Universal indicator
paper is comprised of absorbent paper that has had a universal indicator injected into it.
What is the design of the electrode
The design of the electrodes is significant; they are road-like structures, commonly built of
glass, with a bulb at the bottom carrying the sensor. The reference electrode is insensitive
to the pH of the solution, and it is made up of a metallic conductor that connects to the
display, which is made up of a voltmeter that displays voltage in pH units.
What is an ideal requirement of buffer solutions

buffer salts of requisite purity can be obtained from the national institute of
standards and technology other national authorities buffer solutions should be stored in
appropriate containers that ensure the stability of the pH through the expiry date and fitted
with tight closure.
How many methods to calibrate the pH meter

pH meter calibration shall be performed by two-point calibration multiple-point calibration

and multiple segment calibration.
What precautions should be taken while using a test sample that is sensitive to
carbon dioxide in the air?

If the Test sample’s pH is sensitive to carbon dioxide in the air, use pure water that has
recently been boiled and then stored in a container designed to keep carbon dioxide out.
Can we measure the pH of a gas

The only way to determine the pH of a gas is to dissolve it in distilled water and measure
the result.

What Is Karl Fischer Titration?

Karl Fischer titration is a titration method that uses volumetric or coulometric titration to
determine the quantity of water present in a given analyte. This method for quantitative
chemical analysis was developed by the German chemist Karl Fischer in the year 1935,
Today, specialized titrators (known as Karl Fischer titrators) are available to carry out such
titrations.

Principle of Karl Fischer Titration

The principle of Karl Fischer titration is based on the oxidation reaction between iodine
and sulphur dioxide. Water reacts with iodine and sulphur dioxide to form sulphur trioxide
and hydrogen iodide. An endpoint is reached when all the water is consumed. The
chemical equation for the reaction between sulphur dioxide, iodine, and water (which is
employed during Karl Fischer titration) is provided below.
Karl Fischer Titration Equipment
Drying tube, sample injection cap, electrode analysis, Drain cook, a cathode chamber,
detection electrode, rotor, anode chamber, KF reagent.
Ingredients of KF reagent:
Iodine, Buffer (Imidazole), sulphur dioxide, solvent (methanol).
Karl Fischer Titration Procedure
The Karl Fischer titration experiment can be performed in two different methods. They are:

 Volumetric determination – This technique is suitable to determine water content

down to 1% of water. The sample is dissolved in KF methanol and the iodine is
added to KF Reagent. The endpoint is detected potentiometrically.
 Coulometric determination – The endpoint is detected in this experiment
electrochemically. Iodine required for KF reaction is obtained by anodic oxidation of
iodide from solution.

Karl Fischer Titration Applications

 It is used in technical products such as plastics, oils, gases.
 It is used in pharmaceutical products.
 It is used in cosmetic products.
 It is used in the industry.

Advantages of Karl Fischer Titration

 It is fitted for determining water in gases, liquids and solids.
 The coulometric titrator helps in detecting free water, dissolved water, and emulsified water.
 It is a swift process which demands a minimal amount of sample preparation.
 Extremely accurate method.

Limitations of Karl Fischer Titration

 It is a destructive technique.
 The solvent consumption is high as the manual volumetric titration demands reloading during each
determination.
 Coulometric titration is fitted only for samples that contain a small amount of water.
 Coulometric titration takes extremely long periods to determine.

How can the KF reagent be prepared?

Make a solution by mixing 170 mL of pyridine and 670 mL of methanol. Add 125 g of iodine to
the solution and cool it. Take a 250 mL graduated cylinder and add 100 mL of pyridine. Keep it on
an ice bath. Pass in sulphur dioxide (dry) till its volume reaches 200 mL.
What is a major difference between volumetric and coulometric titration?
The main difference between them is that:
Volumetric method – the titrant is directly added to the sample with the help of a burette.
Coulometric method – the titrant is produced electrochemically inside the titration cell.

