Instrumental Technique for Environmental Analysis Chapter 6 GC

Rezaul Karim Environmental Science and Technology Jessore University of Science and Technology

Chapter content and

Gas Chromatography (GC) Principles Instrumentation Detectors Applications of GC

Reference
James W Robinson, 1995. Undergraduate instrumental analysis, Marcel Dekker, Inc. NY 2. Skoog, Holler & Crouch 2007, Instrumental Analysis, Brooks Cole Cengage Learning, USA. 3. Daniel C. Harris , 2010, Quantitative Chemical Analysis , 8th edition, W. H. Freeman and Company 41 Madison Avenue New York. 4. S. Ahuja and N. Jespersen (Eds), 2006, Comprehensive Analytical Chemistry, Volume 47, Elsevier B.V.
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INTRODUCTION

The development of column chromatography did not progress until the publication of a most valuable paper by Martin and Synge in 1941, for which they were later awarded the Nobel Prize. In that paper they introduced liquidliquid chromatography plate theory, a first model that
could describe column efficiency. they first suggested the possibility of using gas as the mobile phase in a chromatographic system.

Ten years later, James and Martin introduced the first gas chromatography apparatus - suitable
only for the detection and determination of acids and bases. The first commercial instrument was delivered by Griffin

and George (London) in late 1954

In 1955 Glueckauf derived the first comprehensive equation describing the relationship between HETP and particle size, particle diffusion and film diffusion ion exchange. The Dutch scientists van Deemter, Zuiderweg and Klinkenberg developed the rate theory, an alternate to the plate theory, describing the chromatographic process in terms of kinetics and mass transfer. By the early 1980s,
fused-silica capillary columns, selective and sensitive detectors, fully automated systems, and sophisticated techniques such as gas chromatography-mass spectrometer (GC-MS) and gas chromatography-infrared spectrometer (GC-IR)

gas chromatography by this time was a well-established, well-known analytical technique.

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The major advantages of GC

High resolution
A 50 meter capillary column can easily generate 100,000 theoretic al plates

Speed:

Sensitivity
the measureme nt of partspermillio n (ppm) level of almost all volatile organic

Precision and accuracy

analysts to perform quantitative analysis accurately under a variety of condition s.
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Most chromato grams are complete in a matter of minutes.

Gas Chromatography

Gas chromatography has been widely used in

foods, petroleum products, pesticide and pesticide residues, pharmaceutical products, environmental monitoring, clinical chemistry and a number of other fields.
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GC

Principles
Gas-liquid chromatography (often called gas chromatography) is a powerful tool in analysis. The basis for gas chromatographic separation is the distribution of an analyte between two phases. All forms of chromatography involve a stationary phase and a mobile phase. In all the other chromatography,

the mobile phase is a liquid.

In gas-liquid chromatography,
the mobile phase is a gas such as helium the stationary phase is a high boiling point liquid absorbed onto a solid.
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Instrumentation

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Instrumentation

A gas chromatographic system is composed of four major components:

Carrier gas source, sample introduction system, column and Detector

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Carrier gas source

The mobilc-phase gas in GC is called the carrier gas and must be chemically inert. However, a number of restrictions must be placed on the selection of a carrier gas. the carrier gas must be inert, not reacting with analytes and

hazardous. Helium is the most common mobile-phase gas used, although argon, nitrogen, and hydrogen are used. In addition, the carrier gas system often contains a molecular sieve to remove impurities and water.
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with any components of the GC system. the carrier gas must be suitable with the detectors. the carrier gas must be of high-purity grade and not

A two stage regulator is used at the outlet of the carrier gas cylinder to set an appropriate cylinder outlet pressure to the GC system and to monitor the residual pressure in the cylinder. Most GC instruments are equipped with a

The flow controller is used to ensure obtaining a constant flow despite changes in pressure and pressure drops through the GC column. The flow meter is used to set a carrier gas flow rate to a desirable level and to monitor the stability of the carrier gas flow

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flow controller flow meter

Sample introduction system

GC samples are generally introduced onto GC column through an injection port, in which there is a self-sealing septum to ensure no leaking will occur. Gaseous samples are best handled by on-column introduction, which eliminates band broadening due to the dead volume of the injection port. Liquid samples are usually introduced with a syringe. Sample injection sizes depend on

the concentrations of the sample components being analysed, the capacity of the column, and the sensitivity of the detector

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The solvent-flush method

1. 2. 3.

