A.

Latar Belakang

Kromatografi merupakan suatu teknik analisis yang mencakup metoda

pemisahan dan metoda penentuan baik secara kualitatif maupun kuantitatif.
Bentuk analisis lengkap ini merupakan keunggulan utama dari kromatografi.
Terdapat dua klasifikasi besar dalam kromatografi yaitu kromatografi gas
dimana fasa gerak adalah gas dan kromatografi cairan yang mempunyai fasa
gerak berbentuk cairan.
Pada awalnya kromatografi gas hanya digunakan dalam analisis gas,
tetapi dengan kemajuan teknologi, kromatografi gas dapat digunakan untuk
analisis bahan cair dan padat dengan syarat bahwa bahan yang akan dianalisis
mudah menguap atau bisa diderivatisasi terlebih dahulu menjadi bahan yang
mudah menguap. Awal perkembangan kromatografi gas (GC) difokuskan
pada kolomnya, yaitu isi kolom (fasa diam) dan ukuran kolom, sehingga
lahirlah kolom kapiler GC. Perkembangan selanjutnya yaitu penggabungan
dari GC kolom kapiler dengan berbagai jenis detektor yang spesifik, salah
satunya adalah penggabungan dengan spektrometri massa, yang dikenal
sebagai GC-MS. Dengan memanfaatkan spektrometer massa sebagai detektor,
identifikasi kualitatif menjadi lebih akurat karena melalui detektor ini dapat
dihasilkan spektrum massa dari puncak kromatogram yang dipakai untuk
keperluan konfirmasi puncak.

B. Tujuan

Setelah melaksanakan praktikum ini, praktikan dapat mengetahui cara

membuat parfum dengan benar dan mengetahui cara menguji mutu parfum
yang benar.
 II. HIGH PERFORMANCE LIQUID CHROMATOGRAPY

High performance liquid

chromatography
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High performance liquid chromatography

A HPLC. From left to right: A pumping device generating a

gradient of two different solvents, a steel enforced column
and an apparatus for measuring the absorbance.
Acronym HPLC
Classification Chromatography
organic molecules
biomolecules
Analytes
ions
polymers
Agilent Technologies
Beckman Coulter, Inc.
Hitachi
PerkinElmer, Inc.
Manufacturers
Shimadzu Scientific Instruments
Thermo Electron Corporation
Varian, Inc.
Waters Corporation
Other Techniques
Chromatography
Aqueous Normal Phase
Chromatography
Related
Ion exchange chromatography
Size exclusion chromatography
Micellar liquid chromatography
 Liquid chromatography-mass
Hyphenated
spectrometry
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High-performance liquid chromatography (or High pressure liquid

chromatography, HPLC) is a form of column chromatography used frequently
in biochemistry and analytical chemistry to separate, identify, and quantify
compounds. HPLC utilizes a column that holds chromatographic packing material
(stationary phase), a pump that moves the mobile phase(s) through the column,
and a detector that shows the retention times of the molecules. Retention time
varies depending on the interactions between the stationary phase, molecules
being analyzed, and the solvent(s) used.

Contents
[hide] [hide]
 1 Operation
 2 Types of HPLC
o 2.1 Normal phase chromatography
o 2.2 Reversed phase chromatography
o 2.3 Size exclusion chromatography
o 2.4 Ion exchange chromatography
o 2.5 Bioaffinity chromatography
o 2.6 Isocratic flow and gradient elution
 3 Parameters
o 3.1 Internal diameter
o 3.2 Particle size
o 3.3 Pore size
o 3.4 Pump pressure
 4 Manufacturers of HPLC chromatographs
 5 Manufacturers of HPLC columns and accessories
 6 See also
 7 References

 8 External links

[edit] Operation
The sample to be analyzed is introduced in small volume to the stream of mobile
phase and is retarded by specific chemical or physical interactions with the
stationary phase as it traverses the length of the column. The amount of
retardation depends on the nature of the analyte, stationary phase and mobile
phase composition. The time at which a specific analyte elutes (comes out of the
end of the column) is called the retention time and is considered a reasonably
unique identifying characteristic of a given analyte. The use of pressure increases
the linear velocity (speed) giving the components less time to diffuse within the
column, leading to improved resolution in the resulting chromatogram. Common
solvents used include any miscible combinations of water or various organic
liquids (the most common are methanol and acetonitrile). Water may contain
buffers or salts to assist in the separation of the analyte components, or
compounds such as Trifluoroacetic acid which acts as an ion pairing agent.

A further refinement to HPLC has been to vary the mobile phase composition
during the analysis, this is known as gradient elution. A normal gradient for
reversed phase chromatography might start at 5 % methanol and progress linearly
to 50 % methanol over 25 minutes, depending on how hydrophobic the analyte is.
The gradient separates the analyte mixtures as a function of the affinity of the
analyte for the current mobile phase composition relative to the stationary phase.
This partitioning process is similar to that which occurs during a liquid-liquid
extraction but is continuous, not step-wise. In this example, using a
water/methanol gradient, the more hydrophobic components will elute (come off
the column) under conditions of relatively high methanol (which is hydrophobic);
whereas the more hydrophilic compounds will elute under conditions of relatively
low methanol/high water. The choice of solvents, additives and gradient depend
on the nature of the stationary phase and the analyte. Often a series of tests are
performed on the analyte and a number of generic runs may be processed in order
to find the optimum HPLC method for the analyte - the method which gives the
best separation of peaks.

[edit] Types of HPLC

[edit] Normal phase chromatography

For more details on this topic, see aqueous normal phase

chromatography.

Also known Normal phase HPLC (NP-HPLC), this method separates analytes
based on polarity; it was the first kind of HPLC chemists developed. NP-HPLC
uses a polar stationary phase and a non-polar mobile phase, and works effectively
for relatively polar analytes. The polar analyte associates with and is retained by
the polar stationary phase. Adsorption strengths increase with increased analyte
polarity, and the interaction between the polar analyte and the polar stationary
phase (relative to the mobile phase) increases the elution time. The interaction
strength not only depends on the functional groups in the analyte molecule, but
also on steric factors. The affect of sterics on interaction strength allows this
method to resolve (separate) structural isomers.

Use of more polar solvents in the mobile phase will decrease the retention time of
the analytes while more hydrophobic solvents tend to increase retention times.
Very polar solvents in a mixture tend to deactivate the column by occupying the
stationary phase surface. This is somewhat particular to normal phase because it is
most purely an adsorptive mechanism (the interactions are with a hard surface
rather than a soft layer on a surface).
NP-HPLC had fallen out of favor in the 1970's with the development of reversed-
phase HPLC because of a lack of reproducibility of retention times as water or
protic organic solvents changed the hydration state of the silica or alumina
chromatographic media. Recently it has become useful again with the
development of HILIC bonded phases which utilize a partition mechanism which
provides reproducibility.

[edit] Reversed phase chromatography

Reversed phase HPLC (RP-HPLC or RPC) has a non-polar stationary phase and
an aqueous, moderately polar mobile phase. One common stationary phase is a
silica which has been treated with RMe2SiCl, where R is a straight chain alkyl
group such as C18H37 or C8H17. With these stationary phases, retention time is
longer for molecules which are more non-polar, while polar molecules elute more
readily. An investigator can also increase retention time by adding a polar solvent
to the mobile phase, or decrease retention time by adding a more hydrophobic
solvent. Reversed phase chromatography (RPC) is so commonly used that it is not
uncommon for it to be incorrectly referred to as "HPLC" without further
specification. The pharmaceutical industry regularly employs RPC to qualify
drugs before their release.

RPC operates on the principle of hydrophobic interactions, which result from

repulsive forces between a polar eluent, the relatively non-polar analyte, and the
non-polar stationary phase. The binding of the analyte to the stationary phase is
proportional to the contact surface area around the non-polar segment of the
analyte molecule upon association with the ligand in the aqueous eluent. This
solvophobic effect is dominated by the force of water for "cavity-reduction"
around the analyte and the C18-chain versus the complex of both. The energy
released in this process is proportional to the surface tension of the eluent (water:
7.3 × 10-6 J/cm², methanol: 2.2 × 10-6 J/cm²) and to the hydrophobic surface of the
analyte and the ligand respectively. The retention can be decreased by adding less-
polar solvent (MeOH, ACN) into the mobile phase to reduce the surface tension
of water. Gradient elution uses this effect by automatically changing the polarity
of the mobile phase during the course of the analysis.

Structural properties of the analyte molecule play an important role in its retention
characteristics. In general, an analyte with a larger hydrophobic surface area (C-H,
C-C, and generally non-polar atomic bonds, such as S-S and others) results in a
longer retention time because it increases the molecule's non-polar surface area,
which is non-interacting with the water structure. On the other hand, polar groups,
such as -OH, -NH2, COO- or -NH3+ reduce retention as they are well integrated
into water. Very large molecules, however, can result in an incomplete interaction
between the large analyte surface and the ligands alkyl chains and can have
problems entering the pores of the stationary phase.

Retention time increases with hydrophobic - non-polar - surface area. Branched

chain compounds elute more rapidly than their corresponding linear isomers
because the overall surface area is decreased. Similarly organic compounds with
single C-C-bonds elute later than the ones with a C=C or C-C-triple bond, as the
double or triple bond is shorter than a single C-C-bond.

Aside from mobile phase surface tension (organizational strength in eluent

structure), other mobile phase modifiers can affect analyte retention. For example,
the addition of inorganic salts causes a moderate linear increase in the surface
tension of aqueous solutions (ca. 1.5 × 10-7 J/cm² per Mol for NaCl, 2.5 × 10-
7
 J/cm² per Mol for (NH4)2SO4), and because the entropy of the analyte-solvent
interface is controlled by surface tension, the addition of salts tend to increase the
retention time. This technique is used for mild separation and recovery of proteins
and protection of their biological activity in protein analysis (hydrophobic
interaction chromatography, HIC).