What is coulometric Karl Fischer?

The Karl Fischer titration is merely a means of measuring sample water content. Modern
instruments, such as the Aquamax KF, apply the coulometric principle, whereby the water present
in the sample is coulometrically titrated to a predefined endpoint at which free iodine exceeds a
minute.

Why we use sodium tartrate in Karl Fischer?

The volumetric standard for Karl Fischer titration is sodium tartrate dihydrate. It is stable and non-
hygroscopic, under normal conditions. Sodium tartrate dihydrate has a 15.66 percent stoichiometric
water content and is primarily used in volumetry to measure the titer.

What electrode is KF titration?

The reaction has reached its termination point because the iodine is in abundance. For
electrochemical indication of the end point, the most complex KF titration technology uses a
double platinum electrode, but visual and photometric indications are still used.

How do you calculate Karl Fischer factor?

The water equivalence factor F is determined according to the formula 0.1566 x w / v in mgs of
H2O per ml of reagent, where W is the sodium tartrate weight in mgs, and V is the reagent volume
in ml.

How do you make Karl Fischer reagent?

A solvent alcohol (ROH), an established concentration of iodine (I2), a base (RN) and sulphur
dioxide (SO2) are the reagents. In an aqueous environment, the Bunsen reaction between iodine
and sulphur dioxide is the basis for the reactions of Karl Fischer Reagents.

Density Meter / Digital Density Meters

Density meters are instruments that measure the density of a sample liquid or gas. Digital
density meters are used in the pharmaceutical, petroleum, chemical, and food and beverage
industries for quality control and in research and development.

Modern digital density meters are based on the oscillating U-tube principle. The tube, usually a
U shaped glass tube, is excited and starts to oscillate at a certain frequency depending on the
filled-in sample. Through determination of the corresponding frequency the density of the
sample can be calculated.

There are digital density meters that can measure specific gravity and refraction index as well
as the density of a sample. There are density meters available that are portable or bench top
models, that can measure liquid, gas, or both, and have a built in thermostat or need a water
bath to maintain temperature. Some other variations include sample volume needed, speed of
testing, and the ability to take and store images.

Differences Between Normality and Molarity

Here are some key differences between normality and molarity.

Normality Molarity

Also known as equivalent concentration. Known as molar concentration.

It is defined as the number of gram It is defined as the number of moles per litre
equivalent per litre of solution. of solution.

It is used in measuring the gram It is used in measuring the ratio between the
equivalent in relation to the total volume number of moles in the total volume of the
of the solution. solution.

The units of normality are N or eq L-1 The unit of molarity is M or Moles L-1

Q1. What is Gas Chromatography ?

Ans. : Gas chromatography is a process used to separate substances in to its components. In this
process, a Mixture is vaporized and injected into stream of carrier gas with system.
Q2. What is Basic Principle of Gas Chromatography Separation?
Ans. : The basis for Gas Chromatographic separation is the distribution of a sample between two
phases. One of these phases is a stationary bed of large surface area and other phase is a gas, which
percolates through the stationery bed.
Q3. What is Basic Part of Gas Chromatography ?
Ans. : The basic parts of a gas chromatograph are :

1. Carrier Gas Cylinder.

2. Pressure regulator and flow controller (Pneumatic system)
3. Injection Port (Sample inlet).
4. Column.
5. Detector.
6. Recorder
Q4. What are the Advantages of Gas Chromatography ?
Ans. : 1. It is a micro method and only a few sample is enough for analysis.
2. Sensitivity of the method is very high.
 3. Gives good precision and accuracy.
4. It involves relatively simple instrumentation. Operation of a GC and related calculation
do not require highly skilled personnel.
5.The cost of equipment is relatively low and its life is large.
Q5. What are the Limitations of Gas Chromatography ?
Ans. : 1. Analysis of solid samples cannot be done directly.
2. Samples having the boiling point more than 700 o C cannot be analysed.
3. Different types of functional group required different columns, so requirements of columns are
high.
4. Components which are decomposed / sublimed at operating temperature, can not be analysed.
Q6. What is Baseline ?
Ans. : Any part of the chromatogram where only mobile phase is emerging from the column.
Q7. What is the function of Carrier Gas ?
Ans. : Main function of carrier gas is transportation of components through column.
Q8. What is the function of Column ?
Ans. : Column is the HEART OF GC system where remarkable separation takes place.
Q9. What is the function of Detector ?
Ans. : The detector is the BRAIN of GC, indicates the presence and measures the amount of components
in the column effluent.