4.

5.
6.

rinsing the syringe with solvent, completely filling and expelling the syringe several times; wiping excess solvent from the syringe needle; drawing about IL of solvent into the syringe, followed by about lpL of air, then followed by drawing in excess sample; positioning the syringe plunger for the required injection volume, and wipe excess sample from the needle; drawing in air until the sample is entirely within the syringe barrel; and Inserting the syringe into the injection port, rapidly depressing the plunger, and after a delay of about 1 second quickly and smoothly withdrawing the syringe.
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The gas chromatograph comprises an oven with sufficient volume to hold one or two columns easily and which can heat up to more than 400C. A weak thermal inertia permits a rapid but controlled temperature climb (gradient able to attain 100C/ min). The temperature must be controlled to within 0.1C in order to get reproducible separations in isothermal or temperature programmed modes. By installation of a cryogenic valve fed with N2 or CO2 in the liquid state, the oven can be regulated at low temperature.

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Thermostatically controlled oven

Columns : two general classifications

packed Columns
Packed columns are tubes of copper, stainless steel, glass or other materials formed in any shape that will fit the GC oven.

Capillary or open tubular columns

The vast majority of analyses use long, narrow open tubular columns made of fused silica (SiO2) and coated with polyimide.

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Packed columns

Common forms are U-shape, W-shape, and coiled tubes). They are typically 36 mm in diameter and 15 m in length. Packed columns are prepared by filling them with finely divided stationary-phase-coated support. The solid support is often silica that is silanized to reduce hydrogen bonding to polar solutes. For tenaciously binding solutes, Teflon is a useful support, but it is limited to 200C. The support particle size range from 40 to 60 mesh for coarse particles, and from 100 to 160 mesh for fine particles.
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Open tubular columns

The vast majority of analyses use long, narrow open tubular columns made of fused silica (SiO2) and coated with polyimide (a plastic capable of withstanding 350C) for support and protection from atmospheric moisture. Open tubular columns offer

higher resolution, shorter analysis time, and greater sensitivity than packed columns, but they have less sample capacity.
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the type and form of the coating, capillary column

The wall-coated column features a 0.1- to 5 mthick film of stationary liquid phase on the inner wall of the column.
The most important and widely used type If it is not specified, the capillary column is generally of the WCOT type

A support-coated column has solid particles coated with stationary liquid phase and attached to
the inner wall.

In the porous-layer column, solid particles are the active stationary phase.
With their high surface area, support-coated columns can handle larger samples than can wall-coated columns.
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Fused-silica wall-coated (FSWC)

Fused-silica capillaries are drawn from specially purified silica that contains minimal amounts of metal oxides. These capillaries have much thinner walls than glass columns. The tubes arc given added strength by an outside protective polyimide coating, which is applied as the capillary tubing is drawn. The resulting columns are quite flexible and can be bent into coils with diameters of a few inches. Silica open tubular columns are available commercially and offer several important advantages:
flexibility and inertness.
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Capillary columns are typically coated by a dynamic

In the dynamic method, a dilute coating solution is passed slowly through the column at a controlled rate, followed by nitrogen drying. a static method In the static method, the column is filled with the coating solution, which is then evaporated in a laminar fashion using a special oven, leaving a thin film deposition of the coating on the
internal wall of the column.

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Compares the performances of columns.

Property Column length, L, m Inside diameter, mm FSWC 10-100 0.1-0.3 WCOT 10-100 0.25-0.75 SCOT 10-100 0.5 Packed 1-6 2-4

Efficiency,plates/m
Sample size, ng Relative pressure Relative speed

2000-4000
10-75 low fast

1000-4000
10-1000 low fast

600-1200
10-1000 low fast

500-1000
10-106 high Slow

Flexibility
Chemical ineretness

Yes
Best

Yes
poorest

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How separation works on the column: One of three things might happen

It may condense on the stationary phase. :

A compound with a boiling point higher than the temperature of the column will obviously tend to condense at the start of the column. However, some of it will evaporate again in the same way that water evaporates on a warm day - even though the temperature is well below 100C. The chances are that it will then condense again a little further along the column.
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How separation works on the column: One of three things might happen