Another important component is the influence of the pH since this can change the
hydrophobicity of the analyte. For this reason most methods use a buffering agent,
such as sodium phosphate, to control the pH. A volatile organic acid such as
formic acid or most commonly trifluoroacetic acid is often added to the mobile
phase, if mass spectrometry is applied to the eluent fractions. The buffers serve
multiple purposes: they control pH, neutralize the charge on any residual exposed
silica on the stationary phase and act as ion pairing agents to neutralize charge on
the analyte. The effect varies depending on use but generally improve the
chromatography.

Reversed phase columns are quite difficult to damage compared with normal
silica columns, however, many reversed phase columns consist of alkyl
derivatized silica particles and should never be used with aqueous bases as these
will destroy the underlying silica particle. They can be used with aqueous acid,
but the column should not be exposed to the acid for too long, as it can corrode
the metal parts of the HPLC equipment. The metal content of HPLC columns
must be kept low if the best possible ability to separate substances is to be
retained. A good test for the metal content of a column is to inject a sample which
is a mixture of 2,2'- and 4,4'- bipyridine. Because the 2,2'-bipy can chelate the
metal, the shape of the peak for the 2,2'-bipy will be distorted (tailed) when metal
ions are present on the surface of the silica.[citation needed]

[edit] Size exclusion chromatography

For more details on this topic, see size exclusion chromatography.

Size exclusion chromatography (SEC), also known as gel permeation

chromatography or gel filtration chromatography, separates particles on the basis
of size. It is generally a low resolution chromatography and thus it is often
reserved for the final, "polishing" step of a purification. It is also useful for
determining the tertiary structure and quaternary structure of purified proteins.

This technique is widely used for the molecular weight determination of

polysaccharides. SEC is the official technique (suggested by European
pharmacopeia) for the molecular weight comparison of different commercially
available low-molecular weight heparins.

[edit] Ion exchange chromatography

For more details on this topic, see Ion exchange chromatography.

In Ion-exchange chromatography, retention is based on the attraction between

solute ions and charged sites bound to the stationary phase. Ions of the same
charge are excluded. Some types of ion exchangers include: (1) Polystyrene
resins- allows cross linkage which increases the stability of the chain. Higher
cross linkage reduces swerving, which increases the equilibration time and
ultimately improves selectivity. (2) Cellulose and dextran ion exchangers (gels)-
These possess larger pore sizes and low charge densities making them suitable for
protein separation. (3) Controlled-pore glass or porous silica.

In general, ion exchangers favor the binding of ions of higher charge and smaller
radius.

An increase in counter ion (with respect to the functional groups in resins)

concentration reduces the retention time. An increase in pH reduces the retention
time in cation exchange while a decrease in pH reduces the retention time in anion
exchange.

This form of chromatography is widely used in the following applications: In

purifying water, preconcentration of trace components, Ligand-exchange
chromatography, Ion-exchange chromatography of proteins, High-pH anion-
exchange chromatography of carbohydrates and oligosaccharides, etc.

[edit] Bioaffinity chromatography

This chromatographic process relies on the property of biologically active

substances to form stable, specific, and reversible complexes. The formation of
these complexes involves the participation of common molecular forces such as
the Van der Waals interaction, electrostatic interaction, dipole-dipole interaction,
hydrophobic interaction, and the hydrogen bond. An efficient, biospecific bond is
formed by a simultaneous and concerted action of several of these forces in the
complementary binding sites.

[edit] Isocratic flow and gradient elution

With regard to the mobile phase, a composition of the mobile phase that remains
constant throughout the procedure is termed isocratic.

In contrast to this is the so called "gradient elution", which is a separation where

the mobile phase changes its composition during a separation process. One
example is a gradient in 20 min starting from 10 % methanol and ending up with
30 % methanol. Such a gradient can be increasing or decreasing. The benefit of
gradient elution is that it helps speed up elution by allowing components that elute
more quickly to come off the column under different conditions than components
which are more readily retained by the column. By changing the composition of
the solvent, components that are to be resolved can be selectively more or less
associated with the mobile phase. As a result, at equilibrium they spend more time
in the solvent and less time in the stationary phase, and therefore they elute faster.

[edit] Parameters
[edit] Internal diameter

The internal diameter (ID) of an HPLC column is a critical aspect that determines
quantity of analyte that can be loaded onto the column and also influences
sensitivity. Larger columns are usually seen in industrial applications such as the
purification of a drug product for later use. Low ID columns have improved
sensitivity and lower solvent consumption at the expense of loading capacity.

 Larger ID columns (over 10 mm) are used to purify usable amounts of

material because of their large loading capacity.
 Analytical scale columns (4.6 mm) have been the most common type of
columns, though smaller columns are rapidly gaining in popularity. They
are used in traditional quantitative analysis of samples and often use a UV-
Vis absorbance detector.
 Narrow-bore columns (1-2 mm) are used for applications when more
sensitivity is desired either with special UV-vis detectors, fluorescence
detection or with other detection methods like liquid chromatography-
mass spectrometry
 Capillary columns (under 0.3 mm) which are used almost exclusively with
alternative detection means such as mass spectrometry. They are usually
made from fused silica capillaries, rather than the stainless steel tubing that
larger columns employ.

[edit] Particle size

Most traditional HPLC is performed with the stationary phase attached to the
outside of small spherical silica particles (very small beads). These particles come
in a variety of sizes with 5μm beads being the most common. Smaller particles
generally provide more surface area and better separations, but the pressure
required for optimum linear velocity increases by the inverse of the particle
diameter squared.[1][2][3]

This means that changing to particles that are half as big, keeping the size of the
column the same, will double the performance, but increase the required pressure
by a factor of four. Larger particles are more often used in non-HPLC applications
such as solid-phase extraction.

[edit] Pore size

Many stationary phases are porous to provide greater surface area. Small pores
provide greater surface area while larger pore size has better kinetics especially
for larger analytes. For example a protein which is only slightly smaller than a
pore might enter the pore but not easily leave once inside.

[edit] Pump pressure

Pumps vary in pressure capacity, but their performance is measured on their

ability to yield a consistent and reproducible flow rate. Pressure may reach as high
as 40 MPa (6000 lbf/in2), or about 400 atmospheres. Modern HPLC systems have
been improved to work at much higher pressures, and therefore be able to use
much smaller particle sizes in the columns (< 2 micrometres). These "Ultra High
Performance Liquid Chromatography" systems or UHPLCs can work at up to 100
MPa (15,000 lbf/in²), or about 1000 atmospheres. Note that the term "UPLC",
sometimes found instead is a trademark of Waters Corporation and not the name
for the technique in general.

[edit] Manufacturers of HPLC chromatographs

 Agilent Technologies
 Beckman Coulter, Inc.
 Hitachi
 Jasco Corporation, JAPAN
 Knauer
 PerkinElmer, Inc.
 Shimadzu Scientific Instruments
 Thermo Fisher Scientific
 Varian, Inc.
 Waters Corporation

[edit] Manufacturers of HPLC columns and

accessories
 Agilent Technologies
 AkzoNobel
 Beckman Coulter, Inc.
 Merck KGaA
 Phenomenex
 Restek
 SGE Analytical Science
 Shimadzu Scientific Instruments
 Sigma-Aldrich
 Thermo Fisher Scientific
 Tosoh Corporation
 Varian, Inc.
 Waters Corporation
High Performance Liquid Chromatography
(HPLC)
The Protein Facility utilizes three Beckman Instruments System Gold high
performance liquid chromatography systems for the separation of peptides and
proteins. One system is devoted to analytical scale separations, one is for micro-
analytical purposes such as trypsin digest separations and the last system can
handle preparative and semi-preparative scale separations. Users are also trained
to run the HPLC systems themselves at a reduced charge. Below is a short
description of reversed phase chromatography.

Reversed Phase Chromatography

In reverse phase chromatography, the packing is nonpolar and the solvent is polar
with respect to the sample. Retention is the result of the interaction of the
nonpolar components of the solutes and the nonpolar stationary phase. Typical
stationary phases are nonpolar hydrocarbons, waxy liquids or bonded
hydrocarbons (such as C18, C8, C4, etc.) and the solvents are polar aqueous-organic
mixtures such as methanol-water or acetonitrile-water.

The C18, C8, and phenyl bonded phases are most often used in the reverse phase
mode. It has been estimated that 60-90% of all analytical LC separations are done
on bonded phases in the reverse phase mode. Bonded phases made by covalently
bonding a molecule onto a solid stationary phase are intended to prepare "liquid
coatings" which will be permanent. Silica is a reactive substrate to which various
functionalities can be attached or bonded. The functionalities most widely bonded
to silica are the alkyl (C18 and C8), aromatic phenyl, and cyano and amino groups.