Q10. What are the main characteristics of Carrier Gas ?

Ans. : Following are main characteristics of carrier

1. It should be inert so that its activity with stationery phase or hardware does not exist.
2. It should be suitable for the detector employed and the type of sample analysed.
3. It should give best column performance constant with desired speed of analysis.
4. It should be readily available in high purity.
5. It should not be cause the risk of fire or explosion hazard.
6. It should be cheaper.
Q11. What is the full form of “EPC” ?

Ans. : Electronic Pneumatic Control

Q12. How many types of Columns ?
Ans. : There are mainly two types of columns
1. Packed Column
2. Capillary Column
Q13. How many types of Detectors commonly used in Gas Chromatography ?
Ans. : There are many types of detectors but mainly following two are basic detectors
1. Thermal Conductivity Detector (TCD)
2. Flame Ionisation Detector (FID)

While other are selective detectors

3. Flame Photometer Detector

4. Electron capture Detector
5. Hall Electrolyte Detector
Q14. What is the full form of “FID” ?
Ans. : Flame Ionization Detector
Working Principle : A tiny flame of hydrogen is maintained at a capillary jet, air is
introduced through a side for supporting combustion. Column effluent are led into the flame
where ionisation of components take place. On combustion of the compound. An electrode
system located close by picks up the ionisation current which is then amplified and fed in to
recorder. When only carrier gas passing through the flame, there is no or very small and
constant ionisation current flowing across the electrode and when no sample component is
eluting, the steady base line will be on recorder when sample components elute and passing
through the flame, its molecules are ionised and the resulting ionisation current which is
amplified and fed to suitable recorder. Ionisation current is proportionate to the concentration
of components
Q15. What is the full form of “FPD” ?
Ans. : Flame Photometer Detector
Q16. What is the full form of “TCD” ?
Ans. : Thermal Conductivity Detector
Q17. What is the full form of “ECD” ?
Ans. : Electron Capture Detector
It is also called selective detector. A typical ECD is shown in Figure

Working Principle : The carrier gas stream, usually ultra pure N2 or Ar with 5% CH4 enters
through inlet A and leaves through exit B. A tritium or Ni63 foil, placed inside the cell, ionises
the carrier gas molecule to form electrons that move slowly towards anode under a fixed
voltage. A standing current is produced which is amplified when a component having affinity
for electrons elute from column and enter the detector, it captures some electrons causing a
drop in standing current. This loss of current is fed to a recorder.

Q18. What are four types of quantitative calculations in Gas Chromatography analysis ?
Ans. : There are four methods for quantitative analysis
1. Area Percentage (Area Normalization)
2. Area Normalisation with Response Factor.
3. External Standard
4. Internal Standard
Q19. What is the function of pressure regulators ?
Ans. : The Function of pressure regulator are
1. To convert gas stored at higher pressure inside cylinder, to a lower operating pressure range.
2. The regulator maintain constant working pressure for most of the cylinder volume.
3. It prevent any dangerous pressure built up in the system.
Q20. What are the Injection technique used in Gas Chromatography?
Ans. : There are mainly four type of injection technique used in Gas Chromatography
1. On Column Injection (By Syringe)
2. Gas Sampling Valve
3. Head space
4. Thermal Desorption
Q21. What is Response Factor ?
Ans. : Response factor is a ratio of area of component Vs. Amount of component.
Q22. What is the function of Oven ?
Ans. : The main function of Oven is to maintain the temperature of Column.
Q23. What is the Sample Loop?
Ans. :Sample Loop is a specially design tube connected located in sampling valve to control accurate sample
injection volume.
Q24. What is on spec product ?
Ans. : If product analysis meets the specification, this product is called on spec product. The specification
for all products are available in QAD website.
Q25. What is called off spec product?