It may dissolve in the liquid on the surface of the stationary phase.:

some compounds will be more soluble in the liquid than others. The more soluble ones will spend more of their time absorbed into the stationary phase; the less soluble ones will spend more of their time in the gas

It may remain in the gas phase:

The process where a substance divides itself between two immiscible solvents because it is more soluble in one than the other is known as partition.
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Solid Support Materials

The packing, or solid support in a packed column, holds the liquid stationary phase in place so that as large a surface area as possible is exposed to the mobile phase. The ideal support consists of small, uniform, spherical particles with good mechanical strength and a specific surface area of at least 1m'/g. In addition, the material should be inert at elevated temperatures and be uniformly wetted by the liquid phase. No material is yet available that meets all of these criteria perfectly.
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The two ends are fitted with glass wool plugs, to retain packing materials. Most packed column supports are prepared from diatomaceous earth, which is composed of skeletons of diatoms. The diatomite is basically amorphous hydrous silica.

It consists of the skeletons of thousands of species of single-celled plants that once inhabited ancient Such plants received their nutrients and disposed of their wastes via molecular diffusion through their pores.

lakes and seas.

As a result, their remains are well-suited as support materials because GC is also based on the same kind of molecular diffusion.
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Particle Size of Supports

The efficiency of a gas chromatographic column increases rapidly with decreasing particle diameter of the packing. The pressure difference required to maintain an acceptable flow rate of carrier gas, however, varies inversely as the square of the particle diameter. the latter relationship has placed lower limits on the size of particles used in GC because it is not convenient to use pressure differences that are greater than about 50 psi. As a result, the usual support particles are 60 to 80 mesh (250 to 170 m) or 80 to 100 mesh (170 to 149 m).

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The Stationary liquid Phase

Desirable properties for the immobilized liquid phase

low volatility (ideally. the boiling point of the liquid should be at least 100C higher than the maximum operating temperature for the column); thermal stability; chemical inertness; solvent characteristics such that k and values for the solutes to be resolved fall within a suitable range

Many liquids have been proposed as stationary phases in the development of GLC. The proper choice of stationary phase is often crucial to the success of a separation.

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To have a reasonable residence time in the column, an analyte must show some degree of compatibility (solubility) with the stationary phase. The choice of liquid stationary phase is based on the rule like dissolves like.

Non-polar columns are best for non-polar solutes. Columns of intermediate polarity are best for intermediate polarity solutes, and strongly polar columns are best for strongly polar solutes.

Generally, the polarity of the stationary phase should match that of the sample components. When the match is good, the order of elution is determined by the boiling point of the eluents.

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Polar stationary phases

Contain functional groups e.g. -CN, -CO, & OH. polyester phases are highly polar. Polar analytics include alcohols, acids, and amines;

Non-polar, stationary phases

dialkyl siloxanes and Saturated hydrocarbons

Solutes of medium polarity

ethers, ketones, and aldehydes

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Widely Used Stationary Phases

Stationary Phase

Polydimethyl siloxane
5% Phenylpolydimethyl siloxane 50% Phenylpolydimethyl siloxane 50% Tritluoropropylpolydimethyl siloxane Polyethylene glycol

Common trade Maximum name temp., c OY-l, SE-30 350

Common Applications

OY-3,SE-52

350

OY-17 OY-2l0

250 200

General-purpose nonpolar phase, hydrocarbons, polynuclear aromatics, steroids, PCBs General-purpose nonpolar phase, hydrocarbons, polynuclear aromatics, steroids, PCBs Drugs, steroids, pesticides, glycols
Drugs, steroids, pesticides, glycols

Carbowax20M

250

50% Cyanopropyl polydimethyl siloxane

OY-275

- 240

Free acids, alcohols, ethers, essential oils, glycols Polyunsaturated fatty acids, rosin acids, free acids, alcohols

The current phases correspond in principle to two families: the polysiloxanes and the polyethylene glycols
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Polysiloxanes Polysiloxanes /silicone oils and gums are based upon a repetitive backbone that consists of two hydrocarbon chains per silicon atom. There are about 20 different compositions of alkyl or aryl chains (methyl or phenyl) to which can be incorporated further functional groups (e.g. cyanopropyl, trifluropropyl). Monomers combined in variable proportions also convey changes in the properties of stationary phases (polarity, extended stability from 50 to 300/325C, for the dimethyl polysiloxanes, depending on the column).
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Polyethyleneglycols (PEG) The best known representative of this family is Carbowax. These polar polymers (Mr=1500 to 20 000 for the Carbowax 20M) can be used for deposition, impregnation or as bonded phases (40<T<240/260 _C, depending on column diameter and film thickness).