General characteristics of reversed phase chromatography

 Broad scope which allows sample types with a wide range of polarities
and molecular weights to be separated.
  General rapidity of mobile phase column equilibration during methods
development and gradient regeneration.
 General ease of use.
 Applicability to separation of ionic or ionizable compounds by
manipulating secondary chemical equilibrium such as ionization control
and ion pairing in the aqueous mobile phase.
o Buffering the mobile phase in the pH range from 2 to 5 with one of
the common buffers, the ionization of the weak acids can be
suppressed or controlled allowing them to be retained in their
neutral form. Similarly weak bases can be retained in their neutral
form at pH 7-7.5.
o For strong acids and bases ionization control cannot be employed
because the stability of alkyl bonded phases is diminished below
pH 2 and above pH 7.5. Highly hydrophilic weak acids and bases
often remain difficult to retain with ionization control. In such
cases ion pair reversed phase chromatography can be used. In this
method, counterions (species of opposite charge to the solutes)
thereby regulate the retention. Typically alkyl amines or tetra alkyl
amines are added to ion pair with acids whereas alkyl sulfates,
sulfonates, or phosphates are used to ion pair with bases. The
technique is an alternative to ion exchange chromatography for
analysis of ionic compounds.
 The possibility of special selectivity such as structural or steric are
achievable by specific mobile phase additives:
o Metal ions are capable of binding to organic compounds in a very
selective method which is used for ligand exchange
chromatography. The selectivity generated in these metal ion phase
systems is based in part on differences of the solute (ligand)
binding strength to the metal ion. An alternate approach is the
addition of various chelating agents (4-dodecyldiethylene-triamine
- C12 dien) in combination with a metal ion. The type and strength
of the metal chelate complex-solute binding can be greatly varied
depending upon the chemical environment surrounding the metal
ion as determined by the chelating agent added.

 
 III. GAS CHROMATOGRAPHY

Kromatografi Gas-Cair
Kromatografi gas-cair (biasa disebut kromatografi gas) merupakan analisis yang
sangat bermanfaat.

Pelaksanaan kromatografi gas-cair

Pengantar

Seluruh bentuk kromatografi terdiri dari fase diam dan fase gerak. Dalam seluruh
bentuk kromatografi yang lain, anda akan menemui fase gerak adalah cairan.
Dalam kromatografi gas-cair, fase gerak adalah gas seperti helium dan fase diam
adalah cairan yang mempunyai titik didih yang tinggi diserap pada padatan.

Bagaimana kecepatan suatu senyawa tertentu bergerak melalui mesin, akan

tergantung pada seberapa lama waktu yang dihabiskan untuk bergerak dengan gas
dan sebaliknya melekat pada cairan dengan jalan yang sama.

Diagram alir kromatografi gas-cair

Injeksi sampel

Sejumlah kecil sampel yang akan dianalisis diinjeksikan pada mesin

menggunakan semprit kecil. Jarum semprit menembus lempengan karet tebal
(Lempengan karet ini disebut septum) yang mana akan mengubah bentuknya
kembali secara otomatis ketika semprit ditarik keluar dari lempengan karet
tersebut.

Injektor berada dalam oven yang mana temperaturnya dapat dikontrol. Oven
tersebut cukup panas sehingga sampel dapat mendidih dan diangkut ke kolom
oleh gas pembawa misalnya helium atau gas lainnya.

Bagaimana kerja kolom?�@

Material padatan

Ada dua tipe utama kolom dalam kromatografi gas-cair. Tipe pertama, tube
panjang dan tipis berisi material padatan; Tipe kedua, lebih tipis dan memiliki fase
diam yang berikatan dengan pada bagian terdalam permukaannya.

Untuk menyederhanakan, kita akan melihat pada kolom terpadatkan.

Kolom biasanya dibuat dari baja tak berkarat dengan panjang antara 1 sampai 4
meter, dengan diameter internal sampai 4 mm. Kolom digulung sehingga dapat
disesuakan dengan oven yang terkontrol secara termostatis.

Kolom dipadatkan dengan tanah diatomae, yang merupakan batu yang sangat
berpori. Tanah ini dilapisis dengan cairan bertitik didih tinggi, biasanya polimer
lilin.

Temperatur kolom

Temperatur kolom dapat bervariasi antara 50 oC sampai 250 oC. Temperatur

kolom lebih rendah daripada gerbang injeksi pada oven, sehingga beberapa
komponen campuran dapat berkondensasi pada awal kolom.

Dalam beberapa kasus, seperti yang anda akan lihat pada bagian bawah, kolom
memulai pada temperatur rendah dan kemudian terus menerus menjadi lebih
panas dibawah pengawasan komputer saat analisis berlangsung.

Bagaimana pemisahan berlangsung pada kolom?

Ada tiga hal yang dapat berlangsung pada molekul tertentu dalam campuran yang
diinjeksikan pada kolom:
 Molekul dapat berkondensasi pada fase diam.
 Molekul dapat larut dalam cairan pada permukaan fase diam
 Molekul dapat tetap pada fase gas

Dari ketiga kemungkinan itu, tak satupun yang bersifat permanen.

Senyawa yang mempunyai titik didih yang lebih tinggi dari temperatur kolom
secara jelas cenderung akan berkondensasi pada bagian awal kolom. Namun,
beberapa bagian dari senyawa tersebut akan menguap kembali dengan dengan
jalan yang sama seperti air yang menguap saat udara panas, meskipun temperatur
dibawah 100 oC. Peluangnya akan berkondensasi lebih sedikit selama berada
didalam kolom.

Sama halnya untuk beberapa molekul dapat larut dalam fase diam cair. Beberapa
senyawa akan lebih mudah larut dalam cairan dibanding yang lainnya. Senyawa
yang lebih mudah larut akan menghabiskan waktunya untuk diserap pada fase
diam: sedangkan senyawa yang suka larut akan menghabiskan waktunya lebih
banyak dalam fase gas.

Proses dimana zat membagi dirinya menjadi dua pelarut yang tidak bercampurkan
karena perbedaan kelarutan, dimana kelarutan dalam satu pelarut satu lebih mudah
dibanding dengan pelarut lainnya disebut sebagai partisi. Sekarang, anda bisa
beralasan untuk memperdebatkan bahwa gas seperti helium tidak dapat dijelaskan
sebagai �gpelarut�h. Tetapi, istilah partisi masih dapat digunakan dalam
kromatografi gas-cair.

Anda dapat mengatakan bahwa substansi antara fase diam cair dan gas. Beberapa
molekul dalam substansi menghabiskan waktu untuk larut dalam cairan dan
beberapa lainnya menghabiskan waktu untuk bergerak bersama-sama dengan gas.

Waktu retensi

Waktu yang digunakan oleh senyawa tertentu untuk bergerak melalui kolom
menuju ke detektor disebut sebagi waktu retensi. Waktu ini diukur berdasarkan
waktu dari saat sampel diinjeksikan pada titik dimana tampilan menunujukkan
tinggi puncak maksimum untuk senyawa itu.

Setiap senyawa memiliki waktu retensi yang berbeda. Untuk senyawa tertentu,
waktu retensi sangat bervariasi dan bergantung pada:
 Titik didih senyawa. Senyawa yang mendidih pada temperatur yang lebih
tinggi daripada temperatur kolom, akan menghabiskan hampir seluruh
waktunya untuk berkondensasi sebagai cairan pada awal kolom. Dengan
demikian, titik didih yang tinggi akan memiliki waktu retensi yang lama.
 Kelarutan dalam fase cair. Senyawa yang lebih mudah larut dalam fase
cair, akan mempunyai waktu lebih singkat untuk dibawa oleh gas
pembawa.. Kelarutan yang tinggi dalam fase cair berarti memiiki waktu
retensi yang lama.
 Temperatur kolom. Temperatur tinggi menyebakan pergerakan molekul-
molekul dalam fase gas; baik karena molekul-molekul lebih mudah
menguap, atau karena energi atraksi yang tinggi cairan dan oleh karena itu
tidak lama tertambatkan. Temperatur kolom yang tinggi mempersingkat
waktu retensi untuk segala sesuatunya di dalam kolom.

Untuk memberikan sampel dan kolom, tidak ada banyak yang bisa dikerjakan
menggunakan titik didih senyawa atau kelarutannya dalam fase cair, tetapi anda
dapat mempunyai pengatur temperatur.

Semakin rendah temperatur kolom semakin baik pemisahan yang akan anda
dapatkan, tetapi akan memakan waktu yang lama untuk mendapatkan senyawa
karena kondensasi yang lama pada bagian awal kolom!

Dengan kata lain, menggunakan temperatur tinggi, segala sesuatunya akan melalui
kolom lebih cepat, tetapi pemisihannya kurang baik. Jika segala sesuatunya
melalui kolom dalam waktu yang sangat singkat, tidak akan terdapat jarak antara
puncak-puncak dalam kromatogram.

Jawabannya dimulai dengan kolom dengan suhu yang rendah kemudian perlahan-
lahan secara teratur temperaturnya dinaikkan.

Pada awalnya, senyawa yang menghabiskan lebih banyak waktunya dalam fase
gas akan melalui kolom secara cepat dan dapat dideteksi. Dengan adanya sedikit
pertambahan temperatur akan memperjelas �gperlekatan�h senyawa.
Peningkatan temperatur masih dapat lebih `melekatan` molekul-molekul fase
diam melalui kolom.

Detektor

Ada beberapa tipe detektor yang biasa digunakan. Detektor ionisasi nyala
dijelaskan pada bagian bawah penjelasan ini, merupakan detektor yang umum dan
lebih mudah untuk dijelaskan daripada detektor alternatif lainnya.

Detektor ionisasi nyala

Dalam mekanisme reaksi, pembakaran senyawa organik merupakan hal yang

sangat kompleks. Selama proses, sejumlah ion-ion dan elektron-elektron
dihasilkan dalam nyala. Kehadiran ion dan elektron dapat dideteksi.

Seluruh detektor ditutup dalam oven yang lebih panas dibanding dengan
temperatur kolom. Hal itu menghentikan kondensasi dalam detektor.
Jika tidak terdapat senyawa organik datang dari kolom, anda hanya memiliki
nyala hidrogen yang terbakar dalam air. Sekarang, anggaplah bahwa satu senyawa
dalam campuran anda analisa mulai masuk ke dalam detektor.