Ans.: If product analysis does not meets the specification, this product is called off spec product. The
Specification for all products is available in QAD website.
Q26. Which Detector is used to measure the Hydrogen ?
Ans.:Thermal Conductivity Detector (TCD)
Q27. Which Detectors are used to measure Sulfur Components?
Ans.:Flame Photometer Detector (FPD) or Sulphur Chemiluminesance Detector (SCD)
Q28. What is Calibration?
Ans.: Calibration is the process of evaluating and adjusting the precision and accuracy of measurement
equipments.
Q29. What is Retention Time ?
Ans.:The time elapsed between the moment of sample introduction and the peak maxima.
Q30. What is full form of “GSV” ?
Ans.:Gas Sampling Valve
Q31. What is full form of “LSV” ?
Ans.:Liquid Sampling Valve
Q32. Describe Basic working principle of Flame Ionization Detector ?
Ans.:Column effluent are led into the flame maintain by air and hydrogen where ionisation of components
take place. An electrode system located close by picks up the ionisation current which is then amplified
and fed in to recorder.
Q33. Describe Basic working principle of Thermal Conductivity Detector ?
Ans.:The TCD is based on the principle of thermal conductivity which depends upon the composition of the gas.
Thesample components in the carrier gas pass into the measuring channel. A second channel serves as a
referencechannel where only pure carrier gas flows. Electrically heated resistance wires are located in
both channels. Thedifference in thermal conductivity between the column effluent flow (sample
components in carrier gas) andthe reference flow of carrier gas alone, produces a voltage signal
proportional to this difference. The signal isproportional to the concentration of the sample components.

Q34. What are the utility gas used for Flame Ionization Detector ? What is typical ratio ?
Ans.:Hydrogen & Air, Typical ratio is 1:10
Q35. What is the unit of column flow ?
Ans.:millilitre \ minute
Q36. What is Isothermal Temperature condition ?
Ans.:It means that you keep the temperature constant during the entire run.
Q37. What is the Chromatogram?
Ans.:A chromatogram is a plot of the response of the detector as a function of elution time.
Q38. What is the length of Capillary column ?
Ans.:Normally capillary column length is 10 to 60 meter.
Q39. What is the length of Packed column ?
Ans.:Normally packed column length is 1 to 5 meter.
Q40. What is Headspace Chromatography Technique ?
Ans.:'Headspace' is the gas space above the sample in a chromatography vial. Volatile sample
Components diffuse in to the gas phase, forming the headspace gas. Headspace gas is introduced
in GC by capillary loop or by Syringe.
Q41. Why not use Oxygen gas in Gas chromatography ?
Ans.:Oxygen is a reactive element and can react with substances to be analyzed.

Q42. What is split in gas chromatography ?

Ans.:The sample volume what we are injected is to high for capillary column thus split help to dived the volume
and pass through less amount and remaining sample gone as a waste. This is to minimize the load on
column.
e.g. : Split ratio is 1 : 10 that means whatever the sample injected that portion is divided into 10 parts from that
10 parts only 1 part will go into injector.

Q43.What is the effects results from slow injection of a large sample volume ?

Ans.:1. Decrease the peak resolution

2. Retention time will change

Q44. Which type of column has the greater efficiency and resolution, packed or capillary?