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The column temperature

Column temperature is an important variable that must be controlled to a few tenths of a degree for precise work. Thus, the column is ordinarily housed in a thermo-stated oven. The optimal column temperature depends on the boiling point of the sample and the degree of separation required. Roughly, a temperature equal to or slightly above the average boiling point of a sample results in a reasonable elution time (2 to 30 min). For samples with a broad boiling range, it is often desirable to employ temperature programming, in which the column temperature is increased either continuously or in steps as the separation proceeds.

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Figure 2.5 shows the elution of C5 to C15 linear alkanes from a 3-m-long column.

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At a constant temperature of 30C (not shown), heavier alkanes take so long to be eluted and emerge over such a long time that they would not be detected. The three traces in Figure 2.5 show what happens when column temperature is raised from 30 to 150C at rates of (a) 20C/min, (b) 40C/min, and (c) 60C/min. Broad, late-eluting peaks can be sharpened and eluted in less time with temperature programming. To maintain adequate resolution of earlier eluting peaks, programs often include a period of time at constant, low temperature prior to raising the temperature

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The detector
The GC detector, producing an electrical signal that is proportional in intensity to the concentration or the mass of the eluted analyte. Since the introductions of gas chromatography, over 40 detectors have been developed. Some are designed to respond to most compounds in general, while others are designed to be selective for particular types of substances. There are several different types of detector in use.

the flame-ionisation (FID), the thermal conductivity (TCD) the electron capture detectors (ECD).
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Characteristics of the Ideal Detector

Adequate sensitivity. Just what constitutes adequate sensitivity cannot be described in quantitative terms. For example, the sensitivities of the detectors are described in this section vary by a factor of 107. Good stability and reproducibility. A linear response to solutes that extends over several orders of magnitude. A temperature range from room temperature to at least 400C. A short response time independent of flow rate. High reliability and ease of use. The detector should be foolproof in the hands of inexperienced operators, if possible. Similarity in response toward all solutes The detector should be non-destructive.

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A flame ionization detector

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A flame ionization detector

The flame ionisation detector has become the most popular detector for GC over the last 40 years, and nothing suggests that this position will ever be challenged. The reasons for that come from its simplicity, reliability, relatively high sensitivity for wide variety of organic compounds, and excellent linearity (as high as 10 s).
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The whole detector is enclosed in its own oven which is hotter than the column temperature. That stops anything condensing in the detector. The FID utilises a flame produced by the combustion of hydrogen and air. Very little ions are formed because of the combustion of the hydrogen and air. Large increase in ions will occur when orgamc compounds are introduced into the flame through the FID jet. The ions will be attracted by the FID collector on which a polarising voltage is applied, and produce a current, which is proportional to the quantity of analyte in the flame. For optimal FID operation, the carrier, hydrogen and air flow must be properly set and adjusted.
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Advantages The FID exhibits

a high sensitivity (~ 10-13g/s), large linear response range (~107), and low noise. It is generally rugged and easy to use.

Disadvantages of the flame ionization detector are that

it destroys the sample during the combustion step and requires additional gases and controllers.
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Application of GC

To evaluate the importance of GC, we must distinguish between the two roles the method plays. performing separations.

In this role, GC methods are unsurpassed when applied to complex organic, metal-organic, and biochemical systems made up of volatile species or species that can be derivatized to yield volatile substances. In this role, retention times or volumes are used for

the completion of an analysis.

qualitative identification, and peak heights

or peak areas provide quantitative information.
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Application of GC

For qualitative purposes, GC is much more

limited than most of the spectroscopic

methods considered in earlier chapters. Thus, an important trend in the field has been in the direction of combining the
remarkable separation capabilities of GC with the superior identification properties of such instruments as mass, IR, and nuclear

magnetic resonance spectrometers.

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