Ketika dibakar, itu akan menghasilkan sejumlah ion-ion dan elektron-elektron

dalam nyala. Ion positif akan beratraksi pada katoda silinder. Ion-ion negatif dan
elektron-elektron akan beratraksi pancarannya masing-masing yang mana
merupakan anoda.

Hal ini serupa dengan apa yang terjadi selama elektrolisis normal.

Pada katoda, ion positif akan mendatangi elektron-elektron dari katoda dan
menjadi netral. Pada anoda, beberapa elektron dalam nyala akan dipindahkan pada
elektroda positif; ion-ion negatif akan memberikan elektron-elektronnya pada
elektroda dan menjadi netral.

Kehilangam elektron-elektron dari satu elektroda dan perolehan dari elektroda

lain, akan menghasilkan aliran elektron-elektron dalam sirkuit eksternal dari
anoda ke katoda. Dengan kata lain, anda akan memperoleh arus listrik.

Arus yang diperoleh tidak besar, tetapi dapat diperkuat. Jika senyawa-senyawa
organik lebih banyak dalam nyala, maka akan banyak juga dihasilkan ion-ion, dan
dengan demikian akan terjadi arus listrik yang lebih kuat. Ini adalah pendekatan
yang beralasan, khususnya jka anda berbicara tentang senyawa-senyawa yang
serupa, arus yang anda ukur sebanding dengan jumlah senyawa dalam nyala.

Kekurangan detektor ionisasi nyala

Kekurangan utama dari detektor ini adalah pengrusakan setiap hasil yang keluar
dari kolom sebagaimana yang terdeteksi. Jika anda akan mengrimkan hasil ke
spektrometer massa, misalnya untuk analisa lanjut, anda tidak dapat menggunakan
detektor tipe ini.

Penerjemahan hasil dari detektor

Hasil akan direkam sebagai urutan puncak-puncak; setiap puncak mewakili satu
senyawa dalam campuran yang melalui detektor. Sepanjang anda mengontrol
secara hati-hati kondisi dalam kolom, anda dapat menggunakan waktu retensi
untuk membantu mengidentifikasi senyawa yang tampak-tentu saja anda atau
seseorang lain telah menganalisa senyawa murni dari berbagai senyawa pada
kondisi yang sama.
Area dibawah puncak sebanding dengan jumlah setiap senyawa yang telah
melewati detektor, dan area ini dapat dihitung secara otomatis melalui komputer
yang dihubungkan dengan monitor. Area yang akan diukur tampak sebagai bagian
yang berwarna hijau dalam gambar yang disederhanakan.

Perlu dicatat bahwa tinggi puncak tidak merupakan masalah, tetapi total area
dibawah puncak. Dalam beberapa contoh tertentu, bagian kiri gambar adalah
puncak tertinggi dan memiliki area yang paling luas. Hal ini tidak selalu
merupakan hal seharusnya..

Mungkin saja sejumlah besar satu senyawa dapat tampak, tetapi dapat terbukti
dari kolom dalam jumlah relatif sedikit melalui jumlah yang lama. Pengukuran
area selain tinggi puncak dapat dipergunakan dalam hal ini.

Perangkaian kromatogram gas pada spektrometer massa

Hal ini tidak dapat dillakukan menggunakan detektor ionisasi nyala, karena
detektor dapat merusak senyawa yang melaluinya. Anggaplah anda menggunakan
detektor yang tidak merusak. Senyawa,

Ketika detektor menunjukkan puncak, beberapa diantaranya melalui detektor dan

pada waktu itu dapat dibelokkan pada spektrometer massa. Hal ini akan
memberikan pola fragmentasi yang dapat dibandingkan dengan data dasar
senyawa yang telah diketahui sebelumnya pada komputer. Itu berarti bahwa
identitas senyawa-senyawa dalam jumlah besar dapat dihasilkan tanpa harus
mengetahui waktu retensinya

Gas-liquid chromatography
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 material may be challenged and removed.
Gas-liquid chromatography

A gas chromatograph with a headspace sampler

Acronym GLC, GC
Classification chromatography
organic
Analytes inorganic
must be volatile
Agilent (a spin-off of Hewlett-Packard)
PerkinElmer
Manufacturers Shimadzu
Thermo Electron Corporation
Varian, Inc.
Other Techniques
Thin layer chromatography
Related
High performance liquid chromatography
Hyphenated Gas chromatography-mass spectrometry
This box: view • talk • edit

Gas-liquid chromatography (GLC), or simply gas chromatography (GC), is a

type of chromatography in which the mobile phase is a carrier gas, usually an inert
gas such as helium or an unreactive gas such as nitrogen, and the stationary phase
is a microscopic layer of liquid or polymer on an inert solid support, inside glass
or metal tubing, called a column. The instrument used to perform gas
chromatographic separations is called a gas chromatograph (also: aerograph, gas
separator).

Gas Chromatography is different from other forms of chromatography (HPLC,

TLC, etc.) because the solutions travel through the column in a gas state. The
interactions of these gaseous analytes with the walls of the column (coated by
different stationary phases) causes different compounds to elute at different times
called retention time. The comparsion of these retention times is the analytical
power to GC. This makes it very similar to HPLC.

Contents
[hide] [hide]
 1 History
 2 GC analysis
 3 Physical components
o 3.1 Autosamplers
o 3.2 Inlets
o 3.3 Columns
o 3.4 Detectors
 4 Methods
o 4.1 Carrier gas selection and flow rates
o 4.2 Inlet types and flow rates
o 4.3 Sample size and injection technique
 4.3.1 Sample injection
o 4.4 Column selection
o 4.5 Column temperature and temperature program
 5 Data reduction and analysis
 6 Application
 7 GCs in popular culture
 8 See also
 9 References

 10 External links

[edit] History
Chromatography dates to 1903 in the work of the Russian scientist, Mikhail
Semenovich Tswett. German graduate student Fritz Prior developed solid state
gas chromatography in 1947. Archer John Porter Martin, who was awarded the
Nobel Prize for his work in developing liquid-liquid (1941) and paper (1944)
chromatography, laid the foundation for the development of gas chromatography
and later produced liquid-gas chromatography (1950).
[edit] GC analysis
A gas chromatograph is a chemical analysis instrument for separating chemicals
in a complex sample. A gas chromatograph uses a flow-through narrow tube
known as the column, through which different chemical constituents of a sample
pass in a gas stream (carrier gas, mobile phase) at different rates depending on
their various chemical and physical properties and their interaction with a specific
column filling, called the stationary phase. As the chemicals exit the end of the
column, they are detected and identified electronically. The function of the
stationary phase in the column is to separate different components, causing each
one to exit the column at a different time (retention time). Other parameters that
can be used to alter the order or time of retention are the carrier gas flow rate, and
the temperature.

In a GC analysis, a known volume of gaseous or liquid analyte is injected into the

"entrance" (head) of the column, usually using a microsyringe (or, solid phase
microextraction fibers, or a gas source switching system). As the carrier gas
sweeps the analyte molecules through the column, this motion is inhibited by the
adsorption of the analyte molecules either onto the column walls or onto packing
materials in the column. The rate at which the molecules progress along the
column depends on the strength of adsorption, which in turn depends on the type
of molecule and on the stationary phase materials. Since each type of molecule
has a different rate of progression, the various components of the analyte mixture
are separated as they progress along the column and reach the end of the column
at different times (retention time). A detector is used to monitor the outlet stream
from the column; thus, the time at which each component reaches the outlet and
the amount of that component can be determined. Generally, substances are
identified (qualitatively) by the order in which they emerge (elute) from the
column and by the retention time of the analyte in the column.

[edit] Physical components

Diagram of a gas chromatograph.

[edit] Autosamplers
The autosampler provides the means to introduce automatically a sample into the
inlets. Manual insertion of the sample is possible but is no longer common.
Automatic insertion provides better reproducibility and time-optimization.

Different kinds of autosamplers exist. Autosamplers can be classified in relation

to sample capacity (auto-injectors VS autosamplers, where auto-injectors can
work a small number of samples), to robotic technologies (XYZ robot VS
rotating/SCARA-robot – the most common), or to analysis:

 Liquid
 Static head-space by syringe technology
 Dynamic head-space by transfer-line technology
 SPME

Traditionally autosampler manufactures are different from GC manufactures and

currently no GC manufacture offers a complete range of autosamplers.
Historically, the countries most active in autosampler technology development are
the United States, Italy, and Switzerland.

[edit] Inlets

The column inlet (or injector) provides the means to introduce a sample into a
continuous flow of carrier gas. The inlet is a piece of hardware attached to the
column head.

Common inlet types are:

 S/SL (Split/Splitless) injector; a sample is introduced into a heated small

chamber via a syringe through a septum - the heat facilitates volatilization
of the sample and sample matrix. The carrier gas then either sweeps the
entirety (splitless mode) or a portion (split mode) of the sample into the
column. In split mode, a part of the sample/carrier gas mixture in the
injection chamber is exhausted through the split vent.
 On-column inlet; the sample is here introduced in its entirety without
heat.
 PTV injector; Temperature-programmed sample introduction was first
described by Vogt in 1979. Originally Vogt developed the technique as a
method for the introduction of large sample volumes (up to 250 µL) in
capillary GC. Vogt introduced the sample into the liner at a controlled
injection rate. The temperature of the liner was chosen slightly below the
boiling point of the solvent. The low-boiling solvent was continuously
evaporated and vented through the split line. Based on this technique, Poy
developed the Programmed Temperature Vaporising injector; PTV. By
introducing the sample at a low initial liner temperature many of the
disadvantages of the classic hot injection techniques could be
circumvented.
 Gas source inlet or gas switching valve; gaseous samples in collection
bottles are connected to what is most commonly a six-port switching valve.
 The carrier gas flow is not interrupted while a sample can be expanded
into a previously evacuated sample loop. Upon switching, the contents of
the sample loop are inserted into the carrier gas stream.
 P/T (Purge-and-Trap) system; An inert gas is bubbled through an
aqueous sample causing insoluble volatile chemicals to be purged from the
matrix. The volatiles are 'trapped' on an absorbent column (known as a
trap or concentrator) at ambient temperature. The trap is then heated and
the volatiles are directed into the carrier gas stream. Samples requiring
preconcentration or purification can be introduced via such a system,
usually hooked up to the S/SL port.
 SPME (solid phase microextraction) offers a convenient, low-cost
alternative to P/T systems with the versatility of a syringe and simple use
of the S/SL port.