Ans.:Capillary

Q45. What would be the effect of increasing carrier gas flow on retention times?
Ans.:Retention times will be decrease
Q46. What would be the effect of decreasing carrier gas flow on retention times?
Ans.:Retention times will be increase
Q47. What would be the effect of increasing oven temperature on retention times?
Ans.:Retention times will be decrease
Q48. What would be the effect of decreasing oven temperature on retention times?
Ans.:Retention times will be Increase
Q49. What is an Inert Gas? What is importance of an inert gas in Gas chromatography ?
Ans.:Aninert gas is a gas which does not undergo chemical reactions under a set of given conditions. Usually
an inert gas is used a carrier gas.
Q50. What is the limitations of packed column compare to capillary column?
Ans. : There are mainly three limitations of packed column
1. Poor separation and poor resolution
2. Need frequent regeneration
3. Need more amount of carrier gas
QUANTITAIVE CALCULATIONS :

There are four methods for quantitative analysis

1. Area Percentage (Area Normalization)
2. Area Normalisation with Response Factor.
3. External Standard
4. Internal Standard
Area Normalisation :
By normalising, it mean calculating the percent composite by measuring the area of each peak
and dividing the individual area by the total area.

Area(i)

% Component(i) = -------------- x 100

 Area(i)

Where, Area(i) = Area of component(i) peak.

Area = Sum of total area in the chromatogram.

This method can be used when it is assumed that all peaks are eluted and that each
component has the same detector response.

Limitations : Exact concentration can not be determined.

1. Area Normalization with Response Factor :

Area of the component are not directly proportional to the percent composition i.e. different
components have different detector response. Therefore, it is necessary to determine
response factor. Once determined, these response factors can be used to calculate the
percent composition.

X(i) Area(Ref)

RF(i) = --------- x ------------

Area(i) X(Ref)

Where, X(i) = Amount of component(i) in the calibration sample.

 Area(i) = Area of component(i) peak.

Area(Ref) = Area of component peak related as a reference component.

X(Ref) = Amount of the reference component in calibration sample.

Analysis of each component of interest (i) in the analysis run.

RF(i) x Area(i)

% Component(i) = ----------------------

(RF(i) x Area(i))

Where, Area(i) = Area of component(i) measured in the analysis run.

 = The sum.

RF(i) = Response Factor of component(i)

2. Internal Standard Method :

In this method the standard material which is distinct from any component in the sample is
added ( usually by weight) to each individual calibrations and analysis samples.

The Internal Standard material must be chosen in such a way that :

 It is not present at all in the sample.

 Its concentration should be of same concentration as the peak of interest.
 It should elute distinctly but as close to the peak of interest as possible.
 It should not have any chemical reaction with the sample.

Calibration for each component of interest :

X(i) Area(IS)

RF = --------- x -----------

Area(i) X(IS)
 Where, X(i) = Amount of component(i) in calibration sample.

Area(i) = Area of component(i) peak

Area(IS) = Area of Internal Standard peak.

X(IS) = Amount of Internal Standard in calibration sample.

Analysis of each component of interest(I) in the analysis run.

IS x RF(i) x Area(i)

% Conc. of component = ----------------------------- x 100

SA x Area(IS)

Where, IS = Amount of Internal Standard added to the sample.

SA = Amount of sample material measured.

Area(i) = The peak area of component(i) in the sample run.

Area(IS) = Area of the Internal Standard peak in the sample run.

3.External Standard Method :

External Standard technique is good for single component requirement and to determine
absolute value.

Response Factor of the component is determined by injecting a known concentration blend

and from the Area of this component RF is calculated.

Limitations : Requires constant volume injection each time.

Calibration for each component of interest :

X(i)
 RF(i) = ----------

Area(i)

Where, X(i) = Amount of component(i) (% or ppm) is used in the calibration mixture

Area(i) = Peak area of component(i)

Analysis of each component of interest :

(% or ppm) Conc. of component(i) = Area(i) x RF(i)

Where,

Area(i) = Area of component(i) peak

RF(i) = Response Factor of component(i)