[edit] Columns

Two types of columns are used in GC:

 Packed columns are 1.5 - 10 m in length and have an internal diameter of

2 - 4 mm. The tubing is usually made of stainless steel or glass and
contains a packing of finely divided, inert, solid support material (eg.
diatomaceous earth) that is coated with a liquid or solid stationary phase.
The nature of the coating material determines what type of materials will
be most strongly adsorbed. Thus numerous columns are available that are
designed to separate specific types of compounds.
 Capillary columns have a very small internal diameter, on the order of a
few tenths of millimeters, and lengths between 25-60 meters are common.
The inner column walls are coated with the active materials (WCOT
columns), some columns are quasi solid filled with many parallel
micropores (PLOT columns). Most capillary columns are made of fused-
silica with a polyimide outer coating. These columns are flexible, so a very
long column can be wound into a small coil.
 New developments are sought where stationary phase incompatibilities
lead to geometric solutions of parallel columns within one column. Among
these new developments are:
o Internally heated microFAST columns, where two columns, an
internal heating wire and a temperature sensor are combined within
a common column sheath (microFAST);
o Micropacked columns (1/16" OD) are column-in-column packed
columns where the outer column space has a packing different
from the inner column space, thus providing the separation
behaviour of two columns in one. They can be easily fit to inlets
and detectors of a capillary column instrument.

The temperature-dependence of molecular adsorption and of the rate of

progression along the column necessitates a careful control of the column
temperature to within a few tenths of a degree for precise work. Reducing the
temperature produces the greatest level of separation, but can result in very long
elution times. For some cases temperature is ramped either continuously or in
steps to provide the desired separation. This is referred to as a temperature
program. Electronic pressure control can also be used to modify flow rate during
the analysis, aiding in faster run times while keeping acceptable levels of
separation.

The choice of carrier gas (mobile phase) is important, with hydrogen being the
most efficient and providing the best separation. However, helium has a larger
range of flowrates that are comparable to hydrogen in efficiency, with the added
advantage that helium is non-flammable, and works with a greater number of
detectors. Therefore, helium is the most common carrier gas used.

[edit] Detectors

A number of detectors are used in gas chromatography. The most common are the
flame ionization detector (FID) and the thermal conductivity detector (TCD).
Both are sensitive to a wide range of components, and both work over a wide
range of concentrations. While TCDs are essentially universal and can be used to
detect any component other than the carrier gas (as long as their thermal
conductivities are different than that of the carrier gas, at detector temperature),
FIDs are sensitive primarily to hydrocarbons, and are more sensitive to them than
TCD. However, an FID cannot detect water. Both detectors are also quite robust.
Since TCD is non-destructive, it can be operated in-series before an FID
(destructive), thus providing complementary detection of the same eluents.

Other detectors are sensitive only to specific types of substances, or work well
only in narrower ranges of concentrations. They include:

 discharge ionization detector (DID)

 electron capture detector (ECD)
 flame photometric detector (FPD)
 Hall electrolytic conductivity detector (ElCD)
 helium ionization detector (HID)
 nitrogen phosphorus detector (NPD)
 mass selective detector (MSD)
 photo-ionization detector (PID)
 pulsed discharge ionization detector (PDD)
 thermal energy analyzer (TEA)

Some gas chromatographs are connected to a mass spectrometer which acts as the
detector. The combination is known as GC-MS. Some GC-MS are connected to
an NMR spectrometer which acts as a back up detector. This combination is
known as GC-MS-NMR. Some GC-MS-NMR are connected to an infrared
spectrophotometer which acts as a back up detector. This combination is known
as GC-MS-NMR-IR. It must, however, be stressed this is very rare as most
analyses needed can be concluded via purely GC-MS.
[edit] Methods
The method is the collection of conditions in which the GC operates for a given
analysis. Method development is the process of determining what conditions are
adequate and/or ideal for the analysis required.

Conditions which can be varied to accommodate a required analysis include inlet

temperature, detector temperature, column temperature and temperature program,
carrier gas and carrier gas flow rates, the column's stationary phase, diameter and
length, inlet type and flow rates, sample size and injection technique. Depending
on the detector(s) (see below) installed on the GC, there may be a number of
detector conditions that can also be varied. Some GCs also include valves which
can change the route of sample and carrier flow, and the timing of the turning of
these valves can be important to method development.

[edit] Carrier gas selection and flow rates

Typical carrier gases include helium, nitrogen, argon, hydrogen and air. Which
gas to use is usually determined by the detector being used, for example, a DID
requires helium as the carrier gas. When analyzing gas samples, however, the
carrier is sometimes selected based on the sample's matrix, for example, when
analyzing a mixture in argon, an argon carrier is preferred, because the argon in
the sample does not show up on the chromatogram. Safety and availability can
also influence carrier selection, for example, hydrogen is flammable, and high-
purity helium can be difficult to obtain in some areas of the world. (See: Helium--
occurrence and production.)

The purity of the carrier gas is also frequently determined by the detector, though
the level of sensitivity needed can also play a significant role. Typically, purities
of 99.995% or higher are used. Trade names for typical purities include "Zero
Grade," "Ultra-High Purity (UHP) Grade," "4.5 Grade" and "5.0 Grade."

The carrier gas flow rate affects the analysis in the same way that temperature
does (see above). The higher the flow rate the faster the analysis, but the lower the
separation between analytes. Selecting the flow rate is therefore the same
compromise between the level of separation and length of analysis as selecting the
column temperature.

With GCs made before the 1990s, carrier flow rate was controlled indirectly by
controlling the carrier inlet pressure, or "column head pressure." The actual flow
rate was measured at the outlet of the column or the detector with an electronic
flow meter, or a bubble flow meter, and could be an involved, time consuming,
and frustrating process. The pressure setting was not able to be varied during the
run, and thus the flow was essentially constant during the analysis.

Many modern GCs, however, electronically measure the flow rate, and
electronically control the carrier gas pressure to set the flow rate. Consequently,
carrier pressures and flow rates can be adjusted during the run, creating
pressure/flow programs similar to temperature programs.

[edit] Inlet types and flow rates

The choice of inlet type and injection technique depends on if the sample is in
liquid, gas, adsorbed, or solid form, and on whether a solvent matrix is present
that has to be vaporized. Dissolved samples can be introduced directly onto the
column via a COC injector, if the conditions are well known; if a solvent matrix
has to be vaporized and partially removed, a S/SL injector is used (most common
injection technique); gaseous samples (e.g., air cylinders) are usually injected
using a gas switching valve system; adsorbed samples (e.g., on adsorbent tubes)
are introduced using either an external (on-line or off-line) desorption apparatus
such as a purge-and-trap system, or are desorbed in the S/SL injector (SPME
applications).

[edit] Sample size and injection technique

[edit] Sample injection

The rule of ten in gas chromatography

The real chromatographic analysis starts with the introduction of the sample onto
the column. The development of capillary gas chromatography resulted in many
practical problems with the injection technique. The technique of on-column
injection, often used with packed columns, is usually not possible with capillary
columns. The injection system, in the capillary gas chromatograph, should fulfil
the following two requirements:

1. The amount injected should not overload the column.

2. The width of the injected plug should be small compared to the spreading
due to the chromatographic process. Failure to comply with this
requirement will reduce the separation capability of the column. As a
general rule, the volume injected, Vinj, and the volume of the detecor cell,
 Vdet, should be about 1/10 of the volume occupied by the portion of sample
containing the molecules of interest (analytes) when they exit the column.

Some general requirements, which a good injection technique should fulfill, are:

 It should be possible to obtain the column’s optimum separation

efficiency.
 It should allow accurate and reproducible injections of small amounts of
representative samples.
 It should induce no change in sample composition. It should not exhibit
discrimination based on differences in boiling point, polarity,
concentration or thermal/catalytic stability.
 It should be applicable for trace analysis as well as for undiluted samples.

Please help improve this article or section by expanding it.

Further information might be found on the talk page or at requests for expansion.
(February 2007)

[edit] Column selection

Please help improve this article or section by expanding it.

Further information might be found on the talk page or at requests for expansion.
(February 2007)

[edit] Column temperature and temperature program

A gas chromatography oven, open to show a capillary column

The column(s) in a GC are contained in an oven, the temperature of which is

precisely controlled electronically. (When discussing the "temperature of the
column," an analyst is technically referring to the temperature of the column oven.
The distinction, however, is not important and will not subsequently be made in
this article.)

The rate at which a sample passes through the column is directly proportional to
the temperature of the column. The higher the column temperature, the faster the
sample moves through the column. However, the faster a sample moves through
the column, the less it interacts with the stationary phase, and the less the analytes
are separated.

In general, the column temperature is selected to compromise between the length

of the analysis and the level of separation.

A method which holds the column at the same temperature for the entire analysis
is called "isothermal." Most methods, however, increase the column temperature
during the analysis, the initial temperature, rate of temperature increase (the
temperature "ramp") and final temperature is called the "temperature program."

A temperature program allows analytes that elute early in the analysis to separate
adequately, while shortening the time it takes for late-eluting analytes to pass
through the column.

[edit] Data reduction and analysis

Qualitative analysis:

Generally chromatographic data is presented as a graph of detector response (y-

axis) against retention time (x-axis). This provides a spectrum of peaks for a
sample representing the analytes present in a sample eluting from the column at
different times. Retention time can be used to identify analytes if the method
conditions are constant. Also, the pattern of peaks will be constant for a sample
under constant conditions and can identify complex mixtures of analytes. In most
modern applications however the GC is connected to a mass spectrometer or
similar detector that is capable of identifying the analytes represented by the
peaks.

Quantitive analysis:

The area under a peak is proportional to the amount of analyte present. By

calculating the area of the peak using the mathematical function of integration, the
concentration of an analyte in the original sample can be determined.
Concentration can be calculated using a calibration curve created by finding the
response for a series of concentrations of analyte, or by determining the relative
response factor of an analyte. The relative response factor is the expected ratio of
an analyte to an internal standard (or external standard) and is calculated by
finding the response of a known amount of analyte and a constant amount of
internal standard (a chemical added to the sample at a constant concentration, with
a distinct retention time to the analyte).
In most modern GC-MS systems, computer software is used to draw and integrate
peaks, and match MS spectra to library spectra.

[edit] Application
In general, substances that vaporize below ca. 300 °C (and therefore are stable up
to that temperature) can be measured quantitatively. The samples are also required
to be salt-free; they should not contain ions. Very minute amounts of a substance
can be measured, but it is often required that the sample must be measured in
comparison to a sample containing the pure, suspected substance.

Various temperature programs can be used to make the readings more

meaningful; for example to differentiate between substances that behave similarly
during the GC process.

Professionals working with GC analyze the content of a chemical product, for

example in assuring the quality of products in the chemical industry; or measuring
toxic substances in soil, air or water. GC is very accurate if used properly and can
measure picomoles of a substance in a 1 ml liquid sample, or parts-per-billion
concentrations in gaseous samples.

In practical courses at colleges, students sometimes get acquainted to the GC by

studying the contents of Lavender oil or measuring the ethylene that is secreted by
Nicotiana benthamiana plants after artificially injuring their leaves. These GC
analyses hydrocarbons (C2-C40+). In a typical experiment, a packed column is
used to separate the light gases, which are then detected with a TCD. The
hydrocarbons are separated using a capillary column and detected with an FID. A
complication with light gas analyses that include H2 is that He, which is the most
common and most sensitive inert carrier (sensitivity is proportional to molecular
mass) has an almost identical thermal conductivity to hydrogen (it is the
difference in thermal conductivity between two separate filaments in a
Wheatstone Bridge type arrangement that shows when a component has been
eluted). For this reason, dual TCD instruments are used with a separate channel
for hydrogen that uses nitrogen as a carrier are common. Argon is often used
when analysing gas phase chemistry reactions such as F-T synthesis so that a
single carrier gas can be used rather than 2 separate ones. The sensitivity is less
but this is a tradeoff for simplicity in the gas supply.

[edit] GCs in popular culture

Movies, books and TV shows tend to misrepresent the capabilities of gas
chromatography and the work done with these instruments.

In the U.S. TV show CSI, for example, GCs are used to rapidly identify unknown
samples. "This is gasoline bought at a Chevron station in the past two weeks," the
analyst will say fifteen minutes after receiving the sample.
In fact, a typical GC analysis takes much more time; sometimes a single sample
must be run more than an hour according to the chosen program; and even more
time is needed to "heat out" the column so it is free from the first sample and can
be used for the next. Equally, several runs are needed to confirm the results of a
study - a GC analysis of a single sample may simply yield a result per chance (see
statistical significance).

Also, GC does not positively identify most samples; and not all substances in a
sample will necessarily be detected. All a GC truly tells you is at which relative
time a component eluted from the column and that the detector was sensitive to it.
To make results meaningful, analysts need to know which components at which
concentrations are to be expected; and even then a small amount of a substance
can hide itself behind a substance having both a higher concentration and the same
relative elution time. Last but not least it is often needed to check the results of the
sample against a GC analysis of a reference sample containing only the suspected
substance.

A GC-MS can remove much of this ambiguity, since the mass spectrometer will
identify the component's molecular weight. But this still takes time and skill to do
properly.

Similarly, most GC analyses are not push-button operations. You cannot simply
drop a sample vial into an auto-sampler's tray, push a button and have a computer
tell you everything you need to know about the sample. According to the
substances one expects to find the operating program must be carefully chosen.

A push-button operation can exist for running similar samples repeatedly, such as
in a chemical production environment or for comparing 20 samples from the same
experiment to calculate the mean content of the same substance. However, for the
kind of investigative work portrayed in books, movies and TV shows this is
clearly not the case.

Introduction

Gas chromatography - specifically gas-liquid chromatography - involves a sample

being vapourised and injected onto the head of the chromatographic column. The
sample is transported through the column by the flow of inert, gaseous mobile
phase. The column itself contains a liquid stationary phase which is adsorbed onto
the surface of an inert solid.

Have a look at this schematic diagram of a gas chromatograph:

Instrumental components

Carrier gas
The carrier gas must be chemically inert. Commonly used gases include nitrogen,
helium, argon, and carbon dioxide. The choice of carrier gas is often dependant
upon the type of detector which is used. The carrier gas system also contains a
molecular sieve to remove water and other impurities.

Sample injection port

For optimum column efficiency, the sample should not be too large, and should be
introduced onto the column as a "plug" of vapour - slow injection of large samples
causes band broadening and loss of resolution. The most common injection
method is where a microsyringe is used to inject sample through a rubber septum
into a flash vapouriser port at the head of the column. The temperature of the
sample port is usually about 50C higher than the boiling point of the least volatile
component of the sample. For packed columns, sample size ranges from tenths of
a microliter up to 20 microliters. Capillary columns, on the other hand, need much
less sample, typically around 10-3 L. For capillary GC, split/splitless injection is
used. Have a look at this diagram of a split/splitless injector;
The injector can be used in one of two modes; split or splitless. The injector
contains a heated chamber containing a glass liner into which the sample is
injected through the septum. The carrier gas enters the chamber and can leave by
three routes (when the injector is in split mode). The sample vapourises to form a
mixture of carrier gas, vapourised solvent and vapourised solutes. A proportion of
this mixture passes onto the column, but most exits through the split outlet. The
septum purge outlet prevents septum bleed components from entering the column.

Columns

There are two general types of column, packed and capillary (also known as open
tubular). Packed columns contain a finely divided, inert, solid support material
(commonly based on diatomaceous earth) coated with liquid stationary phase.
Most packed columns are 1.5 - 10m in length and have an internal diameter of 2 -
4mm.

Capillary columns have an internal diameter of a few tenths of a millimeter. They

can be one of two types; wall-coated open tubular (WCOT) or support-coated
open tubular (SCOT). Wall-coated columns consist of a capillary tube whose
walls are coated with liquid stationary phase. In support-coated columns, the inner
wall of the capillary is lined with a thin layer of support material such as
diatomaceous earth, onto which the stationary phase has been adsorbed. SCOT
columns are generally less efficient than WCOT columns. Both types of capillary
column are more efficient than packed columns.
In 1979, a new type of WCOT column was devised - the Fused Silica Open
Tubular (FSOT) column;

These have much thinner walls than the glass capillary columns, and are given
strength by the polyimide coating. These columns are flexible and can be wound
into coils. They have the advantages of physical strength, flexibility and low
reactivity.

Column temperature

For precise work, column temperature must be controlled to within tenths of a

degree. The optimum column temperature is dependant upon the boiling point of
the sample. As a rule of thumb, a temperature slightly above the average boiling
point of the sample results in an elution time of 2 - 30 minutes. Minimal
temperatures give good resolution, but increase elution times. If a sample has a
wide boiling range, then temperature programming can be useful. The column
temperature is increased (either continuously or in steps) as separation proceeds.

Detectors

There are many detectors which can be used in gas chromatography. Different
detectors will give different types of selectivity. A non-selective detector responds
to all compounds except the carrier gas, a selective detector responds to a range of
compounds with a common physical or chemical property and a specific detector
responds to a single chemical compound. Detectors can also be grouped into
concentration dependant detectors and mass flow dependant detectors. The signal
from a concentration dependant detector is related to the concentration of solute in
the detector, and does not usually destroy the sample Dilution of with make-up
gas will lower the detectors response. Mass flow dependant detectors usually
destroy the sample, and the signal is related to the rate at which solute molecules
enter the detector. The response of a mass flow dependant detector is unaffected
by make-up gas. Have a look at this tabular summary of common GC detectors:

Support Dynamic
Detector Type Selectivity Detectability
gases range
Flame
Hydrogen Most organic
ionization Mass flow 100 pg 107
and air cpds.
(FID)
Thermal
conductivity Concentration Reference Universal 1 ng 107
(TCD)
Halides, nitrates,
Electron nitriles,
capture Concentration Make-up peroxides, 50 fg 105
(ECD) anhydrides,
organometallics
Nitrogen- Hydrogen Nitrogen,
Mass flow 10 pg 106
phosphorus and air phosphorus
Sulphur,
Hydrogen phosphorus, tin,
Flame
and air boron, arsenic,
photometric Mass flow 100 pg 103
possibly germanium,
(FPD)
oxygen selenium,
chromium
Aliphatics,
aromatics,
ketones, esters,
Photo- aldehydes,
ionization Concentration Make-up amines, 2 pg 107
(PID) heterocyclics,
organosulphurs,
some
organometallics
Hall Halide, nitrogen,
Hydrogen,
electrolytic Mass flow nitrosamine,    
oxygen
conductivity sulphur
The effluent from the column is mixed with hydrogen and air, and ignited.
Organic compounds burning in the flame produce ions and electrons which can
conduct electricity through the flame. A large electrical potential is applied at the
burner tip, and a collector electrode is located above the flame. The current
resulting from the pyrolysis of any organic compounds is measured. FIDs are
mass sensitive rather than concentration sensitive; this gives the advantage that
changes in mobile phase flow rate do not affect the detector's response. The FID is
a useful general detector for the analysis of organic compounds; it has high
sensitivity, a large linear response range, and low noise. It is also robust and easy
to use, but unfortunately, it destroys the sample.

Kromatogerafi gas adalah satu teknik yang popular untuk

menganalisis sampel yang boleh meruap. Sampel yang sesuai
dianalisis menggunakan teknik ini mestilah boleh larut dalam pelarut
organic. Sampel akan disuntik melalui penyuntik dimana suhu yang
digunakan biasanya lebih tinggi daripada suhu turus. Sampel yang
meruap akan dibawa melalui turus oleh fasa bergerak seperti gas
nitrogen tulen atau helium. Sampel yang terpisah daripada turus
seterusnya dicamkan oleh pengesan (FID). Pengesan FID berfungsi
dengan bantuan gas hidrogen dan udara mampat secara berterusan.
Pemilihan turus yang sesuai dan manipulasi suhu oven pemisahan
yang baik boleh dicapai. Terdapat dua jenis manipulasi suhu oven
secara isotermal dan pemprograman suhu.
 Kromatografi merupakan pemisahan yang terjadi karena adanya
adsorbsi selektif komponen dalam campuran antara fase gerak (fase mobil)
dan fase diam (fase stasioner). Fase mobil (pelarut) dan komponen campuran
akan melewati kolom dan diadsorbsi oleh fase diam (adsorben). Oleh karena
perbedaan dalam adsorpsi dan pelarutan maka komponen kan melewati kolom
dengan laju yang berbeda-beda.
Kromatografi kolom bekerja berdasarkan polaritas dan bobot molekul.
Molekul dengan polaritas dan bobot molekul paling tinggi akan keluar dari
kolom paling akhir dan begitu pula kebalikannya. Umumnya komponen yang
mempunyai struktur yang mirip lebih sulit dipisahkan. Pelarut dalam
menseleksi campuran tergantung kepolaran, mulai dari pelarut nonpolar
(seperti heksan) sampai polar (asam organik). Pelarut terbaik dipilih
berdasarkan percobaan dan pengalaman.
Pada kromatografi kolom (kromatografi adsorpsi), fase diam akan
berinteraksi dengan sampel dan tiap komponen akan diadsorpsi dari pelarut
oleh adsorben dengan tingkat tertentu. Adsorben yang baik mempunyai luas
permukaan yang besar dan mempunyai banyak sisi aktif. Adsorben yang dapat
digunakan yaitu alumina, silika gel, pati, dan sebagainya. Kromatografi ini
dilakukan pada tekanan sekitar 1 atmosfir dengan diameter dalam kolom 1-5
cm. Teknik pemisahan dilakukan dengan menuangkan sampel secara hati-hati
ke dalam kolom dan elusi terjadi secara lambat mulai dari 30 menit sampai
beberpa jam.
 Ukuran panjang kolom sekitar 10-100 kali garis tengah dalam.
Perbandingan ini tergantung dari sukar tidaknya pemisahan. Semakin sukar
memisah, maka perbandingan harus semakin besar. Untuk perbandingan bobot
adsorben dengan sampel biasanya 30:1 untuk pemisahan yang normal
sedangkan untuk pemisahan yang sulit bisa sampai 100:1 atau 300:1. Laju
pelarut harus tetap, yang baik diperoleh kecepatan rata-rata 1 cm/menit. Laju
ini tergantung dari ukuran partikel adsorben, ukuran kolom (panjang dan
diameter dalam), kekentalan cairan, dan tekanan terhadap kolom.

1. Fase Diam (Adsorben)

Daya adsorpsi dari fase diam tergantung dari sifat kimia permukaan,
luas relatif permukaan, dan perlakuan pendahuluan yang dilakukan.
Beberapa adsorben yang dapat digunakan mulai dari daya adsorpsi
terbesar yaitu alumina, charcoal, silika gel, magnesia, kalsium karbonat,
sukrosa, pati dan selulosa serbuk. Dalam kromatografi, yang sering
digunakan yaitu alumina dan silika gel, karena mudah didapatkan dan
paling berguna.
Dalam sistem kromatografi kolom, yang perlu diperhatikan adalah
sifat, derajat atau tingkat keaktifan adsorben, dan ukuran partikel. Sifat
adsorben tergantung pH dan tingkat keaktifannya. Alumina dan silika gel
akan berfungsi melalui titik-titik permukaan teroksigenasi terutama gugus
hidroksil. Gugus ini menarik molekul sampel karena aksi dipol-dipol dan
ikatan hidrogen. Perlu dihindari adanya air dan pelarut berproton seperti
alkohol dan amina, sebab permukaan adsorben tidak dapat berfungsi
dengan baik.
Alumina (Al2O3) mempunyai pusat aktif Al+, Al-OH, Al-OH+,
kadang-kadang Na+ dan H+. Alumina bersifat asam (pH 4) mengandung
ion klorida dan alumina bersifat basa mengandung pusat-pusat aluminat
yang berfungsi sebagai penukar ion. Alumina bersifat asam digunakan
untuk pemisahan asam amino dan asam peptida, sedangkan alumina
bersifat basa dipakai lebih luas terutama untuk senyawa polar yang akan
teradsorpsi sangat kuat, sedangkan alumina bersifat netraldipalai untuk
 pemisahan ketosteroid, glikosid, ketal, lakton, ester, dan untuk
mendehidrasi pelarut. Luas permukaan relatif, sekitar 150 m2/g dan
dipakai umumnya pada kadar air 3 %.
Silika gel ( SiO2)mempunyai luas permukaan relatif sekitar 500 m2/g
dan kurang aktif jika dibandingkan dengan alumina. Biasanya digunakan
untuk pemisahan senyawa organik yang sensitif terhadap sifat mengkatalis
dari permukaan aktif. Akan tetapi, silika gel merupakan adsorben serba
guna dan banyak dipakai. Silika gel juga dapat digunakan hampir pada
semua pelarut, kecuali air, metanol, dan etanol, sebab jika ada pelarut
tersebut, adsorben tidak dapat berfungsi dengan baik.

2. Fase Gerak (Pelarut)

Peristiwa yang terjadi jika sampel bergerak bersama pelarut maka
sampel cenderung melarut di dalam fase gerak dan bergerak bersamanya.
Pelarut yang baik harus dapat melarutkan sampel dan bersaing dengan
daya serap dari adsorben. Hal ini terjadi jika pelarut yang digunakan
bukanlah yang berproton. Hidrokarbon, eter, dan senyawa karbonil dapat
digunkan untuk pemisahan dengan teknik ini. Perisstiwa lain yaitu pelarut
akan bersaing dengan sampel untuk mendapat tempat pada permukaan
adsorben, kemudian sampel didesak dari adsorben oleh pelarut. Pelarut
berproton seperti alkohol dan amina kurang baik dalam mendorong
sampel.
Pada umumnya, senyawa non polar akan didesak dari kolom
sebelum senyawa polar, jika menggunakan pelarut non polar. Jika
menggunakan pelarut kloroform atau etil asetat maka ester keluar lebih
dulu. Untuk mengelusi alkohol dapat juga menggunakan pelarut polar
seperti aseton atau etanol.
Pealrut dalam mengevaluasi sampel diusahakan tidak terlalu cepat
dan tidak terlalu lambat. Jika terlalu cepat, komponen dalam sampel sulit
dipisahkan, dan sebaliknya mengakibatkan waktu retensi (Rt) terlalu lama.
Hal ini dapat mengakibatkan peleebaran pita komponen yang berlebihan
dan pengenceran terhadap pelarut.
 Pelarut yang digunakan dapat 1 jenis atau campuran pelarut. Agar
diperoleh pelarut atau campuran pelarut yang tepat maka dapat
menggunakan deret eluotropik pelarut berdasarkan daya elusi terendah
sampai yang lebih itnggi pada adsorben alumina atau silika gel.
Tabel 1. Deret Eluotropik Beberapa Jenis Pelarut
Jenis Pelarut
- Hidrokarbon seperti heksan, heptan, dan petroleum eter
- Sikloheksan
- Karbon tetraklorid
- Benzen
- Toluen
- Tetra hidrofuran
- Metilen klorid
- Kloroform
- Etil eter (eter)
- Etil asetat
- n-propanol
- Etanol
- Asetonitril
- Metanol
- Air

Dalam pemilihan pelarut pada kromatografi kolom dapat juga

menggunakan pendekatan melalui penelusuran pustaka, yaitu dicari
informasi mengenai senyawa dengan ukuran molekul hampir sama dan
mempunyai gugus fungsi sama. Cara lain dengan menerapkan data
kromatografi lapis tipis pada pemisahan yaitu dalam penggunaan jenis
adsorben sama kecuali ukuran partikel serta yang memberi kepastian
bahwa pemisahan berlangsung pada waktu yang masuk akal dan
pemakaian jumlah pelarut yang logis. Terakhir menggunakan elusi
landaian yaitu susunan pelarut diubah dari medium non polar ke medium
lebih polar.
B. Kromatografi Gas
Dalam tahap-tahap awal perkembangan kromatografi gas, alat ini
diterapkan pada analisis gas dan uap dari komponen-komponen yang sangat
mudah menguap. Setelah itu, Martin dan Synge mengembangkan teori dasar
dalam kromatografi gas-cairan (GLC), semuanya terbuka untuk teknik analisis
kimia. Sebagai suatu alat analitik, kromatografi gas dapat digunakan untuk
analisis dan pemisahan langsung sampel-sampel gas, larutan-larutan, dan
padatan yang bersifat mudah menguap. Jika sampel yang akan dianalisis tidak
menguap, maka dapat digunakan suatu teknik pirolisis dalam kromatografi
gas. Ini merupakan modifikasi dari teknik dimana sampel yang tidak mudah
menguap tersebut mengalami pirolisis sebelum masuk ke kolom.
Kromatografi gas, disamping untuk tujuan analisis, juga sering
digunakan untuk mempelajari struktur suatu komponen kimia, menentukan
mekanisme dan kinetik dari reaksi-reaksi kimia, serta mengukur isotermal,
sifat panas larutan, sifat panas penyerapan, energi bebas dari larutan dan/atau
penyerapan, koefisien aktivitas, dan tetapan difusi. Penerapan kromatografi
gas di berbagai bidang diantaranya adalah di bidang obat-obatan dan farmasi,
lingkungan hidup, industri minyak, kimia klinik, pestisida dan residunya, dan
pangan.
1. Fase Gerak
Fase bergerak merupakan komponen keseluruhan sistem
kromatografi gas yang penting. Komponen-komponen sampel dialirkan
melalui kolom dengan bantuan dorongan gas pengemban yang sesuai.
Penggunaan gas sebagai fase bergerak menyebabkan keseimbangan antara
fase bergerak dan fase stasioner berlangsung dengan cepat, menyebabkan
metode kromatografi gas ini performansinya tinggi. Fase bergerak dari
gas, mempunyai tahanan aliran yang rendah, menyebabkan dengan kolom
yang panjang dapat dihasilkan pemisahan yang baik.
Kondisi yang tetap setiap saat perlu dipertahankan agar hasil
pemisahan memuaskan. Suatu pengatur tekanan digunakan untuk
menjamin tekanan yang seragam pada pemasuk kolom sehingga diperoleh
 laju aliran gas yang tetap. Selain itu, untuk menjaga kondisi aliran yang
baik dan tetap, kromatografi gas juga dilengkapi dengan silinder gas, alat
pemurni gas, pengontrol aliran, meter aliran, dan pemanas awal. Pada
sembarang suhu tertentu, laju aliran tetap akan mengelusi komponen
campuran pada waktu yang khas (waktu tambat). Oleh karena laju aliran
tetap, komponen mempunyai volume gas pembawa yang khas (volum
tambat).
Gas yang biasa dipakai adalah hidrogen, helium, dan nitrogen.
Syarat-syarat gas pembawa adalah
1. Lembam, untuk mencegah interaksi dengan cuplikan (fase diam)
2. Dapat meminimumkan difusi gas
3. Mudah didapat dan murni
4. Cocok untuk detektor yang digunakan

2. Fase Diam (Stasioner)

Salah satu alasn mengapa kromatografi gas-cairan (GLC) diterima
secara luas adalah karena tersedianya bermacam-macam fase cair dengan
sifat yang berbeda-beda. Didalam memilih fase cair untuk suatu analisis
tertentu, perlu diperhatikan beberapa kriteria yang harus dipertimbangkan,
yaitu
1. Apakah fase cair tersebut sifatnya selektif terhadap komponen
sampel yang akan dipisahkan
2. Apakah akan ada reksi-reaksi yang sifatnya tidak kembal;i antara
fase cair dan komponen-komponen campuran yang akan dipisahkan
3. apakah fase cair tersebut mempunyai tekanan uap yang rendah
pada suhu yang digunakan untuk analisa, dan apakah sifatnya stabil
terhadap panas
Dua sifat lainnya dari fase cair yang penting untuk diperhatikan
adalah kekentalan dan kemampuan untuk melapisi penunjang padat yang
merata. Idealnya fase cair mempunyai kekentalan yang rendah dan dengan
kemampuan melapisi yang tinggi.
3. Komponen Kromatografi gas
Pada dasarnya, suatu kromatografi gas terdiri dari enam komponen
utama, yaitu sistem gas pengemban (carrier gas) termasuk tangki
penyuplai gas serta pengatur dan alirannya, sistem penyuntikan sampel,
kolom pemisah, pendeteksian (detektor), elektrometer dan sistem pencatat
(rekorder), dan unit termostat untuk mengatur suhu oven.

1. Sistem Gas Pengemban

Fungsi dari gas pengemban adalah mengangkut sampel melalui
kolom ke detektor. Pemilihan gas pengemban yang tepat sangat
penting karena mempengaruhi kolom dan detektor dalam melakukan
fungsinya. Untuk mjendapatkan hasil analisis terbaik, gas pengemban
harus mempunyai kemurnian paling rendah 99.995 %. Adanya
pengotor seperti udara air dapat menyebabkan penguraian sampel,
disamping mengurangi daya tahan kolom dan detektor. Gas pengmban
juga harus bersifat inert dengan komponen-komponen dari sampel dan
isi kolom.
Gas pengemban dapat diperoleh dari tangki gas bertekanan tinggi
sampai sekitar 150-160 atm. Tangki tersebut dilengkapi dengan
regulator pengatur tekanandua tahap, yaitu tekanan dalam tang ki dan
tekanangas yang keluar dari tangki. Kecepatan aliran gas pengemban
perlu diatur karena akan mempengaruhi hasil pemisahan komponen
dalam kolom. Perubahan dalam kecepatan aliran akan menyebabkan
perubahan dalam waktu retensi.

2. Sistem Penyuntikan Sampel

Fungsi utama dari sistem penyuntikan sampel adalah untuk
menerima sampel, menuapkannya segera jika sampel dalam bentuk
buka gas, dan untuk mengantarkan komponen menguap ke kolom
analitik dalam suatu suntikan sekecil mungkn.disain sistem
penyuntikan sampel beragam tergantung pada jenis kolom yang
digunakan, apakah jenis kolom kapiler atau kolom jejal. Disamping itu
 juga tergantung dari jenis sampel, apakah dalam bentuk gas, cair, atauy
padatan, sebagai contoh sampel yang tidak menguap membutuhkan
alat pirolisis untuk menguapkan.

3. Kolom Pemisah dan Oven Kolom

ada dua jenis kolom yang digunakan dalam kromatografi gas
secara umum, yaitu kolom jejal dan kolom tubuler terbuka. Kolom
yang pertama adalah kolom metal atau gelas yang diisi bahan
pengepak yang terdiri dari penunjang padatan yang dilapisi fase cair
yang tidak menguap atau adsorben. Kolom tubuler terbuka sangat
berbeda dengan kolom jejal, yaitu gas yang mengalir sepanjang kolom
tidak mengalami hambatan, karena kolomnya merupakan tabung tnpa
bahan pengisi.
Kemampuan memisahkan komponen per meter kolom pada
kolom tubuler terbuka tidak jauh berbeda dengan pemisahan pada
kolom jejal. Meskipun demikian, penggunaan kolom yang sangat
panjang bersama-sama dengan waktu analisis yang relatif cepat
merupakan alat penolong yang berharga bagi para ahli kimia untuk
dapat memisahkan komponen-komponen yang perbedaannya kecil
dalam sifat-sifat fisiknya.
Fungsi dari oven kolom adalah untuk memberikan suhu yang
diinginkan pada kolom dimana terjadi proses pemisahan.

4. Detektor
Fungsi detektor adalah untuk mendeteksi dan memberikan respon
dalam bentuk sinyal-sinyal listrik jika komposisi gas yang keluar dari
kolom berubah. Jenis detektor yang digunakan tergantung pada
penerapannya. Detektor yang paling umum digunakan adalah detektor
konduktivitas panas (TCD), ionisasi api (FID), dan penangkap elektron
(ECD)

5. Elektrometer dan Rekorder

 Salah satu aspek dari instrumentasi kromatografi gas yang
menyebabkan dapat dimengertinya analisis baik kualitatif maupun
kuantitatif, adalah bagian elektrometer dan rekorder. Sinyal-sinyal
yang keluar dari detektor ionisasi adalah arus listrik yang sangat
lemah, misalnya aliran elektron. Jika arus elektron yang kecil sebesar
10-11 A mengalir pada tahanan 1010 Ohm. Maka voltase melalui
resistor harus 0.1 Volt. Voltase ini lebih dari cukup untuk
menggerakkan rekorder potensiometer 1mv dengan skala penuh.
Meskipun demikian, rekorder potensiometrik hanya dapat menerima
sinyal sepanjang tahanan dengan nilai beberapa ribu Ohm. Hal inilah
yang menjadikan penguat elektrometer berfungsi untuk
mengembangkan suatu voltese yang identik sepanjang unsur-unsur
atenuator yang tahanannya relatif kecil. Suatu rekorder potensiometer
kemudian dapat dihubungkan pada atenuator untuk pembentukan
grafik dari sinyal keluaran detektor.

DAFTAR PUSTAKA

Fardiaz, Dedi. 1989. Kromatografi Gas dalam Analisis Pangan. Pusat Antar
Universitas Pangan dan Gizi Institut Pertanian Bogor, Bogor.

McNair, H. M. dan E. J. Bonelli. 1988. Dasar Kromatografi Gas. Institut

Teknologi Bandung, Bandung.

Mulyadi. 1994. Peningkatan Kadar Vetiverol Minyak Akar Wangi dengan Cara
Deterpenasi Menggunakan Kromatografi Kolom. Skripsi. Fakultas
Teknologi Pertanian Institut Pertanian Bogor, Bogor.